ISSN 0304-5250 INDIAN JOURNAL OF ECOLOGY Volume 46 Special Issue-8 December 2019

THE INDIAN ECOLOGICAL SOCIETY INDIAN ECOLOGICAL SOCIETY (www.indianecologicalsociety.com) Past President: A.S. Atwal and G.S.Dhaliwal (Founded 1974, Registration No.: 30588-74) Registered Office College of Agriculture, Punjab Agricultural University, Ludhiana – 141 004, Punjab, India (e-mail : [email protected]) Advisory Board Kamal Vatta S.K. Singh S.K. Gupta Chanda Siddo Atwal B. Pateriya K.S. Verma Asha Dhawan A.S. Panwar S. Dam Roy V.P. Singh Executive Council President A.K. Dhawan Vice-Presidents R. Peshin S.K. Bal Murli Dhar G.S. Bhullar General Secretary S.K. Chauhan Joint Secretary-cum-Treasurer Vaneet Inder Kaur Councillors A.K. Sharma A. Shukla S. Chakraborti N.K. Thakur Members Jagdish Chander R.S. Chandel R. Banyal Manjula K. Saxexa Editorial Board Chief-Editor Anil Sood Associate Editor S.S. Walia K. Selvaraj Editors M.A. Bhat K.C. Sharma B.A. Gudae Mukesh K. Meena S. Sarkar Neeraj Gupta Mushtaq A. Wani G.M. Narasimha Rao Sumedha Bhandari Maninder K. Walia Rajinder Kumar Subhra Mishra A.M. Tripathi Harsimran Gill

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Analysis of the Climate Data (Earth Observation Stations and Space Stations) Through Geographic Information System

Balsam Shaker Shnichal Department of Geography and GIS, Faculty of Arts, Iraqia University, Baghdad, Iraq E-mail: [email protected]

Abstract: Earth observation stations' data on meteorological parameters is inadequate for several technical or financial reasons as well as data interpretation under different circumstances. This leads to misleading and inaccuracy for their results, so it was necessary to search for an alternative to achieve accuracy. The digital data of space station in climatic studies such as TIRS, NOAA, GOES, NIMBUS, ERS-1, METEOSAT-9 are the means to fill the shortage of data recorded by earth observation stations. The present study focused on space recordings with earth observations and tested its accuracy extent using the Pearson correlations coefficients, the results showed that there was a strong direct relationship between space and ground data (r=0.931-0.970), proving the accuracy and convergence of its records with the ground data.

Keyword: Satellite data, Climate data, Weather forecasts, GIS, Accuracy

Researchers in the field of climate studies suffer from the rainfall components). problem of the lack of recorded data for the meteorological The analytical approach with quantitative analysis stations, both local and regional, to make their studies semi method to description strength of the relationship between disabled and linked to the available data, so there was a need the accuracy of space data and its results with meteorological to adopt other sources to provide climate data, therefore stations of the documented study, represent the results in the there is need to adopt data provided by specialized satellite form of a cartographical statistical model (Tenget al 1993 and for climate studies (NOAA, TIRS, ERS-1, NIMBUS, GOES Lakshmi and Susskind 2000) and METEOSAT-9) for climate data. But the key question The ground meteorological stations and their problems: remains whether it is possible to rely on these satellite data to The government ground meteorological stations provide replenish the shortage and inaccuracy of their recordings. various climatic data for most of the climatic elements in their The idea of current research aimed to make comparison main stations, with limitation to some elements in the between the results of a documented study that adopted the secondary stations (agricultural). These records date back to data of the meteorological ground stations with the results of 1940s (monthly rates only), some of which date back to the the satellite data by adopting the same mechanism of work beginning of the 20th century, such as Baghdad, Mosul and and the duration of the documented study. Pearson Basra so considering as a source for studies of climatic correlation coefficients were used to prove strength of the changes. Data are available with many measurements of the relationship between climatic data for Geo-meteorological elements (hourly, daily, monthly, yearly), especially after the stations and data provided by the global sites associated with mid-eighties of the last century where access to it are easy, satellites specialized in climate studies. and measured/recorded by specialists at that time with high accuracy than it is in present conditions. The meteorological MATERIAL AND METHODS organization were tabulated and organized electronically Area study: The spatial boundaries were confined to the from the last decade of the last century (Ji et al 2015, Tote et political borders of Iraq, and between the longitudes 38° 45̍- al 2015, Nash and Sutcliffe 1970. Wang et al 2011. Ahmed et 48° 45̍̍ east and the latitudes 29° 5̍-37̍°22̍ (Republic of Iraq, al 2016). 2001). Twelve main space observation stations The most important problems faced by researchers (Sulaymaniyah, Mosul, Kirkuk, Khanaqin, Rutbah, Baghdad, during the adoption of the data of ground stations are: the Al-Hayy, Najaf, Amarah, Nasiriyah and Basrah) were heterogeneity of their distribution, which does not provide selected because they represent the different topographical overall coverage of all regions (Map 2), which gives a area of Iraq with different directions (Table 1 and Map 1). The completeness in their recording and the increase in the temporal boundaries of the study were the study period 1979- percentage of missing observed elements so its 2000. The daily data were used for both temperature and recommended using of experimental criteria to replenish the 2 Balsam Shaker Shnichal

Table 1. Climatic stations included by the study data deficit. The current positions of the observation stations Station Ground meteorological Space meteorological are professionally ineligible for measurement due to the stations stations urban extension, which reduces the accuracy of the recorded Latitude, Longitude, Latitude, Longitude, North East North East data, as well as the personal inflection for the observers. Latelya problem has recently emerged, it's the high financial Sulemanea 35.438 45.3125 35.32 45.27 cost associated to obtain data by students and researchers. Mosel 36.3747 43.125 36.19 43.09 Climatic satellite stations and their problems: The Kirkuk 35.438 44.375 35.28 44.24 satellites specialized in climate studies provide data that their Khanaqin 34.1891 45.3125 34.29 45.23 recording accuracy reaches to observation level for hourly, Rutba 32.9401 40.3125 33.03 40.17 daily, monthly and annually for the climatic elements. They Ana 34.5013 41.875 34.22 41.57 are easily obtained and not interrupted for a relatively long Baghdad 33.2524 44.375 33.18 44.24 period of time, starting from 1970, to be used for climate Alhi 32.0035 45.9375 32.08 46.02 studies and for pursuance of change. In addition, provides a Najaf 32.0035 44.375 31.57 44.19 wide coverage ratio compared to ground stations (Map 2) and with high accuracy observation, which reduces the loss Omara 32.0035 47.1875 31.5 47.1 of recording elements, not related to the privacy of the Nasirea 31.0668 46.25 31.01 46.14 observers (Turab et al 2014, Barril et al 2015, Sumi et al Basra 30.4423 47.8125 30.31 47.47 2013). Capabilities and accuracy in measuring and recording Source (Iraq, Ministry of Transport,G.O.M),https://globalweather.tamu.edu elements, as well as its electronic availability and ease of obtaining solve most of the problems faced by researchers. The most important problems of satellite data are the weakness of communication between the concerned institutions and the research centers (universities and scientific centers), as well as the problem of the weakness of English language and the inability to use the sites provided by most researchers. The most important problem facing satellite data is measurements of climatic elements and connecting with the work of satellite sensors. Statistical processing: The study was based on the selection of a documented scientific study that used the data of the government Meteorological Stations for the study of climate fluctuation and its effect on the limits of the climatic regions in Iraq. Depended on the data of the monthly averages of the ground stations for the period 1970-2000 were obtained for the two elements of temperature and rainfall (Katsanos et al 2016). Koppen classification (Peel et Map 1. Location of search stations al 2007, Kottek et al 2006) were used as a criteria for determining climatic regions. It is possible to compare the data recorded with the satellite data by adopting the same time period of the above study and follow the same steps in the work to achieve the results of the use of satellite data as an alternative to ground stations data. Compared the results of the documented study and using the Pearson correlation (R) which were equal and associated with it and had a strong positive relationship either with the general climate classification or with the climate classification for years chosen from the period studied (Table 2). The ArcGIS geostatistical analyst extension provides the capability for modeling using deterministic and Fig. 2. The decrease of weight with distance illustration Using GIS for Climate Data 3 geostatistical methods. The idw method is adopted and uses the measured values surrounding the prediction location to predict a value for any un sampled location, based on the assumption that things that are close to one another are more alike than those that are farther apart. The points highlighted in the data view show the locations and the weights that will be used for predicting a location at the center of the ellipse (the location of the crosshair). The search neighborhood is limited to the interior of the ellipse. In the eastern sector, one point (brown) will be given a weight between 5 and 10 per cent, the rest of the points in the search neighborhood will receive lower weights. Map 2. Geographical distribution of ground observation The analysis of the climatic space stations data show stations and space stations. Source: Iraqi, Ministry of Transport and Communications, 1981-2018 similarity to the ground stations used in the study of Salman https://globalweather.tamu.edu for different years and the application of the modified Koppen classification, which was based on satellite data, showed that Table 4. Results of the general classification of the Iraq there is anapproachin results in most stations (Tables 3, 4, 5 climate for the period 1981-1982 and 6). Stations Satellite climate stations Ground climate stations Station Region Station Region RESULTS AND DISCUSSION classification description classification description Through using the results of satellite data in draw maps Sulemanea 100.75 Humid 44.66 humid Mosel 66.15 Humid 17.52 Semi arid Kirkuk 34.26 Humid 15.61 Semi arid Table 2. Pearson correlation coefficient between the results of the satellite data and the data recorded for all Khanqin 11.74 Semi arid 14.34 Semi arid studied stations Rutba 9.49 Dry 6.67 Dry Classification years 1982- 1990- 2000- 2000- Ana 12.27 Semi arid 13.4 Semi arid 1981 1989 1999 1970 Baghdad 4.74 Very dry 3.42 Very dry Pearson correlation coefficient 0.93 0.94 0.97 0.95 Alhi 5.01 Very dry 3.27 Very dry Relationship type Strong Strong Very Strong Najaf 2.27 Very dry 4.77 Very dry positive positive Strong positive Omara 6.18 Dry 5.84 dry positive Nasriea 2.27 Very dry 4.89 Very dry Basra 3.74 Very dry 5.12 Dry Table 3. Results of the general classification of the Iraq Source: Salman 2011; https://globalweather.tamu.edu climate for the period 1970-2000 Stations Satellite climate stations Ground climate stations Table 5. Results of the general classification of the Iraq climate for the period 1989-1990 Region Station Region Station description classification description classification Stations Satellite climate stations Ground climate stations

Nasirea Dry 5.03 Very dry 3.8 Station Region Station Region classification description classification description Najaf Very dry 4.34 Very dry 3.48 Sulemanea 61.89 Humid 34.59 Humid Alhi Dry 5.94 Dry 5.59 Mosel 33.95 Humid 16.52 Semi arid Omara Dry 7.36 dry 7.55 Kirkuk 22.26 Humid 17.07 Semi arid Rutba Dry 6.2 dry 6.37 Khanqin 10.67 Semi arid 14.44 Semi arid Khanaqin Semi dry 13.87 dry 8.79 Rutba 4.03 Very dry 4.55 Very dry Ana 2.64 Very dry 6.46 Dry Ana Dry 7.17 dry 7.32 Baghdad 0.56 Very dry 6.23 Dry Sulemanea Humid 37.95 humid 45.4 Alhi 6.3 Dry 3.27 Very dry Baghdad Dry 5.69 Very dry 4.8 Najaf 1.75 Very dry 1.63 Very dry Basra Dry 5.85 dry 5.52 Omara 11.14 Semi arid 3.93 Very dry Kirkuk Semi dry 16.89 Semi dry 19.07 Nasriea 5.54 Dry 3.89 Very dry Mosel Semi arid 18.52 humid 34.69 Basra 7.55 Dry 4.02 Very dry Source: Salman 2011; https://globalweather.tamu.edu Source: Salman 2011; https://globalweather.tamu.edu 4 Balsam Shaker Shnichal

A. Results of ground station data B Results of satellite data A. Results of ground station data B Results of satellite data Source: Table 3 Source: Table 6 Map 3. Results of the comparative study of the general Map 6. Results of the comparative study of the general classification of Iraq for the period,1970-2000 classification of Iraq for the period,1999-2000

A. Results of ground station data B Results of satellite data Source: Table 5 A. Results of ground station data B Results of satellite data Map 5. Results of the comparative study of the general Source: Table 4 classification of Iraq for the period,1989-1990 Map 4. Results of the comparative study of the general classification of Iraq for the period,1981-1982

Map 7. Coincidence map of Map 8. Coincidence map of Map 9. Coincidence map of meteorological stations meteorological stations meteorological stations data with satellite data for data with satellite data for data with satellite data for the years 1981-1982; the years 1989-1990; the years 1999-2000; Source Map 4 . Source Map 5 . Source Map 6 . Using GIS for Climate Data 5

Table 6. Results of the general classification of the Iraq Salman does not know any method of representation climate for the period 1999-2000 adopted, the fact that the researcher did not mention it in the Stations Satellite climate stations Ground climate stations context of her thesis, but the basic stations, their Station Region Station Region representation was coincidence with a large proportion, which classification description classification description shows that maps and tables, while the divergent stations Sulemanea 40.97 Humid 19.53 Semi arid made the representation process is more generalization to Mosel 12.72 Semi arid 8.41 Dry show the difference clearly by the number and distribution of Kirkuk 10.25 Semi arid 7.57 Dry stations. It's possible to depending on climatic satellites data Khanqin 6.01 Dry 4.36 Dry and can be used as an substitute or support data of ground Rutba 2.23 Very dry 2.74 Very dry stations to compensate the losses for any reason. Ana 2.64 Very dry 4.25 Very dry Baghdad 2.77 Very dry 2.67 Very dry CONCLUSIONS The data of the ground stations (Government) despite Alhi 3.78 Very dry 3.11 Very dry their advantages, but suffers from several problems can be Najaf 3.2 Very dry 1.26 Very dry avoided by the adoption of data from the space stations as a Omara 6.43 Dry 4.68 Very dry support to them. There was a strong positive relationship Nasriea 4.93 Very dry 2.7 Very dry between the satellite data and ground data (r value ranged Basra 4.64 Very dry 6.16 Dry between 0.931 - 0.970) proved accuracy. Source: Salman 2011; https://globalweather.tamu.edu REFERENCES Ahmed M, Sultan E, Yan and Wahr J 2016. Assessing and improving land surface model outputs over Africa using GRACE, field, and remote sensing data. Surveys in Geography 37(3): 529- 556. Barril PA, Fumian TM, PrezVE, Gil PI, Martínez LC and Giordano MO 2015. Rotavirus seasonality in urban sewage from Argentina: effect of meteorological variables on the viral load and the genetic diversity. Environmental Research 138(2): 409-415. Hanssen AW and Kuipers WJA 1965. On the relationship between the frequency of rain and various meteorological parameters. Meded., Verh 81(2): 2–15. Ji G, Senay B and Verdin JP 2015. Evaluation of the Global land data assimilation system (GLDAS) air temperature data products. American Meteoric. Society 16(2): 2463-2480. Katsanos A, Retalis S and Michaelides A 2016. Validation of a high- resolution precipitation database (CHIRPS) over Cyprus for a 30 year period. Atmospheric Research 169: 459-464. Kottek M, Grieser J, Beck C, Rudolf B and Rubel F 2006. World map of the Köppen-Geiger climate classification updated. Meteorol. Z., 15(3): 259-263. Lakshmi V and Susskind J 2000. Validation of TOVS land surface parameters using ground observations. Journal of Geophysical Research 105: 2179-2190. Nash JV 1970. Sutcliffe river flow forecasting through conceptual Fig. 1. The relationship between the results of the satellite models part I - A discussion of principles. Journal Hydrology data and the ground stations (Source: Based on Table Elsevier 10(1): 282-290. 2) Noman S 2002. Quantitative Methods in Geography using Computer, 2nd edition, Dar Al Safa Publishing and Distribution, Amman. of climatic regions classification and comparing them with Peel MC, Patricio R, BoogaardH and Van der Wijngaart R 2007. regional classification maps based on meteorological data Updated world map of the Koppen–Geiger climate Maps (3, 4, 5, 6), found that there was a semi similarity within classification. Hydrology and Earth System Sciences 11: 1633-1644. the regions borders for the Maps (7, 8, 9) Republic of Iraq 2001. The Council of Ministers, the Planning The variation in the representation of the above regions is Authority, the Central Statistical Organization, the Annual due to the difference in the methods of representation, which Statistical Group. depended on different statistical mechanisms as Salman HA 2011. Climate fluctuation and its impact on the discrepancy of climate regions borders in Iraq, women College generalization, which represents the satellite data in each of Education, University of Baghdad, pp.158-166. station based on similarity (the great probability). The study of Sumi K, Rajendran T, Ramamurthy T, Krishnan G, Nair B and 6 Balsam Shaker Shnichal

Harigane K 2013. Effect of temperature, relative humidity and and Funk C 2015. Evaluation of satellite rainfall estimates for rainfall on rotavirus infections in Kolkata, India. Epidemiology drought and flood monitoring in Mozambique. MDPI AG 7(2): and .Infection 141(8): 1652-1661. 1758-1776. Teng W, Doraiswamy P and Wang J 1993. Relationship between Wang L, Wang L, Koike T, Zhou H, Yang K and Wang A 2011. satellite microwave radiometric data, antecedent precipitation Evaluation and application of a fine-resolution global data set index, and regional soil moisture. International Journal of in a semiarid mesoscale river basin with a distributed Remote Sensing 14: 2483-2500. biosphere hydrological model. Journal of Geographical Tote D, Patricio R, Boogaard H, Van der Wijngaart R, Tarnavsky E Research 116, Doi.10:1029/2011JD015990.

Received 09 September, 2019; Accepted 20 November, 2019 Manuscript Number: S-243 Indian Journal of Ecology (2020) 46 Special Issue (8): 7-14 NAAS Rating: 4.96

Risks of Rainfall Intensity on Geomorphic Processes of Duhok Governorate in Northwest Iraq using RS and GIS

Ruqaya Ahmed M. Amin and Balsam Shaker Shnichal College of Art, Iraqia University, Iraq E-mail: [email protected]

Abstract: The U.S. satellite data was used to obtain rainfall data for the period (2000-2018) for weather maps of the region's systems, and the analysis of the rainy season (2015-2016) included more than 40 measuring stations. During the rainy season, 68 rainfalls that varied in intensity and quantity were recorded. Geomorphic processes represent natural and chemical changes that affect the preparation of materials for processes and thus alter the form of the Earth through erosion, transportation and deposition. This can measure the impact of rain intensity, which divides the world into morphogenetic regions; type of precipitation and its effect is measured for the effect of rain drops, rills erosion and gully erosion. digital processing of space data for the American satellite Landsat 8 was used for chang detection in land cover change due to rainfalls of rainy season, digital elevation data to determine slopes, and application of EPM coefficient to identify the direct impact of intensities on geomorphic operations in northwestern Iraq Governorate of Dohuk.

Keywords: Risk, Rrainfall intensity, Geomorphic, Duhok, Northwest Iraq, RS & GIS

The region is exposed to varying intensity, frequency from US satellite stations http://chrsdata.eng.uci.edu to rainstorms, the importance of daily precipitation is the main obtain rainfall data for the season (2015-2016). Digital source of water in the region, with the assertion that the elevation data DEM 30 meters http://dwtkns.com/srtm30m/ intensity of rainstorms occurs in a sudden, uneven and to derive heights from sea level. Use ARC Map 10.5 to affects human activities with the massive flooding. The achieve research applications and derivatives and build a measurement of rain intensity to identify any of the pressure geographic information base with the use of the spatial systems is more influential, leading to risks of geomorphic analysis Tools and geospatial Wazard to analyze the digital processes in sensitive areas as a natural event. The region is matrices and design spatial maps for study area. exposed to varying intensity, frequency of rain storms, is reflected in the direct recognition of the tension and its impact RESULTS AND DISUSSION on the conduct of the geomorphic processes in northwest Analysis of the rainy season (2015-2016) per hour: During Iraq (Dohuk governorate as a model). Identification of the September 1, 2015 to 30 June, 2016 the rainy days were 68 direct impact of intensity on geomorphic processes in which vary in intensity and quantity. The total rainfall was northwestern Iraq (Dohuk governorate model) is important to 315.5 mm-1. Rain intensity according to classification study. approved by the general authority of Iraqi Meteorological and the General Directorate of Meteorological and Seismic MATERIAL AND METHODS monitoring (Sahar S 2000), for the amount of water falling by Duhok Governorate, located in the far northwest of Iraq, more than (6 mm h-1), and Table 1 shows rain intensity was has a distinctive geographical location as its lies on political recorded higher than 6 mm, according to the indicator, the border with the Republic of Turkey, from east bordered by total rainy days (7 days), and rainy hours (162 hours), the Erbil governorate and from south Ninawa governorate, total rainfall (216.1 mm), (68%) of the total precipitation mountains surrounded Dohuk city from three sides the White amounts for the rainy season (2015-2016). Mountain in north, Mount Zawa in the south and Mam seen The relationship between the pressure systems and the from the east, while from the west open up on the Simel intensity of rain: Pressure system: When following the Agricultural Plain, its astronomical position (N 15o 41' 36'' – synoptic state of the pressure system for the two days (6- N30o 22' 37'' and E 30o 22' 42''-E 45o 03' 44'') in Figure 1. The 7/11/2015), show beginning of precipitation at 13 GMT, as a analytical approach for tracking phenomena and revealing result of the progress of the Mediterranean depression after the circumstances surrounding the region to reach the the retreat of the impact of the subtropical high extension scientific facts was used. away the area (Baidaweed 2003), thus attracting the humid Tools and methods used in research: Daily rainfall data air mass from the Mediterranean (Form 3 1a-1b). As noted at 8 Ruqaya Ahmed M. Amin and Balsam Shaker Shnichal

the hour (19 GMT) The precipitation increased by the rainfall and continued until the Day (7/11/2015) (Fig. 3) and Sudanese depression, which increased the amount of rainfall the control of the Sudanese depression on Iraq's weather in and humidity and the control of the humid air mass, which general (Fig. 2, 2b), as well as effect of relief factor for increased its influence on the atmosphere of the study area in increasing the condensation operations despite the particular, so acting on the integration (Sudanese- existence of tropical trough extension (Al-Khatib 2001), and Mediterranean) (Al-Dazi, SAK 2010) (Fig. 3, 1C-1D). This its impact on the region in general (Fig. 3, 2c). merge and penetration into the study area has increased the The process of decreasing the rains began from 9 GMT

Table 1. Rainfall data for the rainy season (2015-2016) Number Water quantity mm h-1 Per cent Continuity hour-1 Number Water quantity mm h-1 Per cent Continuity hour-1 1 44.9 14.2 21 35 1.7 0.53 2 2 28.8 9.1 8 36 1.7 0.53 2 3 24.3 7.7 15 37 1.6 0.5 4 4 22.7 7.2 10 38 1.6 0.5 6 5 14.8 4.7 17 39 1.5 0.47 2 6 12 3.8 14 40 1.5 0.47 3 7 11.5 3.6 15 41 1.5 0.47 3 8 10.3 3.3 18 42 1.4 0.44 4 9 9.7 3.1 12 43 1.3 0.41 3 10 9.7 3.1 8 44 1.2 0.38 4 11 7.3 2.3 3 45 1.2 0.38 1 12 7.3 2.3 8 46 1.1 0.34 4 13 6.8 2.2 8 47 1 0.31 3 14 6 1.9 5 48 1 0.31 2 15 5.7 1.8 5 49 1 0.31 2 16 5.1 1.6 8 50 0.9 0.28 2 17 4.9 1.55 5 51 0.8 0.25 1 18 4.8 1.52 5 52 0.8 0.25 2 19 4.8 1.52 9 53 0.8 0.25 2 20 4.7 1.48 8 54 0.7 0.22 2 21 4.1 1.3 4 55 0.7 0.22 2 22 3.7 1.2 5 56 0.7 0.22 2 23 3.6 1.1 5 57 0.7 0.22 1 24 3.4 1.07 5 58 0.7 0.22 1 25 3.3 1.04 4 59 0.7 0.22 2 26 3.1 0.98 4 60 0.6 0.19 3 27 2.6 0.82 5 61 0.6 0.19 1 28 2.5 0.79 4 62 0.3 0.09 1 29 2.5 0.79 3 63 0.3 0.09 1 30 2.3 0.72 6 64 0.3 0.09 1 31 2 0.63 5 65 0.3 0.09 1 32 1.9 0.6 5 66 0.2 0.06 1 33 1.9 0.6 4 67 0.2 0.06 1 34 1.7 0.53 2 68 0.2 0.06 1 Sum 315.5 100 336 Source: Based on http://chrsdata.eng.uci.edu/ Rainfall Intensity on Geomorphic Processes of Duhok Governorate 9

on (7/11/2015) and stopped at (12GMT) by the disintegration region (Kinnell and Risse 1998), which is characterized by a coalescence of the depressions and the progress of the steep slope more than 30 degrees, and the superficial Mediterranean depression towards the east and the retreat formations Varying in hardness, which makes them impact of the Sudanese depression to become an effective vulnerability for sever water erosion, as shown on the eroded on the central and southern Iraq (Form 3, 2d). mounts slopes which facing depression, which trigger creep Intensity of rain: From Table 2 and Figure 4 and to and slide of thin soils to form a Geomorphic hazards of, It also understand the causes of the occurrence of the rain, the accelerates the process of mass moving on the Earth's model was taken for the strongest rain intensity and the total surface to form many landforms, with a long dry season of continuous rains for the daily (6-7/11/2015) reached (44.9 acting to prepare ready-to-transport sediments for the mm) and for a period of 21 hours. accumulation plain unit. From the hydrological point of rain Table 3 and Figure 4 show the control ends of tropical intensity, it supplies the aquifer with water and contributes to high pressures to replace the Mediterranean depression and supplying the area with large quantities of water, and can merge during the middle hours cause intensity of the create ground dams and water storage to benefit from its, Sudanese depression with the Mediterranean so increasing especially its supply the region with quantities exceeding the amount of precipitation strengthened by the dominance (200 mm), 42.6% of water during the rainy season (Table 4). and duration Mediterranean depression (Al Nuaimi et al The Fornier equation (Sheikh1987, Nahal 1984, Bergsma 1993). There is a relationship between the peak rainfall 1982) p2 12 during the hours of intensity and the movement of the frontal å=------AFI depressions over the area. Warm front lasted four hours, with P t=1

its progress and concentration, and at 6 a.m. on the following Where AFI represents: Rainfall erosivity. day the amount of peak rainfall at a value of (4.8mm) as a PI2: quantities of monthly precipitation mm-2. result of the RUF's significant control over the area (Al-Saidi P: annual average rainfall mm-1 2014). The rain density began to decline after (5) hours, at 11 erosion intensity factor Intensity of erosion o'clock in the morning, as the amount of rainfall falling at this Less than 50 Weak hour (1, 2 mm) was reduced as a result of the warm front 50-500 Moderate retreat over the area. 500-1000 High The third theme: the geopolitical impact: The rain density More than 1000 Very high was associated with the entry of air depressions from the north and north-west, which are compatible with the erosion Table 3. Quantities of water falling for rain intensity (6- trend and the retreat of slopes at Highland in the north of the 7/9/2015) Hour Rainfall Hour Rainfall Hour Rainfall (mm h-1) (mm h-1) (mm h-1) Table 2. Rain intensity of the season (2015-2016) 13 0.97 22 3.19 5 4.09 Rain Date Water quantity Per cent Continuity 14 1.2 23 2.22 6 4.83 intensity mm h-1 hour-1 15 0.21 0 0.28 7 3.36 1 6-7/9/2015 44.9 14.2 21 18 0.85 1 1.87 8 1.17 2 2-3/10/2015 28.8 9.1 8 19 3.12 2 2.18 9 0.56 3 28/10/2015 12 3.8 14 20 4.33 3 3.84 10 0.7 4 16-17/11/2015 14.8 4.7 17 21 1.84 4 3.51 11 0.58 5 14/12/2015 9.7 3.1 12 Source: Based on http://chrsdata.eng.uci.edu/ 6 29/12/2015 6 1.9 5 Table 4. Estimates of water erosion indicators Measurement Forniya 2 Dogluas 3 Bergsem 4 measuring 7 22/2/2016 11.5 3.6 15 of the Erosion rain measuring the type of nodular 8 25-26/3/2016 10.3 3.3 18 index erosion the size of erosion the rain erosion 9 11/4/2016 22.7 7.2 10 North of 184.2 7.4 4700 Very severe 10 12/4/2016 9.7 3.1 8 Dohuk erosion range 11 16/4/2016 7.3 2.3 3 East of Dohuk 196.1 7.6 3700 Extreme Erosion Range 12 17/5/2016 7.3 2.3 8 South of 144.5 6.2 2700 High Erosion 13 16/6/2016 6.8 2.2 8 Dohuk Range Sum 216.1 68.5 162 West of 136.8 4.8 2700 High Erosion Source: Based on http://chrsdata.eng.uci.edu/ Dohuk Range 10 Ruqaya Ahmed M. Amin and Balsam Shaker Shnichal

3= S=1.65(0.03937P) 2.3/1+0.0007(0.03937P)3.3 maps i.e., slope, geological and lithological formations, land S= Erosion volume (m3 km-2 year-1) uses, density of vegetation are depicted in Figure (5-8) and P= Effective precipitation of Thornthite. Table (4-7). Accordingly other parameters are recorded in Fig. 9-12 and Table 9-11. The relationship between the lithology T otal length of gullies in the basin (m) 4. Degree of erosion= and the protection of the soil (lack of vegetation density) basin area (km2) recorded negative weak values. With increasing inclination of From the indicators adopted in the measurement of slope and decrease the soil protection, the greater of erosivity, water erosion, we conclude the following: The direct Table 6. Slope categories impact on the north and east of the region due to the rainfall No. Slop type Class (%) intensity and value of the indicators, and a strong correlation 1 Flat 0-3 of chemical weathering throughout the research area. An average relationship of freezing and thawing effect in the 2 Simple slope 12-March north of the region represented by mounts. The strong 3 Average slope 20-December relationship of rainfall and weak effect of wind can be 4 Slope 20-35 observed in general. Erosion by creep, fall and hazards of 5 Very steep 30 - More rocks and materials to the domination of general weathering, Source: Digital Space Data (DEM). (2003), p65-71 (Jack E. Ingels (Fanetti, D. which lies within the morphogenic regions (Table 5) for the and L. Vezzoli 2007) north and east of the region in the sub glacial and cold Table 7. Categories of rock resistance map for erosion territory of the rest of the parts. Class Slope type Qualitative assessment of erosion: Water erosion was 1 Solid hard rock extracted on the basis of the application of the model (PAP- 2 Rocks with moderate resistance CAR), which relies on several indicators, including rocky 3 Low resistance rocks composition, slope, vegetation condition, soil, surface formations and precipitation, as well as temperature averages, 4 Talus and gravel and this model is unique in extracting erosion. Dependence on 5 Non-resistant soil Source: Geological plate Data (NJ38-13Musel) (NJ38-9Zakio), Iraqi Geological survey and mining exploration, scale 1/250000, 1995. Kostadinov et al (2008)

Table 8. Values ​​of the coefficient of land susceptibility to erosion by slope categories and rock categories Geological and lithological formations 1 2 3 4 5 Slope 1 1 1 1 1 2 2 1 1 2 3 3 3 2 2 3 4 4 4 3 3 4 5 5 Model 1. For extraction mechanisms of the hazard map of 5 4 4 5 5 5 erosion Source: Table (4-5) (Sha'an 2013, Brady 1984)

Table 5. Morphological analysis of the research stations and the type of prevailing climate Region North of Dohuk East of Dohuk South of Dohuk West of Dohuk Climate type

The effect of natural chemical General weathering Very high Very high Very high Mediterranean region weathering together Effect of chemical weathering Strong Strong Strong Strong Mediterranean region Effect of freezing and melting Strong Medium Medium Medium Mediterranean region Effect of rain Strong Strong Strong Strong Mediterranean region Wind effect Weak Weak Weak Weak Mediterranean region Creep material Strong Strong Medium Medium Mediterranean region Dominant processes General weathering General weathering General weathering General weathering Mediterranean region Morphological regions Semi-icing Semi-icing Cool Cool Rainfall Intensity on Geomorphic Processes of Duhok Governorate 11

Fig. 1. Study area Fig. 2. Image Landsat8 2017

Fig. 3a-B-C-D1. Fig. 3. A2-B2-C2-D2. Weather maps for pressure systems Weather maps for pressure systems

Fig. 4. Form of water quantities of rain intensity (6-7 / 9/2015)

Fig. 5. Slope in degree

Table 9. Categories of soil use map Series Type of soil use 1. Pastures 2. Agricultural Land 3. Seasonal plants 4. Barren Lands 5. Cultivated land Fig. 6. Lithology Source: Digital data for the satellite Oli8 Landsat for the year 2016. 12 Ruqaya Ahmed M. Amin and Balsam Shaker Shnichal

Fig. 7. Land use Fig. 8. Density of vegetation (soil protection coefficient)

Fig. 9. Soil protection coefficient Fig. 10. Coefficient of surface susceptibility to erosion

Fig. 11. Potential erosion coefficient pap-car Table 10. Categories of vegetation density map Series Density (%) 1 20 - 2 20-50 3 50-75 4 75-100 Fig. 12. Strength of the statistical relationship with the Source: NDVI = (NIR - VIS) / (NIR + VIS) Pearson coefficient between variables Rainfall Intensity on Geomorphic Processes of Duhok Governorate 13

Table 11. Soil protection coefficient values by soil use and Table 12. Soil erosion map coefficient values vegetation density Surface susceptibility to erosion Cover density 1 2 3 4 1 2 3 4 Soil 1 5 5 4 4 protection Soil uses 1 5 5 4 4 2 5 5 4 4 2 5 5 4 4 3 3 2 1 1 3 3 2 1 1 4 4 3 2 1 4 4 3 2 1 5 5 4 3 2 5 5 4 3 2 Source: Table 4-9 (Sha'an 2013) Source: Table 4-8 (Sha'an 2013, Gallo et al 1993)

Table 13. Correlation matrix between variables in the Pap-car form Slope Rock Vegetation Soil uses Soil protection Soil susceptibility Possible density to erosion erosion

Slope 1 0.131 0.087 0.895 -0.327 0.807 0.398 Rocky 0.131 1 -.819- 0.269 -0.27 -0.271 -0.638 Plant density 0.087 -0.819 1 -0.269 0.039 0.621 0.727 Soil uses 0.895 0.269 -.269- 1 -0.331 0.505 0.23 Soil protection -0.327 -0.27 0.039 -0.331 1 -0.397 -0.516 Soil susceptibility to erosion 0.807 -0.271 0.621 0.505 -.397- 1 0.749 Possible erosion 0.398 -0.638 0.727 0.23 -.516- 0.749 1 Source: Table Data (4-9), using the spss.19 program in contrast with decrease inclination of slope the greater REFERENCES intensity of the protection coefficient, decrease erosivity has Baidaweed JMT 2003. Thunderstorms in the western Iraq plateau, Journal of the Faculty of Arts 64: 499. been reported by Zorn and Komac 2008). Bergsma EI 1982. Rainfall erosion surveys for conservation planning, Jor, ITC, Netherlands. CONCLUSIONS Brady B 1984. The nature and properties of soils. Macmillan The overall erosion of the region is moderate to weak publishing co. USA. except for the height area facing the pressure systems, which Dazi SAK 2010. Practical analysis of the Iraq climate, Baghdad, Dar Al-Farahidi Publishing and Distribution, 1, pp191. increases the precipitation. There is a relationship between Fanetti D and Vezzoli L 2007. Sediment input and evolution of the penetration of the systems and their entry times to lacustrine deltas: The Breggia and Greggio rivers case study increase rainfall. The existence of a relationship between the (Lake Como, Italy). Quaternary International 173: 113–124. Gallo KP 1993. The use of a vegetation index for assessment of the duration of the systems and their types with recurrence of urban heat island effect, International journal of remote Sensing precipitation especially the north-western sides where the 14(11): 2223-2230. processes of effective precipitation increased by Ibrahim N 1984. Water drift in the Syrian country and methods of control of soil and water conservation, Aleppo University environmental effects. The possibility of using the Journal of Research 6: 82-81. degradation protection coefficient (pop-car) for the Jack EI 2004. Landscaping: Principles and Practices, Thomson GAFRLOVEC model showed the results of the relationship learning, United State of America, New York, 6ed. between the increasing slope of the area and the rareness of Jamal S, Ali F, Abdelhamid S and Haytham M 2013. Employment of remote sensing and GIS in quantitative assessment of water vegetation density (soil protection coefficient) and weak degradation in wadi Amzaz basin (Middle Rifle) through the lithological resistance, a strong relationship with potential Gavrilovic model. Morocco Geography Journal 28(1-2): 76. erosion. Predict the rainfall that accompanies depressions, Khatib HM 2001. A comparison of the atmospheric System at a height of 500 millibar between two years of rain and dryness which is becoming more predictable, and develop periodical (1991/1992 - 1998/1999), Journal of humanities and social to protect citizens and users of land from farmers. Provide a sciences, 28, 2 pp,469. program to manage the protection of the area through the use Kinnell PIA and Risse LM 1998. USLE-M: empirical modeling rainfall, erosion through runoff and sediment concentration. Soil of the protective coefficient (vegetation) resistance erosion. Science Society of America Journal 62: 1662-1672. Use of management mechanisms in the investment of slopes Kostadinov S, Dragovic S, Zilatic M and Todosijevic M 2008. Erosion corresponds to protection and lack of degradation such as control works and the intensity of soil erosion in the upper part of the river Toplica drainage basin. Presented at XXIVth contour terraces. Conference of the Danubian countries on the Hydrological 14 Ruqaya Ahmed M. Amin and Balsam Shaker Shnichal

forecasting and Hydrological bases of water management, surface Systems affecting Iraq's climate during the rainy period Bled, Slovenia. (1950-2001), Maysan Research Journal 10: 120. Mohammed IS 1987. On the problem of erosion and soil erosion in William T translated by Wafeeq Hussain Al Khashab and Ali the Syrian coastal mountains, Kuwait Geographical Society Mohammed 1975. foundations of Geomorphology. Iraq, Bulletin, No. 98:12. university of Mosul. Dar Al Kutb for printing and publishing, pp. Naimi AS 1993. Changes in the amount of rain accompanying air 49-50. systems affecting Iraq, Al-Mustansiriya Journal 4(1): 4. Zorn M and Komac B 2008. Response of soil erosion to land use Sahar S 2000. Classification of precipitation intensity, General change with particular reference to the last 200 year (Julian organization for atmospheric and seismic monitoring, Baghdad. Alps,Western Slovenia). Presented at XXIVth Conference of the Danubian Countries on the Hydrological Forecasting and Saidi AG 2014. The pattern of control of extensible extensions of Hydrological Bases of Water Management, Bled, Slovenia.

Received 22 September, 2019; Accepted 20 November, 2019 Manuscript Number: S-244 Indian Journal of Ecology (2020) 46 Special Issue (8): 15-20 NAAS Rating: 4.96

Digital Modelling of Geomorphic Hazards due to Lunar Gravity in Lake of Mosel Dam using Remote Sensing Data and GIS

Ruqaya Amin Department of Geography and Geographic Information Systems, Faculty of Arts, Iraqia University, Baghdad, Iraq E-mail: [email protected]

Abstract: The collapses inside the closed reservoirs frequently result in a reduction in the chronological age of the reservoirs due to several factors, including geological formation, topography and climate, as well as water supplies, and monitoring the records of the daily water levels of the Mosul Lake reservoir. To raise the level of water, with a lack of interest in the dates of releases of water and reflected on the amounts of sediments due to the deterioration of the banks as a result of rising water levels within the rocks increases geomorphic hazards.GIS and Remote Sensing techniques were used to monitor the height of the levels and their compatibility with the field measurements of the reservoir of the Mosul Lake daily. A sophisticated model was made based on the integration of the databases that are depended on the Hijri calendar as a base In the follow-up of the variation of the reservoir levels and their connection with physical variables (Geology - topography - climate - soil - with the movement of the moon (Tides) and by using linear equations that show the nature of the regulation of the phenomenon in the variations of the levels with the apparent moon motion from one side and its relationship of the of the banks deterioration and increase of sediments within the reservoir on the other hand. The results indicated that in the first and third weeks of each lunar month.

Keywords: Geomorphic hazards, Closed water lakes, Remote sensing, GIS, Lake Mosel dam, Digital modelling, Hijri calendar, Tides

The Environmental planning professionals, especially in MATERIAL AND METHODS the field of the aquatic environment, encounter many The reservoir of Mosul Lake, northwest of Iraq, is located problems, including the problems of sedimentation within in the province of Nineveh, about 50 km north of the city of water reservoir and their continuous increase, which means Mosul-Iraq on the Tigris River (Fig. 1) location of study area. the continued need for dredging operations as well as the dam reservoir was built in 1983 and has a length of 3.2 km associated pollution of these sediments.It is obvious that the and a height of 131 meters (Fig. 2). Considers a largest Dam movement of tides due to the apparent moon motion has an in Iraq and the fourth largest dam in the Middle East. It is effective impact in the movement of water, and therefore the located near the Asci mousl. by examine the geological map formations of waves that hit the banks on the one hand to of northern Iraq, will find that the area falls within the affect the prevailing outcrop and therefore constitute factors formation of the lower fars, fatha formation (Fig. 3), which of rocks weakness on the other, all these with the rise and consists of limestone,siltstone in addition to the anhydrite decline of water levels inside the lake coincide with the Moon and saline materials. Due to the flood of rain water and motion, which has been shown to have a significant impact surface water on the surface of Iraq throughout the Eocene on the increase of deterioration of the lake's banks to form era (cainozoic) was composed of sediments, mostly growing sediments,which will reducing the reservoir's limestone, anhydrite, saltstone and limestone (Kelly and temporal life and reducing its filling and quality efficiency.The Wakely 2007). The brown soils with different deeps problem of the study is that the increase of sediments due to prevailing the lake sides, which are mixed with gypsum, mud, the rise of the water level and the decrease due to the water sand and gravel, whose origins are mostly due to the intake of the rivers entering the water reservoir increases the formation of the dominant fatha formation in the region (Fig. deterioration of the banks due to the permeate of water in the 4). rock outcrop of the lake and leading up to destruction inside The maximum temperature in the summer months, the lake so reduces the temporal age of the reservoir and where the average in July (33.8 degrees Celsius), and the pollution within the water lake. lowest in the winter months - especially in the month of The aim of the research is to identify the impact of the January (7.4 Celsius), the region receives the average moon motion (tidal) and connect it to the time and place of rainfall of 365 millimeters in the cold season Of the year. The increasing the sediments. Research methods (regional, concentration of rain, 49.6% in the spring and 15.9% in the historical and analytical) as well as quantitative methods and fall, and the rainfalls on the region generally varies in modern techniques in digital analysis of space data and GIS patterns, but it is mostly dominated by cyclonic, orographic were used. convective inversion rainfalls, and the cyclonic rainfalls are 16 Ruqaya Amin the predominant rains, mainly its mid-latitude cyclones which forming in the northern parts of the Atlantic ocean (Al- Shalash1972). Moon movement (Tides): Tides emerges by moon and sun gravity for water of seas and oceans, Because the moon is closer to the Earth, so the impact of its gravity is greater, despite its small size, conclude that the moon's gravity is the most important factor in the occurrence of tides, but another factor is the centrifugal force resulting from the rotation of the earth around itself. Tides occur twice every day "every 12 hours" because the parts of the Earth's surface pass during its rotation in front of the moon and the tide occurs in the Fig. 1. Location of the study area (General Survey, Iraq places facing the moon, and soon the ebb occur when these Topographic Map, 2007, Google earth) places move away and the rise of the tide varies according to the location of the moon in orbit for each of Earth and sun. Here, it can be emphasized that the water of closed reservoir and lakes is not static but in constant motion, known as wave movement. It is a water disturbance caused by the movement of its molecules up and down in a regular harmonic movement. waves are often created by winds, storms and what comes from a water into the lake, thus forming isolated solitude waves, in shallow and confined water (Le Provost 1995). subsequently, there is a close relationship between the tide and the phases of the moon. The fact of this phenomenon was understood after the discovery of Newton's law of gravity. Scientists attributed it to Fig. 2. Topography of area (Digital Elevation Data differences in the gravitational forces between the sun and www.uweb.ucsb.edu/~nico/comp/get_dem.htm) the moon on land and the water bodies on Earth (Lambeck 1977). Various technologies have been developed for the use of tidal phenomena in the generation and commercial use of electric power. The monitoring of tidal phenomena and the use of arithmetic tables can help to avoid the dangerous caused by them. Techniques used: Rapid eye European satellite data were used with a modified spatial accuracy reach to 5 m 30/1/2014. As well as a comparison of the changes in the level for temporal duration of the data of the US satellite Landsat TM for the years 2010-2011, 2012-2014 with an improved spatial accuracy of 14.25 meters. Digital elevation data (DEM) to derive heights and depths of the reservoir, as well as the recording of lake levels and matching height of the Fig. 3. The prevailing geological outcrops of the region levels with heights in the reservoir (Dagestani 2003). (Ministry of Industry and Minerals, Mineral Survey Using ArcMap 10.2 to make applications and derivatives and Inspection Authority, Baghdad, 2009) concerning with the research and to build a geographic information database with the use of extension spatial environmental degradation of the reservoir through the analysis tools and Geospatial Wizard to analyze digital aggregation of weights Sum for inputs (environmental matrices and design spatial maps of the region. As well as the elements include lithology, soil, climate, topography, levels) use of Overlay in the preparation of weights Weighting and (Saleh 2010). the formation of a weight matrix to detect the amount of SPSS program to build a digital matrix based on the Hijri Lunar Gravity Evaluation by Remote Sensing Data and GIS 17 date of the studied time period, and extract the linear and most of its appear at the south of the lake. Table 3 shows equation between the variables (water intake, higher level, the validity of the model between the levels in the matrix of the lower level) and its correlation with the regular distribution reservoir levels recorded during the Gregorian calendar year and timing of the movement of the moon and its impact in with the space data and their coincidence with the phases of tides and its reflecting its effect on the deterioration of banks the moon in terms of the first week increasing in the and increase sediment and reduce life age for reservoir attractions and its appear a real increase in the level then (Saleh1992). decline in the second week and so on, as explained previously. RESULTS AND DISCUSSION Quantitative analysis of the incoming and levels of the Lake of Mosul reservoir reveals the highest rate of water incoming during the days of the Hijri calendar from 5-10 of each month and due to rise from 17-21 for the period 2010- 2014 as in (Fig. 5). The linear equation shows the amount of regularity and the absence of it during the movement of the waves daily for the studied period, and this is reflected also at the highest level (Fig. 6) and the lowest level (Fig. 7). Figure 8 shows the fluctuation of the lake's difference between the highest and lowest levels and its decline below the average in the days 1-3, at the beginning of the Fig. 4. The prevailing soils of the region (Buringh 1960) appearance of the moon and the days 26-28 again, at the end of the month and the disappearance of the moon. Which means that the level is directly affected by moon depends upon the linear equation. Note the variation in the level during the first and last quadrature, which indicates the great impact of the moon during this period. The temporal and spatial analysis of the reservoir levels of Mosel Lake: Remote sensing is one of the means used to detect the difference in water level. Here, a model was taken for the lake at the year 2013 with dates for the Gregorian year (6/6, 8/7, 9/8, 26/9/19/10) corresponding to the Hijri year 1434 Jumada Al-Akhra 24, Rajab 26, Shaaban 28, Shawwal 16, Zul-Qa'da 8), respectively. And processing them by the method of the supervised classification and the statistical Fig. 5. Average of daily water incoming m3/s of Mousel lake processing (Maximum Likelihood Classification) and the reservoir for the period 2010-2014 (Based on Table 1) extraction of results after the conversion of the file from the raster to the vector with an areal and the construction of a temporal and spatial database showing the change in the level and its relationship with the phases of moon in a digital matrix, (Table 2) show the date of the space data and the level of the lake are shown in the Hijri calendar and had conformity with the matrix in (Table 1), (Fig. 10 and Fig. 11) clarify The variation in the level affects the size of the sediments temporally and spatially its appear southern end of the lake at the body of the dam. Consider the shallow areas within the lake is the basis in comparing the difference in the level.The gradient according to the levels in the shallow areas is shown to be in October with a low level and a low level of water income to reach an Fig. 6. Average of maximal daily level for Al-Mosel lake area of (13.329 km2) and a rate of 4.95% of the area of lake reservoir for period 2010-2014 (Based on Table 1) 18 Ruqaya Amin

Fig. 7. Average of minimum daily level for Al-Mosel lake reservoir 2010-2014 (Based on Table 1)

Fig. 8. Moon phases Fig. 11. Sedimentary deterioration inside the dam of Mosel dam (Landsat OLI8 for 2013)

Geomorphic hazards by differing levels (Moon movement): As discussed previously that the apparent movement of the moon acts to raise the level of water, as well as the irregular movement of the waves that hit the banks. It works on the wetness of the prevailing outcrop. The solubility in the soluble rocks due to the permeation of the water and its continuous variation in the level on limestone's and the rocks of evaporators such as gypsum, salt, and anhydrite Which leads to the formation of karstic phenomena (terrestrial Fig. 9. Average of daily differences level for Al-Mosel lake cavities) known as land subsidence, on the one hand, as well reservoir 2010-2014 (Based on Table 1) as leakage of rain water in the streams of surface water networks that pour into the reservoir during the seasons of precipitation (Mahsoub 1991). It is well known that rainwater is a weak acid solution, with a great ability to melt calcareous rocks. The acidity of the water increases when it leaks into the soil. practically since a large part of the organic matter present in the soil melts into these water during leakage. landforms resulting from the effect of groundwater appear to form the topography of karst, one of the most important conditions for the formation of karstic landforms (Abul-Enein 1981). Availability of soluble rock and solubility preferably of limestone. The rock should be dense and rich with fissures to facilitate the work of water permeation in the rock.The Fig. 10. Tides level at sourthern Mosel dam lake (Table (1) availability of heavy or medium rain containing different data and DEM) acids, as well as a relatively warm atmosphere.The most Lunar Gravity Evaluation by Remote Sensing Data and GIS 19

Table 1. The rates of water incoming and the highest and important landforms characteristic of the topography of the lowest level in the reservoir of Mosel Lake for the karst (Al-Banna 2000) are: (sinkhole or doline- caves and period 2010-2014 (Ministry of Water Resources, Dams and Reservoirs Authority, Mosel Dam tunnels - lost rivers - solution valleys), and through Management Authority, unpublished data, 2010- laboratory tests of sandy soils and calcareous soils its have 2014) high permeability (Kelley and Wakeley 2007). Sandy soil also Level Average of Average of Average of Days has large spaces Between sand grains, so high permeability variance cm lowest level highest level incoming water cm cm m3s-1 for rocks can be easily permeate within the dominant -0.29268 252.95 310.8915 509.6981 1 formations of Area (Lillesand and Kiefer 2004), as well as the -1.04878 252.94 310.9096 497.4528 2 availability of calcareous soils that containing fissures and 0.243902 252.94 310.9315 512.566 3 voids due to the melting of rock by water, it can be said that 0.878049 252.93 310.9245 549.1887 4 the sandy soil is highly porosity, so the permeation of water is 0.243902 252.97 310.9413 545.6604 5 fast. The clay soil have minute porous, has a large surface -0.34146 253.06 310.9047 549.8113 6 area, absorbs water, and the rocky structure of the soil 0.536585 252.95 310.9019 543.434 7 becomes impermeable, so permeation is very little.The 0.682927 252.99 310.8908 549.7358 8 calcareous soils have wide cracks due to their response to 2.04878 253 310.9296 542.0943 9 the solution and therefore the penetration of water is 1.121951 252.78 310.9417 524.283 10 quickly.Thus, the process of dissolving the rock of origin and 2.536585 252.83 310.9283 503.3585 11 formation of subsurface cavities is done by dissolving 0.804878 252.91 310.927 479.434 12 limestone, dolomite, gypsum, anhydrite, and salt rocks, the 0.95122 252.89 310.9017 477.2264 13 dominant outcrop of the reservoir when exposed to incoming 3 252.89 310.8509 509.5849 14 water which saturated with carbon dioxide and some acids, 1.658537 252.89 310.9085 498.1698 15 resulting in the permeation of water into rock Separated 1.121951 252.84 310.8885 520.1887 16 granules and dissolves the soluble part,This situation is 1.365854 252.89 310.89 586.0566 17 observedat the southern part near the body of the dam and 0.121951 252.88 310.8975 573.9811 18 the most noticeable effect is observed in (Fig. 8), since the 1.902439 252.84 310.9208 563.6792 19 bottom of the lake is the most receiving for sediments from 2.243902 252.86 310.9347 524.8679 20 one hand and the lowest elevation part, which increased the 1.634146 252.97 310.9355 509.1509 21 weakness of the rocks and break them and activated the 0.292683 252.91 310.9289 509.6226 22 0.780488 252.96 310.9358 510.5849 23 Table 3. Level of the Lake of Mosul Dam Year 2013 (Ministry 0.097561 252.81 310.943 509.5283 24 of Water Resources, Damas and Reservoirs 2.439024 252.96 310.9466 555.9811 25 Authority, Mosul Dam Management Authority, 1.7 252.85 310.9487 529.3208 26 unpublished data, 2010-2014) -0.34146 252.81 310.9613 498.0943 27 June-06 July-08 August-9 Sptember-26 October-19 -0.21951 252.88 310.9762 490.8491 28 318.03 318.91 318.36 314.85 312.19 0.5 252.91 310.9987 491.1698 29 24 Jamadi alakhar 26 28 16 Shawwal 8 Dhu al- 2.45 252.81 311.1008 532.5556 30 Rajab Shaban Qa'da

Table 2. Area depth of the lake of Mosul dam year 2013 area km2 Landsat Operational Land Imager (OLI 8) 2013 (6/6, 8/7, 9/8, 26/919/10) Month June-06 July-08 August-9 Sptember-26 October-19 Type depth 318.03 318.91 318.36 314.85 312.19 Level (m) Very deep 82.246 77.0176 105.7176 118.3518 137.1413 Deep 83.5346 84.7861 82.3122 79.929 61.6062 Moderate depth 64.1856 62.6077 42.3576 42.6996 34.5154 Low depth 27.6606 32.6295 24.9039 20.6442 22.5873 Shallow depth 11.5524 12.1383 13.8879 7.5546 13.329 Sum 269.1792 269.1792 269.1792 269.1792 269.1792 20 Ruqaya Amin

pressure for the phenomenon movement of the Moon, with different level had active effects on the environmental degradation of strategic construction as closed water reservoirs for their correlation with economic activities (electric power generation, agriculture, industry, daily water use by population, tourism, etc.).Preparation of temporally tables that calculate the quantities of releasing water incoming for closed reservoirs for Tigris River with the days of the movement of the Moon to reduce the difference levels and avoid the raising of water and reduce its movements inside the reservoir and thus reduce pressure on the banks, which willincreases the efficiency of the reservoir for a longer period on the one hand and reduce pollution by dissolving limestone On the other hand. Trying to control the water networks which its water ends in the reservoir, especially Fig. 12. 3D model of landsat satellite data at lake of Mosel during periods of rainfall in the region to avoid sediments due dam (Landsat OLI8 for 2013) to the flash floods caused by the orographic rain that prevails in the region. factors of mechanical erosion (Mahsoub and Radi1989), but the presence of water is the main factor in the formation of REFERENCES this phenomenon and the increase in sizes, and over time its Abul-Enein HS 1981. Principle of Geomorphology - Study of the Earth's Surface relief Forms. 6th ed. University House. Beirut. may collapse and fall of these rocks when increasing the p. 580. loads overlying it either being natural or human causes to Al-Banna A 2000. Environmental Problems and Conservation of forms Sediments and destruction within the body of the Natural Resources - Study Models in Applied Geography. Arab Thought House, Cairo. p. 94. reservoir in different sizes of soluble materials causes water Al-Shalash AH 1972. The use of some mathematical criteria in pollution and the variation of solid sediments to make piles in determining climate regions in Iraq. Journal of the Faculty of front of the dam increase the pressure, as well as reduce the Arts. Riyadh University. Allen PA and Allin JR 2005. Basin Analysis, principles and volume of water storage, and all this to reduce the temporally applications. 2nd ed. Blackwell publishing. p.63. age and increase environmental degradation,in addition its Buringh P 1960. Soil and soil conditions in Iraq. Republic of Iraq. an economics wasting In all activities ranked in such like this Ministry of Agriculture. p. 308. vital strategic project (Allen and Allin 2005). Dagestani NS 2003. Remote Sensing: Fundamentals and Applications, Dar Al-Manaheg, Amman, Jordan, p.86. Kelley JR and Wakeley LD 2007. Geologic Setting of Mosul Dam and CONCLUSIONS Its Engineering Implications. U.S. Army Engineer District. Gulf The movement of the moon (tides) has a direct effect on Region. Baghdad. Iraq. pp. 38-39. the difference in the level of the closed reservoir.There is a LambeckK 1977. Tidal Dissipation in the Oceans: Astronomical, Geophysical and Oceanographic Consequences. relationship between the date of the release of water incomes Philosophical Transactions for the Royal Society of London. within the reservoir and the movement of the moon, which Series A. Mathematical and Physical Sciences. pp. 287. increases the deterioration of the banks and increase the Le Provost C, Bennett AF and Cartwright DE 1995. Ocean tides for and from TOPEX/POSEIDON. Science 267 (5198): 639. contaminated sediments on the one hand by dissolving Lillesand TM,Kiefer RW and Chipman JW 2004. Remote Sensing limestone and saltrocks, and increase the solid sediments of and Image Interpretation. 5th ed. John Wiley& Sons. New York. different sizes.The relationship between the type and Mahsoub MS 1991. Geomorphology of coasts. Dar Al-Thaqafa for Publishing and Distribution. Cairo. p. 52. porosity and composition of rocks and the quantities and type Mahsoub MS and Radi MD 1989. Geomorphological processes. of sediments inside the reservoir. The variation in the level Arab Culture House for Publishing and Distribution. Cairo. p.26. affects the size of the sediments temporally and spatially and Saleh MA 1992. Remote sensing image: data collection and its appear in the south end of the lake close to the body of the analysis. Research Center. Faculty of Arts, King Saud University. Riyadh. 27: 117. dam.Considering the shallow areas within the lake is the Saleh MA 2010. Processing digital remote sensing images using the basis in comparing the difference in the level.Imperceptible ILWIS program. Riyadh. (E-book online).

Received 24 September, 2019; Accepted 21 November, 2019 Manuscript Number: S-245 Indian Journal of Ecology (2020) 46 Special Issue (8): 21-27 NAAS Rating: 4.96

Effect of Supplementation with High Levels of Phytase Enzyme Compared with Commercial Dose in Corn-Soybean based Diets on Productive and Physiological Performance in Broiler

Husam H. Nafea, Firas M. Hussein1 and Bahaa I. Hamid2 1College of Agriculture, University of Anbar, Anbar, Iraq 2Office of Agricultural Research, Ministry of Agriculture, Baghdad, Iraq E-mail: [email protected]

Abstract: The experiment was conducted at the Poultry Research Station of the Livestock Research Department/ Agricultural research office/ Ministry of Agriculture (Abu Ghraib), during the period from 20 September 2017 to 31 October 2017. 300 broilers male of the Ross 308 species were used in this study, which were distributed randomly in 5 treatments with 3 replicates for each treatment, where each replicates consist of 20 chicks. They were fed on diets equal in the percentage of protein and represented energy, according to the age requirements that mentioned in the Ross 308 manual. The treatments included a positive control treatment with nutrient requirement diets, negative control treatment (with lack of available phosphorus diet 0.16: AP),the negative control treatment with the addition of 600 FTU kg-1 of the phytase enzyme, the negative control treatment with the addition of 1600 FTU kg-1 of the phytase enzyme and the negative control treatment with the addition of 2600 FTU kg-1 of the phytase enzyme. The results of this study showed a significant increase in the body weight rate and the rate of increasing in weight of PC treatment compared with the NC treatment. In addition to significant increase was observed in the mentioned traits of the additional treatment of 600 FTU kg-1 compared with the NC treatment, while it reaches to approximate level to the PC treatment for the addition of 1600 FTU kg-1. Finally, the additional 2600 FTU kg-1 treatment exceeded the other experimental treatments. The results of the study also showed a significant increase in the rate feed consumption ratio (FCR) for the PC treatment compared with the NC treatment. The results also showed a significant deterioration in the feed conversion ratio FCR for the NC treatment compared with the PC treatment. The two additional treatments 1600 and 2600 FTU kg-1 of phytase enzyme to the negative control treatment enhanced the FCR significantly to an approximate level of the PC treatment, and did not differ significantly from the PC treatment. There was a significant increase regarding the phosphorus level in the blood, a significant decrease was noted in the NC treatment compared with the PC treatment, with a significant increase in the phosphorus level at the blood for the two additional treatments of 1600 and 2600 FTU kg-1 of the enzyme to an approximate level of the PC treatment, and a significant increase for the additional treatment 2600 compared to 1600 FTU kg-1. Finally, there were no side effects for the adding the enzyme in the production and physiological broiler performance.

Keywords: Phytase enzyme, Broilers, Protein, Phosphorus

Phosphorus is an important nutritional element in animal et al 2016). Therefore, the improvement in the digested feed, which plays an important role in the energy metabolism, susceptibility, phosphorus, and the following nutrients (Ca, bone development, and cell building (Tayyab and Mc Lean Na, Mg, Cu, Fe, Zn, starch, protein and fat) absorption in 2015). Global reserves of phosphate rock are expected to be these plant sources has been of great interest, because it is fully used during the 50-100 years, as a result, phosphorus represent the most important and major poultry diets, in prices have risen, causing a high cost of poultry diets addition to the phosphorus importance and correlation of with production (Shastak et al 2012, Rodehutscord and Shastak economic and environmental conditions (Sommerfeld et al 2013). Approximately 60-90% of the total phosphorus found 2018). The aim of this study was to find out the effect of in feedstock's of plant origin that used in poultry diets in the adding two high levels, in addition to the traditional level of form of phytic acid (IP-6 Inositol Hexaphosphate), while its phytase enzyme that produced from Buttiauxella bacteria to mixed salts are called phytate or phytine (Rodehutscord et al maize-soybean meal diets. Moreover, determine the best 2016, Adeola 2018). The using of phosphoric phytate in level of addition, and their impact on the productive and monogastric animals, including poultry are low by 40% due to physiological performance of broiler. the lack of the phytase enzyme that necessary for phytite decomposition Phytic acid also plays an anti-nutritional role MATERIAL AND METHODS through chelate correlation with divalent, trivalent metal ions The study was conducted at the Poultry Research positively charged, and the formation of non-soluble Station of the Livestock Research Department Ministry of complexes with starch, fats and proteins, and the inhibition of Agriculture Abu-Ghraib, during September - October 2017. enzyme efficacy (Pirgozliev et al 2012, Lee et al 2015, Singh Broilers male of the Ross species were used in this study at 22 Husam H. Nafea, Firas M. Hussein and Bahaa I. Hamid the age of one day at an initial weight of 40 g bird-1. The study the temperature stability up to 95ºC. This characteristic was included 5 treatments with 3 replicate, where the single very important during the process of making pellet and this replicate contains 20 chicks that distributed randomly to the enzyme has relatively high efficiency in low pH (2.5-5.5), replicates after the weighing of 20 birds group-1. The chicks which allows it to operate at the beginning of the digestive were breeder in a semi-enclosed breeding halls, where the tract. hall was divided into room (pen) with a dimension of 2 x 3 meters, representing one of the experiment's replicate. The RESULTS AND DISCUSSION treatments of the experiment were: a positive control Productive Traits treatment with full nutrient requirements diets without Average live body weight: The body weight 3-week after enzyme addition, a negative control treatment with diet of treatment was significantly higher T5 (negative control NC + 0.16in available phosphorus compared to the positive control 2600 FTU Kg-1, 774.02 g) than the rest of treatments (Table -1 diet and without enzyme addition. Furthermore, the third 2). However, it was at par with NC +1600 FTUKg (T4). The fourth and fifth treatments were negative control with the body weight was significantly higher in T5 than in other addition of 600, 1600 and 2600 FTU of phytase enzyme treatments followed by T4, T1, T3, T2. There was no significant

Standard procedure was followed for conduct of experiment. differences inT4 and T1. Moreover, a continuous lighting system was used during the Average weight gain: The results of the study that listed in first seven days of bird life and then it was reduced to 20 Table 3 showed a significant increase in the average weight hours day-1, where the lighting was cut for 2 hours every 12 gain of PC by (638.40 g) for the period 1-3 weeks, compared hours using electronic hours from 8 to 39 days of age, while to the negative control treatment NC with (474.82 g). The the feed and water were provided for free during the addition of phytase at levels 1600, and 2600 FTU Kg-1 to NC experimental period. The statistical analysis was carried out treatment has led a significant increase in the average with software SAS, version (SAS 2012). The significant weight gain which reached 570.40, 666.80 g, respectively, differences between the average traits were tested using the and at a closer level of PC treatment with a significant polynomial Duncan test). The method of enzymatic analysis superiority of additional treatment 1600 FTU Kg-1 Compared of glucose was adopted according to (Asatoor and King to 600 FTU Kg-1. A significant increase was observed in the 1954). The cholesterol level was estimated according to the average weight gain of the additional treatment 2600 FTU existing instructions in kit work equipment according to Kg-1 by (733.85) compared to the two positive and negative Richmond (1973) method, and the three-fat concentration in control treatments with or without the addition of 600 FTU Kg- blood plasma was estimated according to (Toro and 1. Similarly, in the period from 4 to 6 weeks, the increase in the Ackerman 1975) method, using the analytical equipment kit, average weight gain of PC treatment was (1528.33 g) that manufactured by Biolabo-Frans company, whichis an compared to NC treatment that achieved (968.22 g). The enzymatic method that reads the Absorption with addition of phytase at levels 600, 1600 FTU Kg-1 to the NC Spectrophotometer at a wave length of 546 Nano-meter. The treatment has led to a significant increase, which reached concentration of total proteins in blood plasma was 1294.67, 1591.93 g, respectively, with a significant decrease determined using the same method and technique that in the average weight gain of additional treatment 600 FTU adopted in three-fat concentration in blood plasma. The Kg-1 compared to the PC treatment and there is no significant Biuret method was used to measure the protein amount in differences between the additional treatment 1600 FTU Kg-1 blood plasma (Varley et al 1980), while the phosphorus and PC treatment. A significant increase was observed in the concentration in blood plasma was measured by (Fisk and average weight gain of additional treatment of 2600 FTU kg-1 Subbarow 1925) method. The activity of the Alkaline (1803.92) compared with the two positive and negative Phosphatases ALP enzyme was also evaluated using the control treatments with or without addition of 600 FTU Kg-1. analytical equipment kit manufactured by Biomerieux- For the total average weight gain, in the period from 1 to 6 France as mentioned previously, according to (King and weeks, PC treatment had the superiority by (2166.87 g) over Armstrong 1943) method. The bacterial phytic enzyme E. the NC treatment with (1443.1 g), while the addition of coli, that named commercially as PHY @ 10000 Axtra was phytase at levels 1600, 2600 FTU kg-1to the NC treatment obtained from the (Danisco UK LTD) company that located in has led to a significant increase in the average weight gain by Amman /Irbid. 1865.17 and 2258.87, respectively. A significant decrease in This enzyme was a product of Buttiauxella bacteria the average weight gain of additional treatment 600 reflecting as Trichoderma Reesei fungi, manufactured as compared to the PC treatment and there were no significant spherical fine grains and the advantages of this product was differences between the additional treatments 1600 FTU Kg-1 Effects of High and Commercial Phytase Doses on Broiler Performance 23 and PC treatment. Furthermore a significant increase was based diets observed a significant increase in the PC observed in the average weight gain of additional treatment treatment over the NC treatments (NC Ap: 0.35, and NC1 Ap1: of 2600 FTU Kg-1 (2537.93) compared with the two positive 0.29) with a significant improvement in weight gain by adding and negative control treatments with or without the addition of the phytase enzyme ( 1600 FTU) to the negative control NC -1 600, 1600 FTU Kg . The results were present a good and NC1. agreement with Gautier et al (2018) with corn- soybeans Pieniazek et al (2017), for the duration of (1-42) days,

Table 1. Feed ingredients and chemical structure calculated for the maize diet that used in the experiment Material Starter diets (1-10) day Growth diets (11-21) day Final diets (22-42) day

PC NC PC NC PC NC

Maize 58 58.5 59.4 62.1 61.4 62.1 soybeans based 35 34.9 32.57 32.4 29.75 29.65 soybean meal diets 1 1 1 1 1 1 Metal and vitamins mix 2.35 2.1 3.65 3.45 4.8 4.52 Vegetative oil 58 58.5 59.4 62.1 61.4 62.1

CaHPO4 2 1.1 1.7 0.82 1.55 0.65 Limestone 1 1.56 0.95 1.5 0.85 1.43 Methionine 0.27 0.26 0.23 0.23 0.19 0.18 Lysine 0.28 0.28 0.2 0.2 0.16 0.17 Salt 0.3 0.3 0.3 0.3 0.3 0.3 Total 100% 100% 100% 100% 100% 100% Crude protein 21.71 21.72 20.68 20.66 19.5 19.51 ME 3002 3000 3113 3114 3215 3211 Meth + Cys 1.05 1.05 0.99 0.99 0.91 0.91 Lys 1.4 1.4 1.27 1.27 1.17 1.17 Ca 0.97 0.97 0.87 0.87 0.79 0.79 P 0.48 0.32 0.43 0.27 0.39 0.23 * The minerals and vitamins mixture used in the manufacture of diets Broiler Premix 1% produced by Intraco LTD Company, the Belgian is free of calcium and phosphorus. ** Soybeans to use from an Argentine source of crude protein percentage of 48%, the energy represented 2440 Kcal/ kg *** Chemical composition of the rest of the feedstock's based on (NRC 1994).

Table 2. Effect of addition phytase enzyme on average live body weight (g) Weeks Treatments

Positive control Negative control (T2) Negative control+600 FTU Negative control+1600 Negative control+2600 -1 -1 -1 (T1) kg (T3) FTU kg (T4) FTU kg (T5)

3 bc678.53 ± 32.27 514.88 d ± 14.26 610.50 ±5.01 c ab706.93 ± 31.16 a774.02 ± 29.89 6 b2206.81 ± 65.92 d1483.10 ± 23.11 c1905.17 ± 116.16 b2298.87 ± 115.57 a2577.93 ± 78.84 * Values ​​represent the average ± standard error. ** Samel letters d within one row indicate a non-significant difference (P≤0.05)

Table 3. Effect of addition at different levels of phytase enzyme an average weight gain (g) Weeks Treatments Positive control Negative control Negative control+600 Negative control+1600 FTU Negative control+2600 FTU kg-1 kg-1 FTU kg-1

3-Jan bc638.40 ± 32.36 d474.82 ± 14.24 c570.40 ± 4.97 ab666.80 ± 31.24 a733.85 ± 29.90 6-Mar b1528.33 ± 40.67 d968.22 ± 16.73 c1294.67 ± 113.00 ab1591.93 ± 90.20 a1803.92 ± 56.56 6-Jan b2166.87 ± 65.92 d1443.10 ± 23.11 c1865.17 ± 116.16 b2258.87 ± 115.57 a2537.93 ± 78.84 * Values ​​represent the average ± standard error. ** Small letters varied within one row indicate that there are a significant difference between treatments, at a significant level (P≤0.05). 24 Husam H. Nafea, Firas M. Hussein and Bahaa I. Hamid observed a significant decrease in the average body weight treatment by 3406.87, 3926.71, 4335.05 g, respectively, for NC treatment (- 0.17%) compared to the PC treatment which did not differ significantly from PC treatment. These which full phosphorus requirements. The addition of phytase results were agreed with Pieniazek et al 2017 for the period of enzyme has led to a significant increase compared to the NC (1-42 days), which a significant increase was observed in the treatment and adding of 2000 FTU to NC had led to feed consumption ratio of the PC treatment compared to the significant improvement) with similar levels with a PC. Walk NC diet decreasing of 0.17 ap. The addition of phytase et al (2018) also observed significant linear increase (in the enzyme at the level of 500 and 2000 FTU Kg-1 to the NC daily weight gain with the increasing of additional of phytase treatment has led to a significant increase in feed enzyme levels) to control diets. Taheri et al (2015) also consumption ratio compared to the NC treatment. The study reported similar results. The results were not Momeneh et al results were also agreed with Walk et al (2014) findings, (2018) observed no significant difference in the average body where a significant increase was noted in the feed weight and the weight gain increase of the broiler at the consumption ratio of the PC treatment compared to the NC addition of phytase enzyme at level upto 2500 FTU Kg-1 in full treatment. The addition of phytase enzyme at levels (1000, dietary requirements diet. 2000, 4000) to the NC treatment has increased significantly Feed consumption ratio: The results in Table 4 showed a with the feed consumption ratio compared to NC treatment to significant increase in the feed consumption ratio (FI) of the a closer level of the PC. The results were not agreed with PC treatment (879.25 g) for 1-3 weeks, compared with NC Walk et al (2018), there were no significant differences in the treatment by (739.02 g). There was no significant difference feed consumption ratio between PC, NC and NC1 with the between the additional treatment of 600 FTU kg-1 (811.98) addition of the enzyme at different levels. As well as, the and the two negative and positive control treatments, while results were not agreed with Momeneh et al (2018) findings the addition of phytase enzyme at levels 1600/2600 FTU kg- for the period from (1 to 28 days),which noted that there no 1to NC treatment has led to a significant increase in feed significant difference in the feed consumption ratio, when consumption ratio which reached (903.66, 986.93 g) adding the phytase enzyme at level 0, 500, 2500 positive respectively, which did not differ significantly between them control diet. or in comparison with PC treatment, a significant increase for Feed conversion coefficient (FCC): The results in Table 5 the additional treatment 2600 compared to the additional showed a significant deterioration in the feed conversion treatment 600 FTU kg-1. As well as a significant increase was coefficient FCC for the NC treatment by (1.56) for the period observed in the feed consumption ratio of the PC treatment 1-3 weeks compared to the PC treatment by (1.38). The by (2899.79 g) for the period of 4-6 weeks compared to the addition of phytase enzyme at levels 600, 1600, 2600 FTU NC treatment of 2001.27 g. The addition of phytase enzyme Kg-1 to NC treatment has improved significantly the FCC, at levels 600, 1600 and 2600 FTU kg-1 to NC treatment has led which reached 1.42, 1.36.1.34, the additional treatment 1600 to a significant increase with 2594.88, 3023.05 and and 2,600 did not differ significantly from PC treatment, with 3348.12 g, respectively, and did not differ significantly from an observation of a significant deterioration. in the FCC of the the PC treatment, a significant increase in the additional additional treatment 600 compared to the PC treatment. The treatment of 2600 compared to the additional treatment of results were similar for the period of 4-6 week, with a 600 FTU kg-1. While in the feed consumption ratio during the significant deterioration was observed in the FCC of NC experiment period (1-6 weeks), The PC treatment (3779.71 treatment (1.90) compared to PC treatment (1.74). The g) was superiority compared to NC treatment by 2740.29 g, addition of phytase at levels 600, 1600 for NC treatment has with a significant improvement with the addition of phytase significantly improved the FCC which reached 1.82, 1.74 and enzyme at levels 600, 1600 and 2600 FTU kg-1 for NC 1.71, respectively. The two additional treatments of 1600 and

Table 4. Effect of addition at different levels of phytase enzyme in Feed consumption ratio (g) Weeks Treatments Positive control Negative control Negative control+600 Negative control+1600 Negative control+2600 FTU / Kg FTU / Kg FTU / Kg

3-Jan ab879.25 ± 38.12 c739.02 ± 18.49 bc811.98 ± 8.15 ab903.66 ± 37.13 a986.93 ± 48.66 6-Mar ab2899.79 ± 90.20 c2001.27 ± 17.39 b2594.88 ± 249.37 ab3023.05 ± 205.02 a3348.12 ± 101.23 6-Jan ab3779.71 ± 117.79 c2740.29 ± 20.90 b3406.87 ± 253.86 ab3926.71 ± 230.18 a4335.05 ± 136.86 * Values ​​represent the average ± standard error. ** Small letters varied within one row indicate that there are a significant difference between treatments, at a significant level (P≤0.05). Effects of High and Commercial Phytase Doses on Broiler Performance 25

2600 did not differ significantly from the PC treatment, with an observed in mortality percentage of NC treatment (decrease observation of a significant deterioration in the FCC of the NPP 0.17) compared to the PC treatment and the addition of additional treatment 600 compared to the PC treatment. The phytase at levels 500, 2000 FTU kg-1 with no significant results present a good agreement with Pieniazek et al (2017) differences between the addition treatments and PC findings in a significant deterioration of the FCC in the NC treatment. The results were also not agreed with Liu et al treatment compared to the full requirements of PC treatment. (2016). There was a significant increase was observed in the The addition of the phytase enzyme by 500, 2000 has led to a mortality percentage in the NC treatment (aP, 0.18) significantly improved compared to the NC treatment and did compared to positive control PC (aP, 0.48). not differ significantly from the PC treatment. Beeson et al Physiological Characters (2017) also pointed a significant improvement in the FCC in The results in Table 7 showed no significant differences the full requirements PC treatment compared to the NC in the concentration of Glucose, Cholesterol and Triglyceride treatment. The addition of the phytase enzyme by 500, 1500 in blood between the experimental treatments, a significant -1 FTU kg to the NC treatment resulted in a significant increase was observed at the total protein level of the PC improvement in the FCC. The study findings also agreed with treatment (3.67) compared to NC treatment (3.10), with no Taheri and Taherkhani (2015), where a significant significant differences in the addition of enzyme at levels 600, degradation was observed in the FCC of the NC treatment 1600, 2600 FTU kg-1 to NC treatment (3.3, 3.3, 3.47, (decrease Npp = 0.2) compared to PC treatment. The respectively), compared to positive and negative control addition of the phytase enzyme at levels (1000, 2000, 4000) treatments. While a significant decrease (P <0.05) was to the NC treatment was significantly improved of the FCC observed in the phosphorus concentration in blood serum compared to the NC treatment without addition, and there for NC treatment (4.03) compared to PC treatment (5.4), the was no differences between the additional and PC adding of 600, 1600, 2600 FTU kg-1 to NC treatment has led to treatments. The results did not agree with Walk (2018) and a significant increase in the blood phosphorus concentration Truong et al (2017) and Wu et al (2015).There were no by (4.53, 5.37 and 5.73) respectively, with a significant significant differences in the FCC between experimental decrease (P <0.05) for the additional treatment of 600 FTU treatments when adding the enzyme. kg-1 compared to the PC treatment, which did not differ Mortality: Table 6 shows there was no significant differences significantly from the additional treatment of 1600 and 2600 in the mortality between the experimental treatments. These FTU kg-1, and there was a significant increase was observed results were agreed with Farhadi et al (2017), where noted in serum phosphorus concentration for 2600 treatments there was no significant difference in the mortality compared to the additional treatment of 1600 FTU kg-1. A percentage during the experimental period between PC and significant increase was observed at the ALP level of the NC NC (decrease 0.15 Ap.) with the addition of different levels of treatment (1620) compared to the PC treatment (1236), while phytase enzymes. The results were not agreed with the addition of enzyme at the levels of 600, 1600, 2600 FTU Pieniazek et al (2017) where significant increase was kg-1to NC treatment has led to a significant decrease in the

Table 5. Effect of addition at different levels of phytase enzyme in feed conversion factor (g) Weeks Treatments Positive control Negative control Negative control+600 Negative control+1600 Negative control+2600 FTU / Kg FTU / Kg FTU / Kg

3-Jan c1.38 ± 0.010 a1.56 ± 0.011 b1.42 ± 0.002 c1.36 ± 0.014 c1.34 ± 0.012 6-Mar c1.90 ± 0.009 a2.07 ± 0.187 b2.00 ± 0.019 c1.90 ± 0.020 c1.86 ± 0.027 6-Jan c1.74 ± 0.005 a1.90 ± 0.016 b1.82 ± 0.023 c1.74 ± 0.012 c1.71 ± 0.001 * Values ​​represent the average ± standard error. ** Small letters varied within one row indicate that there are a significant difference between treatments, at a significant level (P≤0.05).

Table 6. Effect of addition at different levels of phytase enzyme in mortality percentage Weeks Treatments Positive control Negative control Negative control+600 FTU Negative control+1600 Negative control+2600 / Kg FTU / Kg FTU / Kg

Mortality a1.67 ± 1.67 a3.33 ± 1.67 a1.67 ± 1.67 a0.00 ± 0.00 a1.67 ± 1.67 * Values ​​represent the average ± standard error 26 Husam H. Nafea, Firas M. Hussein and Bahaa I. Hamid

Table 7. Effect of addition at different levels of phytase enzyme in physiological characters at 42 days Weeks Treatments Positive control Negative control Negative control+600 Negative control+1600 Negative control+2600 FTU / Kg FTU / Kg FTU / Kg

Glucose Mg /100 ml a180.00 ± 2.31 a182.67 ± 2.40 a179.33 ± 4.67 a178.33 ± 2.40 a180.33 ± 1.85 Cholesterol a201.00 ± 2.64 a202.33 ± 1.76 a200.00 ± 3.60 a201.67 ± 0.88 a203.33 ± 2.90 Triglyceride a74.33 ± 2.33 a72.33 ± 1.45 a71.67 ± 1.67 a74.33 ± 2.33 a76.67 ± 0.88 Total protein a3.67 ± 0.09 b3.10 ± 0.11 ab3.30 ± 0.15 ab3.30 ± 0.20 ab3.47 ± 0.13 phosphorus ab5.40 ± 0.10 d4.03 ± 0.12 c4.53 ± 0.09 b5.37 ± 0.12 a5.73 ± 0.12 ALP IU/L c1236.00 ± 33.31 a1620.00 ± 70.24 b1483.33 ± 40.55 c1276.67 ± 21.85 c1210.00 ± 17.32 * Values ​​represent the average ± standard error. ** Small letters varied within one row indicate that there are a significant difference between treatments, at a significant level (P≤0.05). concentration of ALP by 1483.33, 1276.67, 1210, Also, this result was agreed with Tannay Bilal et al (2015) in respectively, with a significant increase for the additional an experiment Period from 1-42 days that there was a treatment of 600 FTU kg-1 compared to PC treatment, which significant increase at ALP concentration in blood serum in was not significantly different from the two additional the NC treatment (decrease 0.15 point) compared to PC treatments of 1600 and 2600 FTU kg-1. These results were treatment. The addition of 600 FTU kg-1 from phytase enzyme agreed with what Islam et al (2017) pointed out for a diet of for NC treatment had significantly decreased P <0.05) ALP less containing from nPP without or with enzyme addition of concentration in blood serum, with a significant difference -1 1500 FTU kg at 42 days with no significant differences in compared with PC treatment. The results are don't agreed Glucose, Cholesterol and Triglyceride levels. The results with Sommerfeld et al (2018), where there was no significant also agreed with Amer (2014) and for the duration of 14-42 difference in serum phosphorus concentration when adding days of birds age with no significant difference in Glucose, the phytase to the full phosphorus requirements diets with Cholesterol and Triglyceride levels between PC treatment percentages500, 1500, 3000 FTU kg-1. Also not agreed with and two negative control treatment NC and NC1 plus 1000, what Momeneh et al (2018) pointed out for a period of 1-28 -1 2000 FTU kg . As well as, the result agreed with Farhadi et al days that there was no significant differences to full dietary (2017) findings, where there was a significant increase in requirements, diet and the addition of phytase at the total protein level of the blood serum for the addition of percentages of 0, 500, 2500 FTU kg-1. phytase treatments at levels of 3000, 4000, 5000, 6000 FTU -1 kg for NC treatment (decrease 0.15 Ap) compared to NC REFERENCES treatment without addition. A significantly increased was Adeola O and Cowieson AJ 2011. Board-invited review: observed in the blood phosphorus level of the PC treatment Opportunities and challenges in using exogenous enzymes to improve non-ruminant animal production. Journal of Animal compared to the NC treatment (decrease 0.15 AP). Science 89(10): 3189-3218. The following levels were added at (500, 1000, 2000, Amer AM 2014. Effect of phytase enzyme supplementation in 3000, 4000, 5000, 6000) from phytase enzyme has led to a deficient phosphate diet on broiler performance and some blood traits. International Journal of Science and Nature 5: 341- significant increase in the phosphorus concentration in the 345. blood serum, no significant difference was observed Asatoor AM and King EJ 1954. Simplified colorimetric blood sugar compared to the PC treatment, also an increasing in ALP method. The Biochemical Journal 56(325th Meeting): xliv. Beeson LA, Walk CL, Bedford MR and Olukosi OA 2017. Hydrolysis enzyme level was observed in blood serum for phosphorus of phytate to its lower esters can influence the growth decreasing of NC treatment (0.15 Ap) compared with a PC performance and nutrient utilization of broilers with regular or treatment. The addition of phytase (4000, 5000, 6000) were super doses of phytase. Poultry Science 96(7): 2243-3353. Bilal T, Atis S and Keser O 2015. The effects of microbial phytase on significantly decreased the ALP levels at the closer levels of serum calcium and phosphorus levels and alkaline PC treatment. The results were agreed with Taheri and phosphatase activities in broilers fed diets containing different levels of phosphorus. Acta Scientiae Veterinariae 43: 1327. Taherkhani (2015) in the experiment for the period 22-42 Dos Santos TT, Srinongkote S, Bedford MR and Walk CL 2013. days, With a significant increase in serum phosphorus Effect of high phytase inclusion rates on performance of broilers concentration for control treatment (0.43 AP) compared with fed diets not severely limited in available phosphorus. Asian- Australasian Journal of Animal Sciences 26(2): 227. NC treatment (0.15: AP), When adding 4000 FTU kg-1 to NC Duncan DB 1955. Multiple range and multiple F tests. Biometrics treatment has led to a significant increase (P <0.05) in serum 11(1): 1-42. phosphorus concentration and near to PC positive control. Farhadi D, Karimi A, Sadeghi G, Rostamzadeh J and Bedford MR Effects of High and Commercial Phytase Doses on Broiler Performance 27

2017. Effects of a high dose of microbial phytase and myo- assay of total cholesterol in serum. Clinical Chemistry 19(12): inositol supplementation on growth performance, tibia 1350-1356. mineralization, nutrient digestibility, litter moisture content, and Rodehutscord M, Rückert C, Maurer HP, Schenkel H, Schipprack W, foot problems in broiler chickens fed phosphorus-deficient Bach-Knudsen KE and Mosenthin R 2016. Variation in chemical diets. Poultry Science 96(2): 3664-3675. composition and physical characteristics of cereal grains from Fiske CH and Subbarow Y 1925.The colorimetric determination of different genotypes. Archives of Animal Nutrition 70(2): 87-107. phosphorus. The Journal of Biological Chemistry 66(2): 375- SAS Institute INC 2012. SAS/STAT User's Guide: Version 9.1. (Cary, 400. NC, SAS Institute, Inc.). Food and Agriculture Organisation (FAO) 2009. Maize, Rice and Scholey DV, Morgan NK, Riemensperger A, Hardy R and Burton EJ wheat, area harvested, production quantity and yield. 2018. Effect of supplementation of phytase to diets low in http://nue.okstate.edu /crop_ information/world_wheat_produ. inorganic phosphorus on growth performance and htm. Date acces. 15/10/10. mineralization of broilers. Poultry Science 97(7): 2435-2440. Gautier AE, Walk CL and Dilger RN 2017. Effects of a high level of Shastak Y and Rodehutscord M 2013. Determination and estimation phytase on broiler performance, bone ash, phosphorus of phosphorus availability in growing poultry and their historical utilization, and phytatedephosphorylation to inositol. Poultry development. World's Poultry Science Journal 69(3): 569-586. Science 97(1): 211-218. Shastak Y, Witzig M, Hartung K and Rodehutscord M 2012. HR T 2015. Effect of phytasesuperdoses and citric acid on growth Comparison of retention and prececal digestibility performance, plasma phosphorus and tibia ash in broilers fed measurements in evaluating mineral phosphorus sources in canola meal-based diets severely limited in available broilers. Poultry Science 91(9): 2201-2209. phosphorus. Poultry Science Journal 3(1): 27-36. Singh N, Kumari A, Gakhar SK and Singh B 2015.Enhanced cost- Islam R, Ideris A, Kasim A, Omar AR, Hussin ASM, Yasmin F and effective phytase production by Aspergillus Niger and its González-Redondo P 2017. Evaluation of humoral immune applicability in dephytinization of food ingredients. Microblog response, body weight and blood constituents of broilers Journal 84(2), 219-226. supplemented with phytase on infectious bursal disease vaccination. Cogent Food & Agriculture 3(1): 1306933. Sommerfeld V, Schollenberger M, Kühn I. and Rodehutscord M 2018. Interactive effects of phosphorus, calcium, and phytase John MK 1970. Colorimetric determination of phosphorus in soil and supplements on products of phytate degradation in the plant materials with ascorbic acid.Soil Science 109(4): 214-220. digestive tract of broiler chickens. Poultry Science 97(4): 1177- King EJ and Armstrong R 1943. Determination of Serum alkaline and 1188. acid Phosphatase using of Spectrophotometry. Canadian Taheri HR, Heidari A and Shahir MH 2015. Effect of high-dose Medical Association Journal 31: 276. phytase supplementation in broilers from 22 to 42 days post- Kumar V, Yadav AN, Saxena A, Sangwan P and Dhaliwal HS 2016. hatch given diets severely limited in available phosphorus. Unravellingrhizospheric diversity and potential of phytase British Poultry Science 56(3): 330-336. producing microbes.SM Journal of Biology 2(1): 1009. Tayyab U and McLean FA 2015. Phosphorus losses and on-farm Lee SH, Cho J, Bok J, Kang S, Choi Y, and Lee PC 2015. mitigation options for dairy farming systems: A review. JAPS: Characterization, gene cloning, and sequencing of a fungal Journal of Animal & Plant Sciences 25(2): 26-29. phytase, PhyA, from Penicillium oxalicum PJ3. Preparative Truong HH, Yu S, Moss AF, Partridge GG, Liu SY and Selle PH 2017. Biochemistry and Biotechnology 45(4): 336-347. Phytase inclusions of 500 and 2000 FTU/kg in maize-based Liu SB, Liao XD, Lu L, Li SF, Wang L, Zhang LY and Luo XG 2016. broiler diets impact on growth performance, nutrient utilisation, Dietary non-phytate phosphorus requirement of broilers fed a digestive dynamics of starch, protein (N), sodium and IP6 conventional corn-soybean meal diet from 1 to 21 d of age. phytate degradation in the gizzard and four small intestinal Poultry Science 96(1): 151-159. segments. Animal Feed Science and Technology 223: 13-22. Momeneh T, Karimi A, Sadeghi G, Vaziry A and Bedford MR 2018. Walk CL, Bedford MR and Olukosi OA 2018. Effect of phytase on Evaluation of dietary calcium level and source and phytase on growth performance, phytate degradation and gene expression growth performance, serum metabolites, and ileum mineral of myo-inositol transporters in the small intestine, liver and contents in broiler chicks fed adequate phosphorus diets from kidney of 21 day old broilers.Poultry Science 97(4): 1155-1162. one to 28 days of age. Poultry Science 97(4): 1283-1289. Walk CL, Santos TT and Bedford MR 2014. Influence of superdoses NRC. 1994. Nutrient Requirements of Poultry. 9th rev. ed. National of a novel microbial phytase on growth performance, tibia ash, Academy of Sciences, Natl. Acad. Press, Washington, DC. and gizzard phytate and inositol in young broilers. Poultry Pieniazek J, Smith KA, Williams MP, Manangi MK, Vazquez-Anon M, Science 93(5): 1172-1177. Solbak A and Lee JT 2017. Evaluation of increasing levels of a Wang ZP, Deng LH, Weng LS, Deng XY, FU XQ, Xin YY and Xiao GY microbial phytase in phosphorus deficient broiler diets via live 2017.Transgenic rice expressing a novel phytase-lactoferric in broiler performance, tibia bone ash, apparent metabolizable fusion gene to improve phosphorus availability and antibacterial energy, and amino acid digestibility. Poultry Science 96(2): 370- activity. Journal of Integrative Agriculture 16(4): 774-788 382. Wealleans AL, Barnard LP, Romero LF and Kwakernaak C 2016. A Pirgozliev V, Bedford MR, Oduguwa O, Acamovic T and Allymehr M value based approach to determine optimal phytase dose: A 2012.The effect of supplementary bacterial phytase on dietary case study in turkey pouts. Animal Feed Science and metabolisable energy, nutrient retention and endogenous Technology 216: 288-295 losses in precision fed broiler chickens. Journal of Animal Wu D, Wu SB, Choct M and Swick RA 2015. Comparison of 3 Physiology and Animal Nutrition 96(1): 52-57. phytases on energy utilization of a nutritionally marginal wheat- Richmond W 1973. Preparation and properties of a cholesterol soybean meal broiler diet. Poultry Science 94(11): 2670-2676. oxidase from Nocardia sp. and its application to the enzymatic

Received 29 August, 2019; Accepted 04 November, 2019 Manuscript Number: S-246 Indian Journal of Ecology (2020) 46 Special Issue (8): 28-30 NAAS Rating: 4.96

Evaluation of Heavy Metals in Nassiryah City, Iraq

Shroog Hameed Hlail and Sanaa Talib Jaweed Department of Biology, College of Education for Pure Sciences, University of Thi-Qar, Iraq E-mail: [email protected]

Abstract: The objective of this study was to evaluate the content of heavy metals in the water, plants (Ceratophyllum demersum and Phragmites australi) and sediment of Euphrates River in Nasiriyah city. The samples were collected from of three stations during the autumn and winter season. Heavy metals (Cd, Cu, Pb, Fe and Zn) were detected using an Atomic Absorption Spectrophotometer. The heavy metals concentrations in the water, plants and sediments were detected during the autumn and winter season. Ceratophyllum showed higher heavy metals concentration of Cd, Cu, Pb, Fe and Zn in station 3 than Phragmites during the autumn season. High amount of heavy metals (Fe, Cu and Zn) in sediment samples were also in station3 during winter season. However, Season and location play a significant role in the heavy metals content of water, plants and sediment. The Ceratophyllum had the highest heavy metals content while Phragmites had the lowest for both the seasons.

Keyword: Euphrates River, Heavy metals, Sediment, Ceratophyllum, Phragmites

Water is the main source of the life, is essential for all 3).The samples of water plants and sediment were collected living organism. The most serious problem that appeared in from three stations of Euphrates River in Nassiryah city the last two centuries is the water pollution due to increment during the autumn (September) and winter (December) of urbanization, population growth, industrial production season. economic evolution, and developments, which reflected on Preparations of plant and sediment: Plant samples were the quality of surface and groundwater. Heavy metals are the washed with tap water to remove soil particles followed by one of the most important pollutant for the aquatic three times of washing, twice with distilled water and once environment and pose a great potential threat to the with distilled deionized water. Samples were then dried with environment. Some of heavy metals are necessary for tissue paper and cut into small pieces. Finally, samples were animals and plants due to their role as micronutrients and oven dried at 60ºC for two days until they reach a constant physiological consequences (Aktar et al 2010). The heavy weight (AOAC 1984). Collected sediment samples were air- metal could enter the ecosystem naturally and anthropogenic dried in the laboratory before being ground and sieved using sources (Chanpiwat et al 2010) and have the ability to a 250µm mash (Badri 1984). accumulate and persist in the environment. Furthermore, Plant and soil digestion: The heavy metals extractions they are transformed from one oxidation state or organic were performed by wet digestion according to the AOAC complex to another therefore, they cannot be destroyed methods(AOAC 1984). Dried samples were weighed (1g) in biologically (Gisbert et al 2003). Heavy metals could be very a conical flask with HNO3HClO (2:1) for 3-6hon a sand bath at danger due to their non-degradable nature and their harmful a temperature of 100°C. Five ml of HCl acid was added to the effect for the biological organism by accumulation in their sample to digest the inorganic and oxide salts. Digested bodies. There are many activities that increase the dispersal samples were filtered with a 0.45 µm pore size cellulose of heavy metals and can cause serious damage through nitrate membrane filter paper, and the volume was made up worsening of water quality and this effect human in addition to to 50 ml with deionized water. Heavy metals concentrations biodiversity of aquatic organism. The aim of this study was to were determined by Atomic Absorption Spectrophotometer. evaluate the accumulation of some heavy metals in water, sediment and plant in Euphrates river water in Nassiryah city. RESULTS AND DISCUSSION Heavy metals content in water: The concentration of heavy MATERIAL AND METHODS metals concentrations in water was in the following order of The water, plants and sediment samples were obtained Fe > Zn>Pb> Cu > Cd. The concentration of heavy metals from three locations, i.e., the beginning of the entry of the increased in winter more than autumn. The station 3 had Euphrates River in Nasiriyah city (Station 1); after thermal more heavy metal then station 2 and 1 especially the Fe energy on Euphrates River in Nasiriyah city (Station 2) and (Table 1). This may be due to high population density near sewage water with Euphrates water in Nasiriyah city (Station this station and the municipal waste was directly discharged Heavy Metals in Nassiryah City, Iraq 29 to the river by the main pipes without treatment. The effluents Table 2. Heavy metals content of sediment (µg g-1 DW) of municipal and industrial waste contain trace metals among Season Station Cd Cu Pb Fe Zn their constituent. There were many industrials discharged Autumn S1 3.20 e 10.21 e 32.91 b 1395.01 c 76.01 c that throughout to the river without treatment especially in S2 6.40 c 14.01 d 30.10 c 1616.20 b 80.79 b station 2. The concentration of many trace metals was S3 8.70 a 18.16 b 35.31 a 1745.01 a 85.50 a greater in particulate phase than dissolved phase for all Winter S1 4.91 d 14.50 d 0.08 e 721.42 f 23.22 f studied stations except in Cd and Pb. S2 5.36 d 16.82 c 0.26 e 7 08.31 e 26.53 e Heavy metals content in sediment: The order of S3 7.83 b 19.42 a 1.03 d 749.63 d 28.74 d abundance of these metals in sediments for the a–fDifferent letters within each column indicate significant difference (p<0.05 exchangeable and residual phase of the three station were in the order of Fe >Mn> Zn > Cu >Pb> Cd (Table 3). Our result features was Sediment texture. The percentage for this revealed that the amount of heavy metal in residual phase contents (sand, silt and clay) were 23.06, 32.6, 45.5%; 30.6, more than in exchangeable phase. This is due to the ability of 34, 35.7 and 42.06, 43, 24% in three station, respectively. heavy metal to absorb on particular part in sediment. The There was variation for the sediment texture between studied suspend matter that has heavy metal tend to be deposited in stations. Station 3 consists of high percentage of clay bottom then absorbed by the organic matter and clay. All of comparing to the other station. Our results were near to the these sources of heavy metal came from anthropogenic results of Al-Hussainy (2005). sources. Station 3 has high mean of heavy metals The heavy metals content in plants: The heavy metals concentration and this could due to wastewater discharges of concentration could change the water feature and their ability leather facilities, which are located nearby the river. The to accumulate could make potential harmful to the aquatic result showed that the sediment of Euphrates river contains organism (Fernandes et al 2007). The Ceratophyllum has from sand, silt and clay. One of the essential physical highest concentration of heavy metal than Phragmites. The

Table 1. Heavy metals content of water (µg l-1) from three sampling station Heavy metals Season

Autumn - September Winter – December

S1 S2 S3 S1 S2 S3

Cd 0.06 e 28.06 b 14.59 d 0.08 e 16.60 c 30.60 a Cu 0.12 d 25.03 c 28.06 b 0.14 d 27.25 b 30.70 a Pb 30.01 c 40.30 b 69.84 a 32.03 c 42.41 b 71.95 a Fe 1005.01 c 2400.72 b 3196.14 a 1007.60 c 2402.99 b 3198.15 a Zn 70.36 c 73.02 b 99.03 a 72.04 c 74.10 b 101.99 a a–e Different letters within each raw indicate significant difference (p<0.05)

Table 3. The heavy metals content of plants (µg g-1 DW) from three sampling stations Season Heavy S1 S2 S3 metals Ceratophyllum Phagmites Ceratophyllum Phagmites Ceratophyllum Phagmites Autumn Cd 0.95 c 0.83 c 3.02 a 2.78 b 2.56 b 2.30 b Cu 0.99 b 0.91 b 3.79 a 3.61 a 3.00 a 2.87 a Pb 0.96 b 0.84 b 3.24 a 2.90 a 2.80 a 2.79 a Fe 80.87 e 81.91 e 190.56 a 171.14 b 132.04 c 108.01 d Zn 13.12 c 12.11 d 29.78 a 25.87 b 4.14 e 14.09 c Winter Cd 0.60 b 0.52 b 1.79 a 0.56 a 1.00 b 0.80 b Cu 0.54 b 0.42 b 1.99 a 1.70 a 1.02 b 1.00 b Pb 0.44 b 0.20 b 1.26 a 1.00 a 1.01 a 0.98 a Fe 70.03 d 62.03 e 160.01 a 140.21 b 145.02 b 132.03 c Zn 9.01 e 7.10 f 24.12 a 18.00 c 20.00 b 14.12 d a–fDifferent letters within each raw indicate significant difference (p<0.05) 30 Shroog Hameed Hlail and Sanaa Talib Jaweed

highest mean was 190.56 µg g-1 for Fe in autumn, while the Sciences. Univ. of Baghdad, Iraq, 138 pp. lowest was 0.52 for cd in winter in Phragmites. There were Aktar M, Paramasivam M, Ganguly M, Purkait S and Sengupta D 2010. Assessment and occurrence of various heavy metals in many factors that affect the concentration of heavy metal in surface water of Ganga river around Kolkata: A study for toxicity plant like soil texture and depend on the evaluation of plant and ecological impact. Environment Monitoring and Assessment 160: 207-213. therefore; among these that Phragmites has the low ability to AOAC 1984. Official Method of Analysis. 14 Ed., the Association of absorption of heavy metal (Skorbilowicz 2009). The plant in Official Analytical Chemist, Arlington, Verginia, USA. station 2,3 had high concentration in trace metal due to the Badri MA 1984. Identification of heavy metal toxicity levels in solid huge density of the disgorge and waste and drainage wastes by chemical speciation. Conservation and Recycling 7(2): 257-269. channels (Station 3) power station disgorge. Chanpiwat P, Sthiannopkao S and Kim K 2010. Metal content variation in wastewater and biosludge from Bangkok's central CONCLUSION wastewater treatment plants, Microchemical Journal 95: 326- 332. Heavy metals may change the properties of water, Fernandes C, Fontainhas-Fernandes A, Peixoto F and Salgado MR sediment and biology of plants. Heavy metals can uptake by 2007. Bioaccumulation of heavy metals in Liza saliens from the Esomriz-Paramos coastal lagoon, Portugal. Ecotoxicology and soil and accumulate in plant tissue. In the current study, it Environmental Safety 66: 426-431. was found that the heavy metals in sediment were connected Gisbert C, Ros R, de Haro A, Walker DJ, Pilar Bernal M, Serrano R with fine grain size. Many of heavy metal concentration are and Avino JNA 2003. plant genetically modified that accumulates Pb is especially promising for phytoremediation. acceptable, while there were minor increase of some metal. Biochemical and Biophysical Research Communication 303(2): The increasing of concentration of some trace metals 440–445. showed a potential accumulation of trace metals. This Griscom SB, Fisher NS and Luoma SN 2000. Geochemical influences on assimilation of sediment-bound metals in clams happened due to the discharge the pollutants from sewage and mussels. Environment Science Technology 34: 91-99. water and thermal energy station runoff. Nolting RF 1986. Copper, zinc, cadmium, nickel, iron and manganese in the southern Bight of the North Sea. Marine Pollution Bulletin 17(3): 113-117. REFERENCES Skorbilowicz E 2009. Aquatic plants as bio indicators of contamination Al-Hussainy AK 2005. Study for characteristics of soil from some of upper Narew river and some of its tributaries with heavy dried areas in Al-Hammar marsh, Iraq. M.Sc. Thesis, College of metals. Environment Protection Engineer 35(1): 65-77. Received 17 September, 2019; Accepted 04 November, 2019 Manuscript Number: S-247 Indian Journal of Ecology (2020) 46 Special Issue (8): 31-35 NAAS Rating: 4.96

Effect of Copper Concentrations on Some Biochemical Properties of the Cladophora

Wameedh A.K. AL-Yasari*, Rasool I. Al-Nafie and Bassam M. Al-Yaseen Department of Biology, College of Science, University of Babylon, Babylon, Iraq *E-mail: [email protected]

Abstract: Green algae, especially Cladophora species are generally considered as the best bioindicator of aquatic bodies contamination by nutrients as well as by heavy metals such as copper. The object of this study was to investigate the effects of different concentrations of copper on some biochemical properties of Cladophora isolates from different areas of Al-Hilla river and cultured in a modified Chu-No.10 medium, which supplemented with 0.1, 0.3 and 0.5 mg. L-1 of copper for two different periods (3 and 6 days). Some biochemical properties such as the content of chlorophyll, antioxidant enzymes such as Catalase (CAT) and superoxide dismutase (SOD), Metallothionein and Cytochrome were estimated. The results of current study indicated to presence of variations in the effects of different concentrations of copper on these properties compared to the control, and these effects were clear through increasing concentrations with increasing exposure time.

Keywords: Antioxidant enzymes, Cladophora, Copper

Chlorophyta, commonly known as green algae, are metals can be divided into two categories, which are responsible for most of the primary productivities in some essential and non-essential. However, essential, which have water bodies. They subsist on inorganic nutrient and produce been reported to be toxic at high concentration, for example organic matter from carbon dioxide by photosynthesis. In the some heavy metals, including cobalt, copper, zinc and absence of photosynthesis, the metabolic process causing manganese are essential for growth at very low oxygen depletion in the aquatic system (Förstner and Prosi concentration but toxic at slightly higher levels (Al-Hejuje 1979). The role of algae in waste-water treatment and their 1997, El-Sheekh et al 2003, Awasthiand Das 2005,Okmen et affinity for heavy metals captions based on high negative al 2007). Whereas, the other category has been reported to surface charge, has been recognized for several decades be toxic even at low concentration such as silver, lead, (Rao 1986). Algae accumulate heavy metals from their mercury and cadmium (Lee et al 2005, Al-Hejuje 1997). In the aquatic environment. Using living algae to remove toxic aquatic ecosystem, the primary producers play an important metals from contaminated water could be advantageous. role in the structure and function of the ecosystem. Any Since they are ubiquitous and have colonized almost all parts negative effects on them will affect upper trophic levels (Li et of the world. They can be grown easily have very simple al 2006). Copper is essential redox-active transition metals, growth requirements. Advantage of using living organisms which acts as a structural element in regulatory proteins and over dead biomass is that they have fast growth rate and participates in electron transport in photosynthesis, hence produce a regenerating supply of metal-removal mitochondrial respiration, oxidative stress responses, cell material (Sobhan and Sternberg 1999). There is evidence wall metabolism and hormone signaling (Moreno 2003, that algae accumulate heavy metals in their tissues when Srivastava et al 2006). The metals as other environmental grown in polluted water (Magdaleno et al 1997, Kim et al factors (ultraviolet, extreme temperature, radiation, 1999). Algae are generally considered as the best environmental pollution, etc.) induce Reactive Oxygen bioindicator of aquatic bodies contamination by heavy metals Species (ROS) formation inside the cell (Marshall and (Chmielewská and Medved 2001).The ubiquity of heavy Newman 2002). These ROS are formed under physiological metals in the biosphere results in the introduction of the high conditions in manageable proportions, while maintaining a amount of toxic metals into the food chain from various balance due to the defensive cellular enzyme (Dröge 2003). sources, including municipal, agricultural and industrial The breakdown of this balance is known as oxidative stress, effluents, and they modify the structure and productivity of which results in alteration of the structure-function of organs, aquatic ecosystem (Magdaleno et al 1997). Metals such as systems or specialized transport (Ames et al 1993). The copper and zinc are released into the aquatic ecosystem by damage caused by oxidative stress in fatty acid rich anthropogenic activities such as agricultural, industrial and structures such as cell membrane, results in lose rigidity, mining (Yruela 2005). From a biological point of view, heavy integrity and permeability. This process known as 32 Wameedh A.K. AL-Yasari, Rasool I. Al-Nafie and Bassam M. Al-Yaseen

Lipidperoxidation, produce carbonylic compounds such as Statistical analysis: SPSS 17.0 program used for least malondialdehyde. Aerobic organisms have developed significance differences (LSD ≤0.05) and analysis of antioxidative defence mechanisms to prevent cell damage by variance test (ANOVA). ROS (Finkel and Holbrook 2000, Sohal et al 2002). The enzyme catalase has an active participation in the reaction RESULTS AND DISCUSSION that reduces the intracellular levels of H2O2 formed from the Algae expose to different stressors and can develop superoxide radical (De Zwart et al 1999). Oxidative stress is tolerance to a wide range of heavy metals. Heavy metals characterized by the increase in ROS as a result of have two levels of impacts; first by inhibiting the algal insufficient antioxidant defense (Mittler 2002). Responses of physiological and biochemical activities, second in changing organisms to oxidative stress include the use of antioxidant the composition and function of the algal community in enzymes, water-soluble antioxidant compounds and lipid- general. Algae are very sensitive to the metals toxicity, soluble antioxidant molecules (Burritt et al 2002). Enzymatic therefore, it can function as bio-indicators to indicate the and non-enzymatic reactions of free radical scavengers potential toxic impacts for the heavy metals. The results of minimize cellular oxidation: (SOD), (CAT). Peroxidase (POD) this study presented in tables below shown differences of the and anti-compounds such as Glutathione (GSH) and copper concentrations' impact compare to the control group Carotenoids (CAR) can play a key role in controlling cellular on the biochemical parameters which included Chlorophyll levels of metal-induced ROS (Weckx and Clijsters 1996). determination, SOD and Catalase enzymes, Metallothionein, The importance of antioxidant enzymes in preventing and Cytochrome measured during this study. The results in oxidative stress by scavenging ROS is generally emphasized (Table 1) shown the impact of the different levels of the

(Dawes 2000). SOD dismutase O2 to H2O2 and this is copper concentrations on the chlorophyll content with a decomposed to H2O by POD and CAT, so that the significant difference in compare with the control group accumulation of O2 and H2O2 is effectively prevented (Yan et during the initial study period (3 days) and that might be a al 1997 and Liu et al 2002). result of the copper impact on the chloroplast causing disorders and finally lead to a reduction of photosynthetic MATERIAL AND METHODS pigments content in addition to chlorophyll inhibition and Algae cultivation: The Cladophora collected from AL-Hilla decline in its content (Påhlsson 1989). The results also River in Babylon province, Iraq, was isolated and cultivated in shown significant differences among the values of the modified chu-10 medium that contained different studied enzymes, Metallothionein, and Cytochrome in concentrations of copper (0.1, 0.3 and 0.5 mg l-1) for different compare with the control group and this is due to the osmosis periods (3 days and 6 days) according to Cao et al (2015). of elements that enter the algae body on the sites for Metal solutions: Three concentrations of copper (0.1, 0.3 connecting protein with active enzymes and resulting in and 0.5 mg l-1) were prepared using the standard solution of obstruct the natural way of metabolism (Arunakumara et al Cu+2 (1000 mg l-1) according to Cao et al (2015). 2006). Biochemical analysis: Many biochemical tests were The results in (Table 2) shown that exposure time for (6 determined which include determine the SOD using the days) affected clearly on the differences among the study method described by Marklund and Marklund (1974), criteria with significant difference where the highest value determine the CAT using the method described by Aebi was recorded at the concentration of (0.3) ml l-1 in chlorophyll (1974) Metallothionein and Cytochrome according to Burritt content determination and SOD enzyme compare to control and Larkindale (2002) and total chlorophyll using the method group, while there was a significant difference at all level of described by Lichtenthaler (1987). concentrations for the rest of parameters.

Table 1. Comparison among control group and the concentrations (0.1, 0.3, 0.5) for the biochemical parameters of the Chladophora during exposure period (3 days) Parameters Control Con. 0.1 Con. 0.3 Con. 0.5

Chlorophyll 1.276 ± 0.03 D 0.353 ± 0.02 A 1.070 ± 0.00 c 0.50 ± 0.01 B SOD 1.863 ± 0.11 A 1.973 ± 0.12 A 2.573 ± 0.12 b 2.433 ± 0.20 B CAT 6.093 ± 0.02 B 4.083 ± 0.04 A 4.06 ± 0.01 a 6.056 ± 0.02 B Metallothionin 1.08 ± 0.01 A 1.633 ± 0.02 C 1.373 ± 0.06 b 1.63 ± 0.02 C Cytochrom 1.364 ± 0.93 B 1.422 ± 0.36 C 1.518 ± 0.11 d 1.334 ± 0.42 A Similar letters for the studied parameters mean there is no significant difference, p ≥ 0.05 Cladophora Effectiveness by Copper 33

All significant differences for all parameters compare to ions from bio-molecules and cellular work units. This may the control group in previous tables indicate that all lead to the modification and disruption of enzymes, disrupting physiological changes have impacted by external conditions (rupture) cellular membranes and organs membranes and including heavy metals such as copper. Decrease of their integrity. Thus toxic metals have the ability to interfere chlorophyll content in (Table 1 and 2) indicates evident impact with a wide range of activities in living organisms. It may of copper as a stressor on algae as the concentrations are not impact organisms metabolism, growth and differentiation the optimum for the metabolic activity for the cell where the (Awasthi and Das 2005). magnesium in the chlorophyll being replaced by the elements ions and therefore the cells work on protoporphyrin Table 3. The biochemical parameters comparison during the accumulation and the chlorophyll synthesis stops as a result of two exposure period* for the control group inhibit the reduction process in the biosynthesis pathways of Parameters G3 G6 Sig this pigment (El-Sheekh et al 2003). In contrast, there is a high Chlorophyll 1.276 ± 0.037 0.280 ± 0.026 0.000 activity for the antioxidant enzymes SOD and CAT which SOD 1.863 ± 0.115 2.156 ± 0.200 0.110 reduce the activity of ROS resulting from increased stresses or CAT 6.093 ± 0.028 4.08 ± 0.036 0.000 exposure to external stimuli (Converti et al 2006). It is known Metallothionin 1.080 ± 0.017 1.213 ± 0.011 0.001 that copper is an essential element to be involved in the Cytochrom 1.364 ± 0.938 1.410 ± 0.10 0.013 synthetic of defense enzymes, but at the excess *G3: Exposure period 3 days, G6: Exposure period 6 days concentration, the effects could cause an inhibition as it is evident from (Tables 1 and 2) where the activity of SOD and Table 4. The biochemical parameters comparison during the CAT increased when increase the exposure period for two exposure period* for the concentration group oxidative stress resulted from the activity of ROS. (0.1) The results in (Table 3, 4, 5 and 6) showed the significant Parameters G3 G6 Sig differences for the studied parameters for the two exposure Chlorophyll 0.353 ± 0.020 0.483 ± 0.030 0.005 periods which are 3 days (G3) and 6 days (G6). Where the SOD 1.973 ± 0.127 2.430 ± 0.207 0.041 significant difference was noticed for the activity of SOD CAT 4.083 ± 0.045 2.053 ± 0.030 0.000 enzyme, metallothionein, Cytochrome and control group Metallothionin 1.633 ± 0.023 0.953 ± 0.051 0.000 (Table 3). Cytochrom 1.422 ± 0.360 1.388 ± 0.850 0.011 Significant differences of SOD, CAT, Cytochrome and *G3: Exposure period 3 days, G6: Exposure period 6 days chlorophyll content at concentration of (0.1) mgl-1 (Table 4). While all parameters except Cytochrome were showed a Table 5. The biochemical parameters comparison during the significant difference at concentration of (0.3) mg l-1 (Table 5). two exposure period* for the concentration group Similar results were recorded at the concentrations of (0.5) (0.3) -1 Parameters G3 G6 Sig mgl (Table 6). The above results, in addition to the heterogeneity of the Chlorophyll 1.070 ± 0.000 0.903 ± 0.025 0.008 effectiveness of the active enzymes, indicate a difference in SOD 2.573 ± 0.120 2.753 ± 0.387 0.511 Cytochrome electron transport systems. This may be due to CAT 4.060 ± 0.017 4.04 ± 0.000 0.184 might be resulted from the disruption of functional groups of Metallothionin 1.373 ± 0.060 1.043 ± 0.020 0.006 vital bio-molecules, such as enzymes and transport systems Cytochrom 1.518 ± 0.116 1.288 ± 0.542 0.000 for essential nutrients and ions, or replacement of basic metal *G3: Exposure period 3 days, G6: Exposure period 6 days

Table 2. Comparison among control group and the concentrations (0.1, 0.3, 0.5) for the biochemical parameters for the Chladophora during exposure period (6 days) Parameters Control Con. 0.1 Con. 0.3 Con. 0.5

Chlorophyll 0.28 ± 0.02 a 0.483 ± 0.03 B 0.903 ± 0.02 C 0.616 ± 0.01 D SOD 2.156 ± 0.200 a 2.43 ± 0.207 a,b 2.753 ± 0.387 B 2.513 ± 0.115 a,b CAT 4.080 ± 0.03 b 2.053 ± 0.03 A 4.040 ± 0.00 B 6.096 ± 0.04 C Metallothionin 1.213 ± 0.011 c 0.953 ± 0.05 A 1.043 ± 0.02 B 1.666 ± 0.03 D Cytochrom 1.41 ± 0.10 c 1.388 ± 0.85 B 1.288 ± 0.542 A 1.404 ± 0.216 C Similar letters for the studied parameters mean there is no significant difference, p ≥ 0.05 34 Wameedh A.K. AL-Yasari, Rasool I. Al-Nafie and Bassam M. Al-Yaseen

This study also showed the correlation between some of obvious, especially during the exposure period (6 days) and the studied parameters. The results (Table 7) exhibited a this is evident by the action of the SOD enzyme on the root of negative correlation between the impact of SOD and OH to give hydrogen peroxide, which changes the base chlorophyll, as well as a negative correlation between material of the activity of the CAT enzyme into a water chlorophyll and Metallothionein. Positive correlations were molecule (Liu et al 2002, Yan et al 1997). The activity of these recorded between chlorophyll and CAT, and between two enzymes are increased by the increasing of the chlorophyll and Cytochrome, and between SOD and CAT electronic transport compounds such as Cytochrome and during exposure period (3 days). Metallothionein, as the living organisms have a general Positive correlation was observed among chlorophyll, mechanism for removing the mineral toxins inside the cells, SOD and CAT, and between SOD and CAT, and between which is the formation of peptides and proteins binding CAT and Cytochrome, and between CAT and Metallothionein metals such as Phytochelatins and Metallothioneins (Burritt during exposure period (6 days). et al 2002). To understand the harmony and consistency among the studied criteria, (Table 7 and 8) proved that there are evident REFERENCES impacts of copper on the biochemical parameters of the Aebi H 1974. Catalase. In: Hans Ulrich Bergmeyer (eds). Methods of Enzymatic Analysis. Academic Press,pp 673-684. Chladophora. Where the correlation between the chlorophyll Al-Hejuje MM 1997. Distribution of heavy elements in water and content and the antioxidant enzymes indicates an increase in sediments from Al-Ashar and Al-Khandak canals connected these enzymes activity to maintain the chlorophyll quantity (Li with Shatt Al-Arab river and their effects on algae MSc. thesis, University of Basrah, Basrah, Iraq. et al 2006). Also, the correlation between SOD and CAT was Ames BN, Shigenaga MK, and Hagen TM 1993. Oxidants, antioxidants, and the degenerative diseases of aging, pp. 7915- 7922. In: Proceedings of the National Academy of Sciences of Table 6. The biochemical parameters comparison during the the United States of America.September 1993, 90 (17). two exposure period* for the concentration group (0.5) Arunakumara K, Xeucheng Z and Song X 2006. A comparative study on bioaccumulation of lead and cadmium by cyanobacterium Parameters G3 G6 Sig Synechocystis sp. PCC 6803 under laboratory conditions. Ciencias Marinas 33(3): 271-280. Chlorophyll 0.50 ± 0.017 0.616 ± 0.015 0.001 Awasthi M and Das DN 2005. Heavy metal stress on growth, SOD 2.433 ± 0.206 2.513 ± 0.115 0.597 photosynthesis and enzymatic activities of free and immobilizedChlorella vulgaris. Annals of Microbiology 55(1): 1- CAT 6.056 ± 0.025 6.096 ± 0.040 0.232 7. Metallothionin 1.630 ± 0.020 1.666 ± 0.035 0.209 Burritt DJ, Larkindale J and Hurd CL 2002. Antioxidant metabolism in Cytochrom 1.334 ± 0.428 1.404 ± 0.216 0.000 the intertidal red seaweed Stictosiphona arbuscula following desiccation. Planta 215(5): 829-838. *G3: Exposure period 3 days, G6: Exposure period 6 days Cao DJ, Shi XD, Li H, Xie PP, Zhang HM, Deng JW and Liang YG 2015. Effects of lead on tolerance, bioaccumulation, and antioxidative defense system of green algae, Cladophora. Table 7. Correlation relationship among the studied Ecotoxicology and Environmental Safety 112: 231-237. parameters during exposure period (3 days) Chmielewská E and Medved' J 2001. Bioaccumulation of Heavy Chlorop- SOD CAT Cytochrom Metallothionin Metals by Green Algae Cladophora glomerata in a Refinery hyll Sewage Lagoon. Croatica Chemica Acta 74(1): 135-145. SOD -0.050 Converti A, Lodi A, Solisio C, Soletto D, Del Borghi M and Carvalho JCM 2006. Spirulina platensis biomass as adsorbent for copper CAT 0.233 -0.297 removal. Journal of Food 5(2): 85-88. Cytochrom 0.231 0.393 -0.862** Dawes IW 2000. Response of eukaryotic cells to oxidative stress. Agricultural and Biological Chemistry 43: 211–217. Metallothionin -0.944** 0.355 -0.331 -0.044 De Zwart LL, Meerman JH, Commandeur JN and Vermeulen NP Significant at 1% level of significance 1999. Biomarkers of free radical damage. Applications in experimental animals and in humans. Free Radical Biology and Medicine 26(1-2): 202-226. Table 8. Correlation relationship among the studied Dröge W 2003. Free radicals in the physiological control of cell parameters during exposure period (6 days) function. Physiological Reviews 82(1): 47-95. Chlorop- SOD CAT Cytochrom Metallothionin El-Sheekh MM, El-Naggar AH, Osman MEH and El-Mazaly E 2003. hyll Effect of cobalt on growth, pigments and the photosynthetic electron transport in Monoraphidium minutum and SOD 0.692* Nitzchiaperminuta. Brazilian Journal of Plant Physiology 15(3): CAT 0.198 0.090 159-166. Finkel T and Holbrook NJ 2000. Oxidants, oxidative stress and the Cytochrom -0.864** -0.603* 0.121 biology of aging. Nature 408(6809): 239-247. Metallothionin 0.049 -0.063 0.918** 0.451 Förstner U and Prosi F 1979. Heavy metals pollution in freshwater Significant at 5 and 1% level of significanceecosystems. In: O. Ravera (Ed.) Biological aspects of Cladophora Effectiveness by Copper 35

freshwater pollution, Pergamon Press, Oxford, New York,pp. Trends in Plant Science 7(9): 405-410. 129-161. Moreno MD 2003. Toxicologia Ambiental: Evaluacion de Riesgopara Kim Y, Park J, Yoo J and Kwak J 1999. Removal of lead using la Salud Humana (Environmental Toxicology: Human Risk xanthated marine brown alga, Undaria pinnatifida. Process Assessment), McGraw-Hill, Madrid, Spain, p.348. Biochemistry 34(6-7): 647-652. Okmen G, Donmez G and Donmez S 2007. Influence of osmotic and Lee DY, Fortin C and Campbell PGC 2005. Contrasting effects of metal stresses on nitrogenase activity of cyanobacteria isolated chloride on the toxicity of silver to two green algae, from paddy fields. African Journal of Biotechnology 6(15): Pseudokirchneriella subcapitata and Chlamydomonas 1828–1832. reinhardtii. Aquatic Toxicology 75(2):127-135. Påhlsson AMB 1989. Toxicity of heavy metals (Zn, Cu, Cd, Pb) to Li M, Hu C, Zhu Q, Chen L, Kong Z and Liu Z 2006. Copper and zinc vascular plants. Water, Air, and Soil Pollution 47(3-4): 287. induction of lipid peroxidation and effects on antioxidant Rao S 1986. Adaptation of algae to heavy metals toxicity. enzyme activities in the microalga Pavlova viridis International Journal of Environmental Studies 27: 210-223. (Prymnesiophyceae). Chemosphere 62(4): 565-572. Sobhan R and Sternberg SP1999.Cadmium removal using Lichtenthaler HK 1987. Chlorophyll and carotenoids: Pigments of cladophora. Journal of Environmental Science and Health Part photosynthetic biomembranes. Methods in Enzymology 148: A 34(1): 34-53. 331-382. Sohal RS, Mockett RJ and Orr WC 2002. Mechanisms of aging: an Liu DY, Wang YB, Zhang XX and Si Q 2002. Effect of sewage appraisal of the oxidative stress hypothesis. Free Radicals irrigation on wheat growth and its activated oxygen metabolism. Biology and Medicine 33(5): 575-86. Journal of Applied Ecology 13(10): 1319-1322. Srivastava SS, Mishra RD,Tripathi S, Dwivedi D and GuptaK 2006. Magdaleno A, Gómez CE, Vélez CG and Accorinti J 1997. Copper-induced oxidative stress and responses of antioxidants Preliminary toxicity tests using the green alga Ankistrodesmus and phytochelatins in Hydrilla verticillata (L.F.) Royle.Aquatic falcatus. Environmental Toxicology and Water Quarterly Toxicology80: 405-415. 12(1):11-14. Weckx JEJ and Clijsters HMM 1996. Oxidative damage and defense Marklund S and Marklund G 1974. Involvement of the superoxide mechanisms in primary leaves of Phaseolus vulgaris as a result anion radical in the autoxidation of pyrogallol and a convenient of root assimilation of toxic amounts of copper. assay for superoxide dismutase. European Journal of PhysiologiaPlantarum 96(3): 506-512. Biochemistry 47(3): 469-474. Yan C, Hong Y, Fu S, Fang C, Lei J and Shen Q1997. Effect of Cd, Marshall JA and Newman S 2002. Differences in photoprotective Pb stress on the activated oxygen scavenging system in pigment production between Japanese and Australian strains tobacco leaves. Chinese Journal of Geochemistry 17(4): 372- of Chattonella marina (Raphidophyceae). Journal of 378. Experimental Marine Biology and Ecology 272(1): 13-27. Yruela I 2005. Copper in plants. Brazilian Journal of Plant Physiology Mittler R 2002. Oxidative stress, antioxidants and stress tolerance. 17(1): 145-156.

Received 20 August, 2019; Accepted 04 November, 2019 Manuscript Number: S-248 Indian Journal of Ecology (2019) 46 Special Issue (8): 36-39 NAAS Rating: 4.96

Ameliorative Effect of Ascorbic Acid on Hematological Parameters Due to Cadmium Toxicity in Cyprinus carpio

Hamza G. Al-Agidi, Kadhim O.M. AL-Humairi and Sadeq M. Al-Haider1 Al-FuratAL-Awsat Technical University, AL- Musaib Technical College, 51009, Babylon, Iraq 1College of Veterinary Medicine, Al-Qasim Green University, Iraq E-mail: [email protected], [email protected]

Abstract: The present study showed the protective effects of Vitamin Cby reducing the toxicity of cadmium on the hematological parameters in common carp during exposure period. The results of present study showed significant differences at P≤0.05 between treatments with cadmium and control, ascorbic acid only and ascorbic acidic cadmium on hematological parameters. The results also showed the effect of Vitamin C

(ascorbic acid) to reduce cadmium toxicity after 7 to 14 days after chronic exposure to CdCl2 increased white blood cells number, mean corpuscular volume, Mean Corpuscular Haemoglobin and decreased RBC, Hb content, PCV and the mean corpuscular hemoglobin concentration as compared to control were found. The exposure of cadmium with ascorbic acid the WBC, RBC decreased and Hb contents, RBC, the packed cell volume PCV increase values of these parameters as that in control groups when compared with cadmium exposure groups.

Keywords: Ascorbic acid, Cadmium toxicity, Common carp, Hematological parameters

The Heavy metals concentration trend to increase more other chemicals (Adekola et al 2007). Therefore, heavy and more in the all environment spheres especially in the metals monitoring in aquatic environment are very important, aquatic environment. Fishes have been recognized as good especially for breeding of fish. Increased loads of toxic metals accumulators of organic and inorganic pollutants (Eneji et al in waste water may increase the risk of water contamination 2011). Aquatic organisms such as fish accumulate metals in the current study, the hematological parameters were from water or sediment with concentrations higher many studied to evaluate the cadmium chloride impacts on carp times than the concentrations of these metals in the water, fish. also they can concentrate metals at different levels in Red blood corpuscle RBC and hemoglobin can be used different organs, and these heavy metals acquired by the as an indicator foranemia, f1uid volume disturbances. In chain of food as a result of pollution will be posed chemical general, heavy metals have acute effect in hematological hazards and threatening to human. However, low levels of parameters and cause reduction in RBC and Hb. Further, heavy metals, such as copper, cobalt, zinc, iron and anemia could be happened due to plasma increasing volume manganese are crucial for activity of enzyme activity and caused by disturbed water balance and will decrease RBC many other biological processes. Other heavy metals, for production (Saraswathi et al 2003). In addition, as the example cadmium and lead are not recognized to have haemoglobin (Hb) is the main constituents of blood, which essential role in living organisms and they are toxic even at transport of oxygen throughout the blood capillaries, low concentrations, while the essential metals also decreasing the haemoglobin level might reduce the developed to become toxic at high level of concentrations, biological activity of fishes due to stress syndrome. the consequences of heavy metal pollution can be hazardous Researchers have reported alteration in RBC and Hb level to humans through food (Sani 2011). (Garg et al 1989, Magare and Patil 2000), and also in the Fish is the most susceptible aquatic animals to the heavy present investigation on the protective effect of ascorbic acid metals. The physical and chemical parameters of the to reduce the toxicity of cadmium on hematological environment in which the fish resides appeared to affect the parameters in Cyprinu scarpio. rate of bioaccumulation of trace elements (Cao et al 2010). Normally, fish may be considered as one of the healthiest and MATERIAL AND METHODS cheapest source of protein due to it's higher cysteine amino Experimental fishes: The freshwater common carp Cyprinu acid composition than most other sources of protein. The scarpio with a size range of 16-20cm and, weighing 54 ±4 g effects of exposure to any hazardous constituent depend on irrespective of their sex, have been chosen as the test the dose, exposure time, exposure mode, and presence of organism in the present study. Before the exposure Ascorbic Acid on Hematological Parameters Due to Cadmium Toxicity in Cyprinus carpio 37 experiment fishes were acclimatized to the laboratory Red Blood Corpuscular (RBC) measurement: Counting of conditions in large fiber glass tanks with unchlorinated RBC was done with a Neubauer chamber, Sohn and Henry ground water for 3 days at a room temperature of 28 ± 20Cᵒ. (1969). Red glass bead pipette was used for indicting the In order to avoiding the carp fishes overcrowding as its counting chamber (triplicate). benthic in nature, five fishes putted in each tank. After every Haemoglobin (Hb) content: Cyanmethemoglobin method 24h, water was changed completely. To prevent the escaping was performed for determination of Hb (Dacie and Lewis of fishes, tanks were covered with net. No feed used during 1968). 0.02 ml of blood sample was pipetted into Darbkinp's the experiment period (14 days). reagent (5ml) and shaken then allowed to settle for 10 min.

Chemicals and reagents: Cadmium chloride (CdCl2) and after that, absorbance was measured at 540 nm wavelength vitamin C (L-Ascorbic acid) were obtained from the with spectrophotometer against a reagent blank. environmental laboratory at the Collage of Science, Babylon Estimation of Packed Cell Volume (PCV): Evaluation of -1 University, then the CdCl2 was dissolved in H2O (1 mg l ) and PVC was achieved using micro-haematocrit method the L-Ascorbic acid was added with a ratio of 50 mg l-1 (Al (Snieszko 1960).Non-clotted blood specimens (Heparinized) Maamouri 2011). The other chemicals (reagent grade) were were collected in even bored capillaries. It was indorsed to obtained from local scientific distributors in Iraq. run 1/2 - 3/4 lengths of capillary tube closed with sealing wax Animals and experimental design: 60 healthy adult on opposite sides and then assigned to a high speed micro- individuals of common carp Cyprinu scarpio (60 ± 5 g) were haematocrit centrifuge (12000 rpm) for 5 min. obtained from a registered fish farm in Babylon province. 3 White Blood Corpuscles (WBC) measurement: WBCs days period of time was used for fish acclimation. Then it were measure during the method described by Donald spread in glass aquaria with (dimensions, 90 × 30 × 50 cm) Hunter and Bomford (1963). filled with 120 L fresh water(de-chlorinated); the carp fishes Statistical analysis: Data analysis was done using SPSS were kept in a laboratory, setting with a photoperiod, light (12 ver. 23, and present as mean SD. It was considered as hours) and dark cycle (12 hours). Furthermore, temperature significant when P value was 0.05. was maintained of 28 ± 2°C and then water aquarium was aerated. Water dissolved oxygen level (6.5 mg l-1), pH (6.9 ± RESULTS AND DISCUSSION 0.4), and electrical conductivity (219 ± 2 μm ho cm-1) of The results of presents study were to determine the aquaria water which measured using multi meter; conditions impact of cadmium toxicity on C. carpio and ameliorative were closely monitored and kept stable during the effect by L-ascorbic acid on the levels of hematological experiments. They were subdivided into four groups with 15 indices, the hematological indices in presents study showed individuals for each one. Control group were given tap water significant differences at p≤0.05 among the treatments. only .Second group were given Vitamin C at a dose 50 mg l-1. Ascorbic acid alone has no effect on all hematological Third groups were given cadmium at a dose 1 mgl-1 and fourth markers as compared with treatments of cadmium groups were (Vitamin C and cadmium) given Vitamin C 50 RBCs: Red blood cells number in control was 3.5 and 3.3x mg l-1 prior three hours of cadmium in concentration 1 mg l-1 106 mm-3 at 7 and 14 days respectively. While in cadmium administration. Cyprinu scarpio was exposed to sub-lethal exposure, its number was significantly decreased concentration of Cadmium for 14 days. For sub-lethal (2.62–1.46) 106 mm-3 at 7 and 14 days, respectively as exposure experiment, three replicates were maintained. The compared with treatment groups with ascorbic acid which its water was changed every day, so that the concentration of numbers significantly increased at 7 and 14 days (3.12-3.3) cadmium chloride remained the same during the 106 mm-3, respectively (Fig. 1). experimental period. WBC: The number of white blood cells in cadmium exposure Haematological studies: Blood was taken at the end of 7 significantly increased at 7 and 14 days and reached to day and 14 day by catching each fish very gently using a 4.32–5.87x 106 mm-3. While it was decreased significantly in small net, one by one with minimal disturbances. Each fish treatment with ascorbic acid along with cadmium exposure to wrapped in a clean and dry towel was trapped, the posterior be (3.87 – 3.11 x 106/ mm3). The WBC for the control half of its body was cleaned with coarse clean paper. The treatment was (3.22- 3.17x 106 mm-3) (Fig. 2). blood samples were collected using severance of caudal Hb: The Hb level in treatment with ascorbic acid with peduncle and placed in Eppendorf tubes holding 1% (EDTA) cadmium exposure reached at 7 and 14 days (9.1-8.9 g 100 (Mgbenka et al 2003). Haematological parameters were ml-1) as compared with control groups at 7 and 14 days (9.6- evaluated using the methods of Hesser (1960) and Blaxhall 10.8) g 100 ml-1, respectively. The level of hemoglobin and Daisley (1973). percent in cadmium exposure was significantly decreased at 38 Hamza G. Al-Agidi, Kadhim O.M. AL-Humairi and Sadeq M. Al-Haider

7 and 14 days to reach (8.3-7.4) g 100 ml-1 (Fig. 3). increased at 7 and 14 days and reached 28.23 and 16.85%, PCV: The packed cell volume PCV% in the control was (34.6- respectively. While its concentration in treatment with 33.6%) at 7 and 14 days and its percentage in cadmium ascorbic acid plus cadmium exposure decreased at 7 and 14 treatment were significantly decreased from 29.4%-26% days (26.92-26.23%) as compared with control groups at 7 and vitamin C+ cadmium treatments were from 32.9-33.8% and 14 days (31.21-25.8%), respectively (Fig. 7). at 7 and 14 days, respectively (Fig. 4). The percent of MCV in cadmium exposure was 40 7 day significantly increased at 7 and 14 days and reached to 35 112.21-178.08 µm3 While its concentration in treatment with 3 30 14 day 25 ascorbic acid plus cadmium exposure was significantly /mm 6 20 3 decreased at 7 and 14 days (108.33-97.57 µm ) as 10 15 10 compared with control groups at 7 and 14 days (98.85- PCV 3 5 108.33 µm ), respectively (Fig. 5). 0 The MCH in cadmium exposure was 31.67-44.8 pg, Control A.A Cd Cd+A.A while in treatment with vitamin C+ cadmium MCH was Fig. 4. PCV 106 mm-3 in bloods of C. carpio during period of reached 29.16-31.98 pg and in control was ranged between exposure 3085-30.35 pg at 7 and 14 days exposure respectively (Fig. 6). The MCHC in cadmium exposure was significantly 200 7 day

4 3 150 14 day 3.5

7 day /mm 3 3 6 100

14 day 10 2.5 /mm 6 2

MCV 50 10 1.5

RBC 1 0 0.5 Control A.A Cd Cd+A.A 0 6 -3 Control A.A Cd Cd+A.A Fig. 5. MCV 10 mm in bloods of C. carpio during period of exposure Fig. 1. Red blood corpuscular counts 106 mm-3 in bloods of C. carpio during period of exposure 50 7 day

7 3 40 14 day 7 day 6 3

/mm 30 5 14 day 6 10

/mm 4 20 6

10 3 MCH 10

CB 2 W 1 0 Control A.A Cd Cd+A.A 0 Control A.A Cd Cd+A.A Fig. 6. MCH 106 mm-3 in bloods of C. carpio during period of 6 -3 exposure Fig. 2. White blood cells counts 10 mm in bloods of C. carpio during exposure period

7 day 35 12 14 day 7 day 30 3

3 10 14 day 25

8 /mm

6 20 /mm 6

6 10 15 10 4 10 Hb 2 MCHC 5 0 0 Control A.A Cd Cd+A.A Control A.A Cd Cd+A.A Fig. 3. Hb 106 mm-3 in bloods of C. carpio during period of Fig. 7. MCHC% 106 mm-3 in bloods of C. carpio during period exposure of exposure Ascorbic Acid on Hematological Parameters Due to Cadmium Toxicity in Cyprinus carpio 39

REFERENCES Progressive Fish Culturist 22(4): 164-171. Abbas HHH 2006. Acute toxicity of ammonia to common carp Kori-Siakpere O, Ake JEG and Idoge E 2005. Haematological fingerlings (Cyprinus carpio) at different pH levels. Pakistan characteristic as of the African snakehead, Parachacnna Journal of Biological Sciences 9: 2215-2221. obscura. African Journal of Biotechnology 4: 527–530. Adekola FA and Eletta OAA 2007. A study of heavy metal pollution of Kumar R and Agarwal VP 1993. Effect of sublethal value of LC50 of Asa River, Ilorin. Nigeria; trace metal monitoring and mercury chloride on haematological parameters on Clarius geochemistry. Environmental Monitoring and Assessment batrachus. Environment and Ecology 2: 23-25. 125(1-3): 157-163. Magare SR and Patil HT 2000. Effect of pesticides on oxygen Al Maamouri and Anas M Mohamad 2011. Comparative study of the consumption, red blood cell count and metabolites of fish, effects of zinc and chromium elements on common carp fish. Puntius ticto. Environment and Ecology 18(4): 891-894. M.Sc. Thesis, Collage of Science- University of Babel. Mc Donald MD and Grosell M 2006. Maintaining osmotic balance Agarwal RA 1983. Pesticides and environmental pollution. Indian with an aglomerular kidney. Comparative Biochemistry and Farming 33(5): 21-25. Physiology 143: 447-458. Blaxhall PC and Daisley KW 1973. Routine haematological methods Mgbenka BO, Oluah NS and Umeike I 2003. Effect of Gammalin 20 for use with fish blood. Journal of Fish Biology 5: 771-781. (lindane) on differential white blood cell counts of the African catfish, Clarias albopunctatus. Bulletin of Environmental Blaxhall PC 1972. The hematological assessment of the health of Contamination and Toxicology 71(2): 248-254. freshwater fishes: A review of selected literatures. Journal of Fish Biology 4: 593–604. Sani U 2011. Determination of some heavy metals concentration in the tissues of Tilapia and Catfishes. Biokemistri 23(2): 73-80. Chauhan RRS, Saxena RK and Kumar S 1994. Rogor induced haematogical alterations in Cyprinu scarpio. Adv. Bios. 13: 57. Sarswathi K, Ramesh M, Jayalakshmi K, Revathy B and Noortheen A 2003. Haematological responses of fingerlings of Cyprinus Christensen GM, Mickim JM, Brings WA and Hunt EP 1972. carpioto sublethal toxicity of sodium nitrite exposure. Changes in the blood of brown bull head, (Ictalurus nebuloris) Aquaculture 4(1): 49-56. following short and long term exposure to copper (II). Toxicology and Applied Pharmacology 23: 417-427. Sen G, Bahera MK, Behera R and Patel PN 1993. Toxic effect of zinc on freshwater fish, Channa punctatus. Environment and Donald Hunter and Bomford 1963. A dangerous misprint. British Ecology 11: 735-737. Medical Journal 1: 1024. Shalaby AM, Abbas HHH and Khatab YA 2005. The effect of EnejiI S, Sha'Ato R and Annune PA 2011. Bioaccumulation of heavy sublethal doses of cadmium on the disposition of some trace metals in fish (Tillapia zilli and Clarias gariepinus) organs from elements and liver function in common carp Cyprinu scarpio L. river Benue, North Central. Pakistan Journal of Analytical Envt. Egyptian Journal of Basic and Applied Physiology 4: 383–395. 12(1&2): 25-31. Snieszko SF 1960. Microliacmatocrit as a tool in fisheries Garg VK, Garg SK and Tyagi SK 1989. Manganese induced management. Special Scientific Reports-Fisheries. No. 3l4. U. haematological and biochemical abnormalities in S. Dept. Inter. Fish and Fisheries WildLife Special Sci-Report, Heteropneustes fossilis. Journal of Environmental Biology 10: 15. 349-353. Sohn ID and Henry IB 1969. In: Todd-Sunfordclinica diagnosis by Gill TS, Pant SC and Pant J 1988. Gill, liver and kidney lesions Laboratatory methods, 14th edn., W. B. Saunders company, associated with experimental exposure to carbaryl and Philadelphia, London, Toronto. 19: 139-143. diemethoate in the fish, Puntius conchonius. Bulletin of Zikić RV, Stajn S, Pavlović SZ, Ognjanović BI and Saićić ZS 2001. Environmental Contamination and Toxicology 41: 71-78. Activities of superoxide dismutase and catalasein erythrocyte Gupta A, Rai DK, Pandey RS and Sharma B 2009. Analysis of some and plasma transaminases of goldfish (Carassius auratus heavy metals in the riverine water, sediments and fish from river gibelio Bloch) exposed to cadmium. Physiological Research 50: Ganges at Allahabad. Environment Monitoring Assessment 105–111. 157: 449-458. Web JL 1966. Enzymes and metabolic inhibitor. Vol. 3. Academic Hesser EF 1960. Methods for routine fish hematology. The press. New York.

Received 07 October, 2019; Accepted 20 November, 2019 Manuscript Number: S-249 Indian Journal of Ecology (2019) 46 Special Issue (8): 40-43 NAAS Rating: 4.96

Effect of Crude Oil Pollution and Water Strom on Some of the Physical and Chemical Properties of Soil

Mohannad I. Khuder, Ehab M. Amen and Noor Fanar Abdulbaqi Natural Resources Research Center (NRRC), University of Tikrit, Tikrit, Iraq E-mail: [email protected]

Abstract: The current research aimedat studying some of the geotechnical properties of soil polluted with crude oil flowing from wellsin Al- Alam city, southwest Allas oil field.Three stations were selected to represent the study area and two samples were taken from each station. Onesample was polluted and the other one was non-polluted. The physical tests of the polluted and non-polluted samples of soil showed that they were sand soil and well-sorted. The specific gravity values of the non-polluted soil were lower for the polluted soil. The moisture content of the non-polluted soil was 2.14-2.91%, while it was decreased by 1.01-1.53% in the polluted soil. The Atterberg test indicated that the polluted and non-polluted samples of soil were non-plastic. The electrical conductivity of the non-polluted soil was 1.21-2.03 and was a little higher for the polluted soil.The pH values were between 8.14-8.73 for the non-polluted soil, against 7.84-8.11 for the polluted soil. The gypsum content of the non-polluted soil ranged between 6.9-9.3%, whereas, the polluted soil had a lower gypsum contentas 5.3-8.2%. The percent organic content of non-polluted soil was lower as compared polluted soil.

Keywords: Chemical properties, Physical properties, Crude Oil, Water Strom, Health problem

The ecosystem is disturbed by some surrounding siltstone with various rates. The sand deposits are often have factorsas they affect it adversely. The most dangerous factor high porous, forming natural groundwater aquifers besides is pollution by organic material. Soil forms the upper part of its usefulness in different industrial fields.Thedeposits the earth surface.Soil pollution occurs due to imbalance in soil study area cover wide spaces with different thicknesses and components. This happens when unusual materials are were formed due to erosion since the oldest formations that mixed with the natural components of soil,thus affectingits consisted of sandstone, silt and clay.They were formed chemical and physical composition. Oil pollution causes a during the Pleistocene and Holocene ages.Tectonically, the great threat to soil and a major global problem in both study area is located at unstable plate within foot hill zone, industrial and developing countries because of affecting the which is considered as a boundary between the range of soil features (Chorom 2010). Sixteen organic-rich lower folds and the non-trigonometric zone formed by alpine argillaceous limestone and black shale samples from three movements (Jassimand Goff 2006). wellslocated at the Middle–Upper Jurassic Naokelekan The study area is considered as an important area Formation in the west Zagros Basin of northern Iraq were because it contains raw materials suitable for different evaluated as source rocks (El Diasty et al 2018). engineering purposes (raw materials for various construction The study area is located within the boundaries of industries) before being polluted with crude oil flowing from Salahaddin province between longitude (435400-445700) Allas oil field that was destroyed by the terrorist. The and latitude (345000-344000) to thenortheast of Tikrit city in significance of this study lies in raising awareness of people the distance of almost (21) kilometers within the northern living nearby the study area in order to take all precautions to Himreen Anticline (Fig. 1). avoid the pollution risks and its health problems. Some Fatha (middle Miocene) is located at the core and limbs physical and chemical properties of soil have been studied to of northern Himreen Anticline and it is the largest outcrop in understand the effect of crude oil pollution on them. the Anticline.It consists of upper and lower areas separated by a layer of gypsum mass.The lower part consists of several MATERIAL AND METHODS strands of yellowish green or grayish green limestone with The survey wasconductedat three stations representing successive gypsum and limestone; whereas the upper part the polluted area. Two samples were taken from each station, consists of red limestone layered with successive gypsum one sample is polluted, while the other one is non-polluted and calcareous rocks (Georges et al 2010). Injana formation (the distance between the polluted and non-polluted samples is considered as the most widespread formation in is 7 meters) at depth (0-30) cm. By using GPS, the Iraq.Itconsists of successions of sandstone, mudstone and coordinates of each station were determined (Table 1). The Diversity of Fresh Water Caridea and Brachyura 41

Fig. 2. Effect of oil pollution on the agriculture in the study area

Fig. 1. Site map of the study area Table 1. GPS location of stations Station no. Easting Northing samples were kept in bags marked with the station number 1 343507 4433127 and sample location to conduct the necessary tests on these 2 343710 4439001 samples.These procedures were made during April 2017. 3 353800 4410100 Physical tests: Moisture content (ASTM-D-221-698-2004), specific gravity and consistency (Atterberg) limits was done estimated according to US specification. The grain size Table 2. Moisture content of polluted and non-polluted soils in the study area analysis or Sieve Analysis test has been done according to Station no. Polluted soil Non-polluted soil US specification (ASTM-D-422-63-2004). Size analysis test was achieved by employing sieve analysis through using a 1 1.53 (2.17) 2.72 (2.64) number of sieves. 2 1.32 (2.19) 2.91 (2.67) Chemical analysis: All chemical analyses have been done 3 1.01 (2.12) 2.14 (2.68) at Agriculture college Tikrit University. Organic material contenttest has been done according occurrence as a result of a change in the moisture content to standard specification (ICRDA 2003). (Scott 1974). Gypsum content: the standard method for determining Theatterberg limits showed that soil in the study area is the gypsum content for the soils in the study area is based on of non-plastic type containing high level of sand content. The the study of Richards (1954) which contains sedimentation specific gravity variedbetween 2.12-2.17 in the polluted by acetone solution. soil;,while in the non-polluted soil the rangewas between pH value test has been done according to International 2.64-2.68 (Table 2). Based on observing specific gravity Standard Specifications (ICARDA-2003). results of polluted soil shown in Table 3, they are close to Electrical conductivity tests have been done according values of organic soil suggested by Das (1982). This is to the International standard specifications (ICARDA-2003). because the polluted soil has high organic content that leads It is obvious that the oil pollution affects the physical and to decrease its value of specific gravity compared withnon- chemical properties of soil. Accordingly, a lot of grass has polluted soil as the water density is higher than oil density. died because of this pollution which works as a physical Since the transit from sieve average is 200 less than effect to prevent or reduce the water and oxygen from 10%, so, after calculating the grains diameters and the reaching the grass, as shown in Figure 2. percentage of different soil weight sizes, the relation between grain diameter and these percentages has been illustrated in RESULTS AND DISCUSSION a figure on a half logarithmic paper, as shown in Figures (3, 4 Moisture content values of polluted soil ranged between and 5). 1.01-1.53%, but they were higher in non-polluted soil 2.14- Also, the size analysis results of polluted and non- 2.91% (Table 2). The soil has relatively high organic content, polluted soils were identical, and the percentage of small size leading to the probability of swelling and shrinkage (silt and clay) was less than 10% of the sample total weight. 42 Mohannad I. Khuder, Ehab M. Amen and Noor Fanar Abdulbaqi

The percentages of the polluted and non-polluted soils in Table 3. Real values of specific gravity of different soil (Das three stations under study are shown in Table 4. 1982) The results of organic content are shown in Table 5. The Soil types Real values of specific gravity limits organic content in polluted soil ranged between 1.7-2.8, but it From To was lower in non-polluted soil represented as 0.33-0.78. It is Sand 2.63 2.67 noteworthythat the organic material ratio in polluted soil had Silt 2.65 2.7 Clay and Silty clay 2.67 2.9 Organic soil Less than 2

Table 4. Size analysis results of soils (polluted and non- polluted) Station No. Clay, silt (%) Sand (%) Soil type according to unified classification

1 8 92 SW 2 9 91 SW Fig. 3. Grain diameter and percentage of soil weight size for 3 5 95 SW station 1

exceeded the value of 1%, which is high value as0.5% can be a sufficient rate to cause problems in soil strength (Azlan et al 2012). The organic content increased in the polluted soil compared with non-polluted soil because it is saturated with carbohydrates. Besides, the organic contents are type of material that help to reduce soil strength due to many reasons,such as increasing soil permeability and organic soil degradation.Whereasdrying and shrinking occur because of containing very high gaps rate and high compressibility (Dunn et al 1980). Fig. 4. Grain diameter and percentage of soil weight size for station 2 Results of chemical tests showed that the rate of gypsum content of polluted soil ranged between 5.3- 8.2;while non-polluted soil ranged between 6.9-9.3, as shown in table5. A decreasing rate of gypsum in polluted soil was observed when compared withnon-polluted soilbecause of low moisture content in polluted soil.Since the increase of moisture content will dissolve gypsum, there is an inverse relationship between the gypsum content and the moisture content. The existence of gypsum reduces soil resistance (Al-Dewani and Abdulla 1993). PH value results of non-polluted soil ranged between Fig. 5. Grain diameter and percentage of soil weight size for 7.84-8.11, but it rangedbetween 8.14-8.37 for polluted soil. station 3 As shown in Table 5, the pH values of non-polluted soil

Table 5. Chemical analysis tests for polluted and non-polluted soil samples No. station Polluted soil Non-polluted soil

Organic matter Gypsum PH (%) EC ms Organic matter Gypsum PH (%) EC ms content (%) content (%) content (%) content (%)

1 1.7 2.03 8.73 9.3 0.33 2.16 8.11 8.2 2 2.8 1.39 8.27 7.2 0.78 1.47 7.84 6.4 3 2.3 1.21 8.14 6.9 0.64 1.33 7.94 5.3 Diversity of Fresh Water Caridea and Brachyura 43 samples were lowerthan those of polluted soil samples.The ASTM-D-221-698 2004. Standard Test Method for Laboratory pH value affects the soil.Thus, when itis increased, the soil Determination of Water (Moisture) Content of Soil & Rock by Mass. alkaline is increased too;whereas, the solubility of gypsum ASTM-D-422-63 2004. Standard Test Method for Particle-Size and calcium carbonate is reduced. The pH value is affected Analysis of Soils. by the concentration of dissolved carbonate and carbonate in Azlan A, Awng ER, Ibrahim CO and Noorhaidah A 2012. Correlation between Soil Organic Matter, Total Organic Matter and Water water (Collins 1975). Content, with Climate and Depths of Soil at Different Land use in Results of electrical conductivity tests showed that Kelantan, Malaysia. values of polluted soil ranged between 6.9-9.3; but they were Chorom M, Hosseini SS and Motamedi H 2010. Bioremediation of crude oil polluted soil as affected by sewage- sludge.19th World lower for non-polluted soil represented as 5.3-8.2, as shown Congress of Soil Science, Soil Solutions for Changing World, in Table 5. The increaseinvalues of electrical conductivity in pp. 4-7. polluted soil was obvious compared with non-polluted soil. Collins AG 1975. Geochemistry of oil field, water developments, in This isbecause exotic materials, such as carbohydrates, in petroleum science. vol. 1 Amsterdam, Holland. Das MB 1982. Soil Mechanics Laboratory Manual. The University of soil helped to increase the electrical conductivity. Texas at El paso, p.83. Dunn I, Anderson L and Kiefer F 1980. Fundamental of Geotechnical CONCLUSIONS analyses, John Wiely & sons Inc., USA, 414 P. Edition, New York, John Wiely and sons, Inc. London Engineering vol. 1, 2nd Content and specific gravity of the polluted soil were ed., Surrey University press. lower thanthose of non-polluted soil.However, the El-Diasty WS, El-Beialy SY, Peters KE, Batten DJ, Al-Beyati FM, pollutanthad no effect on the results of volumetric analysis Mahdie AQ and Haseeb MT 2018. Organic geochemistry of the middle-upper Jurassic Naokelekan Formation in the Ajil and and Atterberg limits. Hence, their values were identical forall Balad oil fields, northern Iraq. Journal of Petroleum Science and the samples of polluted and non-polluted soils.On the other Engineering 166: 350-362. hand, oil pollution had a clear impact on pH, organic content Georges Stoops, Vera Marcelino and Florias Mees 2010. Interpretation of Micro-morphological Features of Soils and OC and electrical conductivity EC. Unlike the samples of non- Regolith, 2nd edition. polluted soil, the values of pH, OC and EC werehigher in the ICARDA. 2004. ICARDA Annual Report 2003. International Center polluted soil samples.On contrary, the values of gypsum for Agricultural Research in the Dry Areas, Aleppo, Syria.iv+128 pp. content were lower in samples of polluted soilthan those of Jassim SZ and Goff CJ 2006. Geology of Iraq, Published by Dolin, non-polluted soil. Prague and Moravian. Museum, Brno, 341p. Richards LA 1954. Diagnosis and improvement of saline and alkali REFERENCES soils. USDA-SCS Agric. Handbook No.60 U.S. Govt. print. Office Washington, DC. Al-Dewani S and Abdulla F 1993. Swelling properties of Gypsiferous soil 1st, Conference of Soil Mechanics and Foundation, Engin, Scott CR 1974. An Introduction to Soil Mechanics and Foundation. Nccl, Iraq. pp. 350-357. 2nd. ed. Applied science. Ripple Road, Essex. England.

Received 29 August, 2019; Accepted 10 October, 2019 Manuscript Number: S-250 Indian Journal of Ecology (2019) 46 Special Issue (8): 44-49 NAAS Rating: 4.96

Detection of Organochlorine Pesticides in Typha domingensis in the Euphrates River - Middle of Iraq

Amirah Imran, H. AL-Janabi, Hala Jumaah Asree, Ali Abed Gatea, Nawar AL-Janabi and Ahmed Ali Khirallah1 Department of Biotechnology, College of Biotechnology, University of AL-Qasim Green, Babylon, Iraq 1 College of Medicine, University of AL-Ameed, Karbala, Iraq E-mail: amirah.imran77@ gmail.com

Abstract: The specimens of this plant, were collected from five different sites in the Euphrates river. The examination of these specimens revealed, a high percentage (25.058%) of OC compounds (Endrin Ketone), in Typha domingensis, as a compared with lowest percentage (0.385%) of (TCMX Compound) at the 5thsite, followed by highest percentage (24.851%) for (Endrin Ketone) compound at the 4th site, 24.825% in the 3rd site, 24.79% in the 1stsite, and the lowest 24.557% recorded in the2ndsite. These results demonstrate differences between lowest and highest concentration in most pesticide isolation from Typha domingensis This study revealed a significant concentration of the component of the residues of chlorinated hydrocarbon in the sample of water aquatic plant (typha domingenisis).

Keywords: Typha domingensis, Organochlorine (OC) pesticide, Euphrates river, Iraq

The pesticides are important component of integrated pest characterized by high water level of the river and the management. The indiscriminate use resulted in the presence of some plant species. The third site, located on the development of resistance to pesticides, resurgence of new Euphrates river after passing through the Al-Saddah city pest and pesticide residue. Manu studies are conducted by about 1 km and 15km away from the first site and the river earlier workers and estimated chlorinated pesticides water in this location, under the influence of the human residues in living organisms, (Ali 2012) identified pesticide activity, where the heavy water that directly pour to the river residues in some aquatic organisms Fish in the (Hammar leading to deposit in the middle of river, which allow the Marshes), and other studies on the affected of OC pesticides growth of some aquatic plants. The fourth site was located on that studied the bio-accumulated in plants and animals which the Euphrates river within the agricultural area to the north of leads to contamination of food consumed by humans (Kaiser the city of Al-Hendia about 15 km with intensive agricultural 2000, Meeker et al 2007, Rusiecki et al 2008, Briz et al 2011). activity of this region and the river was characterized by an Typha domingensis has been used as a vital indicator to increase in supply and the presence of few small rivulets that detect the accumulation of pollutants in the aquatic pour into river from neighboring farmland. The fifth site was ecosystem because of their ability to accumulate pollutants in located on the Euphrates river in the Al-Hendiacity before their tissues as well as being a good biomarker because they branching to the (Al-Kufa and Al-Abbasiyah) rivers(1 km), are immobile to detect pollutants in water bodies by the where the emergence of islands was noticed, which was organic pesticides chlorinated in the papyrus plant in the composed of sediments in most seasons and the spread of Euphrates River and its use as a vital indicator of pollution of some aquatic plants. The samples of the papyrus were the river with pesticides. collected from sites, placed in a sterile bags and stored in a box until analysis was done in the laboratory. For extraction MATERIAL AND METHODS and estimation of pesticides from papyrus 5 g of the sample The study area on the Euphrates river included five sites was extracted and mixed with 5 g NaOSO4 and using cold (Fig. 1) from the Al-Musayyib to the Al-Kefal because of their solvents (ether: acetone) by 1:1 (Steinwandter 1992), pre- environmental diversity, first site was located on the concentration of extract, and clean-up of extract was done Euphrates river president in the area of ​​Musayyib near the Al- according to USEPA, (1996). The standard solution was Musayyib bridge and features the high level of river water and prepared and stored under -4°C in the dark. The solutions are the presence of some types of plant intensively. The second prepared daily and diluted with ethyl acetate for being site was located on the Euphrates river president before perishable.Used a GC-MS device with the Varian 3800 branching at the edge of the Al-Hendia river about [2 km], electron microscope with the American Ion-trap mass against the cement factory of Al-Saddah, that was spectrometer (Varian Instruments, Sunnyvale, CA, USA). Detection Organochlorine Pesticides in Typha domingensis 45

The samples were injected into the apparatus directly with injection technique and automatic injector temperature programming. The concentration of samples was calculated according to the following equation: (A) – (Vt) Concentration (µg/L)= (Vi) (Vs) A =Amount of material injected (ng),Vi=Volume of extract injected (μL),Vt= Volume of total extract (μL),Vs =Volume of water extracted (μL) Chlorinated organic compounds Fig. 3. Percentage of total annual total concentrations of TCMX (Tetrachloro meta xylene): The highest percentage chlorinated organic pesticides in papyrus T. of TCMX concentrations in the papyrus plant was at the 2nd domingensis from site 2 site, compared to the lowest percentage of the 5th site. As for the concentration of seasons, the highest concentration during winter and loswetdurinf summer was recorded at all sites. This increased winter concentrations in the aquatic body was mainly due to water temperature. Alpha-BHC, gamma-BHC, beta-BHC and delta-BHC: The percentage of pesticides was higher in 2ndsiteand the lowest

Chlorinated organic compounds Fig. 4. Percentage of total annual total concentrations of chlorinated organic pesticides in papyrus T. domingensis from site 3

Chlorinated organic compounds

Fig. 5. Percentage of total annual total concentrations of Fig. 1. Study sites on the Euphrates river (http://maps. chlorinated organic pesticides in papyrus T. google.ca/maps ) domingensis from site 4

Chlorinated organic compounds Chlorinated organic compounds Fig. 2. Percentage of total annual total concentrations of Fig. 6. Percentage of total annual total concentrations of chlorinated organic pesticides in papyrus T. chlorinated organic pesticides in papyrus T. domingensis from site 1 domingensis from site 5 46 Amirah Imran, H. AL-Janabi, Hala Jumaah Asree, Ali Abed Gatea, Nawar AL-Janabi and Ahmed Ali Khirallah percentage in the 5th site. The highest concentration of these therefore the pesticides are consumed by plants, fish and pesticides was during the winter at all sites and lowest other organisms in the surrounding environment and concentration in summer, indicating that the temperature accumulate in their bodies (Hilfi 2005). increases the proportion of these two compounds as well as Chlordane: It is a pesticide called Cyclodiene derived from other compounds in which the strength of the correlation is Hexachlorocyclopentadiene. The concentrations of very high. chlordane are identified in both Trans and Cis. This may be Aldrin and dieldrin: Aldrin Alicyclic compounds are less due to the repeated use of these pesticides in the agricultural, oxidative than aromatics and oxidize rapidly in the fishing and bird operations as well as the currents from other environment to the results of the present study showing the areas and from the nearby agricultural lands. These presence of these two pesticides at all the study sites . The compounds (which are characterized by their resistance to highest concentration of papyrus aldrin and dieldrin was the degradation and susceptibility) gluing to molecules and even highest value on site 2 compared to the lowest values long-distance transport to areas where such pesticides have ​​identified at 5th site. The same is true for global studies where not been used (in water, sediment and organisms within that pesticide concentrations are lower than in the present study, range). with dildrin concentrations ranging from 0.07 to 7.22 μgl-1 in Endrin, ketone, aldrin and aldehyde: These compounds the water of the Mans River (Mansingh et al 2000). This is due were highest at 5th site. The reason for increase in the winter is to the spraying of agricultural land near the water with the that they are inversely associated with temperatures. The pesticide, causing rain to drift into the nearby waterway flow velocity and pH that have a negative impact on them are (Mallin et al 2011). With regard to the increase of these increasing during the winter due to the erosion of water from compounds, there is a recent source of introduction of aldrin the nearby agricultural lands due to rainfall and these results into water through fishing, agriculture or air sedimentation. are higher than reported by Malik et al (2009), Xue and Xue The increase in dieldrin concentrations at the expense of (2006). The increase in the concentration of these pesticides aldrin indicates the high conversion of aldrin to dieldrin and in the different study sites can be attributed to the dust borne

Table 1. Concentrations of chlorinated organic pesticides in aquatic T. domingensis (μg kg-1) from site 1 during different seasons, 2017-2018 Compounds Summer 2017 Autumn 2017 Winter 2017 Spring 2018 Seasonal mean ± SD

TCMX 4.334 6.865 11.991 9.747 8.234±2.893 Alpha-BHC 7.703 11.639 19.611 16.122 13.768±4.500 Gamma-BHC 5.469 8.472 14.556 11.894 10.097±3.434 Beta-BHC 48.759 69.817 112.466 93.798 81.210±24.074 Delta-BHC 39.317 56.437 91.111 75.934 65.699±19.572 Heptachlor 31.086 44.774 72.496 60.361 52.179±15.648 Aldrin 5.357 8.315 14.305 11.683 9.915±3.381 Heptachlor-epoxide 35.669 51.267 82.858 69.030 59.706±17.832 Trans-Chordane 36.519 52.471 84.781 70.639 61.102±18.238 Cis-Chordane 24.428 35.339 57.436 47.764 41.241±12.473 P,P'-DDE 25.857 37.364 60.670 50.468 43.589±13.155 Endosulfan I 27.794 40.108 65.049 54.132 46.770±14.078 Dieldrin 14.713 21.572 35.464 29.383 25.283±7.842 Endrin 54.818 78.403 126.171 105.261 91.163±26.963 P,P'-DDD 29.824 42.985 69.641 57.973 50.105±15.046 Endosulfan II 31.053 44.727 72.422 60.299 52.125±15.633 P,P'-DDT 12.733 18.767 30.988 25.638 22.031±6.898 Endrin aldehyde 256.290 363.895 581.831 486.434 422.112±123.018 Methoxychlor 78.551 112.033 179.845 150.162 130.147±38.278 Endosulfan sulfate 32.383 46.611 75.428 62.814 54.309±16.266 Endrin ketone 282.547 401.102 641.215 536.111 465.243±135.536 Decachlorobiphenyl 42.195 60.515 97.620 81.378 70.427±20.944 Mean ± SD 51.245±69.673 73.739±98.729 118.088±157.577 98.501±131.818 LSD (0.05) Seasons 13.976 = compounds= 32.776 interferences=65.551 Detection Organochlorine Pesticides in Typha domingensis 47

Table 2. Concentrations of chlorinated organic pesticides in aquatic T. domingensis (μg kg-1) from site 2017-2018 Compounds Summer 2017 Autumn 2017 Winter 2017 Spring 2018 Seasonal mean±SD TCMX 8.233 12.134 20.035 16.576 14.244±4.460 Alpha-BHC 13.426 19.492 31.780 26.402 22.775±6.936 Gamma-BHC 9.982 14.612 23.988 19.885 17.116±5.293 Beta-BHC 76.705 109.161 174.895 146.122 126.720±37.105 Delta-BHC 62.153 88.539 141.982 118.589 102.815±30.167 Heptachlor 49.466 70.563 113.290 94.587 81.976±24.118 Aldrin 9.810 14.369 23.601 19.560 16.835±5.211 Heptachlor-epoxide 56.529 80.570 129.261 107.948 93.577±27.485 Trans-Chordane 57.839 82.426 132.225 110.427 95.729±28.110 Cis-Chordane 39.204 56.021 90.079 75.171 65.118±19.225 P,P'-DDE 41.406 59.142 95.063 79.339 68.737±20.276 Endosulfan I 44.392 63.372 101.812 84.986 73.640±21.698 Dieldrin 24.229 34.802 56.214 46.841 40.521±12.087 Endrin 86.044 122.395 196.018 163.791 142.062±41.558 P,P'-DDD 47.521 67.806 108.890 90.905 78.780±23.190 Endosulfan II 49.415 70.491 113.176 94.491 81.893±24.094 P,P'-DDT 21.179 30.479 49.314 41.069 35.510±10.632 Endrin aldehyde 396.569 562.419 898.320 751.287 652.148±189.606 Methoxychlor 122.622 174.227 278.745 232.995 202.147±58.997 Endosulfan sulfate 51.464 73.394 117.809 98.368 85.258±25.071 Endrin ketone 437.039 619.766 989.848 827.853 718.626±208.900 Decachlorobiphenyl 66.587 94.825 152.014 126.980 110.101±32.282 Mean ± SD 80.537±107.386 114.591±152.170 183.561±242.871 153.371±203.169 LSD (0.05) Seasons= 21.540= compounds= 50.516 interferences=101.03

Table 3. Concentrations of chlorinated organic pesticides in aquatic T. domingensis (μg kg-1) from site (3) during different seasons, 2017-2018 Compounds Summer 2017 Autumn 2017 Winter 2017 Spring 2018 Seasonal mean±SD TCMX 4.728 7.576 13.343 10.818 9.116±3.255 Alpha-BHC 8.518 12.947 21.916 17.989 15.342±5.063 Gamma-BHC 6.004 9.384 16.229 13.232 11.212±3.864 Beta-BHC 54.709 78.400 126.382 105.379 91.217±27.084 Delta-BHC 44.086 63.347 102.357 85.281 73.767±22.020 Heptachlor 34.826 50.225 81.413 67.762 58.556±17.605 Aldrin 5.879 9.206 15.945 12.995 11.006±3.804 Heptachlor-epoxide 39.980 57.530 93.072 77.514 67.024±20.063 Trans-Chordane 40.937 58.885 95.235 79.324 68.595±20.519 Cis-Chordane 27.335 39.609 64.471 53.588 46.250±14.033 P,P'-DDE 28.943 41.889 68.109 56.632 48.893±14.800 Endosulfan I 31.121 44.976 73.036 60.753 52.471±15.839 Dieldrin 16.405 24.122 39.751 32.910 28.297±8.822 Endrin 61.526 88.060 141.800 118.276 102.415±30.334 P,P'-DDD 33.405 48.212 78.202 65.074 56.223±16.928 Endosulfan II 34.789 50.172 81.330 67.691 58.495±17.587 P,P'-DDT 14.177 20.966 34.715 28.696 24.638±7.761 Endrin aldehyde 288.192 409.254 654.443 547.117 474.751±138.402 Methoxychlor 88.226 125.894 202.186 168.792 146.274±43.064 Endosulfan sulfate 36.284 52.292 84.712 70.521 60.952±18.300 Endrin ketone 317.733 451.113 721.253 603.006 523.276±152.485 Decachlorobiphenyl 47.324 67.935 109.679 91.407 79.086±3.255 Mean ± SD 82.363±111.076 132.708±177.283 110.670±148.302 LSD (0.05) Seasons= 15.723= compounds= 36.874 interferences=73.748 48 Amirah Imran, H. AL-Janabi, Hala Jumaah Asree, Ali Abed Gatea, Nawar AL-Janabi and Ahmed Ali Khirallah

Table 4. Concentrations of chlorinated organic pesticides in aquatic T. domingensis (μg kg-1) from site (4) during the different seasons, 2017-2018 Compounds Summer 2017 Autumn 2017 Winter 2017 Spring 2018 Seasonal mean±SD TCMX 3.282 5.312 9.423 7.623 2.321±6.410 Alpha-BHC 5.984 9.141 15.535 12.737 3.609±10.849 Gamma-BHC 4.192 6.601 11.481 9.345 2.754±7.904 Beta-BHC 38.915 55.805 90.014 75.040 19.310±64.943 Delta-BHC 31.342 45.073 72.885 60.712 15.699±52.503 Heptachlor 24.740 35.719 57.954 48.221 12.551±41.658 Aldrin 4.102 6.475 11.279 9.176 2.712±7.758 Heptachlor-epoxide 28.415 40.926 66.265 55.174 14.303±47.695 Trans-Chordane 29.097 41.892 67.808 56.464 14.628±48.815 Cis-Chordane 19.399 28.151 45.875 38.116 10.005±32.885 P,P'-DDE 20.545 29.775 48.469 40.286 10.552±34.768 Endosulfan I 22.099 31.976 51.981 43.224 11.292±37.320 Dieldrin 11.606 17.108 28.251 23.373 6.290±20.084 Endrin 43.775 62.692 101.006 84.235 21.627±72.927 P,P'-DDD 23.727 34.284 55.664 46.305 12.069±39.995 Endosulfan II 24.713 35.681 57.895 48.171 12.539±41.615 P,P'-DDT 10.019 14.858 24.660 20.370 5.533±17.476 Endrin aldehyde 205.375 291.684 466.490 389.973 98.672±338.380 Methoxychlor 62.811 89.666 144.058 120.250 30.702±104.196 Endosulfan sulfate 25.779 37.192 60.306 50.188 13.047±43.366 Endrin ketone 226.435 321.528 514.122 429.819 108.713±372.976 Decachlorobiphenyl 33.650 48.344 78.106 65.079 16.800±56.294 Mean ± SD 40.909±55.885 58.631±79.190 94.542±126.392 78.812±105.731 LSD (0.05) Seasons= 11.210= compounds= 26.289 interferences=52.578

Table 5. Concentrations of chlorinated organic pesticides in aquatic T. domingensis (μg kg-1) from site (5) during different seasons, 2017-2018 Seasonal mean± SD ٢٠١٨ Compounds Summer 2017 Autumn 2017 Winter 2017 Spring TCMX 2.159 3.847 7.267 5.770 1.930±4.760 Alpha-BHC 4.407 7.032 12.350 10.023 3.002±8.453 Gamma-BHC 2.916 4.920 8.978 7.202 2.291±6.004 Beta-BHC 31.795 45.843 74.294 61.841 16.060±53.443 Delta-BHC 25.497 36.917 60.049 49.924 13.057±43.096 Heptachlor 20.006 29.137 47.630 39.535 10.439±34.077 Aldrin 2.841 4.815 8.811 7.061 2.256±5.882 Heptachlor-epoxide 23.062 33.468 54.543 45.318 11.896±39.097 Trans-Chordane 23.630 34.272 55.825 46.391 12.166±40.029 Cis-Chordane 15.564 22.842 37.584 31.131 8.321±26.780 P,P'-DDE 16.517 24.194 39.741 32.935 8.776±28.346 Endosulfan I 17.810 26.024 42.662 35.379 9.391±30.468 Dieldrin 9.083 13.659 22.926 18.869 5.231±16.134 Endrin 35.837 51.571 83.437 69.488 17.987±60.083 P,P'-DDD 19.164 27.944 45.725 37.941 10.037±32.693 Endosulfan II 19.983 29.106 47.581 39.494 10.429±34.041 P,P'-DDT 7.762 11.788 19.940 16.371 4.602±13.965 Endrin aldehyde 170.240 242.023 387.409 323.770 82.066±280.860 Methoxychlor 51.669 74.005 119.243 99.441 25.535±86.089 Endosulfan sulfate 20.870 30.362 49.586 41.171 10.851±35.497 Endrin ketone 187.756 266.844 427.025 356.910 90.417±309.633 Decachlorobiphenyl 27.416 39.638 64.390 53.556 13.972±46.250 Mean ± SD 33.453±46.479 78.045±105.120 64.978±87.936 LSD (0.05) Seasons= 9.323 compounds= 21.865 interferences= 43..729 Detection Organochlorine Pesticides in Typha domingensis 49 by the residues of chlorinated organic pesticides from Environment Monitoring and Assessment 177: 1-21. polluted areas especially during summer and autumn Doong RA, Peng CK, Sun YC and Liao PL 2002. Composition and distribution of organochlorine pesticide residues in surface periods, where the currents settle and the precipitation sediments from Wu-Shi River Estuary, Taiwan. Marine Pollution increases. Bulletin 45: 246-253. Heptachlor and Heptachlor-epoxide: The contents were Halfi Mushtaq Abdul Mahdi Aziz 2005. Effect of insecticides on the pollution of marshlands environment in Southern Iraq. Journal higher than the recorded in 2012 in Iraq and concentration of Mesopotamia for Marine Sciences 20(1): 81-89. very near to the results reported by Turgut et al (2010) in Malik A, Ojha P and Singh K 2009. Levels and distribution of Turkey. This may be due to the entry of these pesticides in persistent organochlorine pesticide residues in water and sediments of Gomti river (India)-A tributary of the Ganges River. Iraqi waters through Turkish (directly deposited in water in Environment Monitoring and Assessment 148(4): 421-435. Turkey). The concentration of these pesticides differ slightly Mallin MA, McIver MR, Fulton M and Wirth ED 2011. Elevated levels in the plant samples where the highest concentrations was of metals and organic pollutants in fish and clams in the Cape Fear River watershed. Archives of Environmental recorded at 2nd site, where the proportion increased in the Contamination and Toxicology 61: 461-471. papyrus plant. Endosulfan compounds recorded the highest Mansingh A, Robinson DE, Henry C and Lawrence V 2000. Pesticide nd contamination of Jamaican environment. II. Insecticide concentrations of endosulfan pesticides in 2 site than other residues in the rivers and shrimps of Rio Cobre Basin, 1982- sites. 1996. Environment Monitoring and Assessment 63: 459-480. DDT (Dichlorodiphenyltrichloroethan): The DDT Meeker JD, Altshul L and Hauser R 2007. Serum PCBs, p,p´ -DDE nd and HCB predictthyroid hormone levels in men. Environmental concentrations increased in 2 site,the increase in Research 104: 296-304. concentrations of DDT compounds in the determinants of the Kaiser J 2000. Endocrine disrupters: Panel cautiously confirms low- current study can be explained by the fact that dose effects. Science 290: 695–697. microorganisms, particularly phytoplankton, thrive in the Rani Manju, Gupta RK, Jyoti Yadav and Sandeep Kumar 2017. A s s e s s m e n t o f o r g a n o p h o s p h a t e s i n d u c e d spring and summer, producing anaerobic degradation of acetylcholinesterase inhibition in Indian major carps. Journal of surface sediments, as a result of the movement of tidal Entomology and Zoology Studies 5(2): 1369-1371. Rusiecki JK, Baccarelli A, Bollati V, Tarantini L, Mooore LE and currents as well as the currents that transport these Bonefeld-Jorgenson EC 2008. Global DNA hypomethylation is compounds (Chen et al 2011). This explains the presence of associated with high serum-persistent organic pollutants in high DDT compounds during winter than in the rest of the Greenlandic unit. Environmental Health Perspectives 116: 1547–1552. seasons. Results of current study agree in terms of increase Salvado V, Quintanta XD and Hidalgo M 2006. Monitoring of with Doong et al (2002), Sundar et al (2010), Chen et al nutrients, pesticides and metals in waters, sediments and fish of a wetland. Archives of Environmental Contamination and (2011), Al-Ali (2012) in different countries and locations. Toxicology 51: 377-386. Decachlorobiphenyl: The highest percentage of pesticide Sharma N, Sumarwar S and Pandey J 2016. Enzymatic responses to concentration in Decachlorobiphenyl was highest at the 2nd pesticide chlorpyrifos exposure in kidney of fish Gambusia th affinis. International Journal of Pure and Applied Bioscience site compared to the lowest percentage in the 5 site. The 4(3): 136-143. results were higher than those of Salvado et al (2006), with Steel RGD and Torrie JH 1980. Principles and Procedures of concentrations of Decachlorobiphenyl in water of 1.05 ngl-1 Statistics. A Biometrical Approach. New York, USA, 633p. -1 Steinwandter H 1992. Development of Microextraction Methods in and sediment of 2.12 ng g , while the concentration of the Residue Analysis. Cairns T and Sherma J, eds. Emerging pesticide was due to the introduction of the pesticide into Strategies for Pesticide Analysis, 3-50.CRC. Boca Raton, F.L. plants through its use in fishing and birds (Hilfi 2005). Sundar G, Selvarani J, Gopalakrishnan S and Ramachandran S 2010. Occurrence of organochlorine pesticide residues in green mussel (Perna viridis L.) and water from Ennore creek, REFERENCES Chennai, India. Environment Monitoring and Assessment 160: 593-604. Ali BSA 2012. Residues of some insecticides in water, sediments and neighborhoods from eastern Hor al-Hammar, Iraq. Ph.D. thesis, Turgut C, Atatanir L and Cutright TJ 2010. Evaluation of pesticide Faculty of Agriculture, University of Basra, Iraq. contamination in Dilek National Park, Turkey. Environment Monitoring and Assessment 170: 671-679. Briz V, Molina-Molina JM, Sánchez-Redondo S, Fernández MF, Grimalt JO,Olea N, Rodríguez-Farré E and Suñol C 2011. United Nations of Environment Program, UNEP 1999. Guidelines for Differential estrogenic effects of the persistent organochlorine the Identification of PCBs and Materials Containing PCBs. P: 2. pesticides dieldrin, endosulfan and lindane in primary neuronal US Environmental Protection Agency (US EPA) 2013. Health Effects cultures. Toxicological Sciences 120(2): 413–427. of PCBs. 31 January 2013. Chen W, Jing M, Bu J, Burnet JE, Qi S, Song Q,Ke Y, Miao J, Liu M Xue N and Xue X 2006. Composition, distribution and and Yang C 2011. Organochlorine pesticide in surface water characterization of suspected endocrine–disrupting pesticides and sediments from the Peacock river drainage basin in in Beijing Guan Ting Reservoir (GTR). Archives of Xinjiang, China: A study of an arid zone in Central Asia. Environmental Contamination and Toxicology 50: 463-473.

Received 14 September, 2019; Accepted 04 November, 2019 Manuscript Number: S-251 Indian Journal of Ecology (2019) 46 Special Issue (8): 50-53 NAAS Rating: 4.96

Effect of Organic and Chemical Fertilizers on Growth and Yield of Cucumber, Cucumis sativus L.

Nasser Jubair Radhi Alhasnawi, Amer Neama Kareem1 and Zina Mohsin Abdullah Department of Horticulture and Landscaping, College of Agriculture, University of Kufa, Kufa, Iraq 1Department of Agronomy, Directorate of Agriculture in Babylon, Province of Babylon, Iraq E-mail: [email protected]

Abstract: An experiment was conducted at the organic fertilizers producing section, to study the effects of different treatments of organic and chemical fertilizers and spraying with microelements liquor on growth and yield of cucumber plants. The experiment included four organic fertilizer treatments (palm fronds compost, corncobs compost, faba bean remain compost, sheep manure and a chemical fertilizer treatment), while the microelements were sprayed at three concentration levels,0.0 ml l-1 (distilled water only), 0.6 ml l-1 and 1.2 ml l-1. Results showed that the faba bean remain compost treatment had the highest rate of leaf chlorophyll content (100.26 spad) with significant difference only from palm fronds compost (76.79 spad). Fertilizer treatments did not differ in fruit length and diameter except the sheep manure, which recorded the highest fruit diameter with significant difference only from palm fronds compost. Faba bean remain compost on the other hand had the highest values of number of fruits plant-1 (56.11) with significant difference only from chemical fertilizer (49.91 fruits plant-1). Generally, fertilizers treatments did not differ among each other in case of the yieldplant-1. Relative to spraying concentrations, treatments of 1.2 ml l-1 resulted in significantly highest values of leaf content of total chlorophyll (93.16 spad), fruit length (14.9 cm) and diameter (2.87 cm), number of fruits plant-1 (57.47) and total yield plant-1 (5.52 kg).

Keywords: Organic fertilizers, Micronutrients liquor, Cucumber

Cucumber (Cucumis sativus L.) in the cucurbitaceae system to ensure the availability of nutrients at sufficient plant family is important summer vegetable crop and widely quantities is of significance to obtain high production with grown in most world countries including Iraq. It is believed to good quality. Moreover, not only safe/effective fertilization be globally spread from its origin from India and Africa. method is required but low cost and environmental friendly Cucumber in Iraq is grown either in open fields during the fertilizers are also necessary. spring and autumn or in protected conditions (plastic houses) Equilibrium of nutrients in plant roots environment plays during early spring and winter. According to (FAO 2012) the vital role in plant growth and high yield crop (Saidi 2005). It is total area occupied by cucumber cultivation was estimated to noted that cultivation of cucumber under protected conditions be 34850 hectare with yield average of 9599 kg ha-1. is usually in company with frequent use of chemical fertilizers Cucumber is cultivated for its fruits to be consumed in fresh added to the soil or sprayed on foliage. Beside the risk of use as salad, cooked or pickled (Matloob et al 1989). environmental pollution, over application of chemical Cucumber is valuable with its nutritional and medical values fertilizers to the soil negatively affect the chemical properties as it is rich in Carbohydrates, P, K, protein, carbs, vitamins C, of the soil, which leads to declining in total production. B1, B2 and Niacin (Hassan 2001). Cultivation of cucumber in Bayoumi (2006) noted that the fertilizing cucumber with protected environment (plastic houses) is one of newly organic fertilizers in combination with marine seaweed adapted agricultural forms in Iraq with increased attention. extracts resulted in a significant increase in plant length, This because protected cultivation can produce and maintain number of leaves, leaf area, total chlorophyll content, early high yield when a crop cannot be grown in open field during and total yield. It was also found that treating cucumber cold weather of early spring and winter. plants with animal and plant manure led to significant The increase in agricultural production is required to increase in plant dry weight and fruit weight (Manmoud meet the needs of the growing population through various 2009). Olaniyi et al (2009) reported that the addition of types of cultivations and practices, including type and organic fertilizers at 4 t ha-1 to three varieties of cucumber method of fertilization. Such increase can be made by significantly increased plant length, number of leaves and focusing on improving the agricultural processes by fruits per plant, fruit weight, total yield and fruit length and increasing production per area unit. As fertilization plays a diameter of the three varieties compared to untreated control major role in compensating the necessary elements for plant plants. growth in the field soil. Thus, an appropriate fertilization The aim of the study, therefore, is to evaluate the effect Organic and Chemical Fertilizers on Growth in Cucumis sativus L 51 of organic fertilizers and spraying with micro-elements liquor covered with soil. Cucumber seeds cv Sayff were planted in on growth and yield of cucumber plants under plastic house 25/12/2015 in holes 45 cm apart with three seeds in each conditions. hole. The chemical fertilizer was added at rate of 100g uria+150 DAP for each line at time of planting as first MATERIAL AND METHODS application. The second application was added 30 days post An experiment was carried out to study the effect of planting and the applications continued every 20 days different treatments of organic and chemical fertilizers and interval. As for microelements treatments, foliar spray liquor foliar spraying with microelements on growth and yield of containing Fe 0.4%, Copper 0.08, Manganese 0.4%, Zinc cucumber cultivar Sayff. The experiment was carried out 0.3%, Magnesium 0.2%, Calcium 0.35%, Boron 0.45%, during the spring 2016 growing season in a plastic house at Molybdenum 0.05% was applied at three formerly mentioned the organic fertilizer factory-Babylon site, Department of concentration levels, the first spray was applied 30 days post Plant Protection. The experiment was conducted in a split planting and additional applications were sprayed every 15 plot design based on complete randomized block design days interval during the experiment period. RCBD with three replicates. Main plots were the At the end of the experiment, vegetative and yield microelement treatments, which were sprayed at three parameters components were measured, including, leaf concentration levels (treatments) (B1; 0 ml l-1-distilled water content of total chlorophyll which was measured for all only, B2; 0.6 ml l-1 and B3; 1.2 ml l-1), while the subplots were experimental units of each treatment using SPAD-502 four organic fertilizer treatments (T1; palm fronds compost, chlorophyll meter, fruit length, fruit diameter, number of fruits T2; corncobs compost, T3; faba bean remain compost T4; plant-1 (total number of fruits in an experimental unit/number sheep manure and T5; a chemical fertilizer treatment). of plants in that unit) and plant yield average (accumulative Before planting, samples were taken from the plastic house yield, kg per experimental unit/number of plants). for soil chemical and physical properties analysis (Table 1). The experimental data were analyzed and analysis of The organic fertilizers under study were also subjected to variance ANOVA was performed using GenStat, 2012, 2nd analysis of contents (Table2). The soil of the plastic house Edition. Means were compared by the least significant was prepared and divided into five furrows of 15.5 m length difference at P≤0.05. and 0.65 m width, which included 15 experimental units per replicate with 1mbetween replicates. A drip line irrigation RESULTS AND DISCUSSION system was performed on the top of each furrow (line). The It can be noticed from Table 3 that the treatment of faba organic fertilizers of each treatment were added at a rate of beans compost recorded the highest rate of chlorophyll in 60 kg line-1 (10 m2) in tunnels along each furrow and then leaves (100.26 SPAD) with significant difference from the palm fronds compost which resulted the lowest rate (76.79 Table 1. Physical and chemical characteristics of the SPAD) but did not differ from other treatments. The reason experiment greenhouse soil may be due to the high nitrogen and phosphorus ratio in this pH EC Per cent N P K (dSm-1) organic matter (mg kg-1) (mg kg-1) (mm ch l-1) treatment (Table 2), which play essential role in plant growth processes, especially nitrogen, which participate in most 7.53 3 1.827 0.68 0.0036 1.050 biological activities including cells division and thus reflected Claygl-1 Siltgl-1 Sandgl-1 Soil texture C C:N on increasing plant vegetative growth, photosynthesis 1.067 1.5588 416 448 136 Sandy silt efficiency and chlorophyll content. Relative to foliar spray treatments with microelements, Table 2. Physical and chemical characteristics of the organic plants treated with the concentration of 1.2 ml l-1 had fertilizers used in the experiment significantly higher chlorophyll content than the two other Fertilizer Character type concentrations. This can be attributed to the presence and pH EC N N C C:N OM positive effects of micro elements in the process of Dcm-1 (%) (%) (%) (%) photosynthesis, especially the magnesium that is involved in Faba beans 7.59 4.67 1.69 0.0956 3.81 1:32 75.72 compost the construction of chlorophyll molecule. Similar effect was -1 Palm fronds 7.46 4.73 1.29 0.099 44.42 1:38 76.6 also recorded in the interaction treatments, the 1.2 ml l compost interacted with faba bean compost resulted in the highest Corncobs 7.92 3.14 1.16 0.095 42.98 1:26 74.12 value of total chlorophyll (104.70 SPAD), which significantly compost differed from the lowest value (73.23 SPAD) from combination Sheep 8.89 3.19 1.40 0.047 49.7 1:35 85.86 manure of spraying of distilled water and palm fronds compost. 52 Nasser Jubair Radhi Alhasnawi, Amer Neama Kareem and Zina Mohsin Abdullah

Both organic and microelement fertilizers increased highest rate of fruit diameter (2.898 cm) with only a significant nutrient availability and plant uptake and absorption difference from the lowest (2.754 cm) in T1. But, there were efficiency, and thus significantly affecting vegetative growth, significant differences among spraying with microelements photosynthesis efficiency and chlorophyll content. This from treatments. The highest fruit diameter was recorded in one hand increases nitrogen abundance and availability to treatment of B3 while the lowest with significant difference plant roots and magnesium among sprayed microelements was in B1 treatment. This might be related to the from another, which have a significant role as they are microelements effect in cells division. One of these is zinc, present in chlorophyll molecules (Addiscott 1974). which is important element in the synthesis of tryptophan that Although T5 recorded the highest mean of fruit length essential in synthesis of IAA which increased cell division as (14.844 cm), but did not significantly differ from other reflected by the increase in fruit diameter (Ali et al 2012). The treatments (Table 4). This may be explained to the equilibrium interaction between fertilizer and spray concentrations effect of maximum limit of this character among treatments. showed significant effect among different interaction However, values of the same character differed among treatments. The combination (T4 - B3) recorded the highest treatments of microelements concentration levels, where the fruit diameter of 2.993 cm, while the combination (B1 - T1) highest fruit length (14.908 cm) was in B3 treatment compared gave the lowest rate of 2.687 cm. This may be due to the to the lowest (14.657 cm) resulted from B1 treatment. The combined effect of the microelements when interacted with reason is mostly due to the role of the microelements and their the components of the fertilizer which is reflected on this effect in increasing fruit growth and length. parameter. Table 4 also showed similar effect in case of interaction It is clear from the results (Table 5) that the treatment T3 treatments. The combination of T5 with B3 treatment had the was only significantly higher than the treatment T5 in case of highest fruit length rate of 15.117 cm while T4/B1 combination number of fruitsplant-1. The other treatments did not differ resulted in the lowest rate that of 14.26 cm. This is a common from each other. Fertilizer treatments were close to each positive effect due tomicro-elements roles within fertilizer other in their effect on number of fruit plant-1. Nutrients in combination, especially nitrogen activities in cell. these fertilizers, in general, contributed in to increase plant Division and nucleic acids (RNA and DNA) synthesis (Al- enzymes activities and enhance flowering and number of Sahaf et al 2011). These results are in line with findings of flowers, or increase vegetative growth by increasing number former studies by Muhammad (2009) Ali et al (2012). of leaves which reflects the increase in the number of flowers The same behavior was observed in case of fruit and in turn the number of fruits. diameter (Table 4). Fertilizers treatments did not differ among In case of comparing the three spraying treatments, each other except for the treatment T4, which gave the results showed that B3 significantly differed from the two

Table 3. Effect of different types of fertilizer and foliar spray Table 4. Effect of different types of fertilizer and foliar spray with microelements and leaf content of total with microelements on fruit length and diameter* of chlorophyll of cucumber plants under plastic house cucumber plants under plastic house conditions conditions Fertilizer type Fruit length (cm) Fertilizer type Leaf chlorophyll content (SPAD) Distilled 0.6 ml l-1 1.2 ml l-1 Average Spraying with microelements concentrations water only B1 B2 B3 Average Palm fronds 14.947 14.693 14.790 14.810 compost (2.687) (2.780) (2.797) (2.754) T1 73.23 74.27 82.87 76.79 corncobs 14.607 14.567 14.823 14.666 T2 77.17 88.20 89.20 84.86 compost (2.793) (2.803) (2.877) (2.824) T3 102.93 93.13 104.70 100.26 Faba bean 14.790 14.823 14.893 14.836 remain compost (2.847) (2.843) (2.800) (2.830) T4 98.77 99.47 100.97 99.73 Sheep manure 14.263 14.750 14.917 14.643 T5 85.07 86.83 88.07 86.66 (2.833) (2.867) (2.993) (2.898) Average 87.43 88.38 93.16 Conventional 14.677 14.740 15.117 14.844 chemical fertilizer (2.750) (2.737) (2.913) (2.800) LSD (P≤0.05) Fertilizers 22.613 Average 14.657 14.715 14.908 Spraying concentrations 4.247 (2.782) (2.806) (2.876) Interaction 23.268 LSD (P≤0.05) Fertilizers 0.376 (0.133) Values are means of 3 replicates. T1; palm fronds compost, T2; corncobs Spraying concentrations 0.235 (0.053) compost, T3; faba bean remain compost, T4; sheep manure and T5; conventional chemical fertilizer. Foliar spray concentrations are distilled water Interaction 0.539 (0.155) only (B1), 0.6 ml/L (B2) and 1.2 ml/L (B3) *Fruit diameter in parentheses Organic and Chemical Fertilizers on Growth in Cucumis sativus L 53

Table 5. Effect of different types of fertilizer and foliar spray Our findings showed the positive response of cucumber -1 with microelements on number of fruits plant and to organic fertilization which often exceeded the treatment of plant yield (kg)* of cucumber plants under plastic house conditions chemical fertilization in some of studied parameters. In -1 general, organic fertilizers used in this study were similar in Fertilizer type Number of fruits plant their effect in improving plant growth and yield. The spraying Distilled water 0.6 ml l-1 1.2 ml l-1 Average only of the micro-elements had a clear effect on all the studied traits as a result of their association with the fertilization Palm fronds 46.83 50.70 56.43 51.32 compost (4.610) (4.910) (5.503) (5.008) system which increased the efficiency of the plant response corncobs compost 44.00 49.20 58.90 50.70 to these elements. Therefore, the study recommends the (4.280) (4.903) (5.573) (4.919) application of the organic fertilization practice because of its Faba bean remain 51.70 54.60 62.03 56.11 compost (4.96) (5.437) (6.047) (5.481) substantial effect on plant and crop. Reducing the use of Sheep manure 47.70 51.10 56.10 51.63 chemical fertilizers and encouraging the adoption of organic (4.453) (4.833) (5.393) (4.893) fertilization trend are highly recommended for its importance Conventional 47.47 48.40 53.87 49.91 chemical fertilizer (4.553) (4.790) (5.100) (4.814) for plants, soil and the environment in general. Average 47.54 50.80 57.47 (4.571) (4.975) (5.523) REFERENCES LSD (P≤0.05) Fertilizers 6.123 (0.829) Addiscott TM 1974. Potassium and the distribution of calcium and Spraying 2.930 (0.287) magnesium in potato plants. Journal of the Science of Food and concentrations Agriculture 25:1173-1183. Interaction 6.551 (0.929) Al-Rubaie BJH, Abu Tilisha JJ and Dueni HK 2011. Effect of foliar

-1 nutrients and method of cultivation in growth and yield of *Yield plant (kg) in parentheses cucumber (Cucumis sativus L.) cultivar Rami grown in plastic houses. Al - Qadisiyah Journal of Agricultural Sciences 1(1): other treatments resulting in the highest average number of 42-51. fruits plant-1 of 57.47 followed by 50.80 and 47.54 from Ali JY, Kamel AN, Abbas AJ and Saleh ZK 2012. Effect of locally treatments of B2 and B1, respectively. This may be due to the isolated cyanobacteria and seaweed extracts on vegetative growth, flowering and yield characteristics of cucumber. Tikrit role of micronutrients, which lead to increase in leaf content Journal of Agricultural Sciences 12(3): 148-154. of chlorophyll, beside the presence of boron with its Al-Sahaf FH, Almuharib MZ and Al-Saadi FMJ 2011. Cucumber importance to increases fertilization, number of flowers/plant hybrids response to chemical and organic fertilizers. Iraqi Journal of Agricultural Sciences 42(4): 52-62. and fruit formation ratio and thus increasing the number of Bayoumi YA and Hafez YM 2006. Effect of organic fertilizers fruits (Al-Rubaie et al 2011). Similar to former findings, c o m b i n e d w i t h b e n z o ( 1 , 2 , 3 ) t h i a d i a z o l e _ 7 _ carpothioics_methy ester (pth) on the cucumber powdery interaction treatments had always significant effect on mildew .and the yield production. Acta Biologica Szagegdionsis measured parameters. Interaction of T3 and B3 recorded the 50(3-4): 131-136. highest rate of 62.03 fruits plant-1, while T2- B1 resulted in the FAO 2009. UN Food and Agriculture Organization. http:Faostat.Ao.Org. lowest that of 44.00 fruits plant-1. This can be attributed to Hassan AAM 2001. The Cucurbits. Al-Dar Al-Arabyia for Publishing combined effect of fertilizer treatments and spraying of and Distribution, 1st Edition, Cairo University, Egypt, pp. 497. microelements by increasing the total vegetation, flowering, Laniyi GO and Algbe DD 2009. Effect of organic- mineral fertilizers on number of flowers and fruit formation, and thus reflected on growth, yield and mineral nutrients uptake in cucumber. Journal of Animal and Plant Sciences 5(1): 437- 442. the increase in the number of fruits per plant. Mahmoud EN, Akader and Robin P 2009. Effect of different organic The results of Table 5 showed that fertilizers treatments and inorganic fertilizers on cucumber yield and some properties. World Journal of Agricultural Sciences 5(4): 408-414. did not significantly differ in their effect on plant yield while Matloob AN, Mohamed EES and Abdul KS 1989. Vegetable microelements spraying treatment did. B3 had the highest Production, Volume 2, 2nd revised edition, Ministry of Higher rate of plant yield (5.523 kg plant-1) while the lowest (4.571 kg Education and Scientific Research. University of Al- Mosul, Iraq. plant-1) was recorded in B1. This is logically attributed to the Mohammed ARS 2009. Effect of nitrogen fertilization and spraying role of nutrients in increasing the number of flowers and fruit with seaweed extracts on growth and yield of cucumber. Diyala formation to be, therefore, reflected on the increase of the Journal of Agricultural Sciences 1(2): 134-145. plant yield. Plant yield is also associated with the number of Rogers GS, Littla SH and Silcockand GN 2002. Extradition of a liquid organic fertilizer from poultry Manure for seize in fenland -1 fruits plant , which is affected by many factors including sheltered horticultural production systems. ISHS Acta availability of some microelements such as boron and zinc. Horticultural 642(XXVLHC): 40-1440. On same trend, the interaction between treatments had Saidi AH 2005. Cultivation of plants under different stress conditions and resources (low input) and their geological basis. Dar- significant positive effects on plant yield. Alnasher for Universities, Egypt.

Received 12 September, 2019; Accepted 04 November, 2019 Manuscript Number: S-252 Indian Journal of Ecology (2019) 46 Special Issue (8): 54-59 NAAS Rating: 4.96

Effect of Application of Pseudomonas fluorescens and Nitrogen Fertilizer on the Growth and Yields of Cultivars of Bread Wheat

Ali Salim Hussein, Hussein Razzaq Nayyef and Bassim Hassan Abdulrudha1 Shatra Technical Institute, Southern Technical University, ThiQar Governorate,64007, Iraq 1College of Agriculture, University of Basrah, Al-Qurnah,Basrah, 61016, Iraq E-mail:[email protected]

Abstract: This work was carried out for two Agricultural seasons from (2016 and2017) to investigate the influence of the Pseudomonas fluorescens and nitrogen fertilizer ratios in some growth and yield traits and components of four varieties of soft wheat (Ashur, Fateh, Abu Ghraib 3, Research 22). The design of the complete random segments was used in the experiment of splinters with four replicates. The main panels represented the four wheat varieties. The secondary plates represented bacterial pollutant and nitrogen fertilizer ratios (bacterial vaccine + 50% nitrogen recommended, 100% nitrogen recommended, no vaccine and no nitrogen).The highest rate of grain yield was in the treatment of bacterial vaccine (4.73 and 4.40 t ha-1), while the rate of production of treatment without vaccine and without fertilizer was 1.45, 1.39 t ha-1. The fertilized treatment was significantly higher than the 100% N fertilized treatment with a relative increase of 28.53 and 27.90 for the 2016 and 2017 seasons, respectively. The results showed that the superiority of the cultivar was significantly higher than the rest of the varieties. The highest yield obtained for the grain yield was about 3.68 and 3.56 ha-1, while the lowest yield was 2.68 and 2.63 t ha-1. Category Research-22.It is recommended that the use of bacterial pollination of seeds during the cultivation of wheat varieties referred to in the study to increase their production and reduce the cost of production derived from the use of high rates of chemical fertilizers.

Keywords: Pseudomonas fluorescens, Bread wheat, Nitrogen

Wheat Triticum aestivum is the first crop in the world in plant height and number of grains in spike. Also, Al-Rifai terms of total cultivated area and production. It also comes at (2001) found that the cultivars were significantly superior to the forefront of basic food crops (rice maize and barley) in the cultivars (99%) and the number of grains in the spike. terms of direct consumption by humans for the high The importance of stabilizing the associated nitrogen nutritional value of carbohydrate and protein compounds of with the wheat plant has led to the study of the amount of the its grains Custis (1986) and Bakunovsky, Ustimenko (1988). netting of bacteria from the mineral fertilizers in wheat plants However, the productivity of this crop is still below the has been shown how important the economic stabilization of required level, especially in Iraq as it does not exceed the nitrogen in wheat and studies conducted by Wong and average productivity of one dunum (650 kg dunum-1). And the Stenber (1985), Millet and Fieldman (1987), Khammas et al importance of economic and food for this crop has become (1992) and Amara et al (2002) reported that bacteria attention to and increase production is very important to contributes to increase the production rate of various crops, achieve self-sufficiency and that is seeking those concerned including wheat, rice and maize. all their efforts to search for new ways to achieve this goal. Okon and Hader (1991) found that the use of the The varieties are one of the important factors that lead to Azospirillum bacterium with the chemical fertilizers (such as the increase of production of good varieties with high ability to nitrogen, phosphorus and potassium) was highly effective in invest the primary resources lead to increase the productive increasing production. In other studies by Omar et al (1995), capacity of the crop where Reynold and others (2000) found Kennedy (1997), Wong and Stenberg (2002) observed large out the differences between the varieties of wheat in the numbers of different types of bacteria on the roots of these qualities of growth and yield components. Other varieties in crops. In this work, a practical study was carried out to the number of spikes m-2 reached 615 spikes. The value of determine the effect of Pseudomonas fluorescens on four Abu Ghraib exceeded in the number of ears m-2 and the class varieties of soft wheat grown in Iraq. of fathers (99) Spike and Assyria in the form of grain, which is no different from Abu Ghraib (3), while Adnanieh gave the MATERIAL AND METHODS highest percentage of protein in cereals which reached 23%. A field experiment was conducted inShattra fields during Siddigble et al (1999) pointed out the difference in the the agricultural seasons from (2016 and 2017) to study the number of leaves, the number of ears m-2, the weight of 1000 influence of the Pseudomonas fluorescens and the grains, the grain yield and the biological yield on the ratio of percentage of nitrogen fertilizer in the growth characteristics, Growth and Yields of Cultivars of Bread Wheat 55 yield components and the biological yield for four varieties of Table 1. Chemical and physical properties for soil soft wheat. The experimental soil, whose characteristics are Soil characteristics 2016 2017 described in Table 1, were two orthogonal plows, and then Soil reaction (pH) 7.8 7.6 the process of smoothing, settling and splitting was carried Electrical conductivity 3.78 3.60 out. R and four replicates. The experimental unit included five Organic materials 1.85 % 1.63 % lines of length (4) meters and a distance of (25) cm between Nitrogen 0.163 1.168 line and another. The seeds were planted by hand in the lines Phosphorus 165.12 ppm 163.18 ppm after the seeds were treated with bacterial vaccine and Potassium 1165 1145 according to the transactions. The isolation of bacteria was Sand 280 275 done in the laboratories of the Department of Plant Slime 385.4 374.6 Protection, Faculty of Agriculture Baghdad. They were multiplied to the center of liquid KB. The pollination was Clay 334.6 350.4 carried out at a temperature of 26-28m for six days and the Soil tissue Green clay mix Green clay mix vaccine was reduced to 1 × 10 concentration. The surface sterilization of the seeds was then carried out with sodium of the bacterial vaccine with 50% nitrogen fertilizer in the hypochloride solution. Then was washed with sterile distilled plant height compared to other treatments, which water and then left to dry on the filter paper. After that, the significantly exceeded the other treatments and gave the seeds of the varieties to be vaccinated for three hours were highest rate of plant height was 88.13 and 87.15 cm for the suspended in the bacterial suspension and then dried on the seasons 2016 -2017 and a relative increase of 35.73 and leaves to be prepared for planting. Phosphate fertilizer was 34.22 compared to the comparison treatment and the added at a rate of 25 kg dunum-1. The addition of nitrogen seasons 2016-2017, respectively. And the highest rate of fertilizer was as follows: 50% of the recommended quantity plant height was 80.31 and 79.66cm for the seasons 2016- and 100% of the nitrogen fertilizer recommended, while the 2017, respectively, with a relative increase of 4.42 and third treatment without nitrogen fertilizer and without fertilizer. 3.95cm in comparison to the research class 22, which gave The seeds were planted for all the transactions on November the lowest rate of plant height of 76.91 and 76.59, 15 (Ansari 1982). Then the soil of the experiment was respectively. These results agree with the conclusion of irrigated and the irrigation and weeding and fighting were Amara et al (1999) that the use of these bacteria works to conducted as needed. Plant height, number of branches, stimulate the plant to produce and regulate the secretion of number of ears m-2, length of the spike, number of seeds growth organizations, which have a significant role in spike-1andgrain yield (t ha-1). increasing the growth and height of plants. Number of Plant Branches RESULTS AND DISCUSSION The results shown in Table 3 showed that the treatment Growth Characteristics of the bacterial vaccine and 50% nitrogen fertilizer was Plant height: The results of the statistical analysis shown in significantly higher in the number of branches of the plant and Table 2 indicate that there is a significant effect of treatment gave the highest rate of this characteristic was 2.73 and 2.55

Table 2. Effect of bacterial and nitrogen rates on plant height for four wheat varieties of agricultural dates 2016-2017 Treatment Ashor Fatih Abu Ghraib 3 Research 22 Average injection

2016 2017 2016 2017 2016 2017 2016 2017 2016 2017

Bacterial 88.32 87.23 9.48 88.48 88.49 87.38 86.23 85.49 88.13 87.15 injection + 50 per cent nitrogen 100 per cent 82.58 83.48 84.58 84.73 81.78 83.18 81.33 81.73 82.56 82.56 recommended nitrogen Without 65.38 64.48 66.88 65.78 64.28 63.38 63.18 62.58 64.93 64.93 ainjection and without nitrogen Average items 78.76 78.39 80.31 79.66 78.18 77.94 76.91 76.59 Value less significant difference below the level of 0.05 Average vaccine = 1.16 (1.56) Average Items = Inflammation of the vaccine × varieties = 0.93 (1.02) 56 Ali Salim Hussein, Hussein Razzaq Nayyef and Bassim Hassan Abdulrudha for the seasons 2016 and 2017 and a relative increase of the comparison treatment (without fertilization and without 125.61, 147.57 compared to the treatment without vaccine vaccine). The results showed that the cultivar superiority without fertilization. The treatment of the bacterial vaccine significantly increased the other cultivars and gave the was significantly higher than the 100% nitrogen fertilization highest rate of spike length of 11.41 and 1082 cm for the recommended and a relative increase of 27.57 and 22.59 for seasons 2016 - 2017 respectively and a relative increase of the two seasons, respectively. 108, 38 and 113.46 compared to the 22-research category, The results showed that the cultivar was significantly which gave the lowest rate of spike length and similar results. superior to other cultivars and gave the highest number of Number of ears m-2: The results shown in Table 5 showed branches plants-1 at 2.19 and 2.03 for the seasons 2016- that the treatment of the bacterial vaccine with 50% nitrogen 2017, respectively. The research gave 22 the lowest number fertilizer ha-1 was significantly higher than the other of branches plants-1 at 1.86 and 1.8. These results are with treatments, with a relative increase of 86.65 and 93.46 for the the results of both Khammas et al (1992). years 2016 and 2017, respectively compared to the Length of Spike cm-1: The results of the statistical analysis comparison treatment (without vaccine and without fertilizer). showed that the bacterial treatment + 50% of the nitrogen It also significantly exceeded the toxic treatment of 100% kg fertilizer significantly exceeded the other parameters in the N and a relative increase of 27.68 and 20.86 for the years length of the spike and gave the highest rate of 12.92 and 2016 and 2017. This is due to the fact that the treatment of the 12.68 cm for the seasons 2016 - 2017 respectively and a bacterial vaccine + 50% fertilizer worked to improve the relative increase of 108, 38 and 113.46 in comparison with qualities of growth and created a source of efficient work in

Table 3. Effect of bacterial pollination and nitrogen fertilizer ratios in number of branches / plants for four varieties of soft wheat for the seasons 2016 - 2017 Treatment Ashor Fatih Abu Ghraib 3 Research 22 Average injection

2016 2017 2016 2017 2016 2017 2016 2017 2016 2017

Bacterial 2.74 2.52 2.96 2.83 2.73 2.46 2.5 2.42 2.73 2.55 injection + 50 per cent nitrogen 100 per cent 2.14 2.9 2.25 2.12 2.11 2.08 2.07 2.06 2.14 2.08 recommended nitrogen Without 1.28 1.01 1.38 1.16 2.18 1.00 1.01 1.06 1.21 1.03 ainjection and without nitrogen Average items 2.05 1.87 2.19 2.03 2.00 1.84 1.86 1.82 Value less significant difference below the level of 0.05 Average vaccine = 0.52 (0.40) Average items = 0.12 (0.10) Inflammation of the vaccine × varieties = 0.15 (0.18)

Table 4. Effect of bacterial and nitrogen fertilizer in spike length for four soft wheat varieties for the seasons 2016-2017 Treatment Ashor Fatih Abu Ghraib 3 Research 22 Average injection

2016 2017 2016 2017 2016 2017 2016 2017 2016 2017

Bacterial 13.09 12.89 13.82 13.30 12.43 12.24 12.36 12.30 12.92 12.68 injection + 50 per cent nitrogen 100 per cent 13.00 12.03 13.14 12.14 11.63 12.33 11.38 11.38 12.28 11.72 recommended nitrogen Without injection 6.18 5.88 7.28 7.03 6.00 5.68 5.38 5.18 6.20 5.94 and without nitrogen Average items 10.75 10.26 11.41 10.82 11.01 9.75 9.70 9.62 Value less significant difference below the level of 0.05 Average vaccine = 0.42 (0.56) Average items = 0.33 (0.23) Inflammation of the vaccine × varieties = 0.67 (0.99) Growth and Yields of Cultivars of Bread Wheat 57 turn to the formation of an efficient mouth is the number of exceeded other treatments and gave the highest number of ears m-2 and some components of the outcome and similar grains in the spike of 67,08 and 65,48 for the seasons 2016 results of those reached by Omar and others (1995) and 2017, respectively, with a relative increase of 105.57 and Khammas et al (1992). In the same table, there was a 107.67 compared to the treatment comparison. significant difference between the studied varieties in the This is due to the fact that the bacterial vaccine research, where the superiority of the cultivar was contributed to the increase of nutrients and then increase the significantly higher than the other varieties, achieving a supply of the substances represented to the spike and the relative increase of 20.08 and 20.86 compared to the 22- completion of the growth of the grain also notes from the research class, which gave the lowest number of snails m2 for same table that there are significant differences between the the seasons 2016-2017. The cultivar was given with the varieties where the superiority of the category open on the treatment of bacterial pollination + 50 N% pike m2 reached rest of the other items involved in the experiment, achieving a 445,00 and 435 for the seasons 2016-2017, respectively, and relative increase of 8 , 68 and 8.2 compared to the this is of course due to the nature of genetic composition of comparison coefficients for 2016 and 2017. These results the varieties and these results agree with the findings of both were consistent with the results of Siddigue et al (1999) and Reynold et al (2000). Riffae (2001). Number of grains spikes-1: The results indicated in Table 6 Weight of a thousand tablets (1000) g-1: The results of the show that the treatment of the bacterial vaccine significantly statistical analysis shown in Table 7 showed that the bacterial

Table 5 . Effect of bacterial pollution and nitrogen fertilizer rate in the number of snails m-2 for four soft wheat varieties for the seasons 2016-2017 Treatment Ashor Fatih Abu Ghraib 3 Research 22 Average injection

2016 2017 2016 2017 2016 2017 2016 2017 2016 2017

Bacterial 417.00 412.00 445.00 435.00 379.18 370.38 356.88 347.68 399.45 390.75 injection + 50 per cent nitrogean 100 per cent 365.00 363.00 373.00 368.00 352.58 336.60 334.58 307.70 356.28 343.50 recommended nitrogen Without injection 211.78 205.00 244.70 219.00 206.28 197.60 193.48 190.30 214.00 201.97 and without nitrogen Average items 331.00 326.00 354.23 342.66 112.68 301.52 294.98 283.50 Value less significant difference below the level of 0.05 Average vaccine = 22.24 (20.88) Average items = 19.28 (19.21) Inflammation of the vaccine × varieties = 22.50 (21.88)

Table 6. Effect of bacterial pollen in the number of grains in spike for four varieties of soft wheat for the seasons 2016-2017 Treatment Ashor Fatih Abu Ghraib 3 Research 22 Average injection

2016 2017 2016 2017 2016 2017 2016 2017 2016 2017

Bacterial 67.68 66.78 68.88 67.38 66.28 64.48 65.48 63.28 67.08 65.48 injection + 50 per cent nitrogen 100 per cent 57.28 56.18 58.58 56.88 55.38 54.58 54.78 53.38 56.50 55.25 recommended nitrogen Without 34.58 33.18 35.18 33.28 31.38 30.49 29.38 29.18 32.63 31.53 ainjection and without nitrogen Average items 53.18 52.06 54.21 52.51 51.01 49.84 49.88 48.61 Value less significant difference below the level of 0.05 Average vaccine = 2.25 (1.65) Average items = 0.97 (0.88) Inflammation of the vaccine × varieties = 1.17 (0.99) 58 Ali Salim Hussein, Hussein Razzaq Nayyef and Bassim Hassan Abdulrudha treatment and 50% of the nitrogen fertilizer were superior to Grain yield t ha-1: The results indicated in Table 8 that both the other treatments in the weight of 1000 grains g-1. The the bacterial vaccine and the varieties significantly affected highest weight of 1000 tablets were 25.73 and 26.6 g for the the grain content of the t ha-1. The treatment of the bacterial seasons 2016 and–2017, respectively and a relative vaccine significantly exceeded the other two treatments and increase of 140,91 and 120.56 in comparison with the gave the highest rate of grain yield of 4.73 and 4.40 t ha-1 for comparison treatment (without vaccine without fertilizer) for the seasons 2016. Respectively, with a relative increase of seasons and respectively. These results are consistent with 226.20 and 207.58 compared to the comparison treatment the results of Hader and Okon (1991).The table also showed (without vaccine and fertilizer). significant differences between the cultivars in this class. The The results showed that the effect of the bacterial cultivar was significantly superior to the other varieties and vaccine was 50% with a significant increase of 100% N and a gave the highest weight of 1000 grains at 21.34 and 22.45 g relative increase of 28.53 and 27.90 for the seasons 2016 for the two seasons, respectively, with a relative increase of and 2017, respectively. These results were identical with 10, 68 and 9.56, (22), which gave the lowest rate of weight of those of Okon and Hader (1991) that the use of bacterial one gram/gram. The reason for this is due to the difference in vaccine with chemical fertilizers such as nitrogen and genotypes. These results are in line with what Khammas et al phosphorus showed a high effectiveness in increasing (1992) and Hader (1991) have shown. production. Table 8 shows that the yield of the cultivar was

Table 7. The effect of bacterial pollination and nitrogen fertilizer ratios in the weight of 1000 g g-1 for four varieties of soft wheat for the seasons 2016 and 2017 Treatment Ashor Fatih Abu Ghraib 3 Research 22 Average injection

2016 2017 2016 2017 2016 2017 2016 2017 2016 2017

Bacterial 25.58 26.79 27.38 27.89 25.48 25.78 24.48 25.93 25.73 26.60 injection + 50 per cent nitrogen 100 per cent 23.88 25.09 25.48 26.29 24.18 24.58 23.28 24.34 24.20 25.07 recommended nitrogen Without 10.58 12.49 11.18 13.18 10.88 11.38 10.10 11.18 10.68 12.06 ainjection and without nitrogen Average items 20.01 21.63 21.34 22.45 20.18 20.58 19.28 20.49

Value less significant difference below the level of 0.05 Average vaccine = 1.14 (1.28) Average items = 1.07 (0.80) Inflammation of the vaccine × varieties = 1.37 (1.30)

Table 8. Effect of bacterial pollination and nitrogen fertilizer ratios in grain yield per ton ha-1 for four soft wheat varieties for the seasons 2016-2017 Treatment Ashor Fatih Abu Ghraib 3 Research 22 Average injection

2016 2017 2016 2017 2016 2017 2016 2017 2016 2017

Bacterial 5.02 4.82 5.26 5.10 4.30 4.05 3.88 3.65 4.73 4.40 injection + 50 per cnet nitrogen 100 per cent 3.64 3.54 4.15 4.02 3.25 3.12 2.95 3.09 3.68 3.44 recommended nitrogen Without 1.52 1.47 1.63 1.58 1.45 1.36 1.22 1.15 1.45 1.39 ainjection and without nitrogen Average items 3.39 3.27 3.68 3.56 3.00 2.84 2.68 2.63 Value less significant difference below the level of 0.05 The average vaccine = 0.58 (0.53) Average Items = 0.32 (0.26) Inflammation of the vaccine × Categories = 0.25 (0.22) Growth and Yields of Cultivars of Bread Wheat 59 significantly higher than the other three varieties. The highest varieties of soft wheat in the Basrah. Master Thesis - Faculty of -1 Agriculture - University of Basra. cereal yield was 3.68 and 3.56 t ha and a relative increase of Al-Rawi, Khasha Mahmoud and Abdel-Aziz Mohamed Khalafallah 37.31 and 38.91 for 2016 and 2017, respectively. As shown in 1980. Design and analysis of agricultural experiments. Ministry the table itself there is a significant overlap between the of Higher Education and Scientific Research - Faculty of Agriculture and Forestry - University of Mosul - the edition of bacterial vaccine and varieties, given that the bacterial Dar al-Kutub for printing and publishing - Mosul - p. vaccine with the class opened the highest rate of grain yield Al-Ansari, Majid Mohammed, Abdul Hamid Al-Yunis, Ghanem of 5.26 and 5.10 t ha-1. While the treatment (without vaccine Saadallah Hasawi and Al-Shamaa 1982. Field Crops Production, Ministry of Higher Education and Scientific and without fertilizer) was given with research 22, the lowest Research, University of Baghdad. -1 rate of grain yield was 1.22 and 1.15 t ha for the two Omar, Sarhan Anam Abdo 2004. Response of some varieties of seasons, respectively. bread wheat for water stress under the conditions of the field. Ph.D. Thesis, Faculty of Agriculture, University of Baghdad. These results are consistent with the findings of Amara MAK, Robie A and Talkhan A 2002. Activity of Pseudomonas Reynolds (2000) and Siddigne et al (1999), who emphasized fluorescenmulant in relation to growth regulators production the different varieties among them on the investment of and biological control in wheat plan. Hnnal of Agriculture Science 14(1): 116 -129. primary resources, which lead to increase the productivity of Bhuiya ZHMS, Islam A, Habem KM, Begum MA and Rahinan ZN the crop, as some varieties outperform others in the product 1998. Effect of blue greealgaen as bio fertilizer on rice. and its components and some other qualities. Bangladesh Journal of Agriculture 9(2): 57-68. Curtis BC 1986. Potential for yield increase in wheat proc – wheat research conf Washington,S-2S . CONCLUSIONS Khammas KME, Ageron PAD and Kasser P 1992. Azospirilliumi This study concludes that the use of Pseudomonas rakense sponovarot and Rhizosphere. Soil Res Microbiol Les 140: 79 – 94. fluorescens. It has a significant role in increasing the yield Millet E and Feldman 1987. Yield response of common spring wheat capacity of the wheat crop, in addition to the ease of Azospirillum brasilens eat various level nitrogen fertilization. application where the seeds can be treated with bacteria Plant and Soil 90: 156-164 . before planting and easily. In addition, the use of the New B and Kennedy 1997. Regional distribution and PH sensitivity of spirillum associated with wheat root in eastern Australia. bacterial vaccine reduces the cost of production of the crop Microbial Ecology 17: 297-304. and the use of low rates of nitrogen fertilizer up to 50% of the Okon Y and Hader Y 1991. Microbial inoculates crop yield enhancer amount of fertilizer recommended. Therefore, the paker E-for soil. Linscolorado 6: 71-95 . researcher recommends the use of bacterial pollen Ustimenko M and Bakumovsky GV 1988. Plant growing in the tropics and sub tropic Mirpubi Moscow 85-109. techniques, which in turn increases the productivity of the Siddigue KHKK, Belford MW, Perry and Tennant D 1999. Growth wheat crop in the unit area. Therefore, reducing the amount Development and light interception of old and modern wheat cultivars in a mediterranean (I type environ ment). Australia of nitrogen fertilizer used to 50% of the recommended rate is Journal of Agricultural Research 30: 374-388. considered important, which can reduce the cost of Reynolds MP, Rajaram S and Ayre KPS 2000. Physiological and production. genetic changes of irrigated wheat in the post grain revolution period and approaches for meeting projected global demand. Crop Science 49: 1312-1326. REFERENCES Wong P and Stenbery NE 1985. Characterization of azosperillium Rifai, Shaima Ibrahim Abboud 2000. Effect of dates of agriculture in form Nitrogen fixing root of harvested sorghum plant. Applied some traits of growth and yield and its components of four and Environmental Microbiology 30: 1180-1196.

Received 19 September, 2019; Accepted 04 November, 2019 Manuscript Number: S-253 Indian Journal of Ecology (2020) 46 Special Issue (8): 60-64 NAAS Rating: 4.96

Evaluation of Hydrochemical Properties of Groundwater and Suitability to Irrigation Purpose and Fixed Sand Dunes in Al- Barsa Area, Eastern Al-Dour City, Iraq

Ehab Mohammad Amen, Mayada Ahmed Abdullah and K. MohannadIssa Natural Resources Research Center (NRRC), Tikrit University, Titrit, Iraq E-mail: [email protected]

Abstract: A study was conducted to evaluate the groundwater in Al-Barsa area, eastern of Salah al-Din Governorate, which sand dunes covers large areas. The water samples were taken from all the 11 wells along the path in the study area and then the physiochemical and chemical properties were analyzed. The results of physiochemical tests of water samples showed: pH 6.99 to 7.68, E.C. 2010 to 3610 µmohscm-1, TDS 1536 to 2267 ppm, T.H. 1232 to 1633 ppm, and SAR 0.79 to 2.12. The chemical analysis results Ca++ 309 to 461ppm, Mg++ 72 to 184ppm, Na+ 68 + - - - to 191ppm, K 9 to 27ppm,SO4=1001 to 1599ppm, Cl 97 to 514ppm, HCO3 112 to 225ppm, NO3 14 to 20ppm.The results showed a difference in the characteristics and quality of the groundwater in the study area and after comparing them with the international standards established by the limits and specifications. Ground water in the area is generally suitable for irrigation uses, which requires exploitation to manage the sand dunes and reduce their environmental, economic and social problems. This result will be reflected in improving the living conditions in the study area and in neighboring villages.

Keywords: Al-Barsa, Groundwater quality, Hydrochemistry, Cations, Anions, Sand dunes

Hydrochemistry is of great importance in the process of The study area covers the rocky outcrops of the period estimating the quality of groundwater sources. Water quality (Middle Miocene-recent), as are the formations: Fatha' reflects the geology of the study area, as well as the Formation, Injana, Mukdadiya and Quaternary Deposits. composition of source rocks and minerals, as these main These Formations may be the main source of sand dunes in factors control the chemical structure of the system (Zoran the study area which may be formed by wind erosion or and Adrian 2008). moved from other places. The quality of groundwater differs from basin to another according different factors that form the aquifer. The salinity MATERIAL AND METHODS and chemical composition of water vary depending on Water was pumped from the well for at least half an hour groundwater sources, type of the rocks passing by its and the to ensure that the sample represents the water quality of the speed of the movement of groundwater. In order to evaluate aquifer. The amount of the sample must be enough for all the the groundwater sources, it is necessary to study the required analysis, it was 2 liters. The temperature of the water hydrochemistry of groundwater. There is need to analyzes in the field was recorded because it contains important quantity and quality of the salts depending on the purpose for indicators, the samples were numbered, and the day and which the water is to be used(Al-Manmi 2008). hour at which the samples were recorded to identify and The study aims at evaluation the quality of groundwater distinguish the sample without suffering in the laboratory. The in Al-Barsa Area eastern Al-Dour city through chemical sample was preserved so as not to change the analyzes of the main ions (cations and anions), minor hydrochemical properties of water before testing component, trace elements, saline and hardness, and The geological, geomorphological and hydrological comparing them with the international standards to information available of the study area was collected and determine their possibility use for irrigation purposes and then prepare of topographic maps. Field work included a first thus fixing the sand dunes, and reducing environmental, survey tour for the study area to identify the economic and social problems. geomorphological effect and the rock outcrops of the Location of thestudy area: The study area is located in Al- geological formations and determine the different field Barsa Village\ Eastern Al-Dour City\ Salahaldin Governorate measurements. A second field tour was conducted to locate between longitudinal lines (43º 59 30") (44º 53 00") and two and coordinate wells by using Global Positioning System latitudes (34º 24 30") (34º 40 00"), and dunes cover large (GPS). More than nineteen groundwater samples were areas of this village (Fig. 1). collected from the wells and along the sand dunes depending Evaluation of Hydrochemical Properties of Groundwater 61 on degree of electrical conductivity (Ec), 11 wells were selected on the basis of differences between them. Laboratory tests: Physical and chemical analysis of water samples collected from the wells of the study area were carried out in the laboratories of the Chemical Engineering Department, Engineering College, Water & Soil Science Department, Agriculture College, and the Central Automated Analysis Laboratory in Tikrit University (Table 1). Accuracy: It is a statistical process which consider as a measure of the appropriateness and nearness of the results from their true values, was measured by (Ionic balance), this was done by measuring the relative difference, (Hem 1989). Ionic balance was used to verify the accuracy of the analysis results and showing its high accuracy (Table 2). E% = ( r. Cat - r. Ani\ r.Cat + r. Ani)* 100. Where Concentrations of ions in epm A = 100 – E% Physiochemical Measurement Electrical conductivity (EC): The electrical conductivity was measured by a Conductivity Meter, type: Con, 200, American company (Lovibond), and at temperature of 25°C.

Fig. 1. Map of Iraq, explaining location of the study area

Total dissolved solid (TDS): TDS was measured using TDS measurement device, of Romania company (HANNA) and at laboratory temperature. pH was measured with Multimeter, total hardness (TH) was measured method of Al-Hamdani (2009): TH = 2.5 Ca + 4.1 Mg, where concentrations of Ca and Mg in ppm. Sodium adsorption ratio (SAR): SAR was by the equation of Todd (2005): rNa SAR = r(Ca + Mg)/2

Where Concentrations of Na, Ca and Mg in epm. Chemical Analyses Major components: They include finding the amount of main ions and major roots in water, as per standard methods (APHA, AWWA and WPCF 1975). The main ions include Mg+2, Ca+2, Na+, K+ and were measured by Atomic Absorption -2 - - Spectrophotometer type, (AA-6200), and SO4 ,Cl , HCO3 , as 62 Ehab Mohammad Amen, Mayada Ahmed Abdullah and K. MohannadIssa

anions, SO4 was measured by ultraviolet spectra photo meter supply and treatment, (Baskaran et al 2009).The pH value - - (UV) at wave length of 420 nm, while Cl , HCO3 by titration. was between 6.99-7.68. Minor components: Nitrate was estimated by Ultraviolet Total hardness: TH depends mainly on the presence of spectra photo meter (UV) at wave length of 220nm and dissolved calcium and magnesium salts, (Deborah 1996). TH 275nm. values ​​ranged from 1206-2704 ppm. Heavy components: This group includes many elements, Total dissolved solid: TDS, are the total dissolved salts in but in this study Fe, Co, Zn, Cu, Mn, Cd, and Ni, estimated a solution, whether ionized or non-ionized and do not with the method as of cations. include suspended solids, and dissolved gases in that RESULTS AND DISCUSSION solution (Todd 2005). TDS values ​​ranged between 1536- Physiochemical characteristics: The results of the 2267ppm. physiochemical characteristics are tabulated in Table 1. Sodium Adsorption Ratio (SAR): It is one of the most Temperature: The temperature of water directly affects important physiochemical properties of irrigation water, many of its physical and chemical characteristics. The which is calculated based on the ratio of sodium to total temperature values ​​of water samples in the study area concentrations of calcium and magnesium (Todd 2005). SAR ranged between 8-25ºC. was 0.79-2.12. pH: pH is an important variable in water quality assessment Chemical analyses as it influences many biological and chemical processes The results of the chemical analyses (Cations and within a water body and all processes associated with water Anions) are tabulated in Table 2.

Table 1.Physiochemical properties for groundwater in the study area o Well No. Easting Northing Elevation pH EC ms/cm TDS ppm T c TH SAR 1 407733 3838574 107 7.23 3610 2267 28 1235 1.37 2 409225 383978 113 7.29 3470 2135 27 1370 1.55 3 409048 3838388 100 7.27 2846 1984 27.5 1633 0.97 4 410713 3839248 106 7.33 2620 1986 26 1410 0.79 5 413565 3839075 112 7.35 2730 1973 26.5 1153 1.45 6 413085 3838882 127 7.4 3000 1898 26 1448 1.38 7 416425 3838317 117 7.3 2570 1776 26 1291 1.45 8 411942 3838649 111 7.56 2420 1770 25.5 1530 2.12 9 413426 3839178 112 7.45 2430 1536 25 1371 1.3 10 414563 3839498 120 6.99 2999 2001 25.5 1467 1.18 11 408925 3838767 107 7.68 2010 1889 25 1232 1.46

Table 2. Chemical analysis for groundwater in the study area No. of well Cations Anions Accuracy % ++ ++ + + -2 - - - Ca Mg Na K SO4 HCO3 Cl NO3 1 319.81 106.7 110.5 27 1001.51 179.4 150.8 16.98 96.8 2 386.79 98.2 131.6 17 998.35 265.3 135 19 93.17 3 425.7 139 90.2 12 1417.89 225.7 97.6 16.13 99.11 4 399.87 100 68.3 18 969.12 201.3 121.6 17.92 92.49 5 334 77.46 113 9 934.71 112.1 140 18.52 95.09 6 461.22 72 121 16 977.65 195.2 180.4 20 91.12 7 349 102 120 12 899.67 164 186.33 15.37 92.43 8 309.9 184.3 191 9 1331.43 152.5 214.7 18 96.57 9 402.33 89 111 15 954.12 254.9 199.53 14 95.66 10 381 125.4 104.2 13 1184 128.8 385.23 15 94.9 11 317.4 107 118 26 1004.1 209.8 166 13.9 98.01 Evaluation of Hydrochemical Properties of Groundwater 63

Cations main source is gypsum in addition to clay minerals. Calcium (Ca+2): Calcium values in water samples of the Sodium (Na+1): It's values was 68-191 ppm. Evaporative rock study area ranged between 309-461 ppm, The main source is the most source of sodium (Al-Hasnawi 2009). of calcium ion in groundwater is the dissolution of some Potassium (K+1): It's concentrations was 9-27 ppm. sedimentary minerals (Helstrup and Yakubo 2007). Potassium ion in ground water is usually low as it is much less Magnesium (Mg+2): It ranged between 72-184 ppm. It is a than other cations (Helstrup and Yakubo 2007). Anions Chloride (Cl-1): Its values between 97-514 ppm. The most Table 3. Heavy components for wells water in the study area important sources of groundwater are Halite and Sylvite or Well No. Fe Co Zn Cu Mn Cd Ni from rainwater (Al-Hasnawi 2009). 1 0.01 0.001 0.04 0.008 0.001 0.02 0.01 -2 Sulfate (SO4 ): It's value ranged between1001-1599ppm, Its 2 0.01 Nil 0.021 0.007 0.002 0.033 0.011 sulfate source is caused by the dissolution of evaporates 3 0.01 Nil 0.029 0.002 0.002 0.03 0.001 such as: Gypsum and anhydrite, the decomposition of 4 0.01 0.002 0.02 Nil 0.007 0.031 0.01 organic materials which used in agriculture fertilizers (Al- 5 0.021 Nil 0.01 Nil 0.001 0.03 0.01 Zaibe 2000). - 6 0.1 Nil 0.02 0.001 0.006 0.044 0.02 Bicarbonate (HCO3 ): It's concentrations ranged between 7 0.01 Nil 0.01 Nil 0.009 0.03 0.02 112-225 ppm. This ion is found in groundwater due to the dissolution of limestone rocks and salt deposits of geological 8 0.01 Nil 0.04 0.002 0.005 0.021 0.02 formation (Al-Zaibe 2000). 9 0.01 Nil 0.03 0.001 0.003 0.01 0.01 10 0.02 Nil 0.02 Nil 0.02 0.01 0.01 11 0.02 Nil 0.01 Nil 0.01 0.022 0.02 Table 5. Standard specifications for Irrigation water based on Ayres and Westcot, 1989 Variables Symbols Unit Acceptable limit Table 4. Hydrochemical variables for well water in the study of irrigation water area Variables Parameters Variables Parameters Electrical conductivity EC Μmohs/cm 0-3000 range range Total dissolved solids TDS mg l-1 0-2000

-2 pH 6.99-7.68 SO4 (ppm) 899.67–1417.89 pH pH 14-Jan 6-8.5 EC (µmohs cm-1) 2010–3610 Cl- (ppm) 97.6–385..23 Sodium adsorption ratio SAR mg l-1 0-15

- +2 TDS (ppm) 1536–2267 HCO3 (ppm) 112–265.3 Calcium Ca epm 0-20

ο -2 +2 T (C ) 25–28 NO3 (ppm) 13.9–20 Magnesium Mg epm 0-15 TH(ppm) 1232–1633 Fe (ppm) 0.01- 0.02 Sodium Na+1 epm 0-40 SAR 0.79 - 2.12 Cu (ppm) < 0.01 Potassium K+1 epm 0-2

++ -1 Ca (ppm) 309 – 461 Cd (ppm) 0.03 – 0.01 Bicarbonates HCO3 epm 0-10 Mg++ (ppm) 72 – 184 Zn (ppm) 0.01 - 0.04 Chloride Cl-1 epm 0-30

+ -2 Na (ppm) 68 – 191 Ni (ppm) 0.02– 0.01 Sulfate SO4 epm 0-20

+ -1 K (ppm) 9 – 27 Mn (ppm) 0.001 - 0.02 Nitrate NO3 epm 0-2

Table 6. Crops tolerance to concentrate salts (Todd 1980) Types Crops resistant to low concentrations of Crops resistant to moderate Crops resistant to high concentrations total dissolved salts in water concentrations of total dissolved salts in of total dissolved salts in water water Fruits 0 – 3000 μmohs/cm 3000 – 4000 μmohs/cm 4000 – 10,000 μmohs/cm Lemon, strawberry, peach, apricot, Olives, figs, pomegranates. Palm trees orange, apple Vegetables 3000 – 4000 μmohs/cm 4000 – 10,000 μmohs/cm 10,000-12,000 μmohs/cm Radishes, Celery Cucumbers, basalt, onions, carrots, Spinach - beets potatoes, tomatoes Field crops 4000- 6000 μmohs/cm 6000- 10000 μmohs/cm 10,000 – 16,000 μmohs/cm Field Legumes Corn, rice, wheat Cotton, sugar beet, barley 64 Ehab Mohammad Amen, Mayada Ahmed Abdullah and K. MohannadIssa

- Nitrate (NO3 ): Its concentrations (14-20) ppm. Its sources in REFERENCES groundwater are rainwater, wastewater, soil biological Zoran S and Adrian I 2008. Ground water Management in Northern Iraq. Hydrogeology Journal, No .4. processes, and the result of agricultural activity (Manii 2009). Al-Manmi DA 2008. Water Resources Management in Rania Area Trace elements: The concentrations of trace elements were Sulaimaniyau NE-Iraq. M.Sc. Thesis, University of Baghdad found for the water samples: Iron, Cobalt, Zinc, Copper, (Unpubl), 225p. Manganese, Cadmium and Nickel, Table 3. Its Hem JD 1989. Study and Interpretation of the Chemical Survey. Water Supply Paper 2254, pp. 246. concentrations were very few and within its natural limits. Al-Hamdani JA 2009. Hydrochemical Effect of Groundwater Due to The results of Physiochemical characteristics, (TDS, Irrigation and Drainage Projects in Tawuq Sub-Basin (South of EC, PH, T.H and SAR), and Results of Chemical analysis Kirkuk - North of Iraq). M.Sc. Thesis, University of Baghdad (Unpubl), 121 P. (Cations, Anions, and Trace Elements), Compared with Todd DK 2005. Groundwater hydrology, 3 Edition. John Wiley and standard specifications for Irrigation water(Ayers and Sons, New York, USA, 650p. Westcot 1989) Table 5 and crops tolerance to concentrate APHA AWWA and Wp CF 1975. Standard methods for the examination of water and wastewater. American Public Health salts Table 6 (Todd 1980), and shown the validity these water Association, (14th ed.), Washington, 759p. for irrigation and most of the agricultural crops except three BaskaranSundaram et al 2009. Groundwater sampling and analyses-A field guide. Geoscience Australia, Australia, 105 p. wells (3,8 and 10) because to the increase of SO4 ions, as well as suitability to irrigate plants (Jatropha, Deborah Chapman 1996. Water quality assessments - A guide to use of biota, sediments and water in environmental monitoring. Eucalyptus)(Abou Kheira 2009) (Brenda et al 2015), which Second Edition, Printed in Great Britain at the University Press, can be used in the fixed of sand dunes in the study area and Cambridge, 651 pages. Helstrup T, Jorgensen NO and Yakubo BB 2007. Investigation of disposal of the environmental effects resulting from them in hydrochemi calcharacteristics of groundwater from the addition to the advancement of agricultural reality. cretaceous-eocenelime stone aquifer in Southern Ghana and Southern Togo using hierarchical cluster analyses. Hydrogeology Journal 15: 977-989. CONCLUSIONS Al-Hasnawi SS 2009. Groundwater Quality Index for Dammam After comparing the physiochemical characteristics for Formation in Al- Najaf Area. M.Sc. thesis, Univ. of Baghdad the water samples which are selected from the wells with (unpubl), 189p. Al-Zaibe AH 2000. Geochemical and Environmental Characteristics standard specifications for irrigation water and classification of the Soil and Groundwater Sources in the North Eastern of of crops tolerance to concentrate salts, it was found to be Region of Almafreq. Jordan, Unpubl. Ph.D. Thesis, Univ. of within the permissible limits for irrigation. Depending on the Baghdad, 201p. Manii JK 2009. Hydrochemistry of Al-Hammar Marsh Environment results of the chemical analysis of the water wells and after Southren Iraq. M.Sc. Thesis, University of Baghdad (unpubl), comparing them with the specification, the concentrations of 205p. the main ions (Cations and anions) were within the Ayers RS and Westcot DW 1989. Water Quality for Agriculture, Irrigation and Drai. Paper 29, Rev.1, FAO, Rome, Italy, 174p. permissible limits for irrigation, except three wells (3,8 and Todd DK 1980. Ground water hydrology, 2 nd ed., John wiley& sons 10) because to the increase of sulfate ions. Generally, the New York, USA, 535p. results indicated that the groundwater in the study area AbouKheira AA and Atta NN 2009. Response of Jatrophacurcas L. to suitable for irrigation. Groundwater wells in the study area are water deficit: Yield, water use efficiency and oilseed characteristics. Biomass and Bioenergy 33: 1343 - 1350. suitable for irrigation of plants Jatropha, Eucalyptus that can Brenda Rodrigues de Souza, Igor Alberto Silvestre Freitas, Vinicius be used in sand dune fixation was found in the area, and de Araujo Lopes, Vanessa do Rosario Rosa and Fabio Santos therefore, reduce their environmental, economic and social Matos 2015. Growth of eucalyptus plants irrigated with saline water. African Journal of Agricultural Research 10(10): 1091- problems. 1096.

Received 14 September, 2019; Accepted 04 November, 2019 Manuscript Number: S-254 Indian Journal of Ecology (2019) 46 Special Issue (8): 65-71 NAAS Rating: 4.96

Effect of Heavy Metals on the Protein and Chlorophyll Content of Phragmitus australis and Typha domingensis

Qassim Ammar Ahmood AL-Janabi, Ziad Bassim Hameed1 and Saad Kadhim Ala Allah Al- Kalidy Department of Environment, 1Department of Pollution College of Environmental Science, Al-Qasim Green University, Iraq E-mail: [email protected]

Abstract: This study investigated the phytoremediation potential of two aquatic plants (Typha domingensis and Phragmitus australis), for the removal of three concentration of heavy metal salts; Nickel chloride (NiCl2), Manganese chloride (MnCl2), Cobalt chloride (CoCl2), and Zinc chloride (ZnCl2) and determine concentration of total chlorophyll and protein through exposure of two aquatic plants (Typha domingensis and Phragmitus australis) for heavy metal. The experiment was terminated after 30 days and the samples of the two plants were taken each after 10 days to determine heavy metal, protein and chlorophyll. The results of study demonstrated height various in concentration of heavy metal in tissue of the plant in the end of experiment and the results of study show decrease concentration of total chlorophyll and protein for the aquatic plant.

Keywords: Phytoremediation, Heavy metal, Aquatic plants

Heavy metals are among industrial pollutants, in concentrations of metal in surface water can present a threat contrast to organic materials cannot be degraded in the to human health and the environment. This increase in the environment. Beside the natural activities, nearly all human concentration of heavy metals is a major human and actions can also affect the production of heavy metals as side ecosystem concern (Kabata-Pendias 2011). Due to their biodegradable character, remediation of such pollutedـeffects, accumulating in water, soil, sediment and living non organisms. Migration of these pollutants into no polluted systems requires instant and essential evaluations. areas as dust or leachates through the soil and spreading of Phytoremediation was chosen due to its cost effectiveness, heavy metals include sewage sludge area few examples of ecofriendly nature and easy maintenance of all remediation events contributing towards contamination of the techniques (Uqab et al 2016). ecosystems (Gaur and Abdholeya 2004). Heavy metals are Phytoremediation is an innovative tool with greater clarified as metallic elements that have a comparatively high potential for removing heavy metal pollutants in water bodies density compared to water. Heavy metals are defined as by using plants to clean up soil, sediments and water metallic elements with an atomic number > 20.Cd, Cr, Cu, pollution. This technology is active that is environmentally Hg, Pb, and Zn are the most prevalent heavy metal friendly and possibly coasted. Plants with outstanding ability contaminants. Metals are natural components in soil. Some for metal accumulation are known as hypera ccumulator of these metals are plant-growing micronutrients such as Zn, plants (Cho-Ruk et al 2006). Phytoremediation benefits from Cu, Mn, Ni, and Co, while others have unknown biological the distinctive and selective uptake capacities of plant root features such as Cd, Pb, and Hg. Certain of these are vital to systems, along with the translocation, bioaccumulation and plant growth and are known as micronutrients such as Zn, degradation capabilities of the entire plant body Hinchman et Cu, Mn, Ni, and Co, while others (Cd, Pb, and Hg) have al 1995). Specific plant species can absorb and hyper- unknown biological features. Biological systems are metal accumulate metal contaminants and/or excess nutrients affected and do not present biodegradation but can be from growth in harvestable root and shooting tissue as they accumulated in Living organisms, causing numerous are capable of bioaccumulating large quantities of toxic diseases and disorders even at comparatively low levels metals and nutrients compared to terrestrial plants (Pratas et (Clark1993). Due to various anthropogenic activities such as al 2012). Furthermore, these macrophysics have been burning fossil fuels, release of municipal waste, use of discovered to absorb these pollutants at distinct rates and fertilizers and pesticides, etc., the concentration of heavy efficiencies depending on biochemical structure, habit, metals in the atmosphere has risen.These surplus species, abundance and climate. 66 Qassim Ammar Ahmood AL-Janabi, Ziad Bassim Hameed and Saad Kadhim Ala Allah Al- Kalidy

Studies have discovered that these crops generate according to the following equation. -1 metal binding cysteine rich peptides (phyto-chelatins) during Total chlorophyll (mg g ) = [A645 (20.2) + A 663 (8.02)] v/w x pollutant stress, which detoxify heavy metals by making them 1000 complex(Kinnersely,1993). Plants are able to remove both A645 = the absorbance at 645 nm water and soil contamination from metal. Aquatic plants of all A663 = the absorbance at 663 nm kinds are known to remove heavy metals, whether free W= plant tissues weight floating, submergedor emerging. The aim of this document is Heavy metal concentration: After 30 days of growth, to address the potential of the Phytoremediation method for aquatic plants tissues were collected to determine heavy the treatment of heavy metal contaminated side to provide metals concentrations. Plants tissues were dried at 70◦C, data on the processes taken by crops for heavy metal uptake grounded, sieved through a 40 mm mesh. 0.5 g of the and also to provide an effective short list of aquatic plants for grounded and sieved plants tissues was placed into a Pyrex the remediation of different metals. digestive tube. The samples were left for 16 hours after This study has been conducted to provide some adding 5mL of HNO3, then the samples were digested at overview of the performance of plants to take up heavy 100◦C for an hour. After that, 3mL of 70% perchloric acid was metals and explain the destiny of heavy metals in plant added for each sample and a reflux was done to samples for tissue, particularly arsenic (As), lead (Pb) and mercury 30 minutes at 200◦C until getting a clear solution. The (Hg). samples were centrifuged for 10 minutes at 2000 round minute-1 to get rid of any suspended matter in the sample MATERIAL AND METHODS solution. The samples were filtered through 0.45 µm and the Two aquatic plant species, which are Typha samples completed to 50mL with deionized water. Heavy domingensis and Phragmitus australis were used to study metals concentration in µg g-1 of plant tissues was measured heavy metal effects on total chlorophyll and protein. The by atomic spectrophotometer (Onder et al 2007 and Anton experiment was conducted on a laboratory scale by taking ijevic and Maric 2008). 250 gm wet weight of each plant and adapted in 15 L plastic containers. Five sets of microcosms were set in triplicates, RESULTS AND DISCUSSION which were untreated control, treated with NiCl2, treated with The results of the study showed an increase in the

MnCl2, treated with CoCl2, and treated with ZnCl2 for both concentration of heavy elements in the studied aquatic plants Typha domingensis and Phragmitus australis. Three at the end of the experiment. In the aquatic plant Typha concentrations, 10, 20, 30 mg l-1, were used for each heavy domingensis, 1.47.1.35.1.24 mg g-1 for dry weight of Ni and metal salt. The experiment was terminated after 30days and 3.39, 3.13, 2.86 mg g-1 dry weight of the Zn, 5.71, 5.27, 4.83 the samples of the two plants were taken each 10 days to mg g-1 dry weight of Mn and 3.59, 3.31, 3.03 mg g-1 dry weight determine heavy metal, protein, and chlorophyll. of Co was observed under different concentrations of heavy Protein concentration: Total protein amount in plant tissues metals. The concentrations 30, 20, 10 ppm compared with was determined according to Bradford method. 0.5 g of fresh the control sample are shown in the Figure 1. In Phragmitus plant tissues was smashed by a ceramic mortar and then 1ml australis 1.29, 1.19, 1.09 mg g-1 dry weight of Ni, 3.23, 2.98, of phosphate regular solution and 5 ml of Bradford solution 2.73 mg g-1 dry weight of Zn, 5.55, 5.12, 4.77 mg g-1 dry were added. The spectrophotometer was measured by weight of Mn and 3.39, 3.13, 2.86 mg g-1 dry weight of the spectrophotometer Sp-300 on the wave length 595 nm, and Co was recorded in the concentrations of 30, 20, 10ppm, the protein content was expressed as mg g-1 of plant tissues respectively compared with the control sample (Fig. 2). This (Park 2010). indicates the ability of the studied aquatic plants to Chlorophyll concentration: The total chlorophyll accumulate this element within the plant tissues, or that they concentration in aquatic plants tissues was determined as possess a special mechanism to tolerate high concentrations described by (Aminot and Rey 2000). 0.15 gm of fresh of elements or that they absorb the elements with high tissues of aquatic plants was mashed by a ceramic mortar concentrations, which are transformed into inactive forms of after adding 2 ml of 80% acetone. The mashed plant tissues gaps (Coleman et al 2001). The differences in the were filtered through Watmann No. 1 filter paper, 0.45 µm. concentration of elements accumulated in plant bodies may The extract was gathered, and the size was finalized to 15 ml be due to differences in plant species, plant physiological with 80% acetone. The absorbance was measured by status and receptivity to the element (Forstner and Wittman spectrophotometer Sp-300 on 645nm and 663 nm. The total 1981). The sources show that when exposed to heavy chlorophyll content mg g-1 of plant tissues was calculated elements, plants interfere with the removal of toxicity and Phragmitus australis and Typha domingensis Effectiveness by Heavy Metals 67

1.6 6 1day Ni MN 1.4 10day 1day 5 20day 10day

g 1.2 / 20day

30day g

/ 4 30day 1 tions mg a tions mg tr 0.8 a 3 tr

concen 0.6 concen 2 0.4

1 0.2

0 0 Control 10ppm 20ppm 30ppm Control 10ppm 20ppm 30ppm

4 4

1day Co Zn 3.5 3.5 10day

1day g

20day / 3 g 3 / 10day 30day 2.5 2.5 20day tions mg a

tions mg 30day tr a tr 2 2

concen 1.5

concen 1.5

1 1

0.5 0.5

0 0 Control 10ppm 20ppm 30ppm control 10ppm 20ppm 30ppm

Fig. 1. Three different concentrations of Ni, Co, Zn, and Mn during time periods in Typha domingensis tissues

1.4 Ni 6 MN 1day 1.2 1day 5 10day

g

g 10day / 20day / 1 20day 4 30day 30day tions mg

0.8 tions mg a a tr tr 3 0.6 Concen

Concen 2 0.4

1 0.2

0 0 Control 10ppm 20ppm 30ppm Control 10ppm 20ppm 30ppm

4 3.5 Co Zn 3.5 1day 3 1day 10day 3 10day

2.5 g

20day g / / 20day 2.5 30day 30day 2 tions mg tions mg a a 2 tr tr .5 1.5 Concen Concen 1 1

0.5 0.5

0 0 control 10ppm 20ppm 30ppm Control 10ppm 20ppm 30ppm Fig. 2. Three different concentrations of Ni, Co, Zn and Mn during time periods in Phragmitus australis tissues. 68 Qassim Ammar Ahmood AL-Janabi, Ziad Bassim Hameed and Saad Kadhim Ala Allah Al- Kalidy natural balance of the heavy elements in plants. This is done manganese and 0.87, 1.08, 1.19 mg g-1 dry weight in the by the enzyme Phyto-chelatin Synthase, which activates the presence of the cobalt in the concentrations 30, 20, 10 ppm, presence of heavy element ions using glutathione as a base respectively was recorded when compared with the control material. The response of cells depends on the type of (Fig. 3). Phragmitus australis had a low concentration of element found (Steffen 1990). Favero et al (1996) noted that chlorophyll to Typha domingensis (Fig. 4). The decrease in there are many external factors that influence the chlorophyll concentrations in the experimental plants is due concentration of heavy metals in the tissues of the organism, to the presence of these highly toxic substances and has the including salinity, degree of pH, the effectiveness of complex potential to accumulate in the plant tissue. It inhibits its organic and inorganic molecules, and their effect on physical synthesis by inhibiting the action of the enzymes responsible and chemical processes that control the rate metabolic for its production, such as the amino-levulinic acid processes such as temperature, intensity of light and amount dehydratase and Porphobilinogen deaminase, which is of oxygen. Bioaccumulation also depends on the responsible for the formation of Porphyrin. The studies have concentration of the element in the environmental medium, indicated that some heavy metals are affected by the process the environmental properties of the element, the type of of photosynthesis, chlorophyll production and the synthesis organism and the period of exposure. The results of the study of other dyes such as carotene and efficacy (Prasad and showed a decrease in the total concentration of chlorophyll in Prasad 1990, Saygideger et al 2004, Zengin and Munzuroglu the studied aquatic plants at the end of the experiment. The 2005 and Liu et al 2010). The results showed a significant concentration of chlorophyll in Typha domingensis 1.74, difference at the level of probability (p <0.05) in the total 1.91, 2.53 mg g-1 dry weight in the presence of the nickel, amount of chlorophyll and protein in the tissues of the plants 1.04, 1.15, 1.29 mg g-1 dry weight in the presence of the zinc, used in the experiment and exposed to the different 0.15, 0.21, 0.35 mg g-1 dry weight in the presence of the concentrations of heavy elements used during the duration of

3 Ni 0.6 MN 1day 1day 10day 2.5 0.5 10day 20day 20day g / 30day g

/ 30day 2 0.4 tions mg a tions mg tr a 1.5 .3 tr concen

concen 1 0.2

0.5 0.1

0 0 Control 10ppm 20ppm 30ppm Control 10ppm 20ppm 30ppm

1.4 1.6 1day Co 1day Zn 10day 1.2 10day 1.4 20day 20day

1.2 30day g g /

1 30day /

1 0.8 tions mg tions mg a a tr tr .8 0.6 concen concen .6

0.4 0.4

0.2 0.2

0 0 Control 10ppm 20ppm 30ppm Control 10ppm 20ppm 30ppm Fig. 3. Three different concentrations of Ni, Co, Zn, and Mn effects during the experiment period on the concentration of chlorophyll in Typha domingensis tissues Phragmitus australis and Typha domingensis Effectiveness by Heavy Metals 69

Ni MN 3 0.7 1day 1day 10day 0.6 10day 2.5 20day 20day

30day g g 0.5 / / 30day 2

0.4 tions mg tions mg a .5 a tr tr 0.3 concen 1 concen 0.2

0.5 0.1

0 0 Control 10ppm 20ppm 30ppm Control 10ppm 20ppm 30ppm 1.8 Co 1.8 Zn 1day 1day 1.6 10day 1.6 10day 20day 20day 1.4 1.4

30day g g 30day / / 1.2 1.2

1 1 tions mg tions mg a a tr tr 0.8 0.8 concen concen 0.6 0.6

0.4 0.4

0.2 0.2

0 0 Control 10ppm 20ppm 30ppm Control 10ppm 20ppm 30ppm Fig. 4. Three different concentrations of Ni, Co, Zn, and Mn effects during the experiment period onthe concentration of chlorophyll in Phragmitus australis tissues

1 1.2 1day Ni 1day MN 10day 0.9 10day 1 20day 20day

0.8

g 30day

/

30day g / 0.7 0.8

tions mg 0.6 a tions mg tr a

tr 0.6 0.5 concen 0.4 concen 0.4 0.3

0.2 0.2 0.1

0 0 control 10ppm 20ppm 30ppm control 10ppm 20ppm 30ppm 0.7 Co 1day 0.9 Zn 1day 10day 10day 0.8 0.6 20day 20day

g 30day 0.7 30day

/ g

0.5 / 0.6 tions mg

a 0.4 m tions mg 0.5 tr a tr

0.3 0.4 concen concen 0.3 0.2 0.2 0.1 0.1

0 0 control 10ppm 20ppm 30ppm control 10ppm 20ppm 30ppm Fig. 5. Three different concentrations of Ni, Co, Zn, and Mn effects during the experiment period on protein content in Typha domingensis tissues 70 Qassim Ammar Ahmood AL-Janabi, Ziad Bassim Hameed and Saad Kadhim Ala Allah Al- Kalidy

1 Ni 1day 0.9 MN 1day 10day 10day 0.9 0.8 20day 20day 0.8 30day 0.7 30day

0.7 g g 0.6 / / 0.6 0.5 tions mg

tions mg 0.5 a a tr tr 0.4 0.4 0.3 concen concen 0.3 0.2 0.2

0.1 0.1

0 0 control 10ppm 20ppm 30ppm control 10ppm 20ppm 30ppm

1day 0.7 Co 0.7 Zn 1day 10day 10day 0.6 20day 0.6 20day 30day 30day

0.5 0.5 g

/ g /

0.4 0.4 tions mg a tions mg tr a

0.3 tr 0.3 concen 0.2 concen 0.2

0.1 0.1

0 0 control 10ppm 20ppm 30ppm control 10ppm 20ppm 30ppm

Fig. 6. Three different concentrations of Ni, Co, Zn, and Mn effects during the experiment period on protein content in Phragmitus australis tissues the experiment and this was also found by Keskinkan et al the metabolic processes that occur within it to resist the (2007). This may be attributed to the fact that by increasing concentration of the elements, thus reducing the proportion the concentration of heavy elements in plant tissues, their of protein content in their tissues. chlorophyll content decreases because of its inhibitory effect on the work of enzymes that contribute to the synthesis of REFERENCES chlorophyll and carotene. Nasser enters the installation of Aminot A and Rey F 2000. Standard procedure for determination of chlorophyll by spectroscopic methods. ICES Techniques IN some enzymes contribute to the building of chlorophyll Marine Environ. Sci., Denmark , pp:1-17 (Teisseireet Vernet 2000). The protein content in the aquatic Clark RE 1993. IARC Monographs on the evaluation of carcinogenic plants studied at the end of the experiment also decreased in risks to humans: Chromium, nickel and welding. Quarterly Review of Biology 68: 472. Typha domingensis, 0.55, 0.67, 0.78 mg g-1 dry weight in the Coleman J, Hench K, Garbutt K, Sexstone A, Bissonnete G and -1 presence of the nickel, 0.47, 0.59, 0.68 mg g dry weight in Skousen J 2001. Treatment of domestic wastewater by three the presence of the zinc, 0.66, 0.64, 0.86 mg g-1 dry weight in plant species in constructed wetlands. Water, Air and Soil Pollution 128: 283-295. -1 the presence of the manganese and 0.31, 0.42, 0.54 mg g Favero NC, Bertaggia D and Albergoni V 1996. Metal accumulation dry weight in the presence of the cobalt. in biologicalindicator from lagoon of Venice (Italy). The Phragmitus australis had a low protein content i.e., Environmental Contamination Taxicology 31: 9-18. -1 Forstner U and Wittmann GTW 1981. Metal Pollution in the aquatic 0.55, 0.68, 0.79 mg g dry weight in the presence of the environment. Springer – Verlag. New York, p. 320. -1 nickel, 0.44, 0.51, 0.59 mg g dry weight in the presence of Gaur A and Adholeya A 2004. Prospects of arbuscular mycorrhizal the zinc, 0.56, 0.65, 0.77 mg g-1 dry weight in the presence of fungi in phytoremediation of heavy metal contaminated soils. Current Science 86: 528-534. -1 the manganese and 0.35, 0.44, 0.56 mg g dry weight in the Cho-Ruk K, Kurukote J, Supprung P and Vetayasuporn S 2006. presence of the cobalt with concentrations of 30, 20, 10 ppm, Perennial plants in the phytoremediation of lead contaminated soils, Biotechnology. respectively (Fig. 6). The decrease in the protein content of Kabata-Pendias A 2011. Trace elements in soils and plant. CRC all plants is due to the consumption of the protein content Press, Boca Raton, USA. found in the tissues of these plants in some vital activities or Keskinkan O, Goksu MZL, Yuceer A and Basibuyuk M 2007. Phragmitus australis and Typha domingensis Effectiveness by Heavy Metals 71

Comparison of the adsorption capacities of Myriophyllum and wastewater. Argonne National Laboratory Hinchman, spicatum and Ceratophyllum demersum for zinc, copper and Applied Natural Sciences. lead. Engineering in Life Sciences 7(2): 192-196. Saygideger S, Dogan M and Keser G 2004. Effects of lead and pH on Kinnersely AM 1993. The role of phytochelatins in plant growth and lead uptake chlorophyll and nitrogen content of Typha latifolia L. productivity. Plant Growth Regulation 12: 207-217. and Ceratophyllum demersum L. International Journal of Liu D, Wang X, Chen Z, Xu H and Wang Y 2010. Influence of mercury Agricultural and Biological Engineering 6(1): 168-172. on chlorophyll content in winter wheat and mercury Steffen JC 1990. The heavy metal-binding peptides of plants. Annual bioaccumulation. Plant Soil and Environment 56(3): 139-143. Review of Plant Physiology and Plant Molecular Biology 41: Pak J 2010. Analysis of protein by spectrophotometric and computer 553-575. colour based intensity method form stem of pea (Pisum Teisseire H and Vernet G 2000. Copper induced changes in sativum) at different stages. Anal. Environmental Chemical antioxidant enzymes activities in fronds of duck weed (Lemna 11(2): 63-71. minor). Plant Science 153: 65-72. Prasad MNV, Malek P, Waloszek A, Bojko M and Strazalka K 2001. Uqab B, Mudasir S, Sheikh AQ and Nazir R 2016. Bioremediation: A Physiological responses of Lemna trisulca L. (duck weed) to management tool. Journal of Bioremediation and cadmium & copper accumulation. Plant Science 161: 881- Biodegradation 7: 331. 889. Valliant N, Monnet F, Hitmi A, Sallamon H and Coudret A 2005. Prasad DDK and Prasad ARK 1990. Porphyrin metabolism in lead Comparative study of responses in four Datura species to zinc and mercury treated barja (Pennisetum typhoideum) seedlings. stress Chemosphere 59: 1005-1013. Journal of Biosciences 15(4): 271-279 . Zengin FK and Munzuroglu O 2005. Effects of some heavy metals on Pratas J, Favas PJC, Paulo C, Rodrigues N and Prasad MNV 2012. content of chlorophyll, proline and some antioxidant chemicals Uranium accumulation by aquatic plants from uranium- in bean (Phaseolus vulgaris L.) seedlings. Acta Biologica contaminated water in Central Portugal. International Journal of Cracoviensia 47(2): 157-164. Phytopharmacology 14: 221-234. Zhou X, Li Q, Arita A, Sun H and Costa M 2009. Effects of nickel, Hinchman RR, Negri MC and Gatliff EG 1995. Phytoremediation: chromate and arsenite on histon 3 lysine methylation. using green plants to clean up contaminated soil, groundwater, Toxicology and Applied Pharmacology. In press.

Received 08 October, 2019; Accepted 20 November, 2019 Manuscript Number: S-255 Indian Journal of Ecology (2019) 46 Special Issue (8): 72-76 NAAS Rating: 4.96

Effect of Soil Fertilization with Nutrition Solution (Enraizal) on Fruits Growth and Yield for the Two Hybrids of Cucumber Cultivated in unheated Plastic Houses

Dhafer H. Jawad, Fadhil Jawad Kadhim Altaee, Mohammed J.H. AL- Shareefi and Hussein Ali K. Al-Hasnawi College of Agriculture, Al-Qasim Green University, Iraq E-mail: [email protected]

Abstract: : The experiment was conducted in plastic house for determining the effect of adding four concentrations of nutrition enraizal (0, 1.5, 2, 2.5 ml l-1) water on growth and yield of cucumber hybrids (Saif and Omega). The results showed that the Saif cultivar on Omega cultivar excelled in average fruit length (22.62 cm), fruit diameter (34.88 mm), fruit weight (109.15 g), number of fruits (45.33 plant-1), plant yield (4.975 kg) and total yield (6.965 t house-1). All adding nutrient solution treatments showed a significant excelled in the average fruit growth and yield as compared to the control treatment. The results showed that the Saif cultivar excelled at a concentration 2 ml litre-1 water in fruit length (24.83 cm), fruit diameter (38.67 mm), fruit weight (116.1 g), number of fruits (53.0 fruit plant-1), the plant yield (6.15 kg) and the total yield (8.61 t house-1), while the Omega cultivar gave with non-fertilization the lowest average for those traits accounted 17.67 cm, 30.41 mm, 91.3g, 29.67 fruit plant-1, 2.82 kg and 3.95 t house-1 sequentially.

Keywords: Nutrition, Cucumber, Enraizal, Plastic house Cucumis sativus L. is an important summer vegetable plant, including micronutrients. They also contain a wide crop belonging to the Cucurbitaceae family and one of the range of water-soluble organic compounds such as proteins, most popular vegetables in Iraq. For its economic and sugars and amino acids. Azarmi et al (2009) show that the nutritional importance and its various uses with many foods addition of four levels of organic fertilizer (sheep manure) is 0, and of containing 95-97% water, makes the cucumber plant 10, 20, 30 t ha-1 on the two varieties of option 1Sultan F and the least in terms of calories, so preferred by people Storm F1 cultivated in greenhouses. Where Storm F1 gave interested in diet. It also contains vitamins C, B1 and B2, an increase in total yield amounted 26%, while Sultan F1 niacin, protein, carbohydrates and nutrients, especially iron, gave 25% at the level of 30 tons. Almetoer (2010) showed calcium, phosphorus and potassium (Al Gobaisi 2007, when studying the growth and yield of two hybrids of Arnaout 1980, Hammadi and Mashal 1987). It is useful in high cucumbers (Sayff and Shady) cultivation in greenhouses that and low blood pressure and purifies the body from toxins, as the Shady hybrid was significantly excelled in fruit weight and an analgesic for headache and maintain the vitality of the early greenhouse yield compared to Sayff hybrid, while there epidermis (Sumathi et al 2008). The fruits of cucumbers are were no significant differences between hybrids in average desirable to the consumer, so the demand is high throughput fruit number, single plant yield and total greenhouse yield. AL- the year and in order to meet this growing demand, there has Sahaf et al (2011) reported when using a program of organic been a significant development in the production of fertilizer (Perl humus 200 kg ha-1 + Pow humus 4 kg ha-1 + cucumbers, whether agriculture in the conditions of open Amino power plus 250 ml 100 l-1 water) on cucumber plant fields or under tunnels and plastic house and glasshouse. and for two seasons, they obtained an increase in the yield of The method of cultivation of the cucumber plant under the per plant 967 and 1408.6 g compared to the control treatment conditions of the protected environment approached frequent and for the two seasons respectively. Al-Bayati (2012) studied use of chemical fertilizers added through the soil or spraying the addition of 50 g l-1 of organic fertilizer Roct Dove manure on the plants that weight contain or toxins and have a after fermentation and its addition to the Sayff hybrid negative and long-term impact in humans and the cucumber plant grown in unheated greenhouses led to a environment. Therefore, specialists in the field of agriculture significant increase in the average weight of fruit, the number are to follow the method of organic nutrition as an alternative of fruits, early yield and total yield compared to the treatment to chemical fertilizers (Elia et al 1998). Tiaz and Zieger (2003) of chemical fertilizer (Comparison). Al-Bayati and Kamel showed that the organic fertilizers contain all the nutrients (2015) found that the addition of organic fertilizers (Nitrogen, elements necessary for the growth and development of the org-Vit and Humistar) as well as chemical fertilizer (control) to Soil Fertilization with Enraizal on Fruits Growth 73 two hybrids of cucumbers grown in greenhouses led to a season in one of the unheated greenhouses in Al-Azawiya significant increase in the number of fruits per plant, the total area in the north of Babylon governorate. The experiment yield compared to control treatment. Eifediyi and Remison included eight treatments of two cultivars of hybrid (2010) reported that the use of different levels of compost and cucumbers, Saif and Omega, with the addition of four compound fertilizer for the cucumber plant Ashley cultivars, concentrations of nutrient solution (Enraizal) (0, 1.5, 2, 2.5 The level 10 tons of organic fertilizer with 400 kg ha-1 ml) at the first and second flowers after two weeks from the composite fertilizer with a ratio of 10:10:20 gave highest total first and the third after two weeks from the second. yield, length, diameter and weight of fruit, Rogers et al (2002) Treatments were arranged in Factorial experiment with two found that the addition of an aqueous extract of a mixture of factors and according to Randomized Complete Blocks poultry wastes and aerobic fermented sawdust for 4 hours to Design R.C.B.D and with three replicates. For the first factor, the broccoli and tomato irrigation system in field or cultivars include A1: Saif cultivar produced by the Dutch aquaculture gave the best yield and good quality. Bayoumi company Nunhems and A2: Omega produced by the Dutch and Hafiz (2006) studied that the addition of organic fertilizers company Ergon. and seaweed Extracts resulted in a significant increase in The second factor added concentrations of nutrient total and early yields. Olaniyi et al (2009) showed that the solution Enraizal produced by the Spanish company addition of organic fertilizers in the amount of 4 tons ha-1 to Fabrique (Table 1) includes: B1: without adding three cultivars of cucumbers gave a significant increase in the (comparison); B2: 1.5 ml; B3: 2 ml and B4: 2.5 ml. Random length, diameter, weight of the fruit, the number of fruits, the samples were taken from plastic soil at a depth of (0-30 cm) total yield and for all the three cultivars by control treatment. and the soil was analyzed in the agricultural research Al-Bayati et al (2012) found that the addition of organic laboratory - Directorate of Agriculture of Babylon (Table 2). fertilizer (Atalbolina) at a rate of 200 kg/500 m2 to the The plastic house soil was tilled, smoothed, leveled and cucumber plant cultivar Grass F, has led to a significant divided into terraces and then divided into (24) experimental increase in the total yield and the product yield 29.67 and units of 2 m length and 60 cm wide. Cucumber seeds were 32.81% compared to the control treatment. Hussein (2016) sown in the nursery on 3/1/2019 in cork dishes after filling reported that the addition of the plant nutrient (Karma maxi them with Peat moss as an agricultural medium. The org) at a concentration of 10 and 15 ml l-1 and spraying seedlings were transferred to the plastic house on 61/1, chelated iron at a concentration of 1 and 3 g to the squash where they were cultivated on both sides of the terrace and plant, Razan cultivated in greenhouses, The treatment of the distance between the line and another 50 cm and the (Karma maxi org nutrition additive at a concentration 15 ml distance between the plant and another 40 cm. The nutrient and sprayed chelated iron at a concentration 3 g l-1 was solution was added to plants on flowering time on 25/2 and all significantly excelled in fruit weight ratio, The number of fruits, the recommended agricultural operations were conducted plant yield and total yield as compared to the control during the growth period in the field of irrigation, fertilization, treatment. Al-Tafi et al (2016) showed that the addition of hoeing, weeding and controlling diseases and insects three levels of organic fertilizer A (Homobacter), which is similarly to all the experimental units. The fruits harvesting 3000, 2000 kg ha-1 on the eggplant plant (Solanum begin on 5/3 and lasted until 30/5/2019. melongena) in the greenhouse, has significantly excelled the Studied traits: The average fruit length (cm), fruit diameter level of 3000 kg ha-1 at the level of 2000 kg ha-1 and control (mm), average fruit weight (g), average number of fruits (fruit treatment in fruit weight, number of fruits, per plant yield and plant-1), plant yield (kg) and total yield of the plastic house (t total yield. The aim of this study was to investigate the effect of House-1). The results were analyzed according to the design add-nutrient solution (Enraizal) on the growth and yield of two followed. The least significant difference test was used to hybrids of cucumber Saif and Omega grown in unheated greenhouses. Table 1. The nutrient solution (Enraizal) components

K2O P2O5 Nitric Ueric Organic Total Free amino (%) (%) nitrogen nitrogen nitrogen nitrogen acid (%) MATERIAL AND METHODS (%) (%) (%) (%) The experiment was conducted during the 2019 planting 4.6 2.3 1.1 4.8 0.9 6.8 4.8

Table 2. Some physical and chemical traits of plastic house soil Texture Traits Silt (%) Sand% Clay % Kppm P ppm Nppm Organic matter % Eceds m-1 pH Sandy loam 15.8 80.2 4 360 8.7 52.5 1.13 3.7 7.4 74 Dhafer H. Jawad, Fadhil Jawad Kadhim Altaee, Mohammed J.H. AL- Shareefi and Hussein Ali K. Al-Hasnawi compare averages at a probability level of 0.05 (AlSahoki and Table 3. Effect of the cultivar and adding the nutrient solution Wahib, 1990). Genstat in Statistical Analysis. (Enraizal) and their interaction on the fruit length of the cucumber plant (cm) Adding B1 B2 B3 B4 Average of RESULTS AND DISCUSSION cultivar cultivar Traits of fruit growth: Two cultivars significantly differed in Saif 19.33 22.83 24.83 23.50 22.62 the average length of the fruit. Saif cultivar excelled the (34.88) average in the length of fruit (22.62 cm), compared to Omega Omega 17.67 21.00 23.17 21.33 20.79 cultivar (20.79 cm). Addition of the nutrient solution Enraizal (33.12) Average of 18.50 21.92 24.00 had a significant effect on the above traits. All spraying adding treatments showed significant excelled in average of the fruit LSD 0.05 Cultivar: Adding: Interaction: length, compared to the control treatment. The additive 0.69 0.97 1.37 treatment at a concentration of 2 ml l-1 was achieved the highest average fruit length of (24.00 cm), where it Table 4. Effect of the cultivar and adding the nutrient solution significantly excelled on the other treatments. Also, the (Enraizal) and their interaction on the fruit diameter (mm) of the cucumber plant interaction between the hybrid cultivar and the nutrient Adding B1 B2 B3 B4 Average of solution gave significant effect on the average length of the cultivar cultivar fruit. Where the interaction between Saif and addition the Saif 31.17 33.24 38.67 36.42 34.88 nutrient solution at a concentration of 2 ml 1-1 water was a significant effect on the average of fruit length amounted of Omega 30.41 32.21 35.81 34.07 33.12 (24.83 cm) on other interaction treatments. While the Average of 30.79 32.73 37.24 35.24 interaction treatment between the Omega cultivars and now adding addition which gave the lowest average amounted (17.67 LSD 0.05 Cultivar: Adding: Interaction: cm). 0.64 0.90 1.28 Two cultivars significantly differed in the average diameter of the fruit, Saif cultivar excelled in the average Table 7. Effect of the cultivar and adding the nutrient solution diameter of the fruit (34.88) mm, than in the Omega cultivar (Enraizal) and their interaction on plant yield (kg) of which gave (33.12 mm). The results showed that the addition the cucumber plant Adding B1 B2 B3 B4 Average of of the nutrient solution Enraizal had a significant effect on the cultivar cultivar above traits. All spraying treatments were significant superior Saif 3.935 4.623 6.15 5.192 4.975 in the fruit diameter compared to the control treatment. The -1 additive treatment at a concentration of 2 ml l water was Omega 2.821 3.517 4.975 4.238 3.888 achieved the highest average of fruit diameter (37.24 mm) where it significantly excelled the other additive treatments. Average of 3.378 4.07 5.562 4.715 adding Also, the interaction between the hybrid cultivar and the LSD 0.05 Cultivar: Adding: Interaction: nutrient solution gave a significant effect on the average of 0.246 0.348 0.492 the fruit diameter, where the interaction between Saif and the addition at concentration 2 ml was significantly excelled the proved to be significantly superior the other adding average of the fruit diameter to the 38.67 mm on the other treatments. The interaction between the hybrid cultivar and interaction treatments. While the treatment interaction the adding of the nutrient solution Enraizal had a significant between the Omega cultivar with non-adding treatment gave effect on the average fruit weight. The interaction between the lowest average (30.41 mm). Saif cultivar and the addition of nutrient solution Enraizal (2 Significantly difference in the average of fruit weight was ml-1) was significantly excelled in fruit weight amounted observed among two cultivar, Saif cultivar excelled in the (116.10 g) on the other interaction treatments. While the average of fruit weight (109.15 g) compared to the Omega treatment interaction between the Omega cultivar with non- cultivar (102.36 g). The results showed that the addition of adding treatment which gave the lowest average fruit weight nutrient solution Enraizal was a significant effect on the of cucumber (91.33 g). above traits. All spraying treatments were significant superior Significant difference was observed in the average in the average fruit weight, compared to the control number of fruits plant-1 Saif cultivar excelled in the average of treatment. The adding treatment at a concentration of 2 ml l-1 fruits (45.33) fruit plant-1, while the Omega cultivar gave was recorded with highest average of fruit of (114.09 g) and (37.50 fruit plant-1). The results showed that the addition of Soil Fertilization with Enraizal on Fruits Growth 75 nutrient solution Enraizal was a significant effect on the Table 5. Effect of the cultivar and adding the nutrient solution above traits. All spraying treatments were significant superior (Enraizal) and their interaction on the fruit weight of the cucumber plant (g) in the number of fruits, compared to the control treatment, -1 Adding B1 B2 B3 B4 Average of The adding treatment at a concentration of 2 ml l was cultivar cultivar -1 -1 achieved the highest number of fruit plant (48.67 fruit plant ) Saif 100.05 107.52 116.10 112.93 109.15 and it significantly excelled the other adding treatments. Interaction between the hybrid cultivar and the adding of the Omega 91.33 101.82 112.09 104.1 102.36 nutrient solution Enraizal had a significant effect on the -1 Average of 95.69 104.67 114.09 108.57 average of number of fruits plant . The interaction between adding Saif cultivar and the addition of nutrient solution Enraizal (2 LSD 0.05 Cultivar: Adding: Interaction: ml-1) was significantly excelled in the number of fruits 3.76 5.32 7.53 amounted (53.00 fruit plant-1) on the other interaction treatments, While the treatment interaction between the Table 6. Effect of the cultivar and adding the nutrient solution Omega cultivar with non-adding treatment recorded with the (Enraizal) and their interaction on the number fruit of the cucumber plant (fruit plant-1) lowest number of fruits (29.67 fruit plant-1). Adding B1 B2 B3 B4 Average of Significantly difference in the average the plant yield was cultivar cultivar observed among two cultivars namely Saif and Omega. Saif Saif 39.33 43.00 53.00 46.00 45.33 cultivar excelled in the average of fruits (4.975 kg), while the Omega cultivar gave (3.888 kg). The results showed that the Omega 29.67 35.33 44.33 40.67 37.50 addition of nutrient solution Enraizal was a significant effect Average of 34.50 39.17 48.67 43.33 on the above traits. All spraying treatments showed adding significant excelled in the plant yield, compared to the control LSD 0.05 Cultivar: Adding: Interaction: treatment, The adding treatment at a concentration of 2 ml l-1 1.89 2.67 3.78 was achieved the highest average of this trait amounted (5.562 kg), where it significantly excelled the other adding solution Enraizal (2 ml l-1) was significantly excelled in the treatments, The results of the same table also showed that plant yield amounted 8.61 tons on the other interaction the interaction between the hybrid cultivar and the adding of treatments, While the treatment interaction between the the nutrient solution Enraizal was a significant effect on the Omega cultivar with non-adding treatment which gave the average the plant yield. The interaction between Saif cultivar lowest average amounted (3.950) tons. -1 and the addition of nutrient solution Enraizal (2 ml water) The excelling of the Saif hybrid cultivar in the studied was significantly excelled in the plant yield amounted (6.150 traits compared to the Omega hybrid cultivar is due to genetic kg) on the other interaction treatments, While the treatment differences between hybrids and their different response to interaction between the Omega cultivar with non-adding different environmental conditions during the growing treatment which gave the lowest average amounted (2.821 season (AL-Sahaf et al 2011). The significant excelled of kg). adding the nutrient solution in fruit growth traits as measured The results in Table 8 revealed that the two cultivars by control treatment may be due to the effect of this nutrient significantly differed in the average the total yield. Saif on increasing root growth and Thus increase the absorption cultivar excelled in the average of the total yield of 6.965 tons, of water and necessary nutrients dissolved in it (AL-Sahaf et while the Omega cultivar gave 5.443 tons. The results al 2011). Organic fertilizers also contain a wide range of showed that the adding of nutrient solution Enraizal was a water-soluble organic compounds such as proteins, sugars, significant effect on the above traits. All spraying treatments amino acids, humic and non-organic acids. All these showed significant excelled in the plant yield, compared to compounds contribute directly or indirectly to the growth and the control treatment, The adding treatment at a development of plants. It is either enzymatic or hormonal, concentration of 2 ml l-1 was achieved the highest average of where it contains nutrients that required for the plant or it this trait amounted 7.787 tons, where it significantly excelled affects on the availability of the nutrients already in the soil by the other adding treatments, The results of the same table improving or adding pH to increasing and improving the also showed that the interaction between the hybrid cultivar quality of production (Al-Fartusi 2003). The results showed and the adding of the nutrient solution Enraizal was a significant excelled in the interaction treatment between Saif significant effect on the average the plant yield. The cultivar and adding the adding of nutrient solution Enraizal at interaction between Saif cultivar and the addition of nutrient a concentration of 2 ml-1 water. This may be due to the effect 76 Dhafer H. Jawad, Fadhil Jawad Kadhim Altaee, Mohammed J.H. AL- Shareefi and Hussein Ali K. Al-Hasnawi

Table 8. Effect of the cultivar and adding the nutrient solution AlGobaisi Hassan 2007. Dictionary of herbs and medicinal plants. (Enraizal) and their interaction on the total yield (t Seventh Edition. 347p. house-1) of the cucumber plant Almetour, Aqila Juma Hajim. 2010. Effect of calcium on growth and yield of two cultivars of female hybrid option Cucumis sativus L. Adding B1 B2 B3 B4 Average of cultivated in greenhouses and its effect and temperature cultivar cultivar storage in treasury capacity. Master Thesis. College of Saif 5.509 6.473 8.61 7.268 6.965 Agriculture. AL-Sahaf FH, ALMharib MZK and Jawad FM 2011. Response of Omega 3.95 4.923 6.965 5.934 5.443 cucumber hybirds to chemical and organic fertilizers. The Iraqi Journal of Agricultural Sciences 42(4): 52-62. Average of 4.729 5.698 7.787 6.601 AlSahoki, Medhat and Karima Mohamed Wahib 1990. Applications adding in the design and analysis of experiments. Baghdad University. Ministry of Education and Scientific Research. Iraq. LSD 0.05 Cultivar: Adding: Interaction: 0.344 0.487 0.688 Al-Tafi, Sami Ali Abdul Majeed, Riyad Kazar Kazim, Ali Hassan Ali and Lazim Mohamed Hussein 2016. The effect of organic fertilizer (Humobacter-A) and foliar fertilizer (Top10) on the between the genotype and nutrient solution content of growth and yield of eggplant jewels. Euphrates Journal of nutrients, which has a positive effect on the number of fruits, Agricultural Sciences 4: 107-116. Arnaout, Mohamed El Sayed 1980. Herbs and medicinal plants Food average length, diameter, weight fruits and yield of the plants. and medicine. Egyptian Lebanese House. These results agree with (Al-Bayati 2012, Al-Mutawary Azarmi R, Mousa TG and Bahzad H 2009. The effect of sheep 2010, Al-Bayati Kamel 2015, Al-Bayati 2012, AL-Sahaf et al manure vermicompost on quantitative and qualitative properties of cucumber (Cucumis sativus L.) grown in 2011, Olaniyi 2009, Eifediyi Remison, 2010 and Hussein greenhouse. Africa Journal of Biotech 8(19): 453-457. 2016). We conclude from this study that the Saif hybrid Bayoumi YA and Hafiz YM 2006. Effect of organic fertilizers com- cultivar showed significant excelled in the fruit growth traits bined with benzo (1,2,3) thiadiazole -7- carbothioic acid s- methylester (BTH) on the cucumber powdery mildew and the and the yield on the Omega hybrid cultivar. The addition of yield production. Acta. Biol. Szegediensis 50(3-4): 131-136. -1 nutrient solution (Enraizal) at a concentration of 2 ml l water Eifediyi EK and Remison SU 2010. Growth and yield of cucumber led to a significant increase in the fruit growth and yield of the (Cucumis sativus L.) as influenced by farmyard manure and inorganic fertilizer. Journal of Plant Breeding and Crop Science cucumber plant. It was found that the interaction between the 2(7): 216-220. Saif hybrid cultivar and the addition of nutrient solution Elia AP, Santamaria and Serio F 1998. Nitrogen nutrition yield and (Enraizal) at a concentration of 2 ml l-1 water gave the best quality of Spinach. Journal of Science Food Agriculture 76: 341-346. average of traits during the cultivation season. Hammadi, Fadhel Musleh and Abdul Jabbar Jassem Al-Meshal 1987. Production of vegetables. Ministry of Higher Education REFERENCES and Scientific Research. Baghdad University. Iraq. Al-Bayati, Hussein Jawad Muharram and Tara Jinan Kamel 2015. Hussein, Ahlam Ahmad 2016. Response of squash (Cucurbita pepo) Improving grow and yield of two-hybrids Cucumis sativus L. a Razan cultivar to add plant nutrient Karma maxi org and foliar by covering the soil and adding organic fertilizers under spraying chelated iron inside the greenhouses. Euphrates unheated greenhouse conditions. Euphrates Journal of Journal of Agricultural Sciences 4: 127-134. Agricultural Sciences 7(4): 22-29. Olaniyi JO, Ogunbiyi EM and Alagbe DD 2009. Effect of organic- Al-Bayati, Hussein Jawad Muharram, Mohammed Talal Abdul Salam mineral fertilizers on growth, yield and mineral nutrients uptake Al-Habbar and Walid Badr Al-Din Allila. 2012. Effect of organic in cucumber. Journal of Animal and Plant Sciences 5(1): 437- fertilization on growth and yield of grass F1 cultivated under 442. unheated green houses. Kufa Journal for Agricultural Sciences Rogers GS, Little SA, Silcock SJ and Nunn G 2002. Extraction of a 3(2): 327-336. liquid organic fertilizer from poultry manure for use in field and Al-Bayati, Hussein Jawad Muharram 2012. Effect of different sheltered horticultural production systems. ISHS Acta concentrations of ROCK DOVE MANURE solution on growth Horticultural 642(26): 1340-1440. and yield of female hybrid option Cucumis sativus L. grown in Sumathi T, Ponnuswami V and Selvi BS 2008. Anatomical changes unheated greenhouses. Second Scientific Conference of the Faculty of Agriculture, University of Karbala. 218-224. of cucumber (Cucumis sativus L.) leaves and roots as influenced by shade and fertigation. Research Journal of Al-Fartusi, Beidaa Aboud Jassim 2003. Effect of aqueous extracts of Agriculture and Biological Science 4(6): 630-638. organic wastes on the growth and yield of wheat L. (Triticum rd aestivum). Master Thesis. College of Agriculture, University of Taiz L and Zeiger E 2003. Plant Physiology. 3 ed. Sinauer Baghdad, 117p. Associates, Inc. publisher Sunderland, Massachuts, USA Received 28 September, 2019; Accepted 21 November, 2019 Manuscript Number: S-256 Indian Journal of Ecology (2019) 46 Special Issue (8): 77-81 NAAS Rating: 4.96

Effect of Foliar Application with Novalon on Growth and Grain Yield of Three Genotypes of Maize

Nassr M. Jasim and Basheer H. Abdullah Field Crop Science, College of Agriculture, University of Anbar, Anbar, Iraq E-mail: [email protected]

Abstract: A field trial was carried out to investigate the effect of foliar application with four concentrations (0, 1.5, 3.0 and 4.5 g l-1) of Novalonon growth and grain yield of three maize genotypes (Sarah, Fajer-1 and Maha). Results showed the 21.50 and 52.47% superiority of Novalon (4.5 gl-1) for number of ears plant-1 and grain, respectively compared with the 0 concentration. However, the concentration 3g l-1 gave the highest average 130.34 g of 500grain weight (average of two seasons) compared with 126.06 gm. Maha genotype was superior in the 500 grain weight in both seasons. Fajer-1 with the 4.5 g l-1 gave the highest number of ears (1.33 and 1.36 ear plant-1)in both seasons, compared with 1.00 and 1.03 ears plant-1 for control plants of Sarah genotype in both seasons, respectively.

Keywords: Mineral fertilizer, Genotypes, Chlorophyll, G×E interaction Maize is a major cereal crops in Iraq, grown widely for its belonging to the Qadaa-AL-Ameeriya-AL-Anbar (33.16 nutritive value and great productivity potential. Maize grains longitute and 43.86 altitude). Novalon is Turkish origin consist of 72% starch, 10% protein and 4% fat and each 100 consist of equal quantities of NPK 20% for each and miner gm of grain provide 365 kilocalorie (Ranum et al 2014). Maize elements with the percentages (Boron 0.003%, Copper comes after wheat and rice in terms of acreage and 0.04%, Iron 0.12%, Manganese 0.06%, Molybdenum production. Despite these fact, it's productivity per unit area 0.005%, Zinc 0.06% and Magnesium 0.5%) weight dissolved still low 2.5 t ha-1 compared with the international production in water. Seeds of these cultivars were supplied by the 4.47 t ha-1. This low productivity requires applied researches Directorate of Agricultural Research- Ministry of Agriculture. to investigate and evaluate the causes of such low All necessary seed bed preparation was taken before sowing productivity and proposing solution. Among these causes of from different regions at depth (0-30 cm) to know the crop management, it is essential to provide soil and plant with chemical and physical properties of soil (Table 1). essential nutritive elements for supporting growth and Experiment was conducted in Randomized Complete Block increasing productivity. Foliar application is essential in Design (RCBD) in the arrangement of split-plots with three cases where limiting factors of root absorption does exist like replications (12 experimental units per replication). high pH value or high lime content of soil or high soil salinity Concentrations occupied of the main plots and cultivars in the levels or drought and the fluctuations of temperatures (Kick sub-plots. The dimensions of each experimental unit were 1969, Romhold and EL-fouly 2000), which might increase the 2×3 m, with four rows of 3m width between rows and 0.25 probability of losing such nutritive elements. The foliar between hills in each row for each experimental unit. A application might supply plant requirements for nutritive distance of 0.75 m was left between sub-plots and 2m elements by 80%. Furthermore, foliar application is more between main plots to avoid the drift of spraying between efficient than soil application via soil by 8-20 times when it is plots and to facilitate the spraying. Sowing date was on April applied scientifically according to the plant needs (Kuepper 8, 2018 for spring season and July 28, 2018 for fall season by 2003) and by using responsive genotypes. Maize genotypes placing 2-3 seeds in each hill at depth 3-5cm. Then the plots differ in the growth, leaf angle with stem and leaf area, which were irrigated with continuous watering as necessary. Hand in turn is reflected in the leaves efficiency of nutritive weeding was performed to remove weeds as necessary. absorption. The present study was conducted to investigate Maize stem borer (Sasami acvetico) was controlled by using the effect of foliar application with the major and minor effect granular Diazinone twice, the first after 4-5 leaves stage and on growth and grain yield of three maize genotypes. the second after 15 days from the first application. Foliar application was performed in the early morning up to the full MATERIAL AND METHODS covering by using sprayer (15liter capacity). Foliar A field trial was carried out during the spring and fall application was done twice per each season the first one at seasons of 2018 at private farm in the Ibzabeez region, the beginning of flowering and the second after ten days from 78 Nassr M. Jasim and Basheer H. Abdullah the first spraying. Plants were harvested at full maturity on Table 1. Some chemical and physical properties of soil July 24, 2018 and Nov. 19, 2018 for spring season. The Plant before sowing for spring and fall seasons of 2018 height (cm), Leaf area (m2 plant-1), Chlorophyll index SPAD Character Measure unit Spring season Full season -1 (Jemison and Williams 2006), Number of ears (ears plant-1), EC ds m 2.23 2.28 Weight of 500 grains (15.5% with moisture content) and pH - 7.30 7.20 Grain yield (t ha-1) were measured Available N Mg kg-1 soil 34.0 31.5 Statistical analysis: Statistically analysis for data was Available P = 5.79 5.19 performed according to the design by using Genstat Available K = 97.72 95.48 programme and the using least significant difference (LSD) Available Iron = 5.49 6.13 at 0.05 level to compare between the average of treatment. Available Boron = 0.49 0.42 Total Iron = 274.8 276.5 RESULTS AND DISCUSSION Total Boron = 6.92 6.71 Plant height (cm): It is clear from Table 2 that foliar application with 4.5 l-1 of Novalon recorded the highest Total Zinc = 1.03 1.23 average of plant height (180.14 and177.49 cm) in both Soil Texture -1 seasons, respectively. The percentages increases over other Sand gm kg % 51.0 51.6 concentrations were 10.77 and 10.11%. However, the un- Silt = 24.8 24.2 sprayed plants gave the lowest averages of 162.63 and Clay = 24.7 24.0 161.20 cm, respectively. The presence of major and minor Soil texture Sandy loam Sandy loam nutritive elements in the combination of this fertilizer has improved the plants performance in the most vegetative characters, especially, plant height. This result was in agreement with the findings of Obaid and Sadik (2016) and AL-Shumary et al (2019). Results of Table 1 showed that the Table 2. Effect of foliar application with Novalon on plant plants of Fajer-1 variety gave the highest plant height 174.37 height (cm)* spring and fall seasons of 2018 and 172.36 cm compared with Maha and Sarah, which gave Cultivation (C) Spring season Average the lowest values (169.47 and 166.78 cm) in both seasons, Novalon concentration g ml-1(N) respectively. 0 1.5 3.0 4.5 This varietal differences may be due to their different Sarah 161.0 167.40 174.30 175.17 169.47 genetic-make up which has been reflected in their different (0.45) (0.51) (0.54) (0.56) (0.51) response to the surrounding environmental conditions and Fajer-1 165.60 172.13 175.60 184.13 174.37 hence, different plant height. This result was in agreement (0.47) (0.56) (0.59) (0.62) (0.56) with Shafiullah et al (2018) and AL-Shumary et al (2019). Maha 161.30 169.20 176.80 181.13 172.11 (0.43) (0.54) (0.53) (0.56) (0.51) The significance of the interaction between varieties and Cultivar 162.63 169.58 175.57 180.14 foliar application in the spring season indicated that plant average (0.45) (0.54) (0.55) (0.58) -1 height of Fajir-1 fed with 4.5 gml gave the highest plant LSD 0.05 C: 0.96 N: 1.98 C×N : height 184.13 cm compared with the other interaction where (0.03) (0.04) 2.33 plants of Salah (un-sprayed plants) recorded the lowest plant Cultivation (C) Fall season Average height 161.00 cm. Novalon concentration g ml-1(N) Leaf area (m2 plant-1): Results of Table 2 showed that foliar 0 1.5 3.0 4.5 application with 4.5 gml-1 recorded the highest leaf area 2 -1 Sarah 157.50 165.16 169.33 175.13 166.78 average 0.58 and 0.54 m plant in both seasons, (0.43) (0.48) (0.52) (0.53) (0.49) respectively. Fajer-1 166.16 171.63 172.33 179.30 172.36 No significant differences were found with the (0.46) (0.52) (0.58) (0.58) (0.54) concentration 3.0gml-1 in the both seasons, yet it was superior Maha 159.98 166.17 171.50 178.03 168.92 over the concentration 1.5 gml-1 and control, which gave the (0.45) (0.50) (0.51) (0.52) (0.50) lowest average 0.45 m2 plant in both seasons. The formula of Cultivar 161.20 167.65 171.06 177.49 average (0.45) (0.50) (0.54) (0.54) Novalon fertilizer consists of major and minor elements, LSD 0.05 C: 1.74 N: 2.92 C×N : NS which stimulate effectively the plant growth, besides the role (0.02) (0.03) (NS) of these elements in the activation of enzymes and the *Leaf area (m2 plant-1) Foliar Application with Novalon on Yield of Maize 79 facilitating of assimilates to the leaves sites and to the new Table 3. Effect of foliar application with Novalon on activation reflected in the cells division, increasing their chlorophyll index of leaves (SPAD)* spring and fall seasons of 2018 numbers, their extension and then, increasing the whole Cultivation (C) Spring season Average growth including leaf area. In this context, many researchers Novalon concentration g ml-1(N) found significant effect of foliar application of nutritive 1.5 3.0 4.5 elements on plant growth characters (Khalaf et al 2017, Saleh 0 et al 2017 and AL-Shumary et al 2019). Sarah 42.00 46.67 49.63 51.10 47.35 (1.00) (1.13) (1.20) (1.23) (1.14) It is also clear from Table 2 that the plants of Fajir-1 Fajer-1 40.20 43.33 46.93 49.17 44.91 variety had significantly higher leaf area average 0.56 and (1.10) (1.23) (1.26) (1.33) (1.23) 0.54 m2 plant-1, respectively compared with the plants of Maha 39.83 42.00 45.87 47.83 43.88 Sarah and Maha varieties. These varietal differences may be (1.03) (1.16) (1.23) (1.30) (1.18) due to the different genetic-makeup of these varieties, which Cultivar Average 40.68 44.00 47.48 49.37 (1.04) (1.17) (1.23) (1.29) makes their capabilities in exploiting growth resources LSD 0.05 C: 0.48 N:0.51 C×N : 0.88 different, besides that Fajir-1 gave the highest plant height (0.03) (0.05) (0.07) -1 (Table 2), which in turn, increases the number plant and leaf Cultivation (C) Fall season Average area. This result coincides with the finding of (Al-Fahadand Novalon concertation g ml-1(N) Ubaidi 2017, AL-ObeidiandAL-An, 2017and AL-Shumary et 0 1.5 3.0 4.5 al 2019) who achieved significant differences between Sarah 42.60 46.40 50.53 52.07 47.90 varieties in their leaf area. (1.03) (1.16) (1.16) (1.23) (1.14) Chlorophyll index of leaves (SPAD): It appears from Table Fajer-1 41.10 44.57 49.87 49.27 46.20 3 that increased concentrations of Novalon was (1.13) (1.26) (1.30) (1.36) (1.26) accompanied with significant increases in the chlorophyll Maha 39.33 43.40 46.17 48.00 44.22 (1.10) (1.23) (1.26) (1.33) (1.23) index up to the 4.5 gml-1 which gave the highest average in Cultivar Average 41.01 44.79 48.86 49.78 both seasons (49.37 and 49.78 SPAD, respectively). The (1.09) (1.22) (1.24) (1.31) percentage increase was 21.36 and 21.38%, over control LSD 0.05 C: 0.54 N:0.74 C×N: 0.07 plants which gave the lowest average 40.68 and 41.01 (0.03) (0.06) (0.08) -1 SPAD, respectively. This increase in the chlorophyll index *Number of ears (ear plant ) may be due to the fact that Novalon fertilizer contains Number of ears (ear plant-1): Results of Table 3 showed that essential elements of chlorophyll formation, besides, the increased concentration was accompanied with the availability of daily illumination which is another requirement increased number of ears per plant, where high for formation this character, hence, increased the pigment concentration (4.5 gml-1) gave the highest average of this concentration of leaves. Another researcher found similar character 1.29 and 1.31 in both seasons, respectively, effect of nutritive elements foliar applied on the vegetative compared with 1.04 and 1.09 ear plant-1 for the control plant, parts where chlorophyll content was more (Leilby et al 2017, respectively. The increased ear plant-1 may be due to plant Mreer and Mohamad 2017). height and leaf area (Tables 2) and increased chlorophyll Table 3 shows significant varietal differences, where concentration which in turn increased the exposure of the Sarah gave the highest average 47.35 and 47.90 SPAD, in leave to the light and its absorption and then increases both seasons, respectively, compared with Fajir-1 and Maha photosynthesis efficiency and photo assimilates which which recorded the lowest average 43.88 and 44.22 SPAD, transferred to the new phytomers. Results were accordance respectively. The reason of such discrepancy between with the findings of Khalaf et al (2017). varieties may be due to their different-make up and the effect Table 3 also shows that Fajir-1 had the highest average of genes in the determination and controlling most of this character 1.23 and 1.26 ear plant-1 compared with 1.14 morphological and anatomical characters of plant. In this for Sarah in the both seasons. This varietal difference may be context, Al-Rawi (2018) found that there were significant due to their different genetic-make up, which was reflected in differences of chlorophyll pigment. their different responses to the environmental conditions, The interaction was significant where the highest besides, that Fajier-1 was superior in the leaf area (Table 2). concentration 4.5 gml-1 with the Sarah variety gave the Results were accordance with the findings of Al-Rawi (2018) highest averages 51.10 and 52.07 SPAD compared with and Shafiullah et al (2018), they found that there were 39.83 and 39.33 SPAD for Maha and control plants in the significant differences of between the maize varieties in ear both seasons, respectively. plant-1. 80 Nassr M. Jasim and Basheer H. Abdullah

Concerning the interaction, plants of Fajir-1 sprayed with with 5.64 and 6.52 t ha-1 for control plants in both seasons, 4.5 gml-1 of Novalon gave the highest averages 1.33 and 1.36 respectively. This may be due to the high ears plant. These ear plant-1 in the both seasons, respectively, compared with results are in agreement with the findings of Obaid and Sadik the unsprayed plants of Sarah, which gave 1.00 and 1.03 ear (2016) and Arrak et al (2017). plant-1, respectively. It is also clear from Table 4 that Fajir-1 gave the highest Weight of 500 grains (gm): Table 4 indicates that the plants grain yield 8.24 and 9.33t ha-1 in the both seasons, fed with 3.0 gml-1 significantly gave the highest average of this respectively with 22.62 and 23.41% increases over Sarah character (127.67 and 133.00 gm), respectively with no variety, which gave the lowest 6.72 and 7.56 t ha-1, significant differences with 1.5 gml-1 in the spring season respectively. These results may be due to the superiority of compared with 124.22 g for 4.5 gml-1 spring season and Fajir-1 in the leaf area and number of ears plants-1 (Table 2 127.89 gm fall season. This result is in agreement with the and 3). These findings are in agreement with the results of findings of Arrak et al (2017) and AL-Shumary et al (2019). AL–Heetyet al (2017) and AL-Shumary et al (2019). The Table 3 shows that there were significant differences interaction was not significant. between varieties in the fall season only, where Maha recorded the high weight of 500 grains 131.58 gm, compared REFERENCES with the lowest 129.42 gm for Sarah. This may be due to the Al Rawi AF 2018. Effect of Weed Control Methods in Growth, Yield, prolonged grain filling period for Maha, which increased the and Yield Quality of Three Cultivars of Maize (Zea mays L.) and Combined Weeds. Master Thesis. College of Agriculture- accumulation of photo assimilates in the grains, ultimately University of Anbar. increased their weight. This result was in agreement with the AL–Heety NN, Saif and Al–Kaisy AMA 2017. Response of many findings of Shafiullah et al (2018) and AL-Shumary et al maize cultivars to weed control treatments. Anbar Journal of Agricultural Sciences 5(2): 454-470. (2019). Al-Fahad ACA and Al- Ubaidi MOG 2017. Effect of seed size on some Grain yield (t ha-1): Table 4 indicates that the 4.5 g l-1 gave the growth and yield traits of four synthetic cultivars of maize. AL– highest average 8.64 and 9.90 t ha-1 grain yield compared Anbar Journal of Agricultural Sciences 15(Special Issue): 28- 43. AL-Obeidi NDA and AL-Ani MHI 2017. Response of varieties of maize to bio-fertilization of Bread yeast (Saccharomyces Table 4. Effect of foliar application with Novalon on weight of Cerevisiae L.). AL – Anbar Journal of Agricultural Sciences 500 grains* (gm) spring and fall seasons of 2018 15(2): 471-483. Cultivation (C) Spring season Average AL-Shumary AMJ, Ali HA and Alabdulla SA 2019. Effect of spraying concentrations of integrated nano-fertilizer on growth and yield -1 Novalon concentration g ml (N) of genotypes of corn (Zea mays L.). AL – Muthanna Journal of Agricultural Sciences 7(2): 114-121. 0 1.5 3.0 4.5 Arrak RR and Abdul-Ameer HK 2017. Response of some indicators Sarah 125.30 124.33 127.67 122.67 124.99 of yield and quality of maize (Zea mays L.).vr.Bohuth 106 to (4.96) (6.65) (7.35) (7.93) (6.72) potassium and zinc spray. AL – Kufa Journal for Agricultural Fajer-1 123.07 128.67 125.70 122.33 124.94 Sciences 10(1): 170-183. (6.39) (8.48) (8.87) (9.20) (8.24) Jemison J and Williams M 2006. Potata-Grain Study Project Report Maha 124.33 127.00 129.67 127.67 127.17 Water Quality Office. University of Maine, Cooperation (5.57) (6.96) (8.18) (8.79) (7.38) Extension. Cultivar average 124.23 126.67 127.67 124.22 Kadhim SH and Abduln Nabi RA 2014. The effect of spray stage with (5.64) (7.37) (8.13) (8.64) brosol solution, sowing space and genotype on maize yield. Al- Furat Journal for Agricultural Sciences 6(4): 348-359. LSD 0.05 C: NS N: 2.74 C×N : NS (0.38) (0.29) (NS) Khalaf IT, Assal BA and Araak RR 2017. Effect of ground and foliar fertilization using humzinc compound on some growth and yield Cultivation (C) Fall season Average characters of wheat (Triticum aestivum L.). Karbala Journal for Novalon concentration g ml-1(N) Agricultural Sciences 4(1): 62-73. Kick H 1969. Anorganischestoffe, spurenelment. In: Handbook der 0 1.5 3.0 4.5 pflanzenernahrung von H. Linser 1/1. S-114-121 Springer- Sarah 125.67 128.67 132.00 131.33 129.42 Verlag, Wien New York. (5.41) (7.47) (8.34) (9.02) (7.56) Kuepper G 2003. Foliar fertilization appropriate technology transfer for rural areas (ATTRA). National Sustainable Agriculture Fajer-1 130.67 131.00 132.67 125.67 130.00 (7.65) (8.85) (10.12) (10.71) (9.33) Service. www.attra.ncat.org. Leiby HR, Abd MT and Ali SH 2017. The Effect of spraying fertilizer Maha 127.33 132.33 134.33 132.33 131.58 NPK in the growth properties and yield in three varieties of peas (6.49) (8.20) (8.56) (9.96) (8.30) Pisum sativum L. Al-Qadisiyah Journal of Agricultural Cultivar average 127.89 130.67 133.00 129.78 Science(QJAS) 7(2): 59-66. (6.52) (8.17) (9.01) (9.90) Mreer SS and Mohamad YA 2017. Response of several cultivar of LSD 0.05 C:1.54 N: 1.69 (0.57) C×N: 2.84 maize to soaking seeds wit pyridoxine (Zea mays L.). AL-Anbar (0.35) (NS) Journal of Agricultural Science 15(2): 443-453. *Grain yield (t ha-1) Obaid HM and Sadik SK 2016. Response of some quantity yield Foliar Application with Novalon on Yield of Maize 81

traits of sweet corn (Zea mays L. var. saccharata) for planting Workshop on Foliar Fertilization. Bankok. Thailand, pp.1-32. dates and spraying with bios emulators. Karbala Journal of Saleh BH, Abood NM and AL-Hadeethi MAH 2017. Effect of bio Agricultural Science 3(2): 71-84. fertilizer concentrations on the qualitative and quantitative traits Ranum P, Pena-Rosas JP and Jarcia-Casa lMN 2014. Global maize of three cultivars of sorghum bicolor (L.) Moench. AL-Anbar production, utilization, and consumption. Annals of the New Journal of Agricultural Science 15(2): 430-442. York Academy of Sciences 1312: 105-112. Shafiullah RJ, Basir A, Alam JE, Ali A, Khan GR, Aziz A and KhanI Romhold V and EL-Fouly MM 2000. Foliar nutrient application: 2018. Evaluation of eight maize genotypes for yield and yield challenge and limits in crop production. 2nd International contributing traits. Pure and Applied Biology 7(2): 620-624.

Received 28 September, 2019; Accepted 21 November, 2019 Manuscript Number: S-257 Indian Journal of Ecology (2019) 46 Special Issue (8): 82-84 NAAS Rating: 4.96

Effect of Iron Oxide Nanoparticle and Humic Acid Spray on Growth and Yield of the Fenugreek (Trigonella Foenum Graecum L.) and Fixed Oil Content in Seeds

Qasim A.S. Al-Zyadi and Emad Abud-Alkareem Mohammedreda Al-Thahab College of Agriculture, Al-Muthanna University, Iraq E-mail: [email protected]

Abstract: A field experiment was conducted to study the effect of three concentrations of iron oxide nanoparticle spray (0, 50, 100 mg Fe l-1) and three concentrations of humic acid (0, 100, 200 mg l-1) on growth and yield of Trigonella foenum graecum L. and fixed oil content in seeds. The results showed that the fenugreek plants that have been sprayed with the concentration of 100 mg Fe ha-1 significantly performed better in respect of plant height, shoot dry weight, number of pods per plant, number of seeds per pod, weight of 100 seed, total seed yield and fixed oil percentage (62.01 cm, 7.27 gm plant-1, 19.83 pod plant-1, 15.06 seed pod-1, 1.704 gm, 1023.4 kg ha-1 and 6.84%, respectively). While, plants sprayed with humic acid with concentration (200 mg l-1) gave highest values in plant height, shoot dry weight, number of pods, weight of 100 seed, total seed yield and fixed oil percentage (57.90 cm, 6.79 gm plant-1, 19.23 pod plant-1, 1.594 gm, 921.1 Kg ha-1 and 6.70%, respectively).

Keywords: Fenugreek, Fixed oil, Humic acid, Iron oxide nanoparticle

Fenugreek (Trigonella foenum graecum L.), an annual agricultural operation to ensure the transmission and plant belongs to Fabaceae family, is one of the important distribution of nutrients as the life is the center of chemical medicinal plants used since ancient times and widespread in reactions such as photosynthesis. Humic acid is an most of the world. The importance of the Fenugreek plant to economical commercial product that is quick effective and the seed content of protein, carbohydrates, fats and vitamins harmless to humans, animals and plants (Arancon et al such as vitamin A, B and C (Lakshmi et al 2015) as well as 2004). It contains many nutrients that lead to increased contains many active alkaloids, including trigonelline, productive and vegetative growth (Ogendo et al 2008). For gentianine and carpaine; saponins, including sapogenins, importance of this plant and for improving its cultivation in diosgenin; flavonoids, including vitexin and its glycosides Iraq, an experiment was conducted with the aim to study the and esters; small quantities of essential oil (Khare 2007). The effect of iron oxide nanoparticle/humic acid spray on the seeds also contain about 5% of a stronger swelling, bitter growth parameters and fixed oil content of fenugreek plant. fixed oil (Ram 1993). Fenugreek is characterized with bunch of medicinal uses such as anti-inflammatory, antipyretic, anti- MATERIAL AND METHODS diabetic, diuretic and antibacterial activity (Zhou et al 2012) The experiment was conducted during the 2018-2019 and also has digestive stimulant action, antioxidant potency season in the fields of the research station of the Faculty of and hypocholesterolemic effect (Mehbub et al 2018). Agriculture. Al-Muthanna University in south of Iraq. To study

Nanoparticles have unique features due to their small size the effect of spraying of iron oxide (Fe2O3) nanoparticle 50 nm and large surface area, which increase absorption, increase (US Research Nanomaterial Inc. USA), with three the efficiency of photosynthesis process and increase the concentrations (0, 50, 100 mg Fe l-1) and three concentrations production of active substances in the plant (Singh et al 2016) of humic acid (0, 100, 200 mg l-1) in the growth of the as well as enhance efficiency of fertilizer use, overcome food fenugreek plant and the content of its fixed oil. The enrichment and improve environmental sustainability treatments were arranged in factorial experiment laid out in (Mishra et al 2017). The iron component contributes to a randomized complete block design with three replications. number of biological processes within the plant by stimulating Fenugreek seeds were planted in the field on 20 October of the activity of the enzymes of the transpiration process and 2018. The chemical and physical properties of the soil were the transfer of electrons. It also enters the structure of the estimated prior to planting and soil was silt clay loam (silt chloroplast and many enzymes such as catalase and 63%, sand 23% and clay 14%) with organic matter (0.72%), peroxidase (Barker and Stratton 2015). The spraying of pH (7.4), EC (2.6 dsm), 4.1, 11, 180 and 1.97 ppm available organic nutrients on the vegetative group is an important nitrogen, phosphorous, potassium and iron, respectively. Iron Oxide Nanoparticle and Humic Acid Spray on Yield of the Fenugreek 83

The land was divided into three sectors, each of which Table 1. Effect of iron nanoparticle and humic acid and their included 9 experimental units (plots), each had dimensions interaction in plant height (cm)* -1 3m × 2m and containing 120 plant. Seeds were planted in Iron nano Humic acid (mg l ) particle (mg l-1) experimental unit at rows spacing 30cm and another, and 0 100 200 Mean 10cm between plants. A uniform amount of phosphate 0 43.94 50.16 50.80 52.16 -1 (4.50) (5.27) (5.46) (5.68) fertilization (220 Kg P2O5 ha ) was added to all plants. The iron and humic acid solutions were sprayed in three batches 50 52.40 54.66 59.00 55.60 (5.88) (6.07) (6.76) (6.10) and for 15 days between batch and another. The plants were 100 60.16 62.00 63.88 57.90 harvested on 5 April of 2019. The plant height, shoot dry (6.66) (7.00) (8.14) (6.79) -1 -1 weight, number of seeds pod , number of pods plant , weight Mean 52.16 55.60 62.01 of 100 seed, seed fixed oil percentage and total seed yield (5.16) (6.10) (7.27) were recorded to interpret the results. LSD (0.05) Iron nan. Humic acid Iron nan. x Humic acid 2.73 (0.3) 2.73 (0.3) 4.73 (0.52) The fixed oil was extracted from the samples using a *Shoot dry weight (gm plant-1) Soxhlet apparatus , where 50 g of each sample was taken and put in a thimble (Whatman) after grinding rough with the addition of 250 ml of the solvent (Hexane) and filled the Table 2. Effect of iron nanoparticle and humic acid and their interaction in number of pods (pod plant-1)* device for 4 hours. Then the oil was separated from the Iron nano Humic acid (mg l-1) solvent using a rotary evaporator and weighed, and then particle (mg l-1) 0 100 200 Mean estimated the percentage of fixed oil (AOAC 1980). All statistics analysis was performed by analysis of variance 0 12.66 13.25 16.00 13.97 (12.21) (13.91) (14.25) (13.45) (ANOVA) and least significant difference (LSD) at probability 50 16.33 16.87 18.20 17.13 of 0.05. (15.79) (15.22) (15.24) (15.42) 100 17.00 19.00 23.50 19.83 RESULTS AND DISCUSSION (14.72) (15.77) (14.68) (15.06) Effect of iron oxide nanoparticle spray: The results shown Mean 15.33 (16.37 (19.23 (14.24) (14.97) (14.72) in Table (1-4) showed the significantly effect of iron LSD (0.05) Iron nan. Humic acid Iron nan. x Humic acid nanoparticle spray in growth parameters and oil content of 1.24 (0.97) 1.24 (NS) 2.14 (1.67) fenugreek plant, where the spray exceeded the *Number of seeds (seed pod-1) concentration (100 mg l-1) and gave the highest values of -1 plant height (62.01cm), shoot dry weight (7.27 gm plant ), showed that spray of humic acid with concentration (200 mg -1 number of pods (19.83 pods plant ), number of seeds (15.06 l-1) gave higher value in plant height (57.90 cm), shoot dry -1 seed pod ), weight of 100 seed (1.704 gm), total seed yield weight (6.79 gm plant-1), number of pods (19.23 pod plant-1), -1 (1023.4 kg ha ) and fixed oil percentage (6.84%), Compared weight of 100 seed (1.594 gm), total seed yield (921.1 kg ha-1) to the control treatment (water spray only) that gave the and fixed oil percentage (6.70%), while the lowest values lowest values. The significant superiority of iron oxide were in the control treatment (water spray only). The spraying nanoparticle may be due to the importance of iron in the plant of humic acid did not have any significant effect on the growth. Iron is an essential component of various active number of seeds in bod plant-1. The significant effect of humic compounds in plant cells such as enzymes, cytochromes, acid may be due to its role in stimulating root growth and ferredoxine and in the synthesis of chlorophyll, Therefore, it increasing the absorption of water and nutrients by the plant plays an important role in the metabolism of the plant and that (Cimrin and Yilmas 2005) or by stimulating the production of the processing of iron in conjunction with plant growth and hormones and enzymes within the plant (Sarir et al 2005, efficiency in its use increases its impact on the vegetative Mart 2007). In addition to its role in increasing enzyme qualities and quality of the plant (Barker and Stratton 2015). catalysis and enhances respiration and photosynthesis Nanoparticles also have distinct characteristics due to their process (Nardi et al 2002). high surface area, which leads to increased absorption, Effect of interaction: Results of Tables (1-4) concluded that affects the solubility and spread of nutrients and thus their the interaction between iron nanoparticle spray and humic availability to the plant to increase the process of acid spray was a significant. The highest plant height, shoot photosynthesis, thereby increase the production of active dry weight, number of pods, total seed yield and fixed oil substances in the plant (Singh et al 2016). percentage were recorded by the interaction between iron Effect of humic acid spray: The results of Table (1-4) nanoparticle (100 mg l-1) and humic acid (200 mg l-1). While, 84 Qasim A.S. Al-Zyadi and Emad Abud-Alkareem Mohammedreda Al-Thahab

Table 3. Effect of iron nanoparticle and humic acid and their REFERENCES interaction in weight of 100 seed (g)* AOAC 1980. Official Methods of Analysis. Association of Official Iron nano Humic acid (mg l-1) Agricultural Chemists, 13th Ed. Washington, D.C., USA. particle Arancon NQ, Edward A, Bierman P, Metzger JD, Lee S and Welch C -1 (mg l ) 0 100 200 Mean 2004. Effect of vermicomposts on growth and marketable fruits 0 1.120 1.280 1.363 1.255 of field-grown tomato, pepper and strawberries. Pedoboil 47(6): (366.1) (471.8) (620.0) (486.0) 731-735. 50 1.400 1.510 1.600 1.503 Barker AV and Stratton ML 2015. Iron. Chapter 11. In: Barker AV and (722.0) (775.2) (887.5) (794.9) Pilbeam DJ (eds) Handbook of Plant Nutrition. Second Edition. CRC Press Taylor and Francis Group. London. New York, pp. 100 1.590 1.700 1.820 1.704 399-426. (795.8) (1018.7) (1255.7) (1023.4) Cimrin KM and Yimaz I 2005. Humic acid applications to lettuce do Mean 1.37 1.497 1.594 not improve yield but do improve phosphorus availability. Acta (628.0) (755.3) (921.1) Agriculturae Scandinavica, Section B-Soil and Plant Science LSD (0.05) Iron nan. Humic acid Iron nan. x Humic acid 55: 58-63. 0.134 (45.79) 0.134 (45.79) NS (79.32) Khare CP 2007. Indian Medicinal Plants. An lllustrated Dictionary. *Total seed yield (kg ha-1) Springer Science+Business Media, LLC., 233 Spring Street, New York: pp.675. Lakshmi J, Gowda R, Parashivamurthy S and Shivanandan VN Table 4. Effect of iron nanoparticle and humic acid and their 2015. Influence of pre-flowering pinching and Maleic hydrazide spray on plant growth, seed yield and quality attributes in interaction in fixed oil percentage (%) fenugreek. Legume Research 38(3): 353-357. -1 Iron nano Humic acid (mg l ) Mahbub J, Mou RA, Sikta SA, Rahman A and Dash PR 2018. -1 particle (mg l ) Biological and medicinal significance of Trigonella foenum- 0 100 200 Mean graecum: A review. International Journal of Life Science and 0 5.70 5.89 6.24 5.94 Review 4(2): 15-26. Mart I 2007. Fertilizers, organic fertilizers, plant and agricultural 50 6.43 6.30 6.63 6.45 fertilizers. Agro and Food Business Newsletter, pp. i-iv. 100 6.35 6.95 7.23 6.84 Mishra S, Keswani C, Abhilash PC, Fraceto LF and Singh HB 2017. Mean 6.16 6.38 6.70 Integrated approach of agri-nanotechnology: Challenges and future trends. Frontiers in Plant Science 8(471): 1-12. LSD (0.05) Iron nan. Humic acid Iron nan. x Humic acid Nardi S, Pizzeghello D, Muscolo A and Vianello A 2002. Physiological 0.149 0.149 0.257 effect of humic substances on higher plants. Soil Biology and Biochemistry 34: 1527-1536. -1 the interaction between iron nanoparticle (50 mg l ) and Ogendo RO, Isutsa DK and Sigung DO 2008. Interaction of humic acid with 0 mg l-1 (water spray only) gave the highest Farmyard Manure and plant population density effects on soil value in number of seeds per pod. Interaction between characteristics and productivity of mulched strawberry in a tropical climate. African Journal of Horticultural Science 1: 100- experimental factors did not significantly affect the weight of 115. 100 seeds. Ram PR 1993. Compendium of India Medicinal plants. Vol. 2nd, 3rd, 4th. New Delhi: CDRI, Luck now & National Institute of Science Communication, pp. 659-660, 740. CONCLUSION Sarir MS, Sharif M, Zeb A and Akhlaq M 2005. Influence of different In this study, which was conducted under the levels of humic acid application by various methods on the yield environmental conditions of the province of Al-Muthanna in and yield components of maize. Sahad Journal of Agriculture 21: 75-81. southern Iraq, it was found that the iron nanoparticle spray Singh A, Singh S and Prasad SM 2016. Scope of nanotechnology in with100 mg l-1 was the most appropriate and best for growth crop science: Profit or loss, research and reviews. Journal of and production of fenugreek. Most of the studied traits of Botanical Sciences 5(1): 1-4. Zhuo J, Chau L and Zhou S 2012. Trigonelline: A plant alkaloid with fenugreek increased significantly when spraying the humic therapeutic potential for diabetes and central nervous system acid concentration of 200 mg l-1. disease. Current Medicinal Chemistry 19: 3523-3531.

Received 10 October, 2019; Accepted 21 November, 2019 Manuscript Number: S-258 Indian Journal of Ecology (2019) 46 Special Issue (8): 85-89 NAAS Rating: 4.96

Effect of Spraying with Gibberellic Acid on Growth and Yield of Three Cultivars of Broad Bean (Vicia faba L.)

Ahmed Hasan Fadhil and Mohammed Mehdi Muhsen Almasoody Al- Mussaib Technical College, Al-Furat Al-Awsat Technical University, Babylon, 51001, Iraq E-mail: [email protected]

Abstract: A study of split-plot was conducted for the season 2017-2018 In the fields of Baghdad province, To study the effect of spraying with gibberellic acid and three cultivars in plant height, the number of branches in the plant, the number of pods, the number of seeds in the pod, the weight of 100 g seed, the plant yield (kg) and total plant yield tons ha -1 for peas. The main plot contained the genotypes (local, French and Italian), secondary plots include gibberellic acid concentrations (0, 100, 200 and 300 mg l-1). The results of local genotypes showed the highest average of plant height, number of branches in the plant. The number of pods in the plant, while the Italian genotype gave the highest average of the pod length and the number of seeds per pod, weight of 100 g seed, the plant yield and total plant yield tons ha-1, On the other hand, the spraying with gibberellic acid at a concentration (300 mg l-1) resulted in the highest average in all studied traits. The interaction between the genotypes and the sprayed gibberellin concentrations was significant for some of the studied traits. The interaction (local and 200) gave the highest average in the number of branches of the plant, the interaction (local and 200) gave the highest average in the number of plant pods and interaction (local and 300) gave the highest average in the number of branches of plant, the number of plant pods, the interaction (Italian and 300) gave highest number of seeds per pod, weight of 100 g seed, plant yield, total plant yield, plant height and pod length were not significant for interaction .

Keywords: Broad bean, Genotypes, Gibberellic acid

Vicia faba L. is one of the main vegetable legume plants, affect either the reaction of light or systems of converting which has high protein content. This made it one of the carbon into organic compounds in chloroplasts as well as sources of green protein. They are an important part of their differences in the content of nutrients and protein ratio human food, especially low-income people, it is also and this difference is often due to the nature of the cultivar and important in improving soil fertility through the biological of its genotypes and conditions environmental (Issa 1990, Daur nitrogen fixation in the soil by commensalism with Rhizobium et al 2008). The plant needs growth regulators in order to bacteria of (Kandil 2007). It is a cheap source of protein complete the growth, because of its important role in compared to the protein we get from animals. His plants are improving biological activity, as many researches and studies used as green manure for soils that increase the productivity indicated that the treatment of plants with a specific growth of many crops grown in the field to atmospheric nitrogen regulator leads to the improvement of the plant structure and fixation in the soil (Jasim 2007). The cultivation Broad bean the yield quality and the production of seeds (Khalaf and widespread in Iraq, especially in the province of Baghdad, Rajbo 2006). Gibberellins play a role in balancing the growth Babylon, Kirkuk and Nineveh, with an estimated area of 5100 of internodes and the growth and development of the leaves. hectares in 2010 with a production rate of 2.824 t ha-1) It has been found that plants grow in the long-day or Chilling (Ministry of Agriculture 2012). The cultivar different in the requirement if they remain in the short day or in warm vegetation according to the nature of growth, which differs in conditions, where it remain growing vegetatively and do not large leaves and petiole length, branching and continued flowering. The treatment of these plants with gibberellic acid growth after flowering may lead to a change in the shape of will be compensated by the requirements of the light or cold the plant. Therefore, the distribution and penetration of light period and thus elongate the stems, flowering and the plants through the vegetation is different and is reflected in the where the elongation of the flower stem and flowered They position of the lower leaves, which found that the lower leaves contain more gibberellin compounds than plants that have do not live parasitic on the plant but die gradually, when not elongation of the flower stem and non-flowering stems respiration speed increases on the photosynthesis speed (Saleh 1990). Gibberellin is used to obtain rapid vegetative (Yoshida 1972). Broad bean cultivar differs in their Genotypes growth in leafy vegetable crops and grain feed. The aim of this and also in the nature of their growth and morphological form. study is to evaluate the performance of the genotypes and the They are affected by environmental factors such as light, growth regulator gibberellins (GA3) and their interaction to carbon dioxide, water and nutrient status in the soil, which achieve the integration of production in quantity and quality. 86 Ahmed Hasan Fadhil and Mohammed Mehdi Muhsen Almasoody

MATERIAL AND METHODS differences between the mean of the treatments were tested This experiment was conducted to study Effect of by using the least significant difference (LSD) with a spraying with gibberellic acid on growth and yield of three probability level of 5%. cultivars of Broad bean Vicia faba L. In this experiment used three genotypes of Broad bean obtained from local markets RESULTS AND DISCUSSION (local, French and Italian). Several concentrations of Plant height (cm): Average of the plant height ranged gibberellic acid (0, 100, 200 and 300 mgl-1) were used. The between 80.75 cm in local genotypes, which also significantly broad bean seeds were cultivation on lines of the distance excelled the French and Italian genotypes, which gave the between one line and another 70 cm. The length of the line average amounted 69.33 and 73.50 cm, respectively. As for inside the experimental unit was 3 m. Each line contains 15 the adding of gibberellin, the Table 1 showed that there were pits and the distance between one pit and another 20 cm. The significant differences in the same traits, The concentration seeds were cultivation on 10/10/2017, After the emergence 300 mg l-1 gave the highest average of amounted 78.89 cm of seedlings, the seedling was thinning so that each pit compared to the non-adding treatment which gave the lowest contained one plant (Almayoof 1982). The weed control was average in this trait amounted 70.78 cm. This is due to the manually and the urea fertilizer was added 60 kg in one role of gibberellin in stimulating the process of the plant batch, The harvest was completed at full maturity on elongation, wherever the concentration of gibberellin 17/4/2018. The gibberellin was prepared at a concentration increased, the plant height was increased. These results (100 mg l-1) of acid and prepared by taking a tablet 1g agree with Saleh and Abdul (1980). The increase in plant containing 1g active substance and dissolving it in a liter of height is due to the effect of gibberellic acid in increasing cell distilled water to obtain the stock solution and keeping the division and enlargement, in adding to stimulating growth and solution with a dark bottle in a dark place and take (100 ml) of cell expansion (Abdul 1987 and Saleh 1990). the basic solution and complete the volume to (1000 ml) to The number of branches in the plant: The results in Table 1 obtain a concentration (100 mg l-1) Then spray on the confirmed that there were significant differences in the vegetative part as well as the same way prepare genotypes in the number of leaves in the plant for the season concentrations 200 and 300 was sprayed in the early 2017 – 2018. Local genotypes gave the highest average morning with the use of dishes washing solution to break the amounted of 7.75 on other genotypes. This results agree with surface tension of water The following traits were studied: other researchers have found that there were significant plant height (cm), number of branches per plant, number of differences between the cultivar of broad bean in Average pods, length of the pod, number of seeds per pod, weight of number of branches in the plant Afifi et al 2010, Yucel 2013). 100 g seed, plant yield, total seed yield per tons h-1 The results in Table 1 showed that there were significant Statistical analysis: The statistical analysis was conducted differences between the spraying with concentrations of the split-plot arrangement according to randomized complete growth regulator gibberellin. The results of the same table block design and the main plot contained the genotypes. The were significant for the interaction between (local and 300), secondary plot was contained thiamine acid concentrations which gave the highest average amounted 8.40, compared to and using the statistical program Genstate. The Significant the other interaction, but the difference was not significant

Table 1. Effect of spraying with gibberellic acid, three cultivars and their interaction on average plant height (cm) and number of branches plant-1 (Season 2017-2018) Concentration of Plant height Number of branches plant-1 gibberellins/ Genotypes 0 100 200 300 Average of the 0 100 200 300 Average of the effect of genotypes effect of genotypes

Local 75.33 78.67 81.67 87.33 80.75 6.96 7.46 8.16 8.4 7.75 French 67.67 68.67 69.67 71.33 69.33 5 5.46 5.83 6.46 5.69 Italian 69.33 71.67 75 78 73.5 5.76 6.2 6.73 7.46 6.54 Average of the effect of 70.78 73 75.44 78.89 5.91 6.37 6.91 7.44 concentration for gibberellin LSD0.05 Concentrations Genotypes Concentrations* Concentrations Genotypes Concentrations* 4.363 3.192 genotypes 0.241 0.237 genotypes NS 0.398 Gibberellic Acid on Yield of Broad Bean 87 from the interaction between (local and 200) which also gave genotypes gave the highest average, compared to the other the highest average of the other interaction amounted 8.16. structures amounted (17.17 cm). The results in the same The number of pods in the plant: The results of Table 2 table showed a significant increase of the concentrations 300 showed that the effect of genotypes and gibberellin was mg L ¹ which gave the highest average amounted 18.22 cm significant and that the local genotypes excelled the of all the on the all the concentrations and also on the treatment of non genotypes which it gave the highest average amounted - adding, which gave the lowest average amounted 11.00 cm. 15.58 pods, while the French genotypes gave the lowest The interaction was not significant in this trait. average of 11.00 pods. The reason for excelled of the local The number of seeds in the pod: The results in Table 3 cultivar on other genotypes may be due to excelled in the indicate that there were significant differences between the number of branches plant-1. This result agrees with Al-Jubouri effect of the genotypes and the concentrations and their (2014) and Kubare et al (2014). The results in Table 2 showed interaction. The Italian genotype gave the highest average that there was a significant increase in this trait when added amounted 6.66 seeds on other genotypes. The reason for the gibberellin concentrations. The concentration of 300 mg l-1 different effect of the cultivar in this trait is due to the variance gave the highest average of 16.78 pods, compared to the no of the cultivars in their genetic traits or variation in the nature adding treatment which gave the lowest average amounted of growth and this is reflected in turn on the number of seeds 10.44 pods interaction between (local and 300) and (local per pod. This results agree with Ayed (2012).As for the and 200) gave the highest average of 19.67 and 17.67 pods, concentrations. The concentration of 300 mg l-1 was excelled respectively. and gave the highest average of 6.76 seeds on all the Pod length (cm): The results of Table 2 showed that the concentrations and gave the treatment of non-adding the genotypes had a significant effect in this trait, and the Italian lowest average amounted 5.67 seeds. As for the interaction

Table 2. Effect of spraying with gibberellic acid, three cultivars and their interaction on average the number of pods plant-1 and pod length (Season 2017-2018) Concentration of Number of pods plant-1 Pod length gibberellins/ Genotypes 0 100 200 300 Average of the 0 100 200 300 Average of the effect of genotypes effect of genotypes

Local 11.33 13.67 17.67 19.67 15.58 8.33 9.33 11.67 16.33 7.75 French 10.33 9.33 10.33 14.00 11.00 11.00 12.67 15.00 18.00 5.69 Italian 9.67 11.00 13.67 16.67 12.75 13.67 16.00 18.67 20.33 6.54 Average of the effect of 10.44 11.33 13.89 16.78 11.00 12.67 15.11 18.22 concentration for gibberellin LSD0.05 Concentrations Genotypes Concentrations* Concentrations Genotypes Concentrations* 1.246 2.179 genotypes 2.285 2.267 genotypes 2.525 NS

Table 3. Effect of spraying with gibberellic acid, three cultivars and their interaction on average the number of seed in the pod and weight of 100 g seed (Season 2017-2018) Concentration of Number of seed Pod length gibberellins/ Genotypes 0 100 200 300 Average of the 0 100 200 300 Average of the effect of genotypes effect of genotypes

Local 5.06 5.53 5.85 6.30 5.68 153.67 159.33 162.67 165.67 160.33 French 5.90 6.06 6.33 6.76 6.26 160.67 164.67 168.67 173.67 166.92 Italian 6.06 6.50 6.86 7.23 6.66 164.67 169.67 175.33 182.33 173.00 Average of the effect of 5.67 6.03 6.34 6.76 159.67 164.56 168.89 173.89 concentration for gibberellin LSD0.05 Concentrations Genotypes Concentrations* Concentrations Genotypes Concentrations* 0.172 0.232 genotypes 1.324 1.463 genotypes 0.310 2.247 88 Ahmed Hasan Fadhil and Mohammed Mehdi Muhsen Almasoody

Table 4. Effect of spraying with gibberellic acid, three cultivars and their interaction on average plant yield and total plant yield (Season 2017-2018) Concentration of Plant yield Total plant yield gibberellins/ Genotypes 0 100 200 300 Average of the 0 100 200 300 Average of the effect of genotypes effect of genotypes

Local 261.67 267.00 272.00 277.00 269.42 2.59 2.66 2.79 2.84 2.72 French 269.67 276.00 285.67 292.33 280.92 2.79 2.86 2.89 3.61 3.04 Italian 282.00 288.67 295.00 306.00 292.92 2.90 2.97 3.25 3.70 3.20 Average of the effect of 271.11 277.22 284.22 291.78 2.76 2.83 2.98 3.38 concentration for gibberellin LSD0.05 Concentrations Genotypes Concentrations* Concentrations Genotypes Concentrations* 2.983 3.296 genotypes 2.983 3.296 genotypes 5.063 5.063

was significant, the interaction between (Italian and 300) interaction was significant as the interference between gave the highest average amounted 7.23 seeds on the other (Italian and 300) gave the highest rate of 306.00 g compared interaction. to the other interventions. The interaction between Italian and The weight of 100 g seed: The results in Table 3 showed 300 gave the highest average amounted 306g. that there were significant differences in the weight of 100 g Total plant yield (t ha-1): The results in Table 4 showed that seed, the Italian genotypes gave the highest average there were significant differences in the genotypes. The amounted 173.00 on the other genotypes. The reason for the Italian genotypes gave the highest average of 3.20tha-1 excelled of the Italian cultivar in this trait is due to the number compared to the lowest average of 2.72t ha-1. The reason for of seeds per pod and the pod length and may be due to the the excelled of the Italian cultivar in this trait is due to the different cultivar in the size of seeds, and that the weight of different thermal requirements of the plant in the flowering the seed of the plant is a function of the average of stage, which led to a clear variation in the contract of the photosynthesis and the transfer of its products (Isa 1990). varieties in the study Sau and Minguez (2000). The results in The results in Table 3 showed that there was a significant Table 4 showed that there were significant differences for -1 difference for this trait for the added concentrations. The growth regulators, where The concentration300 mg l concentration 300 mg l-1, gave the highest average amounted excelled on the all other the concentrations and gave the 173.89 g compared to the no adding treatment which gave highest average of 3.38 and The treatment non-spraying -1 the lowest average amounted 159.67 g. The results in Table 3 gave the lowest average amounted 2.76tha . The results in showed that there were significant differences in the Table 4 indicated that there were significant differences for interaction treatments. The interaction between Italian and the interaction, where the interaction between (Italian and 300 gave the highest average of 182.33 g on all interaction 300) and (French and 300) gave the highest average -1 treatments. amounted 3.70 and 3.61 t ha , respectively on the other Plant yield (g): It is clear from the results in Table 4 that there interactions. are significant differences between the genotypes, concentrations and the interaction between them in the REFERENCES average the plant yield. The Italian genotypes of broad bean Abdul Karim Saleh 1987. Plant growth organizations. Parts I and II. Directorate of books for printing and publishing - University of plants achieved the highest average amounted 292.92 g and Mosul - Iraq. the local genotypes gave the lowest average of 269.42 g. Afifi MHM, Mohamed FM and Shaaban SHA 2010. Yield and utrient uptake of some faba bean varieties grown in newly cultivated These results agree with (Alan and Geren 2007). It confirmed soil as affected by foliar application of humic acid. Journal of the excelled of the cultivar and that through its ability to Plant Production 1(1): 77-85. express the genetic nature as plant yield. To find out the Alan O and Geren H 2007. Evaluation of heritability and correlation for seed yield and its components in faba bean Vicia faba L. concentrations of growth regulator used in this research, we Journal of Agronomy 6(3): 484-487. -1 find that the concentration of 300 mg l gave the highest Al-Jubouri Hatem Mohammed Mohsen 2014. The Effect of average of 291.78 g and the treatment of non - spraying gave Distances between furrow on Growth and Gene Genotypes of Pea. M.A. Thesis, faculty of Agriculture .University of Kirkuk. the lowest significant difference for this trait amounted Almayoof Mahmoud Ahmed 1982. The entrance of legumes in Iraq. 271.11g. It is clear from the results in Table 4 that the Dar Al Kutub Printing & Publishing Est. University of Al Mosul. Gibberellic Acid on Yield of Broad Bean 89

Ayed Qutaiba Yusr 2012. Effect of three foliar fertilizers on growth Journal of Life Science 3(4): 287- 305. and yield of two cultivars (Vicia faba L.) under drip irrigation Ministry of Agriculture 2012. Brochure Statistical of Field Crop Data, system. Tikrit University Journal for Agricultural Sciences 12(1) Dept. of Agricultural Economics Res., Directorate of Agric., Res. 131 - 137. pp. 64. 24. DaurI Sepetoglu H, Marwat KB, Hassan G and Khan IA 2008. Effect Mitiku A and Wolde M 2015. Physiological aspects of yield. Annual of different levels of nitrogen on dry matter and grain yield of Review of Plant Physiology 23: 437-464. faba bean Vicia faba L. Pakistan Journal of Botany 40(6): 2453- 2459. Saleh MM and Abdul KS 1980. Effect of gibberellic acid and cycocel on growth, flowering and fruiting of tomato plant. Mesopotamia Issa Taleb Ahmed 1990. Crop plant physiology. Ministry of Higher Journal of Agriculture 15: 137-161. Education and Scientific Research. University of Baghdad, Pp. 496. Saleh Musleh Mohammed Saeed and Caesar Jaafar Abdul 1989. Effect of glycerol and naphthalene acetic acid on onion growth, Jasim Ali Hussein 2007. Effect of foliar fertilization on growth and production and quality. Agriculture Studies 16(9): 39-51. yield of broad bean. Anbar Journal of Agricultural Sciences 5(2): 5-20. Saleh Musleh Mohammed Saeed 1990. Physiology of plant growth regulators. First edition. Ministry of Higher Education and Kandil MO and Hala ZR 2007. Effect of coball fertilizers ongrowth, Scientific Research. Salahaddin University - Iraq. yield and nutrient status of faba bean (Vicia faba L.) Plant Journal of applied Science Research 3(9): 867-872. Sau F and Minguez ML 2000. Adaptation of indeterminate Faba beans to weather and management under a Mediterranean Khalaf Ahmed Saleh and Abdul Sattar Asmir Al-Rajbo 2006. Seed Climate. Field Crops Research 66: 81-99. technology. Dar Ibn Al Atheer Printing & Publishing. University of Mosul, Iraq. Yucel DO 2013. Optimal intra-row spacing for production of local faba bean (Vicia faba L.) major cultivars in the Mediterranean Kubure TE, Cherukuri R, Arvind C and HamzaI 2014. Effect of faba bean (Vicia faba L.) genotypes, plant densities and phosphous conditions. Pakistan Journal of Botany 45(6): 1933-1938. on productivity, nutrients uptake, soil fertility changes and Yoshida S 1972. Physiological aspects of grain yield. Annual Review economics in Central high lands o Ethiopia. International of Plant Physiology 23(1): 437-464. Received 24 September, 2019; Accepted 20 November, 2019 Manuscript Number: S-259 Indian Journal of Ecology (2019) 46 Special Issue (8): 90-92 NAAS Rating: 4.96

Effect of Weed Management on Growth Parameters in Sunflower (Helianthus annuus L )

Mahmoud T. Al-Jayashi and Hayder R. Laibi Agriculture College, Al-Muthanna University, Al-Muthanna, Samawa, Iraq E-mail: [email protected]

Abstract: Field experiment was conducted on chemical control study in three genotypes, Cartage, Tarsan and Flam of sunflower. Treatments included were weedicide Travellan 1 liter and stomp 1.7 liters per 200 liters water per hectare, 2.4.DB 200 mg per kg soil, manual weeding and unweeded control in sub plots. The results showed that weed-free treatment of sunflower crop, gave the highest dry matter yield, plant growth rate, leaf area, index of leaf area, seed in the disk and seed yield. Flama genotype gave the highest yield of dry matter, plant growth rate, leaf area and rate of leaf area index. While Cartage genotype gave the lowest yield of dry matter at 50% flowering, growth rate of the plant, leaf area and the leaf area index.

Keywords: Chemical control, Helianthus annuus L., Sunflower, Manual hoeing, Weed

Sunflower (Helianthus annuus L.) one of the most plot 40 cm, The seeds were planted at March 3, 2016, The important oil seed crop in Iraq. The weed management is experiment was fertilized with NP fertilizer (N 18%, 46% -1 crucial for higher productivity. Sunflower is a weed-sensitive P2O5) before planting at a rate of 240 kg ha .Planting was crop during its early growth stages, despite being a stifling done manually by placing three seeds in the hole, defoliation crop, due to the large leaf size and its rapid growth (Khan at el to one plant when the plants reached the stage of four leaves. 2004). The weed cause a significant loss of sunflower seed Urea (46% N) was added at a rate of 280 kg ha-1 in the first two yield, understanding of genotype under weed competition batches after two to three weeks of germination and 140 kg conditions will inevitably lead to discover its genetic abilities ha-1 and the second at the beginning of the formation of flower by synchronizing the growth and competition of the weed, buds and 140 kg ha-1.The following traits were studied: which leads to decide the appropriate genotype with the right Paper area where the maximum width of the leaves was weed pesticide, Travelan stomp and 2-4-D are used after measured for each plant separately and then the paper area germination to control broad herbs. The aim of this study is to was calculated after El-Sahooki and El-Dabas (1982); leaf investigate the response of three genotypes of sunflower on area index = leaf area of the plant/area occupied by the plant weed management. from the ground; dry matter, which represents the dry weight of green plant parts after drying at 80°C for 48 hours (Al- MATERIAL AND METHODS Chalabi 1988); crop growth rate = dry matter/number of days The experiment was carried out at Muthanna of planting, number of seeds/discand seed yield (g plant-1). Governorate, Warkaa district during the 2016 agricultural season, to study the effect of chemical control and manual RESULTS AND DISCUSSION hoeing on some growth characteristics of three genotypes of Plant growth rate: The differences were significant among sunflower. The experiment was in split -plot and Randomized genotypes, the genotype Flama gave the highest of plant Complete Block Design with three replicates were used. The growth (7.6) but did not differ significantly with Tarsan. main plot had five treatments of chemical control, included Cartage gave the lowest growth rate and did not significantly weed pesticides (Travellan) 1 liter and (stomp)1.7 liters/200 differ with Tarsan (Table 1). Weed-free treatment excelled liters water per hectare, 2.4.-D 200 mg kg-1 soil, manual and gave the highest rate of plant growth, significant control treatment (weed-free) and control. Cartage, Tarsan difference from the other treatments. Significant differences and Flam genotypes constituted sub plots. The field was land in plant growth rate were observed in the interaction between of experiment ploughed in perpendicular tillage with treatments, free-weed *cartage gave highest rate (8.8), mulching plow, the harrowing and leveling process. Each plot cartage * weedy gave the lowest rate of plant growth (3.7). was separated from the other by a distance of 2 m, This may be due to the effectiveness of weedicides in experimental unit area was (3 x 3 m), six lines included the combating the growing of weed with the crop, thus reducing length of the line 3 m, the distance between the line inside the competition for growth elements, provides a better Weed Management on Growth in Sunflower 91 opportunity for the crop to exploit growth elements (Mussavi Table 2. Effect of weed control on leaf area index* of et al 2009). genotypes in sunflower crop at 50% flowering stage Leaf area: The genotypes significantly had affect on the leaf Treatment Genotype Pesticide area, Flama genotype gave the highest rate of leaf area mean Flama Tarsan Cartage (834.7 cm2), superior to the other genotypes, while cartage gave the lowest average (618.5 cm2). Agro-chemical control Travellan 5.5 (7.2) 4.8 (6.8) 3.9 (6.5) 4.7 (6.8) treatments significantly affected the leaf area, weed-free Stomp 4.5 (6.0) 3.7 (5.9) 2.4 (5.5) 3.5 (6.0) gave the highest rate of leaf area (911.6 cm2), a significant 2.4.-DB 4.7(7.5) 3.7 (7.2) 3.9 (7.6) 4.0 (7.4) difference with the rest treatments, weedy - check gave the Weed-Free 6.4 (10.5) 5.9 (8.8) 5.4 (8.8) 5.9 (9.3) 2 lowest leaf area average (525.6 cm ). The superiority of the Control 4.2 (6.3) 3.9 (6.3) 2.5 (3.9) 3.3 (5.5) control treatment (weed-free) was recorded on the leaf area Genotype 5.0 (7.1) 4.2 (7.0) 3.5 (6.4) of the sunflower crop compared to other treatments, due to mean the reduction of competition for the plant by the weed in the CD (p=0.05) Genotype Pesticide Interaction control early stages of crop growth, which allowed the plant to take 0.33 (NS) 0.12 (0.65) 0.60 (1.82) advantage of the light nutrients which affected the increase of *Dry matter yield (t ha-1) in parentheses the leaf area. The interaction between the two factors was found significant in the leaf area, Flama * weed-free gave the between the treatments significantly affected on the leaf area 2 highest leaf area (958.5 cm ), without moral difference with index, Flama* weed-free gave the highest score (6.4), the other, while cartage * weedy check gave the lowest rate without a significant effect in the leaf area index, Flama* 2 (380.6 cm ), without a difference from the rest (Table 1). weed-free gave the highest score (6.4), without moral Leaf area index: Genotypes significantly affected the leaf difference with the rest. Cartage* weedy (check) gave the area index, Flama gave the highest rate of leaf area index lowest rate (2.5), without difference with the other. (5.0), superior to other genotypes, while Cartag recorded the Dry matter yield (t ha-1): There were no significant lowest average index of paper area was (3.58), without differences between the genotypes of sunflower crop in dry significant difference with the Tarsan genotype (Table 2). It matter yield at 50% flowering. The control treatment results was observed in the same table, the pesticide control had a showed significant differences between them, weed-free significant effect on leaf area index, weed-free gave the treatment gave the highest dry matter yield at 50% flowering highest rate (5.9), a significant difference from the rest of the (9.3 tha-1), weedy treatment gave the lowest dry matter yield transactions, weedy (check) gave the lowest rate (3.3), (5.5 tha-1), without significant difference with the treatment of without significant difference with Stomp, 2,4-D BonTarsan. weedicide Stomp, the manual hoeing treatment Increase the leaf area index in a Weed-free treatment, outperformed (Table 2) due to reduced competition of the perhaps this was due to the increase in the leaf area per weed for sunflower plants, resulting in increased plant plant, by reducing the rate of weediness, which reduced height, leaf area and stem diameter. The weed-free *Flama water consumption, increased relative humidity, and gave the highest yield of dry matter at 50% flowering increased the efficiency of photosynthesis. The interaction amounted to 10.5 tha-1, without moral difference with treatments, while the treatment (stomp * cartage) gave the Table 1. Effect of weed control on growth rate of genotypes in lowest average dry matter (5.5 t ha-1), no significant sunflower crop at 50% flowering stage (values of leaf 2 difference with other treatments. area cm in parentheses) Treatment Genotype Pesticide Seeds number in the disk (seed/disk): The genetic mean structure was significantly affected by the number of seeds, Flama Tarsan Cartage the highest number of seeds disk-1 was 1099 (seeds disk-1), Travellan 6.7 (930.6) 7.2 (760.5) 6.5 (630.8) 6.8 (773.5) significantly superior to the other, while cartage gave the Stomp 5.6 (850.3) 5.6 (745.3) 5.5 (540.3) 6.0 (711.9) lowest average number of seeds (723 seeds/disk), without 2.4.-DB 6.0 (748.5) 6.0 (710.7) 7.6 (690.5) 7.4 (716.5) significant difference with the genotype Tarsan (Table 3). It Weed-Free 8.2 (958.5) 7.0 (925.9) 8.8 (580.5) 8.0 (911.6) was noted that control factors significantly affected on the Control 5.2 (685.8) 5.1 (510.6) 3.7 (380.6) 5.2 (525.6) number of seedsdisk-1, The highest weed-free transaction Genotype mean 7.6 (834.7) 7.2 (730.6) 6.5 (618.5) (1099 seedsdisk-1), with a significant difference with the rest CD (p=0.05) Genotype Pesticide Interaction of the treatments, while the the lowest rate reached (733 control seeds disk-1) treatment weedy check. Increase the number of 0.9 (54.1) 0.5 (98.2) 2.5 (101) seedsdisk-1 in free-weed treatment, perhaps was due to the 92 Mahmoud T. Al-Jayashi and Hayder R. Laibi

Table 3. Effect of weed control on seeds number in the disk results showed that the interaction effect had a significant -1 (seed disk ) of genotypes in sunflower crop at 50% effect on seed yield. Flama* Weed-free gave the highest flowering stage seed yield (100.8 g plant-1), no significant difference with the Treatment Genotype Pesticide mean others, Cartage* weedy (check) gave the lowest rate of 47.13 Flama Tarsan Cartage (g plant-1). Travellan 1099 766 759 874.66 (100.8) (80.69) (70.54) (84.10) Stomp 1013 780 764 852.33 CONCLUSION (88.02) (74.33) (62.71) (75.02) The weed-free treatment of sunflower crop, gave the 2.4.-DB 868 882 828 859.3 highest dry matter yield highest plant growth rate, leaf area, (74.70) (60.66) (50.53) (61.96) leaf area index, seed in the disk and seed yield. Flama Weed-Free 1099 777 723 866 (100.8) (71.14) 70.54) (80.82) genotype gave the highest yield of dry matter, plant growth Control 800 790 759 783 rate, leaf area and leaf area index. Cartage genotype gave the (74.01) (80.69) (47.13) (67.27) lowest yield of dry matter at 50% flowering, growth rate of the Genotype 975.80 799 766.6 plant, leaf area and the leaf area index. Weed- free × cartage mean (87.66) (73.50) (63.34) gave the highest growth rate. Flama × weed-free gave the CD (p=0.05) Genotype Pesticide Interaction control highest dry matter yield at 50% flowering, leaf area, leaf area 12.31 (3.70) 4.60 (8.10) 49.62 (18.30) index, the highest seed in the disk and the seed yield. *Seed yield (g plant-1) in parentheses REFERENCES increase in the leaf area per plant-1, by reducing the rate of Al-Chalabi FT 1988. Biological interactions between growth regulating substances and herbicides in weed control. Ph.D. weed, which reduces water consumption, increases the Thesis, University College of North Wales, UK. relative humidity, increases the efficiency of photosynthesis Al-Janabi MA and Younis AQA 1996. Introduction to Field Crop and increases the number of seedsdisk-1 (Nierves and Delos Production. Book House for Printing and Publishing-Mosul University, pp. 133. 1999). The results also showed that the interaction between El-sahooki MM and El-dabas EE 1982. One leaf dimension to the factors significantly affected the number of seeds in the estimate leaf area in sunflower. Journal of Agronomy and Crop disk, Flama * weed-free gave the highest rate of 1099 seeds 15: 199-204. disk-1, without significant difference with the other. Khan MS, Khalil LH and Swati MS 2004. Heterosisfor yield components in sunflower (Helianthus annus L). Asian Journal -1 Seed yield (g plant ): The genotypes significantly affected of Plant Science 3: 207-210. on seed yield, Flama genotypes gave the highest seed yield Munene JT, Kinyamari JL, Holst N and Mworia JK 2008. Compete it (100.8 g plant-1), superior to the other, while the genotype on between cultivated rice (Oryza satival. and wild rice (Oryza punctate l.) in Kenya. African Journal 3(9): 605-611. cartage gave the lowest seed yield (Table 3). The seed yield Mussavi SHK, Alamisacid G, Fathi M,Gharineh H, Morqditel A and was significantly higher in weedicides treated and weed free Siahpoosh A 2009. Optimum rice densiand herbicide application in direct seeding in ahwaz regionfo Iran. Asian than in control. The weed-free treatment gave the highest Journal of Crop Science 1: 58-62. -1 seed yield (100.8 g plant ), with a significant difference with Nierves JCD and Delos AS 1999. Early post-emergencies combined the rest of the treatments. Weedy (check) gave the lowest herbicides and development. Center Builetrin 9(2): 9-14. seed yield (47.13 g plant-1) due to the increase in the leaf area Putnam DH, OPlinger ES, Hicks DR, Dargan BR, Notzel DM, Notzel R, Meronuck A, Dol JD and Schalte EE 2008. Sunflower -1 plant , decrease the rate of presence of weed, which leads to alternative field crops manual. Last up dated. THumar 27(10): reduced water consumption, increase in the soil moisture, 5-10. the increase in photosynthesis efficiency, thus increasing the Safar NH 1990. Oil and sugar crops. Ministry of Higher Education. 450p. area from which the plant derives its food, increase the Steel, RGD and Torrie JH 1960. Principle and Procedure of Statistic. amount of light reaching the plant (Munene et at 2008). The MGGraw-Hill Book Co. Inc., New York, Toronto, London. 312p.

Received 22 September, 2019; Accepted 20 November, 2019 Manuscript Number: S-260 Indian Journal of Ecology (2019) 46 Special Issue (8): 93-96 NAAS Rating: 4.96

Effect of Biofertilizer on Growth and Yield of Wheat Triticum aestivium L

Iktifaa N. Jassim, Ali A.J. Al-Haidary1 and Sophia J. Jassim1 Directorate of Education Dhi-Qar, Iraq 1Agriculture College, Al-Muthanna University, Samawa, Iraq E-mail: [email protected]

Abstract: Laboratory experiments were carried out to isolate and diagnose hydrocarbons decomposer bacteria, from three oil regions (Basra, Dhi-Qar and Muthanna) as bio-fertilizer. The bacterial isolates were diagnosed, based on morphological, microscopic and biochemical properties, six bacterial isolates were found effective in hydrocarbon analysis. The bacterial isolates Citrobacter spp. and Proteus spp. were used, for their superiority to other isolates in the process of hydrocarbon analysis. It was added to the soil individually or double treated with three levels of crude oil (0, 1 and 2%). The bacterial supplementation was superior in plant height, dry weight of total vegetative/grain weight and grains number in spike consecutively were compared to control. The control and all traits reduced when the oil concentration increased significantly.

Keywords: Biofertilizer, Bacteria, Hydrocarbons, Growth, Yield, Wheat Triticum aestivium L, Oil

Environmental concerns are now rising, due to oil spill, MATERIAL AND METHODS as well as chemical waste that is released to the environment, Isolation and diagnosishydrocarbon-decomposing attempts are made to develop techniques to remove oil and bacteria: Soil samples were collected from refineries other pollutants, used many microorganisms capable of (Shuaiba in Basra, DhiQar and Muthanna and were diluted micro-biological decomposition (Dvorak et al 2017). with sterile water. These samples were planted on the brain Biodegradation has emerged as one of the technological and heart infusion broth, to activate bacteria and isolate more economic solutions for recovering oil spilled into the species likely to be present, incubated for 24 hours at 37°C, environment, in areas near the oil fields, the effect on the soil the activated specimens were then cultured on the brain depends on size and type of spill oil, as well as the amount of heart infusion broth on nutrient agar and MacConkey agar, by coverage of the soil and the season in which the leak streaking planning way, then incubated for 24 hours at 37°C, occurred, the toxic effects of crude oil hydrocarbons extend to secondary colonization of developing colonies was carried soil plants and microorganisms, all microorganisms stimulate out on selective media. Each bacteria group had a special metabolic reactions and generally contribute to biological growth media, it had phenotypic qualities that distinguish it activity, each organism had a specific performance in nature from others, microscopic tests and a series of biochemical (Marco-Urrea et al 2015). The level of biodegradation analysis were conducted. depends on the physiological state of microorganisms, the Investigation of the ability of bacterial isolates to analyze most biodegradable microorganisms were fungi and bacteria hydrocarbons: The mineral salts (0.5 g NH4Cl, 4 g NaCl, 0.5 compared to other species, had special features that make it g KH2PO4, 0.5 g MgSO4, 1 g Na2HPO4), were dissolved in 1 more used than others as a biological treatment for oil liter of distilled water and pH was adjusted at 7. The 50 pollution, bacteria differ in their ability to use crude oil, milliliters of mineral solution was distributed in 250 ml conical included various hydrocarbons in its chemical composition as flasks, the media was sterilized and cooled, added 1% crude a source of carbon and energy, and oxidation of petroleum oil to the studied isolates, incubated at 30°C for 42 days, it hydrocarbons (Bao et al 2014). Recent studies confirm that was followed up weekly to observe the decomposition of the microbial receptor can be broken down with many harmful crude oil. compounds, the conversion of hazardous pollutants into Bacterial vaccine preparation to biofertilizers used: Two usable particles is successful, to be used by increasing its isolates of Cytrobacter spp. and Proteus spp. have been stabilization by a biocell or a stimulating cell, which is an selected for use in the field experiment. 250 ml conical flasks important point for the development of modern technologies containing 100 ml of liquid, sterilized with an antiseptic by in bioremediation, The study aimed to investigate the effect of adding (1) ml of the processed liquid culture, sterile pipettes single or double oil-analytic bacteria on the growth of wheat used, incubated at a temperature of 30oC for (2-3) days, after grown in different levels of crude oil. prepared the liquid vaccine and mixed with the carrier (Peat 94 Iktifaa N. Jassim, Ali A.J. Al-Haidary and Sophia J. Jassim moss), wheat grains of IBA99 were contaminated with the first week, increased emulsification and clear turbidity, pollinated peat moss, added 10% Frankincense, an hour and indicates the presence of hydrocarbon decomposing a half leaved, a number of seeds were laid without pollination bacteria in soils contaminated with crude oil (Roling et al (treatments without vaccine). 2002). Golafarin et al (2017) demonstrated the ability of Agricultural experience (Anvils): The soil was air dried, bacteria isolated from contaminated areas to degrade milled and sifted from a 4 mm diameter sieve. The soil was hydrocarbons. Results of the trial were shown after 42 days then autoclaved at 121°C, pressure 15 lb Inch-2, left to cool with weekly follow-up, observe the decomposition of the eye, and contaminated with crude oil, 0, 5 and 10 ml and were inability of microorganisms from unpolluted areas to degrade placed in 5 kg plastic anvils after sterilization with ethyl hydrocarbons, taken from the Muthanna oil refinery, No alcohol. The anvils were planted with sterile wheat seeds, emulsification or turbidity occurred in the media. inoculated with elected bacterial isolates, according to the Biological Parameters treatments and the rate of 15 seeds/anvil. Then it was Wheat Plant height (cm): Table 2 shows that the bacterial reduced to 10 plants after a week of plantation, leaved a vaccine resulted in a significant increase height pollinated portion of the seeds without a vaccine as a comparison and plants with local isolates (Citrobacter spp. and Proteus spp.) without the addition of crude oil. The fertilizer was added as compared to no vaccine. A4 gave the highest rate of plant according to fertilizer recommendation, nitrogen fertilizers height of 39.83 cm (Table 2).This may be due to the ability of (urea 46% N) was applied twice, the first at the beginning of these organisms in the case of synergy to increase the planting and the second in the stage of growth, Phosphate readiness of phosphorus through the production of organic fertilizer in the form of triple superphosphate fertilizer (21% P) acids solvent phosphate, which increases the readiness of in one batch of three days before planting, Potassium phosphorus in the soil, as well as effectiveness in producing fertilizer was added potassium sulphate, 41.5% K (Ali et al growth regulators, such as in dole, acetic acid and gibberellin 2014). that affect cell elongation and division, therefore, protein formation is increasing and this is positively reflected in plant RESULTS AND DISCUSSION height (Hasan 2002,Jat and Shaktwat 2003). Biochemical Tests The uneven effect of crude oil levels added, resulted in

Bacterial isolates and hydrocarbons analysis: The significant differences (P≤0.05), the oil level (O1) exceeded results of the periodic inspection of isolates from the levels (O2 and O3), giving the highest value of plant height contaminated areas for 42 days showed the isolation ability of 49.33cm and an increase of (49.85 and 91.16%) at the of hydrolysis and fracture of hydrocarbons at the beginning of levels O1 and O2, respectively, this confirms the negative

Table 1. Biochemical test results of bacterial isolates for petroleum hydrocarbons Isolate Escherichia (1) Pseudomonas (1) Escherichia (2) Citrobacter Pseudomonas (2) Proteus Test Citrate utilization test _ *+ _ + + _ Catalase test + + + - + - Oxidase test _ + _ + + + Indole test + _ + *+ _ _ Vo.Pro.test _ _ _ + _ _ Gelatin hydrolysis test _ + _ _ + + Motility test + + + + + + Nitrate reduction + + + + + + Methyl red test + _ + _ _ + Starch hydrolysis + _ + + _ + Gas + _ + + _ + TSI

H2S _ _ _ + _ + Urease _ + _ + + _ Gram staining ______(+) Positive interaction of the test (-) Negative reaction for test (*) Weak reaction of the test Effect of Biofertilizer on Growth of Wheat 95

impact of crude oil on plant growth. The second oil level O1 Table 3. Effect of bacterial isolates and oil levels weight of wheat grains (g grain-1)* exceeded the third level O2, the rate gave 32.92cm and an Bacteria Oil levels Mean increase of 27.78% of the third level of oil (O2). The decrease in plant height may be due to increased concentration of O1 O2 O3 crude oil in the soil, inability of the roots to absorb oil A1 1.550 (22.97) 1.140 (9.77) 1.097 (8.10) 1.262 (13.61) compounds, because they are hydrophobic compounds A2 1.747 (30.20) 1.210 (12.87) 1.180 (10.07) 1.379 (17.71) unlike the nature of hydrophilicroots, reduces oxygen from A3 1.800 (30.30) 1.247 (11.93) 0.893 (5.30) 1.313 (15.84) the roots, affecting cell viability and cell division (Vavrek and A4 1.747 (33.63) 1.277 (14.10) 1.257 (11.87) 1.427 (19.87) Camptell 2002). Mean 1.711 (29.28) 1.219 (12.17) 1.107 (8.84) The interaction between bacterial vaccine and oil levels, LSD (0.05) Bacteria Oil levels Interaction was significant A4 (Citrobacter spp. and Proteus spp.) at oil 0.119 (3.28) 0.103 (2.84) 0.206 (5.68) level O gave the height of the plant at 53.50cm, while -1 0 *Number of grains spike isolation A0 gave the lowest mean height at the level of oil O2 (23.33cm). added in a bacterial vaccine, which is important in increasing -1 Dry weight of total vegetative (g plant ): The increase in the growth and, by increased cell division and elongation, crude oil levels led to the reduction of the wheat crop expanding of the root hairs which increases the surface area biological production (Table 2). The O0 oil level gave the of the roots, so nutrient absorption was greater, reflected -1 highest rate of 12.35 g plant , with an increase of 36.77% positively on the increase of various indicators of vegetative from the oil level O1 and 55.35% of the oil level O2, which growth, dry matter composition, increased grain fullness and recorded the lowest average weight of dry wheat plant was high weight (Conejos et al 2009). In addition, it was involved -1 7.95 g plant . The decrease in dry weight by increasing the in the decomposition of the soil, the dismantling of the added concentration of crude oil in the soil is attributed to the effect hydrocarbons, which results in an increase in the nutrient of aromatic compounds involved in the composition of crude supplied to the plant (Das and Kumar 2016). oil which reduce growth and thus negatively affect the dry The increase in oil levels led to a decrease in the weight weight of the plant. of wheat grains, the first oil level (O0) gave the highest The interaction between bacterial vaccine and oil levels, average weight in the grain (1.711 g grain-1), with an increase the isolation treatment of A4 vaccine at oil level O0 gave the of (40.36%) of the oil level (1%), and an increase (49.95%) of 1 highest average dry weight of 13.97 g plant- , while the the oil level (2%), the reason for the low grain weight by isolation of A4 gave the lowest average dry weight at the oil increasing oil levels may be due to the type of crude oil -1 level of O1 (7.71 g plant ). It was attributed to competition components of the Muthanna refinery because it contains between nutrients between plants and bacteria, each of hydrocarbons vary in the speed of disintegration depending which requires similar sources of nitrogen, potassium, on the chemical composition. phosphorus, water, etc. (Vavrek and Camptell 2001). The results indicated the significant effect of interaction Weight of wheat grains (g grain-1): The double bacterial between bacterial vaccine and oil levels. The treatment of A3 vaccine had a significant effect on wheat grain compared at the oil level O0 recorded the highest average of 1000 grains -1 control (A1), A4 (Citrobacter spp. and Proteus spp.) recorded weight of 1.800 g grain , there were no significant differences weight of 1.427 g grain-1 (Table 3). This was due to bacteria with bacterial vaccine A2 and A4 at the oil level O0, A3 at the oil -1 level O2, the lowest average was 0.893 g grain . Table 2. Effect of bacterial isolates and oil levels on wheat Temperature affects the physical state of hydrocarbons. In plant height (cm) cold conditions, liquid hydrocarbons become solid waxy, the Bacteria Oil levels Mean dissolved hydrocarbons precipitate and their solubility

O1 O2 O3 decreases significantly, changes in the physical state reduce

A1 40.33 (12.58) 31.00 (8.10) 23.33 (8.06) 31.55 (9.58) bio-readiness, that temperature has an effect on the effectiveness of bio-emulsions (Ilori et al 2005, Inakollu et al A2 52.00 (13.55) 31.67 (8.06) 26.33 (8.24) 36.67 (9.95) 2004). Feitkenhauer et al (2003) observed that the rate of A3 51.50 (9.30) 32.00 (8.67) 24.33 (7.78) 35.94 (8.58) biodegradation is very slow at low temperatures, it may be A4 53.50 (13.97) 37.00 (11.29) 29.00 (7.71) 39.83 (10.99) due to decreased enzymatic activity, increases as Mean 49.33 (12.35) 32.92 (9.03) 25.75 (7.95) temperature increase, their elevation promotes hydrocarbon LSD (0.05) Bacteria Oil levels Interaction 3.65 (1.425) 3.16 (1.234) 6.32 (2.468) metabolic rates, it was found that the dismantling of *Wheat biological production (g plant-1) naphthalene increases ten times when the temperature 96 Iktifaa N. Jassim, Ali A.J. Al-Haidary and Sophia J. Jassim

increases from 20 to 35°C, which makes a buffer between Collee JG, Fraser AG, Marmion BP and Simmons A 1996. Mackie and McCartney practical medical microbiology.14th ed. water and soil minutes, which leads to lack of water in the root Churchill. Livingston.USA. zone, which adversely affects the growth of the plant. Cornejo HAC, Lourdes MR, Carlos CP and Jose LB 2009. Number of grains spike-1: The effect of bio fertilizer Trichoderma virens, a plant beneficial fungus, enhances -1 biomass production and promotes lateral root growth through application on the number of grains spike , the treatments of an auxin-dependentmechanism in rabidopsis 1. American

A2 and A4 showed a significant increase (P≤0.05) in the Society of Plant Biologists. Plant Physiology 149: 1579-1592. Das AJ and Kumar R 2016. Bioremediation of petroleum number of grains of spike compared to A1, A4 gave the highest -1 contaminated soil to combat toxicity on Withania somnifera number of pills per spike at 19.87 grains spike , followed by through seed priming with biosurfactant producing plant growth -1 promoting rhizobacteria. Journal of Environmental A2 treatment at a rate of 17.71 grains spike , while the Management 174: 79-86. comparison treatment recorded the lowest rate of 13.61 Dvorak P, Nikel PI, Damborsky J and de Lorenzo V 2017. -1 grains spike . This may be due to the increased activity of Bioremediation biology. Doi:10.1016/j.biotechadv.2017.08.001. microorganisms in the Rhizosphere, which increases the Feitkenhauer H, Muller R and Markl H 2003. Degradation of readiness of some nutrients in the form of insoluble polycyclic aromatic hydrocarbons and long chain alkanes at 60- 70°C by Thermus and Bacillus spp. Biodegradation 14: 367-372. compounds, especially phosphorus compounds (Tahir Golafarin C, Abbas A, Mohammad M and Mohmmad D 2017. 2013). The effect of oil level on the number of grains in spike, Bioremediation capability and characterization of bacteria isolated from petroleum contaminated soils in Iran. Sustainable the treatment of the first level O1 exceeded the second level Environment Research 27: 195-202.

O2 and the second level O3, with an increase of 140.59% and Hasan HA 2002. Gibberellin and auxinindole production by plant root 231.22%. The drop in the spike rate of grain in the increase in fungi and their biosynthesis under salinity calcium interaction. Acta Microbiologica et Immunologica Hungarica 49(1):105-118. the concentration of crude oil in the soil may be attributed to Hwaback K, Sikkim H and Mockoh H 2004. Effects of crude oil the chemical nature of crude oil being a complex mixture of components and bioremediation on plant growth. Journal of hydrocarbons with different levels of inorganic sulfur, water Environment Science and Health 39(9): 2465-2472. Ilori MO, Amobi CJ and Odocha AC 2005. Factors effecting and salts. biosurfactant production by oil degrading Aeromonass pp. The interaction between the type of bacterial vaccine isolated from tropical environment. Chemosphere 61: 985-992. and the level of oil indicates significant differences (P≤0.05) Inakollu S, Hung HC and Shreve GS 2004. Bio surfactant enhancement of microbial degradation of various structural in the number of grains in spike, the results showed that the classes of hydrocarbon in mixed water systems. Environment Engineering Science 21: 463-469. interaction treatment between A4 and oil level O1 recorded a rate of 33.63 grains spike-1, followed by interaction treatments Jat BL and Shaktwat MS 2003. Effect of residual phosphorus, sulphur and biofertilizers on productivity, economics and A3 and A2 treatment at oil level O1 at rates of 30.30 tablets nutrient content of pearl millet (Pennisetumglacum L.) in spike-1 and 30.20 grains spike-1, while the interaction between fenugreek (Trigonella foenum-gracceum L.) pear millet cropping sequence. Indian Journal of Agricultural Science A3 and the level of oil O3 recorded the lowest rate of 5.30 73(3): 134-137. -1 grains spike . Macfaddin JF 2000. Biochemical tests for identification of Medical bacteria. Third edition. Williams and willkins company. U.S.A. Marco-Urrea E, Garcia-Romera I and Aranda E 2015. Potential of non- CONCLUSIONS ligninolytic fungi in bioremediation of chlorinated and polycyclic Isolates taken from soil contaminated with hydrocarbons aromatic hydrocarbons. New Biotechnology 32: 620-628. for three different regions (Basra, Dhi-Qar and Muthanna), Nair DR and Schnoor JL1993. Effect of soil condition on model parameters and Atrazine mineralization rates. Water Research important role in the decomposition and fracturing of 28(14): 1199-1205. hydrocarbons, as well as significant improvement in the Roling WFM, Milner MG, Jones DM, Lee K, Daniel F, Swannell RJP biological traits of wheat plant, such as plant height, dry and Head IM 2002. Robusthydrocarbon degradation and dynamics of bacterial communities during nutrient-enhanced oil weight of total vegetative, grain weight and number of grains spill bioremediation applied and environmental microbiology. per spike. American Society for Microbiology 68: 5537-5548. Schnoor JI 1996. Environmental modeling-fate and transport of pollution in water and soil. John Wiley and Sons. NewYork, REFERENCES pp.682. Ali NS, Salman H and Abdul-Razzaq AW 2014. Soil Fertility, Tahir M and Sarwar MA 2013. A Budding complement of synthetic Scientific Books Publishing House. First Edition. Baghdad fertilizers for improving crop production. Pakistan Journal of Life University. PP 423. Science 11(1): 1-7. Atlas RM 2010. Handbook of Microbiological Media, (4thed). CRC Vavrek MC and Camptell WJ 2001. The role of plant –bacterial Press, Taylor & Francis group, Washington, D.C. –fungal interaction in remediation of Oak-Hickory-Pine- Bao M, Pi Y, Wang L, Sun P, LiaY and Cao L 2014. Lipopeptide systems. Baton Rouge. Louisiana. biosurfactant production bacteria Acinetobacter sp. D3-2 and its Vavrek MC and Camptell WJ 2002. Identification of plant traits that biodegradation of crude oil. Environmental Science Processes enhance.http://ipec.utulsa.edu/Ipec/conf/2002/vavrek_campb Impacts 1: 1-7. ell_20.pdf.

Received 14 September, 2019; Accepted 04 November, 2019 Manuscript Number: S-261 Indian Journal of Ecology (2019) 46 Special Issue (8): 97-101 NAAS Rating: 4.96

Effect of Nano Biological and Mineral Fertilizers on Growth and Yield of Wheat (Triticum aestivum L.)

Basim K. Hasan and Turki M. Saad1 College of Agriculture, University of Al-Muthanna, Samawa, Iraq 1College of Agriculture and Marshes, University of Thi–Qar, Iraq E-mail: [email protected]

Abstract: A field experiment was carried out in Al-Gharraf area of Thi–Qargovernorate during the agricultural season winter (2018-2019) insplit- plot arrangement and RCBD design with three replicationses., The main factor included mineral fertilization treatments were 0, 50 and 100% fertilizer recommendation. The secondary factor included bio-fertilization levels 0, 1.5 and 3kg ha-1.The results showed that the treatment of mineral fertilizer exceeded the full recommendation of 50% in all traits, except 1000 grains weight, The results also indicated the superiority of treatment of nanofertilizer (without biofertilizer) in leaf area, spikes number, grains number, 1000 grains weight and grain yield, while 1.5kg ha-1biofertilizer outperformed in plant height and biological yield.

Keywords: Nanoparticles, Nano biological fertilizers, Growth, Yield, Wheat

The high population growth demand for food resources, MATERIAL AND METHODS make agricultural development more economical and Agricultural operations: The field experiment was efficient, as well as modern trends in environmental safety conducted in Thi–Qargovernorate, Al-Gharraf area (30 km from pollution, modern techniques are used to reduce north of Al-Nasiriyah city center) during the agricultural excessive use of chemical fertilizers, reduce nutrient losses season 2018-2019, silty clay texture soil, of 1500 m2 area. in fertilization and increase yield (Ram et al 2017). The land of experiment plowed perpendicular tillage with Sustainable agriculture has focused on reducing the use of mulching plow, the harrowing and leveling process was then common chemical fertilizers, the adoption of new fertilizers carried out, the field was divided into plots, 4m2 dimensions that does not have a negative impact on the environment and (2m * 2m), leaving a separation distance of 3 meters width human health (Parizad et al 2017). The application of bio- between the replicate, 1 meter distance between the fertilization is one of the modern methods that reduce the experimental units within the replicate, 2 meters distance excessive use of nitrogen fertilizers and increase the between the main pieces within the replicate to prevent availability of phosphorus, potassium and other elements (Al- overlap of the experiment factors. Wheat seeds (Bohoth 22) Khafaji 2018). Nanotechnology is one of the important and were planted by 120 kg ha-1 in the lines form, 8 lines per proposed solutions to increase agricultural production, more experimental unit, the distance between the lines is 20 cm, food can be produced at lower costs while reducing energy the necessary irrigation waterways were opened according consumption, environmental pollution, however, many to the need of the plant, taking into account rainfall periods researches have detected toxicity conditions caused by and climatic conditions. At the final ripening stage, the plant nanoparticles of plants ranged This is not about technology was harvested after five months. but because of the materials properties (Al-Ramadi et al Nitrogen fertilizer was added in the amount of 150 kg ha-1 2016). Nanotechnology offers ample scope for new (46% urea fertilizer form) in two batches, the first in the applications in biotechnology, agriculture and fertilizer branching stage and the second in the flowering stage, P2O5 industry, nanoparticles possess special physical and phosphate fertilizer was added in the amount of 80 kg ha-1 chemical properties due to their high surface area, alternative (triple super phosphate fertilizer P2O5% 44 form) at once natural resources can be used to make nanofertilizers to before planting, potassium fertilizer was added 100 kg ha-1 reduce environmental pollution and provide safety and health (potassium sulphate fertilizer 41.5% K form) at once before for humans, as well as the economic costs reduced from the planting, bio fertilizer containing three types of bacteria chemical fertilizers use (Mahmoud et al 2017). The present loaded on nanoparticles (Azospirillium, Pseudomonas and study aimed to evaluate the response of wheat to of Bacillus), it was added at once before planting by dissolving biofertilizers using nanotechnology and their interaction with the required quantity in 100 liters of water and distributing it to levels of mineral fertilization (NPK). the experimental units. 98 Basim K. Hasan and Turki M. Saad

Experiment factors: Mineral fertilization (NPK) treatments Table 1. Effect of nano biological and mineral fertilizer and with three levels (0, 50 and 100% of the fertilizer interaction on wheat plant height (cm)* Nano biological B B B Mean recommendation with took the symbols M , M and M , 0 1 2 0 1 2 mineral respectively). Bio-fertilization treatments with three levels (0, -1 M 79.56 81.43 81.53 80.84 1.5 and 3 kg ha and symbols of B , B and B , respectively. 0 0 1 2 (38.57) (41.45) (40.23) (40.08) Statistical analysis: The data were statistically analyzed M1 83.28 88.17 89.21 86.86 using a split – plot arrangement in Randomized complete (43.38) (48.69) (46.03) (46.03) block design with three replications, the GenStat Twelfth M2 83.64 88.86 88.99 87.16 Edition were used, the averages were compared to (43.58) (48.99) (46.37) (46.31) coefficients using the least significant difference (LSD) at a Mean 82.16 (46.37) 86.55 probability level of 0.05. (41.84) 86.15 (44.21) LSD (0.05) B = 1.203 (0.923) M = 2.372 (1.759) B * M = 2.372 (NS) RESULTS AND DISCUSSION *Flag leaf area (cm) in parentheses Plant height: Significant differences (P≤0.05) among the mean treatments of mineral fertilizer addition, M2 was of various enzymes, stimulate root growth and cell division, superior and gave the best value for this trait, with no regulate osmotic potential, open and close stomata, significant difference from treatment M1, averaging 87.16 and increased vegetative growth rates which lead to an increase 86.86 cm, with an increase of 7.81% and 7.44% sequentially in the number and size of leaves (Valizadeh and Milic 2016, compared to M0 (Table 1). It may be attributed to the increased Bekele 2018). availability of nutrients in the soil solution and the plant The results showed that the addition of bio fertilizer absorption, especially necessary nitrogen in the construction manufactured according to nanotechnology had a significant 2 of chlorophyll, proteins, enzymes, hormones and amino effect(P≤0.05), B1 was significantly higher at 46.37 cm , with acids, enter the cell division, elongation, then increase the an increase of 10.82% compared to B0, due to the growth and height of the plant (Al-Samawi 2012), consistent nanomaterials used as a biofertilizer carrier, they can remain with Mandal et al (2015) noted the importance of nitrogen in in the soil for the longest time, These materials also stabilize increasing plant growth and height. The results also showed the soil and organic soil material, which increased the a significant effect (P≤0.05) of nanobiofertilizer, B2 and B1, vegetative growth of the plant, which in turn reflected on the which did not have a significant difference in plant height, flag leaf area (Ammar 2018), agreed with Kaviani and gave an average of 86.55 and 86.15 cm with an increase of Negahdar (2016) that biofertilizers loaded on nanoparticles

5.34 and 4.56%, respectively, compared to B0, due to the fact increased plant growth by increasing their lifespan in the soil, that the addition of nitrogen-fixing biofertilizers loaded on which allowed the plant to absorb nutrients longer and thus nanoparticles increased soil fertility and enhanced nutrient increase vegetative growth. efficiency by reducing loss, especially nitrogen and thus Spikes number m-2: Table 3 shows that a significant increasing its readiness in the soil solution, which positively differences(p≤0.05) between the fertilizer treatments for the reflected the increased growth and plant height. Di- addition of mineral fertilizer, M2 exceeded and gave the best interaction between mineral fertilization and nanobiofertilizer significant spikes number value, a significant difference from -2 had outperformed the M1B2, M2B2, M2B1 and M1B2 treatments M1, with averaged 461.6 and 458.2 spike m sequentially as compared to the control, which did not significantly differ, compared to control, due to the processing of the plant with

M1B2 gave the highest plant height value (89.12 cm). the essential nutrients NPK, which positively reflected on the Flag leaf area (cm2): The results of Table 1 shows the increase in the spikes number, which agreed with Gul et al superiority (P≤0.05) of fertilizer treatments for the addition of (2011) and Kubar et al (2019).The results in the same table mineral fertilizer, M2 treatment was superior, gave the best showed that the addition of nanobiofertilizer superiority value, followed by M1, with no significant difference,46.22 resulted in B1 and B2, which did not have a significant and 46.13 cm2 with an increase of 15.31% and 15.09%, difference in this characteristic of 430.9 and 427.6 spike m- respectively compared to the control, may be due to the 2compared to control. This can be attributed to the fact that physiological role of NPK mineral nutrients, which play an biofertilizers increase soil fertility, promote plant growth important role in increasing plant effectiveness, nitrogen was through dilution of inhibitory products, increase available of essential for photosynthesis and phosphorus is involved in essential nutrients, especially nitrogen and phosphorus, the formation, division of living cells and cell membranes. thereby increasing growth indicators, including the spikes Potassium is instrumental in plant growth through activation number, this is indicated by Shende et al (2017), which Nano Biological and Mineral Fertilizers on Growth of Wheat 99 showed that the bio fertilizer loaded on nanoparticles has Table 2. Effect of nano biological and mineral fertilizer and -2* achieved a noticeable increase in the overall growth rate of interaction on number of spikes m Nano biological B B B Mean the plant. There was also a significant effect (P≤0.05) of 0 1 2 mineral bilateral interaction between mineral fertilizer and M 262.7 342.2 338.2 314.4 nanobiofertilizer on plant spikes, M B gave the highest mean 0 2 1 (55.13) (55.84) (56.57) (55.85) -2 of 482.9 spike m , While M0B2 recorded the lowest mean of M1 437.3 467.6 469.8 458.2 this trait was 338.2 spike m-2with a decrease of 29.96%, may (70.19) (72.82) (71.07) (71.36) be the nanofertilizers used more efficient than conventional M2 427.1 482.9 474.7 461.6 (70.77) (73.31) (70.51) (71.53) fertilizers because they interfere with the plant and cause Mean 375.7 430.9 427.6 many morphological and physiological changes, depending (65.36) (67.33) (66.05) on chemical composition and size, they enhance the LSD (0.05) B = 7.19 (7.21) M = 6.60 (6.68) efficiency of nutrient use, reduce fertilizer losses by B * M = 11.25 (NS) -1 regulating nutrient release (Mahmoud et al 2017). * Number of grains spike in parentheses Number of grains spike-1: Table 2 showed a significant increase (p≤0.05) in the of mineral fertilization treatments (M2 and M1), a non-significant difference between them, by giving Table 3. Effect of nano biological and mineral fertilizer and the highest average number of spike grains-1, which reached interaction on 1000 grains weight (g)* -1 Nano biological B0 B1 B2 Mean 71.53 and 71.36 spike grains , with an increase of 28.07% mineral and 27.77%, respectively compared to the comparative M0 47.22 47.33 46.44 47.00 treatment, may be attributed to the availability of NPK (3.880) (4.476) (4.547) (4.301) nutrients, which stimulated the vital processes of M1 43.67 46.89 42.78 44.44 (5.120) (7.580) (7.133) (6.611) photosynthesis, the regulation of hormones, which M 42.67 43.56 44.00 43.41 contributed to increasing the percentage of grain hold on the 2 (5.507) (7.876) (7.382) (6.921) spikes, which positively reflected on the increase in the Mean 44.52 45.93 44.41 number of spike grains. The addition of bio-fertilizer loaded (4.836) (6.644) (6.354) on nanoparticles resulted in a significant increase in the spike LSD0.05 B = NS (0.340) M 1.374 (0.197) B*M = NS (0.501) grain number, plants fertilized with treatment B1 produced the *Grains yield (t ha-1) in parentheses highest average number of spike grians at 67.33 grain spike-1, an increase of 3.01%, This may be due to the contribution of biofertilizers to increasing the available and soil content of Table 4. Effect of nano biological and mineral fertilizer and essential nutrients such as nitrogen, phosphorus and interaction on biological yield (t ha-1) potassium, as well as its ability to secrete some of the Nano biological B0 B1 B2 Mean chemical compounds that stimulate growth hormones such mineral M 13.14 14.28 15.58 14.33 as cytokines, gibberellins and indole acetic acid, increased 0 M 17.29 22.32 23.75 21.12 growth indicators and thus increased accumulation of dry 1 matter (Mohamed et al 2019). M2 22.36 26.96 26.77 25.36 1000 grains weight (g): Table 3 indicated that a significant Mean 17.6 21.19 22.03 differences (P≤0.05) between the fertilizer treatments when LSD0.05 B 0.956 M 7.752 B * M = 7.650 adding mineral fertilizer, the comparative treatment (M0) outperformed 47.00 g, while the plants fertilized with fertilizer no significant effects in this characteristic. -1 treatments (M1 and M2), which did not differ significantly, Grains yield (t ha ): The treatment of the addition of mineral produced the lowest grain weight and averages 44.44 and fertilizer M2 significantly superior (p≤0.05) in the grain yield 43.41 g respectively, due to the inverse relationship between trait, it gave the highest grain yield with an average value the grains weight and spikelet number, Tables (3 and 4) (6.921 t ha-1), an increase of 60.91% compared to the shows a decrease in the spikes number and the grains comparison treatment, this is due to the availability and number, which led to the contribution of photosynthesis increased concentration of NPK nutrients in the plant (Al- products to increase the weight of grains (Al-Hassany, 2018), Tahir et al 2013).The treatments of biofertilizer loaded on agreed with Al-Taher (2005).The treatments of nanoparticles, treatment B1 was superior (p≤0.05), which nanobiofertilizer and the interaction between mineral gave an average of 6.644 tha-1, an increase over the fertilizer and the biofertilizer carried on nanoparticles showed comparison of 37.38%, inoculation with bio-fertilizers 100 Basim K. Hasan and Turki M. Saad increased the soil content of organic matter and nitrogen, biological yield when adding nanoparticle fertilizers with NPK improved the availability of nutrients as well as their compound fertilizer to the barley plant. contribution to increasing the absorption capacity of the roots of the nutrients, led to increased vegetative growth, reflected CONCLUSIONS in the increase and redistribution of photosynthesis products, Mineral fertilization with NPK nutrients contributed to the contributed to the increase in grain yield (Mirzaei et al 2010). provision of adequate quantities during the crop growth Mardalipour et al (2014) indicated an increase in the yield stages, which improved growth and yield of wheat plant. The when nanobiofertilizers were added to the wheat plant. The interaction mineral fertilizers with bio-fertilizer loaded on interaction between mineral fertilizer and nanobiofertilizer nanoparticles reduced the amount of mineral fertilizer added levels significantly increased (p≤0.05) the average grain to 50% of the fertilizer recommendation in most studied traits, -1 yield, M2B1 gave the best value amounting to 7.876 tha with a reduces excessive chemical fertilizers uses, soil pollution percentage increase of 102.98%. This is due to the and preserves the environment from pollution. superiority of the aforementioned treatment in the spikes number and the grains number (Tables 2), which came as a REFERENCES result of its superiority in the flag leaf area, supports the direct Abed YM, Abdul-Ratha HA and Hadown HA 2016. Effect of biofertillizer produced from local isolates of pseudomonas relationship between the grain yield, the spikes number and putida and pseudomonas fluorescens on some soil the grains number. characteristics and yield of wheat (Triticum aestivum L.). The Biological yield (t ha-1): Table 4 indicated that a significant Iraqi Journal of Agricultural Sciences 47(6): 1413-1422. Al-Hasany ARK 2018. Effect of foliar nutrition of proline and mixture differences (P≤0.05) between the treatments of mineral of nutrients on growth and yield of different varieties of broad bean (Vicia faba L.). Ph.D. Thesis, College of Agriculture, Al- fertilizer addition, M2 was superior, gave the best value of the Muthanna University. biological yield, a non-significant difference with M1, -1 Al-Khafaji MH 2018. Effect of rhizobia and mycorrhiza averaging 25.36 and 21.12 t ha , an increase of 76.97 and inoculants and concentrations of vitamin B-complex on the 47.38% compared to the comparison treatment. The reason growth and yield of (Vicia faba L.). Ph.D. Thesis, College of Agriculture, Al-Muthanna University. for the increase in the biological yield may be due to the Al-Margani AHF 2005. Effect of soil and spray application of NPK on absorption of nutrients that increased the rate of growth and yield of wheat (Triticum aestivum L.). M.Sc. Thesis, photosynthesis, which was positively reflected in the weight College of Agriculture, Baghdad University. AL-Ramadi HR, Neama AA, Tarek AA, Abdullah HA, Mohammed SA gain of dry matter. Lemraski et al (2017); Jamal and and Tarek AS 2016. Botany and Environmental Muhammad (2007) reported that the addition of mineral Nanotechnology. First Edition, Science and Technological nutrients achieved significant differences in the dry weight of Development Fund, IDF 26613. AL-Samawi HNK 2012. Field Evaluation of some plants water the vegetative total of rice and wheat respectively. The extracts on nitrification inhabitation, ammonia volatilization and barley growth (Hordeum vulgare L.). Ph.D. Thesis, College of results showed that the nanobiofertilizer treatments (B2 and Agriculture, Baghdad University. B ) significantly exceeded (p≤0.05) with B without significant 1 0 Al-Tahir FM 2005. Effect of foliar nutrition of iron, zinc and potassium difference between them, each of 22.03, 21.19 and 17.60 on growth, yield of wheat (Triticm aestivum L.). Ph.D. Thesis, t.ha-1, respectively. This is due to biofertilizers loaded on College of Agriculture, Baghdad University. nanoparticles in improving soil properties as well as Al-Tahir FM, Yahya KJ and Al-Badry SAM 2013. Effect of date and quantity of potassium fertilizer application on growth, yield and increasing an availability of nutrients that the plant benefits in oil quality of sunflower (Helianthus annuus L.) var. flame. IOSR biological processes and the formation of various tissues and Journal of Agriculture and Veterinary Science 4(1): 33-40. Ammar AS 2018. Nanotechnologies associated to floral resources in thus increase the vegetative total, as noted in Table 1.B2 and agri-food sector. Acta Agronomy 67(1): 146-159.

B1 treatments exceeded in the plant height, which was Bekele M 2018. Effects of different levels of potassium fertilization on positively reflected on the increase of biological yield, which yield, quality and storage life of onion (Allium cepa L.) at Jimma, Southwestern Ethiopia. Journal of Food Science and Nutrition agree with Mardalipour et al. (2014) and Mir et al (2015). 1(2): 32-39. They showed that the addition of biofertilizers contributed Burhan MG 2018. Effect of foliar application of nano NPK fertilizer on significantly to increasing the vegetative growth in the barley growth, yield and quality of bread wheat. Ph.D. Thesis, College of Agriculture, Baghdad University. maize crop respectively. There was a significant interaction Gul H, Said A, Saeed B, Mohammad F and Ahmad I 2011. Effect of (p≤0.05) between mineral fertilizer and nanobiofertilizer in foliar application of nitrogen, potassium and zincon wheat biological yield. M B , M B , M B and M B were given the growth. ARPN Journal of Agricultural and Biological Science 2 1 2 2 1 2 1 1 6(4): 56-58. -1 highest mean values of 26.96, 26.77, 23.75 and 22.32 t ha , Jamal Z and Chaudhary MF 2007. Effects of soil and foliar respectively, compared to control and other treatments, this application of different concentrations of NPK and foliar

application of (NH4)2SO4 on growth and yield attributes in wheat result is consistent with the indications Spruogis (2018), (Triticum aestivum L). Pakistan Journal of Plant Science 13(2): which obtained a significant increase in the trait of the 119-128. Nano Biological and Mineral Fertilizers on Growth of Wheat 101

Kaviani B and Negahd N 2016. Effects of biological nanofertilizer on Biosciences 6(4): 157-164. the morphological, physiological and proliferation traits and Mirzaei A, Vazan S and Naseri R 2010. Response of yield quality of Buxus Hyrcana Pojark. Bangladesh Journal of components of safflower (Carthamus tinctorius L.) to seed Botany 45(5): 1135-1142. inoculation with Azotobacter and Azosprilium and different Kubar GM, Talpur KH, Kandhro MN, Khashkhali S, Nizamani MM, nitrogen levels under dry land condition. World Applied Science Kubar MS, Kubar KA and Kubar AA 2019. Effect of potassium Journal 11(10): 1287-1291. (K+) on growth, yield components and macronutrient Mohamed F, Manal A, Thalooth T, Tarek AE and Amal GA 2019. Yield accumulation in wheat crop. Pure and Applied Biology 8(1): and nutrient status of wheat plants (Triticum aestivum L.) as 248-255. affected by sludge, compost, and biofertilizers under newly Lemraski MG, Normohamadi G, Madani H, Sharif Abad HH and reclaimed soil. Bulletin of the National Research Centre 43: 31- Mobasser HR 2017. Two Iranian rice cultivars' response to 36. nitrogen and nano-fertilizer. Open Journal of Ecology 7: 591- Parizad M, Yarnia1 M, Amirnia R, Tarinejad A and Mahmoodi H 2017. 603. Comparison of the effect of nanourea and nanoironfertilizers Mahmoud AWM, El-Attar AB and Mahmoud AA 2017. Economic with common chemical fertilizers on some growth traits and evaluation of nano and organic fertilisers as an alternative essential oil production of (Borago officinalis L.). Dairy and Vet source to chemical fertilizers on (Carum carvi L.) plant yield and Science Journal 2(2):93-97. components. Agriculture (Poľnohospodárstvo) 63(1): 33-49. Ram P, Bhattacharyya A and Nguyen QD 2017. Nanotechnology in Mandal MSH, Ali MH, Amin AKMR, Masum SM and Mehraj H 2015. sustainable agriculture: Recent developments, challenges, and Influence of source of nitrogen on growth and yield of wheat. perspectives. Front in Microbiology 8: 1014-1021. International Journal of Agronomy and Agricultural Research Shende S, Rathod D, Gade A and Rai M 2017. Biogenic copper 6(1): 89-95. nanoparticles promote the growth of pigeon pea (Cajanus cajan Mardalipour M, Zahedi H and Sharghi Y 2014. Evaluation of L.). IET Nanobiotechnology 11(7): 773-781. nanobiofertilizer efficiency on agronomic traits of spring wheat Spruogis V 2018. The influence of bioorganic nanofertilizer on spring at different sowing date. Biological Forum – An International barley and oilseed rape productivity and economical Journal 6(2): 349-356. effectiveness. Žemės Ūkio Mokslai 25(1):18-26. Mir S, Sirousmehr A and Shirmohammadi E 2015. Effect of nano and Valizadeh M and Milic V 2016. The effects of balanced nutrient biological fertilizers on carbohydrate and chlorophyll content of managements and nano-fertilizers effects on crop production in forage sorghum (Speedfeed hybrid). International Journal of semi-arid areas. Current Opinion in Agriculture 5(1): 31-38.

Received 22 September, 2019; Accepted 09 November, 2019 Manuscript Number: S-262 Indian Journal of Ecology (2019) 46 Special Issue (8): 102-107 NAAS Rating: 4.96

Evaluation of Maize Hybrids, their Inbred Lines and Estimation of Genetic Divergence Based on Cluster Analysis

Wajeeha Abed Hassan, Banan Hassan Hadi* and Majid S.H. Hamdalla Crop Sciences Department, College of Agriculture Engineering Sciences, University of Baghdad, Baghdad, Iraq *E-mail: [email protected]

Abstract: Single hybrids were obtained by crossing five inbreeds of maize (ZM49W3E, ZM60, ZM43WIZE, ZM19, CDCN5) in half diallel matting during the spring season 2016. Ten F1s with their inbred lines were evaluated for thirteen characters. The single hybrids surpassed their parents for the most studied characters. The highest superiority for single hybrids was in grain yield and cob weight. The superiority of the single hybrids result in obtaining a high percentage of hybrid vigor, where ZM43WIZE x ZM19 and ZM43WIZE x CDCN5 had highest hybrid vigor in the growth characters, while ZM60x ZM19 possess the highest hybrid vigor in the grain yield and its components. This refers to a genetic diversity between inbred ZM19 and other inbreeds. The agglomerative analysis for grain yield was done. The single hybrids, who exceeded in grain yield, have gathered within the same group. The results showed the availability of genetic variety alleles between the studied inbreeds and the possibility of using this inbreeds as a source of alleles in the future breeding program for improving the local genotypes.

Keywords: Maize, The agglomerative analysis, Single hybrid, Genetic diversity

Maize one of the most important crops in the world as resemblance alleles of the studied genotypes. Cluster food for human, feed for animals and industrial uses (Akundi analysis is an efficient tool for estimating the genetic diversity et al 2006). It is ranked third amongst the cereal crops for among genotypes and assign QTL, in addition, it doesn't area and production and the average world production is need an assumption for data distribution (Yau and Ortize about 5.75 t ha-1 and but in Iraq is low, 1.85t ha-1in (FAO 2019). 1994). Cluster analysis distributes the studied genotypes into There is a high gap between local and world production and analogous groups based on their identical performance that because of the low yield potential of local genotypes. midst the environmental circumstances. The genetic Maize is a highly cross-pollinated crop, therefore hybrid vigor diversity is estimated in term of Euclidean distances which could be an efficacious tool for utilization the potential of describe the dissimilarity between the cases and the inbreeds. The extent for the exploitation of hybrid vigor will ndendrogram is drawn (Yan et al 2001). Zaman and Alam depend on the direction and amount of heterosis and also the (2013) classified thirty-nine crosses of maize into seven type of gene action. The first step in producing the hybrids is groups according to their performances by cluster analysis. the introduction of elite inbreeds, but the critical element of Anees et al (2017) explained that the distribution of maize getting heterosis is testing the diversity between inbreeds crosses into groups based on cluster analysis was attributed and availability favorable genes to improve the local to their heterosis. Hence, the present investigation was genotypes. carried out to evaluate the F1s and their parents under Iraqi Single hybrids were suggested for the first time by Jones conditions and estimate the hybrid vigor and diversity among in 1918 to utilize the heterosis which is defined as a the crosses. superiority of F1s over their parents. Many studies have been conducted to estimate hybrid vigor for several crops. MATERIAL AND METHODS Revealing of the combining ability for the studied genotypes Ten hybrids were generated from five inbreeds viz, can facilitate breeder work in developing new genotypes with ZM49W3E, ZM60, ZM43WIZE, ZM19 and CDCN5, which high potential. Four methods for diallel mating has been set are introduced from Yugoslavia and Italy. The crosses inbred up by Griffing (1956) in order to identify the optimum inbreeds lines were derived from crossing between the inbred lines and evaluate their performances by detecting the resulted according to half diallel matting during spring season 2016. heterosis. Wuhaib and Hadi (2016), Abdulla and Karim The F1 seeds of each cross were bulked together. Ten (2019) found out positive heterosis for the most studied crosses along with five parents were grown in randomized characters and negative heterosis for days to anthesis and complete block design with four replications at the research days to maturity. Classification of the crosses into identical station of Agriculture College/Al-Jadrya Baghdad university groups could be an adequate procedure to identify the during fall season, 2016. A plot dimensions 3*3m consisted of Evaluation of Maize Hybrids and their Inbred Lines 103 five rows , 70 cm between rows and 25 cm within rows, alleles in the offspring does not necessarily mean that their population density was 57143 plants ha-1. The recommended parents have favorite alleles. The highest percentage of package of practices was followed to raise a good crop. heterosis for growth characters occurred in the crosses Harvesting was performed after the grains reached maturity ZM43WIZE x ZM19 and ZM43WIZE x CDCN5, whereas and plants were dried, at which grain moisture was subsisted in the ZM60 x ZM19 cross for yield and its determined. Observations were recorded on ten randomly components. plants for different plant height, leaf area, leaves The hybrids surpassed their parents, where highest numberplant-1, ear height, ear length, number of rows ear-1, dominance was in grain yield and ear weight characters, number of kernels row-1, cob weight, 100-kernel weight and while the lowest superiority was in days to 50 per cent grain yield plant-1. The observations for the characters tasseling (DTT)and days to 50 per cent silking (DTS) (Table namely days to 50 per cent tasselling, days to 50 per cent 2). Heterosis in negative direction is desirable for DDT and silking, days to maturity were recorded on a plot basis. The DTS and indicates the possibilities for breeding of maize for statistical analysis was performed out using GENSTAT earliness. These results are in unison with the results of software. Computation of heterosis for all characters was heterosis (Table 3). Inbred CDCN5 exhibited superiority for carried out as per procedure suggested by Shull (Singh, most of the studied characters over others of inbreeds, while 2012). Cluster analysis was carried out for the hybrids inbred ZM60 showed feeble performance Exist elite alleles in according to agglomerative method (Senath and Soka, 1973, inbreeds don't mean producing excellent hybrids as long it Williams 1976). The dendrogram was performed based on depends on the degree of diversity amongst the parents and Un weight Pair Group Method with Arithmetic Mean the percentage of heterosis. Most of the superiority hybrids (UPGMA). The Euclidean distances were measured from the belong to the inbreed lines ZM43WIZE and ZM19 (Table 4) proximities matrix according to the following formula: and this is supported by the percentage of heterosis (Table Distance (x,y) = {∑i (xi - yi) 2 }½ (Punitha et al 2010) 3), nevertheless, the performance of inbred CDCN5 wasn't poor. The cross CDCN5 x ZM43WIZE has the superiority in RESULTS AND DISCUSSION growth characters while the cross ZM60 x ZM19 was the Single hybrids prevailed on their inbreed lines for all the better in grain yield and the related characters, but the studied characters except ear height (Table 1). These results difference between them didn't reach the significance limits. are in conformity with the findings of earlier inbred lines The crosses ZM49W3E x ZM43WIZE and ZM49W3E x workers (Wuhaib et al 2016, Mussarbat and Aldulaymy 2017, CDCN showed the lowest average of grain yield and this Abdel-Amir and Hadi 2018). reflects the weakness of inbred ZM49W3E who exhibited The results indicated the presence of genetic diversity depression in the grain yield components and this is between the studied inbreeds which reflected on heterosis confirming by the GCA value for this inbred. percentage (Table 3). The highest percentage of heterosis Cluster Analysis appeared for the crosses ZM60 xZM19 and ZM43WIZE The significant variations between the studied xZM19, in addition to the cross ZM43WIZE xCDCN5 and this genotypes (Table 1) are considered an indicator of the is an indicator about existing diverse alleles among the effectiveness of classification of the genotypes. The cluster parents of the previous crosses. The presence of diverse analysis has conducted for the hybrids according to the grain

Table 1. Analysis of variance for characters in single hybrids and their inbreed lines maize S.O.V. D.F Days to Day to Silking Plant height Leaf area Leaves number Ear height Ear length tasseling (cm) (cm2) plant-1 (cm) (cm)

Reps 3 1.31 0.99 310.69 0.0004 0.80 5.74 1.32 Genotypes 14 52.95** 45.95** 4721.14** 0.335** 17.05** 1382.83** 26.42** Error 42 1.30 1.44 216.84 0.0015 0.36 33.07 0.88

S.O.V. D.F Rows number Grains number Ear weight (gm) Cob weight 100 kernel Grain yield ear-1 row-1 (gm) weight (gm) plant-1 (gm)

Reps 3 0.84 18.8 182 49.4 14.4 17.5 Genotypes 14 5.25** 2355.1** **173251 113.5* 74.6** 1061.3** Error 42 0.45 354.7 1729 18.1 2.5 49 104 Wajeeha Abed Hassan, Banan Hassan Hadi and Majid S.H. Hamdalla

Table 2. Mean values of maize genotypes for the studied characters Genotypes Days to Day to silking Plant height Leaf area Leaves number Ear height Ear length (cm) tasseling (cm) (cm2) plant-1 (cm) 1 60.9 66.9 136.3 0.5 12.5 70.8 13.1 2 63.4 68.3 121 0.4 10.8 53.5 14.1 3 68.3 71.5 141.4 0.3 8.4 62.3 13.6 4 69.4 73.2 148.5 0.4 10.6 68.3 12.5 5 60.2 66.5 168.9 0.5 12.2 90.7 14 1x2 63.5 65.6 171.8 0.6 13 84.4 17.9 1x3 58.4 68 171.8 0.6 13.6 107.7 17.8 1x4 66.6 70.9 221.4 0.6 15.2 97.1 18.4 1x5 63.5 65.6 193.7 0.6 13.2 98.1 16.3 2x3 64.6 69 186.9 0.6 15.5 101.9 17.2 2x4 59.5 64.2 170.5 0.5 12.9 76.8 20.3 2x5 60.7 64.4 160 0.6 12.3 78.4 19.7 3x4 58.3 63.7 202.3 0.6 14.3 111.2 17.4 3x5 57.8 60 256.2 0.6 16.7 115.7 19.8 4x5 60.7 65.6 173.5 0.5 13 82.9 16.9 Mean 62.4 66.9 174.9 0.5 12.9 86.6 16.6 LSD 1.6 1.7 21 0.05 0.9 8.2 1.3 Meanof Parents 64.4 69.3 143.2 0.4 10.9 69.1 13.4 Mean of Crosses 61.3 65.7 190.8 0.6 14 59.4 18.2 Percentage of crosses -4% -5% 33% 50% 28% -14% 35% superiority

Genotypes Rows number Grains number Ear weight Cob weight (gm) 100 kernel weight Grain yield plant- ear-1 row-1 (gm) (gm) 1(gm)

1 13.1 24.3 87.2 17.1 24.1 70 2 12.5 21.2 75.3 16.6 20.2 58.6 3 13.8 28.9 98.4 16.9 20.1 81.4 4 13.2 27.8 74.9 14.2 17.4 60.7 5 15 33.2 103.6 16.2 17.6 87.3 1x2 14.8 37.1 169.1 15.6 23.5 153.5 1x3 15.8 37.6 154.7 23.5 23.5 131.1 1x4 15.4 39.7 210.5 22 31.7 188.8 1x5 15.8 35.6 149.7 24 22.1 125.7 2x3 15.1 35.4 193.9 28.6 25.3 165.2 2x4 16.2 42.1 233.8 27.1 30.2 206.7 2x5 15.8 40.9 197.2 23.9 25 173.5 3x4 15.6 39.6 196.4 24.7 24 171.6 3x5 14.9 39.4 226.5 27.7 29.8 198.7 4x5 13.8 40.7 177.6 30.1 26.6 147.5 Mean 14.7 34.9 156.6 21.9 24.1 134.7 LSD 0.96 4.1 9.1 6 2.2 9.9 Mean of Parents 13.5 27.1 87.9 16.2 19.9 71.6 Mean of Crosses 15.3 38.8 190.9 24.7 26.2 166.2 Percentage of crosses 13% 43% 117% 52% 31% 132% superiority Evaluation of Maize Hybrids and their Inbred Lines 105 yield and its components (Fig. 1). The hybrids distributed into two groups; first group included the crosses ZM49W3E x ZM60, ZM49W3E xZM43WIZE, ZM49W3E x CDCN5, ZM60 xZM43WIZE, ZM60 x CDCN5, ZM43WIZE x ZM19, ZM19 x CDCN5, while the second group included the crosses ZM49W3E x ZM19 ZM60 x ZM19 and ZM43WIZE x CDCN5.Second group consisted of the superior hybrids in the grain yield, while the first group covered the rest hybrids. The crosses ZM60 x CDCN5 and ZM43WIZE x ZM19one group due to lowest value of Euclidean coefficients which was 0.72 and 0.96 respectively. Next, the crosses ZM49W3E x ZM43WIZE and ZM49W3E x CDCN5 are gathered in the same group. The distribution of the genotypes in this study was based on the characteristics of the grain yield and its components, which means that this classification took into account the alleles of several characters, and if this classification was based on other characteristics, it would be possible to obtain Fig. 1. The relationship among the single hybrids based on another classification takes into consideration the new cluster analysis

Table 3. Heterosis of maize genotypes for the studied characters Genotypes Days to Days to silking Plant height Leaf area Leaves number Ear height (cm) Ear length (cm) tasseling (cm) (cm2) plant-1

1x2 -1.9 4.4 26 17.5 4.1 19.1 27.1 1x3 1.6 -4.1 21.5 18.5 9.4 52 31.1 1x4 6.1 9.4 49.1 20.1 21.7 37.1 40.4 1x5 -1.3 5.4 14.7 21.4 6.2 8.1 16.7 2x3 1 2 32.2 48.1 43.9 63.6 22.5 2x4 -6 -6.1 14.8 34 20.2 12.3 44.2 2x5 -3.1 0.8 -5.3 30.1 1.4 -13.6 40.3 3x4 -10.9 -14.5 36.2 57.4 34.5 62.7 28 3x5 -9.8 -4.1 51.7 35.6 36.8 27.5 41.1 4x5 -1.3 0.8 2.7 7.5 6.6 -8.5 20.7 Mean -2.56 -0.6 24.36 29.02 18.48 26.03 31.21 S.E. 1.6 2.2 5.9 4.8 4.9 8.7 3.1

Genotypes Rows number Grains number Ear weight (gm) Cob weight (gm) 100 kernel weight Grain yield plant-1 ear-1 row-1 (gm) (gm)

1x2 13.1 52.8 93.9 -9.1 -2.4 119.1 1x3 14.4 29.9 65.7 37.3 -2.6 61.1 1x4 16.4 42.8 141.3 28.5 31.6 169.6 1x5 5.3 7.5 44.5 39.8 -8.3 43.9 2x3 9.6 22.6 107.7 68.5 25.1 103 2x4 22.9 51.6 210.3 62.6 49.7 240.6 2x5 5.1 23.4 90.3 43.5 24 98.6 3x4 13.1 36.9 110.4 45.8 19.4 110.8 3x5 -0.9 18.9 118.5 63.3 48.1 127.4 4x5 -7.8 22.7 71.4 85 51.6 68.8 Mean 9.12 30.91 105.4 46.52 23.62 114.29 S.E. 2.8 4.7 14.6 8.2 7.1 18.1 106 Wajeeha Abed Hassan, Banan Hassan Hadi and Majid S.H. Hamdalla

Table 4. Mean characters for each inbreed lines Traits Inbreed lines

1 2 3 4 5

Day to tasseling 63 62.075 59.775 61.275 60.675 Days to silking 67.525 65.8 65.175 66.1 63.9 Plant height 189.675 172.3 204.3 191.925 195.85 Leaf area (cm2) 0.6 0.575 0.6 0.55 0.575 Leaves no. 13.75 13.425 15.025 13.85 13.8 Ear height (cm) 96.825 85.375 109.125 92 93.775 Ear length (cm) 17.6 18.775 18.05 18.25 18.175 Rows no. 15.45 15.475 15.35 15.25 15.075 Grain no. 37.5 38.875 38 40.525 39.15 Ear weight (cm) 171 198.5 192.875 204.575 187.75 Cob weight (gm) 21.275 23.8 26.125 25.975 26.425 100 kernel weight (gm) 25.2 26 25.65 28.125 25.875 Grain yield per plant (gm) 149.775 174.725 166.65 178.65 161.35

Table 5. Agglomeration schedule maize hybrids, the first is exist genetic divergence among the Stage Cluster Coefficients Stage cluster first Next parents, the second is the parents should own favorable combined appears stage alleles. The availability of these two elements would Cluster 1 Cluster 2 Cluster 1 Cluster 2 determinate the percentage of heterosis. Hence, the current 1 7 8 0.727 0 0 7 study suggests exist genetic divergence among the parents, 2 2 4 0.960 0 0 5 but there is no evidence about exist favorable alleles. 3 3 9 1.006 0 0 6 4 1 5 1.080 0 0 5 REFERENCES 5 1 2 1.855 4 2 7 Abdel-Amer AN and Hadi BH 2018. Evaluate the performance of double, single hybrids and inbreeds of maize under different 6 3 6 2.085 3 0 9 plant population and estimate heterosis and hybrid vigor (some agronomic traits). Anbar Journal of Agricultural Sciences 16(1): 7 1 7 2.272 5 1 8 817-835. 8 1 10 3.106 7 0 9 Abulla AH and Karim AA 2019. Evaluation of F1s, F2s hybrids, 9 1 3 3.223 8 6 0 hetrosis, andinbreeding depression of maize (Zea mays L.). Tikrit Journal of Agricultural Sciences 19(1): 1-17. Akunde SR, Lamid GO and Biotech J 2006. Performance of quality characters. Therefore, the critical factor that determines the protein maize varieties and disease reaction in the derived nature of the character (s) on which the cluster analysis savanna agro ecology of south-west Nigeria. African Journal of Biotechnology 5(19): 1744-1748. would be based, will depend on the goal of the breeder. If it is Anees AHA, AL-rawi WMH and AL-dawoodi SAM 2017. Evaluation to improve a specific character, it is preferable that the lines and their half dialle crosses for phenotypic characteristics classification is based on that attribute only. The goal of the by using cluster analysis of maize (Zea mays L.). Tikrit Journal for Agriculture Sciences 17(3): 33-49. classification is to define the percentage of similar alleles FAO 2019. http://www.fao.org/. among the genotypes, this would provide a consider view for Griffing BA 1956. Concept of general and specific combining ability in the breeder about the possibility of conducting crossing relation to diallel crossing systems.Australian Journal of Biology Sciences 9: 463-493. between the diverse genotypes (Hamdalla 2011). The Hamdalla MS 2011. Genetic diversity of sunflower based on cluster accuracy of the decision to be adopted by the breeder will analysis. Iraqi Journal of Agricultural Science 42(3): 17-23. depend on the efficiency of the assessment of genetic Jeffers JNR 1967. Two case studies in the application of principal divergence. In order for the evaluation to be efficient, it is component analysis. Applied Statistics 16: 225-236. preferable to re-test the field assessment for more than one Mussarbat NA and Aldulaymy HJ 2017. Estimation of heterosis, combining ability and expected genetic advance in maize by season and for more than one site. using half diallel cross. Anbar Journal of Agricultural Sciences 15(2): 408-417. CONCLUSION Punitha B, Vindhiyavarman P and Manivannan N 2010. Genetic divergence study in sunflower (Helianthus annuus L.). There are two important elements for producing elite Electronic Journal of Plant Breeding 1(4): 426-430. Evaluation of Maize Hybrids and their Inbred Lines 107

Shull GH 1910. Hybridization methods in corn breeding. American Agriculture Sciences 47(special issue): 44-55. Breeders Magazine 1: 98-107. Wuhaib KM, Hadi BH and Hassan WA 2016. Hybrid vigor, heterosis, Singh RK and Chaudary BD 2007. Biometrical Methods in and genetic parameters in maize by diallel cross analysis. Quantitative Genetic Analysis. Rev. Ed., Kalyani Publishers International Journal of Applied Agricultural Sciences 2(1): 1- Ludhiana, India, pp.318. 11. Singh BD 2012. Plant Breeding: Principles and Methods. Kalyani Zaman MA and Alam MA 2013. Genetic diversity in exotic maize (Zea Publisher, New Delhi. mays L.) hybrids. Bangladesh. Journal of Agriculture 4(5): 273- Sneath PHA and Sokai RR 1973. Numerical Taxonomy: The 280. Principal and Practice of Numerical Classification, W.H. Yan W, Hunt L, Sheng Q and Szlavnics Z 2000. Cultivar evaluation Freeman and Co., San Francisco. and mega-environment investigation based on the GGE biplot. Williams WT 1976. Pattern Analysis in Agricultural Sciences. Crop Sciences 40: 597-605. Elsevier, Amsterdam. Yau SK and Ortiz G 1994. Regression and analysis of elite wheat Wuhiab KM and Hadi BH 2016. Detection of non-allelic interactions lines grown in rain fed environment. Journal of Genetics and via generation mean analysis in maize. Iraqi Journal of Breeding 48: 183-190.

Received 20 October, 2019; Accepted 24 November, 2019 Manuscript Number: S-263 Indian Journal of Ecology (2019) 46 Special Issue (8): 108-112 NAAS Rating: 4.96

Evaluation of Seed Viability and Vigour of Maize coated with

Local GYPSUM (CASO4 1/2 H2O) and AG-NPS for Resistance of Fusarium verticillioidis

Saddam H. Cheyed, A. Ch. Al-Fahad1, Eman A. Al-Qaisy2 and Aalaa K. Hassan College of Agriculture Engineering Science, University of Baghdad, Iraq 1College of Agriculture, University of Anbar, Iraq 2Ministry of Education, Directorate General of Vocational Education, Second Vocational Section in Karkh E-mail: [email protected]

Abstract: A laboratory experiment consisted of coating maize seeds with local gypsum (lg) and Ag-NPs with the concentration (100, 200, 300 and 400 mg l-1), in contaminated soil with Fusarium verticillioidis was conducted. Results showed significant superiority of coated seeds in the last day of seedling emergence, emergence period, emergence speed and the percentage of field emergence. Non coated seeds were significantly superior in the first day of seedling emergence. Seeds treated with 400 mg l-1 of Ag-NPs were significantly superior in all characters, which included first and last day of seedling emergence and emergence speed. The combination of seeds coated with lg and Ag-NPS (400 mg l-1) was superior over other combinations and control in most studied characters. It can be concluded that using lg and Ag-NPs might protect seeds more against F. verticillioidis compared with single seed coated treatment.

Keywords: Seed-coating, Seedling death, Seed-emergence, Seeds diseases

Using of high quality seeds is considered one of the most Therefore, nanotechnology is getting better solution which important practice to increase production, and hence, significantly reduces the amounts of agrochemicals used for supporting the food security. Agricultural production depends seed protection which in turn protect the environment upon seed quality of the best variety, climate nature in the besides its effects of seeds protection compared with non- region and soil and crop management (Cheyed and nano agents Ag-NPs is considered the most used particles in Elsahookie 2011). High quality seeds coats and absence of the nanotechnology science in the field of foods, industry and insects and diseases (Halmer 2006), which contribute by medicine (Crocetti and Miller 2011, Safavi et al 2011). Ag- almost 30% of total international production. However, seeds NPs act as anti-microbial without harmful effect on the human during their processing subject to many biotic and abiotic tissues (Abdullah and Hamid 2013, Garg et al 2014). stresses which might affect their coats integration. Many Moreover, using seeds against fungi during germination and studies refereed to that fungi of Fusarium genus affect emergence with extra benefits in improving germination various crops among them maize with the species F. percentage and seed emergence, reducing the electrolyte verticillioidis considered the most dominant one in the maize leakages from seeds due to the increasing activity of seed and other crops. This species is one of the pathogenic fungi coat. Also it allow a good adhesion and best protection to the which infect the plant at the early stages (Cumagun et al 2009 compacted compounds with coating materials as fungicides and Choi and Xu 2010). This fungus causes many diseases and insecticides (Pereira et al 2005 and Kunkur et al 2007). among them seed rot and seedling death before and after Seed coating technique was first developed by the emergence, lost of yield quantitatively and qualitatively British Seed Company (German) in 1930. Polymer coating (Moretti et al 2008, Szike et al 2009, Tesso et al 2010). was widely used (i.e. Commercial uses) (Ni 1997). However, Therefore, many solutions were suggested to protect seeds the accurate structure of coating materials is not known and from these diseases via using agrochemical agents on plants the manufacturing companies don'tdeclare the active either foliar application or incorporation with soils by using ingradients of these agents. But, it is known that these agents low concentrations, yet they don't reach the target due to the contain linking materials and matrix like polyvinyl alcohol, infiltration of these chemicals and de cay by light and the polyvinyl acetate, vermiculite and clay. These agents also microorganisms present in soil which require the repeating of contain large amount of acrylic polymers, which is believed treatment or increasing the chemical concentration to that they are carcinogenic and badly affect the soil obtained the full protection of seeds. However, this approach microorganisms (Vyn and Murua 2001). Therefore, the idea will contaminate the water and soil (Gutierrez et al 2012). is to find materials has the characteristic of seed coating Resistance of Maize Seeds to Infection of Fusarium verticillioidis 109 materials, safe to the environment and offering the protection tests were performed to make sure that the agent was of seeds during the process of germination and emergence transformed to the Ag-NPs via using the Transmission on the condition that they are available and cheap. Local Iraqi Electron Microscopy (TEM) at the particles appeared nanoic gypsum (CasO4 ⅟2 H2O) may be a good material for this in size (Fig. 2). purpose. This product results from the burning of gypsum, Seeds were graded by using sieves, seeds of 7-8 mm

CaSO4.2H2O (Equation 1). were taken to facilitate forming seeds template which was

CaSO4 + 2H2O ∆→ CaSO4 + ⅟2 H2O …. (1) manufactured with 9 mm. Templates were manufactured by Local gypsum is characterized as solid under normal using compacted (pressed) cork. The coating thickness was conditions, easy in its decay in water and it's preparation, determined using metal bolls of 9 mm diameter heated under also it may be a good media for stimulant agents, fungicides suitable temperature, then placed on the cork to manufacture and insecticides. Therefore, this study aimed to test the these templates. One hundred seeds of each treatment were suitability of local gypsum as coated materials which might sown in the petri-dishes (40×30 cm dimension contains fixed offer good protection for seeds during germination and amount of field soils after removing all debris, grinding and emergence and also as carrier of nanoparticles under the sieving with 1.8 mm sieve). conditions of polluted soils with F. verticillioidis, which causes Studied characters: The following characters were seedling death and ability to control this disease. estimated according to the Kader (2005) i.e., first counting of field emergence, final counting of field emergence the day of MATERIAL AND METHODS The field emergence experiment and pathological infection test were conducted at seed Technology Lab. Department of Field Crop Sciences and Pathology Lab. Department of Plant Protection-College of Agricultural Engineering Sciences, University of Baghdad during 2019, respectively. This study was carried-out to investigate the effect of seed-coating of maize (synthetic variety 5018)by local gypsum (Lg) (coated and non-coated) and Ag-NPs(100, 200, 300 and 400 mg L-1). The suspension of F. verticillioidis (Fig. 1) was provided by the pathology Lab., Department of Plant Protection where it was diagnosed and make sure that it was pathogenic and propagated according to the method of AL-Juboory (2011) and Juber et al (2014). The surface layer of soil sterilized by Autoclave for 20 minutes under 1.5 bar pressure and 121ºC temperature for two consecutive times Fig. 1. Fungal colony on PDA culture F. verticillioides spores with interval period of 24 hr. was contaminated by fungus spores. They were placed in the petri-dishes of germination, then the pathogen with 1×106 spore ml-1 was added. A random sample of seeds was taken and sent to the pathology Lab. Seeds were free of diseases and sterilized by surface sterilization with hypochlorite sodium 1% (free chore) for two minutes and the washed by distilled sterilized water and dried by using sterilized filter papers. For preparation of Ag-NPs, chemical reduction was used (Rashid et al 2013). AgNO3 and

Sodium citrate (C6H5Na3O7 .2H2O). All reacted solution of agents were prepared in the distilled water where 50ml of Ag

NO3 0.001 Molar was boiled until the boiling point, then 50ml of Sodium citrate 1% was added by dropping them with continuous mixing during the heating until the colour of mixture was turned into pale yellow colour. It was removed from the heating apparatus with continuous stirring until it getting cool under the room temperature. The diagnostic Fig. 2. Illustration of nanoic particles of Ag-NPs by using TEM 110 Saddam H. Cheyed, A. Ch. Al-Fahad, Eman A. Al-Qaisy and Aalaa K. Hassan the last emergence and time required for field emergence gypsum and treated with 400 mgl-1 of Ag-NPs recorded the day. lowest number of days to reach the last day of field Emergence speed(Es) (seedling day-1) represents the emergence (7.75 days), which was not significantly differ with total of emerged seeds for every day proportional to the coated seeds and treated with 200 mgl-1 (8.25 days). number of days (daily emergence) and the character is However, dry seeds required more days to reach last day of considered the most important and accurate criterion of seed emergence (10.75 day) which not significantly different with viability (Yuan-Yuan and Yonglian 2010). It was calculated un-coated seeds treated with only water (10 days) (Table 1). according to the following equation: Results also showed significant superiority of coated seeds Es = ⅀ Ni / Di to reach early the full emergence with an average (8.70 days) Ni = number of seeds emerged per day. Di =Number of compared with un coated seeds which gave the higher days (daily germination). average (9.3 days). The Ag-NPs concentrations (400 and The field emergence percentage (%): The number of 200 mgl-1) gave the lowest average (8.25 and 8.75 days) with no significant differences between them, respectively. Seeds ٧ emerging seedlings was calculated above the soil after days. Results were transformed to percentage according to treated with distilled water recorded the highest average the following equation: (9.88 day). Field emergence (cm) =[NE(after 12 days of Time required for field emergence (day): There were planting)/TNS] ×100 significant differences between combinations of coating with NE= Number of emerged seedling. TNS= Total number gypsum and Ag-NPs treatment compared with the control of seeds (Table 2). Coated seeds and treated with 200 mg l-1 of Ag- Percentage of fungus infection F. verticillioides: At the NPs recorded the lowest average of time required to field end of experiment, five seeds were randomly taken from emergence (4.25 days) with no significant differences each experimental unit of all treatment, sown in petri-dishes 9 between them and Ag-NPs (400 and 100 mg l1-), which gave cm contains 15-20 cm3 of PDA (sterilized by sodium 4.50 day for both combinations. Dry seeds, un-coated seeds hypochlorite for two minutes), and insulated in the incubator and seeds treated with only distilled water and 100mgl-1 of Ag- at 25 + 2oC (Clements et al 2003). NPs recorded the highest average (6.00 days). Table 2 Statistical analysis: ANOVA analysis was performed for all showed significant superiority of coated seeds in giving the characters at two stages. The data were analyzed by using lowest average of required time to the field emergence (4.80 RCBD of treatments without control (dry treatment). Least days) compared with 5.85 day for un-coated seeds. significant difference was used to compare between means Speed of field emergence (seedling day-1): Results of at 5% (Steel and Torrie 1980). Table 2 showed significant differences between the combinations of study factors compared with the control. RESULTS AND DISCUSSION Coated seeds with gypsum and treated with 400 mg l-1 of Ag- The first day of field emergence (day): Combination of un- NPs recorded the highest speed of field emergence (19.96 coated seeds by gypsum and treated with 400 mgl-1 of Ag- NPs gave the lowest number of days to reach the first day of field emergence (3.00 day) which not significantly differ with Table 1. Effect of seed coated with gypsum and Ag-NPs on un-coated seeds and treated with 300 mgl-1 and coated- the first day of field emergence (day) -1 seeds treated with 400 mgl which gave an average of 3.250 Seeds treated Ag-NPs mg l-1 Average day for both treatments, respectively. However, dry seeds 0 100 200 300 400 required more days to reach the first emergence, 4.75 day Coated 4.50 4.25 4.00 3.50 3.25 3.90 (Table 1). Results also indicated that un- coated seeds were (9.75) (8.75) (8.25) (9.00) (7.75) (8.70) significant superior in giving the lowest number days to the uncoated 4.00 3.50 3.50 3.25 3.00 3.45 first field emergence (3.45 days), while the coated seeds give (10.0) (9.50) (9.25) (9.00) (8.75) (9.30) the highest average (3.90 days). Ag-NPs particles with 400 LSD 0.05 NS 0.31 (NS) (0.40) and 300 mgl-1were superior in recoding the lowest average Average 4.25 3.88 3.38 3.38 3.13 (3.150 and 3.38 days) without significant differences (9.88) (9.13) (8.75) (9.00) (8.25) between them, respectively. Seeds treated with distilled LSD 0.05 0.49 (0.63) water recorded the highest average of this character (4.25 Dry seed 4.75 (10.75) days). LSD 0.05 0.33 (0.93) The last day of field emergence (day): Seeds coated with *Last day of field emergence (day) in parentheses Resistance of Maize Seeds to Infection of Fusarium verticillioidis 111 seedling day-1) which not significantly differ with coated seeds Table 2. Effect of seed coated with gypsum and Ag-NPs on treated by Ag-NPs (200 and 100 mg l-1), which gave 19, 39 the required time for field emergence (day) -1 and 17.18 seedling day-1, respectively. Dry seeds gave the Seeds Ag-NPs (mg l ) Average treated lowest speed (8.80 seedling day-1), which not significantly 0 100 200 300 400 different with un-coated and treated seeds with only distilled Coated 5.25 4.50 4.25 5.50 4.50 4.80 water (9.58 seedling day-1) for treated seeds with Ag-NPs. (12.38) (17.18) (19.39) (15.13) (19.96) (16.81) Seeds treated with 400 mg l-1 gave the highest speed (17.19 uncoated 6.00 6.00 5.75 5.75 5.75 5.85 (9.58) (12.04) (13.97) (13.10) (14.42) (12.62) seedling day-1) with no significant differences with seeds -1 -1 LSD 0.05 NS (NS) 0.52 treated with 200 mgl , which gave (16.68 seedling day ). (1.75) While seeds treated only with distilled water gave the lowest Average 5.62 5.25 5.00 5.62 5.12 speed of 10.98 seedling day-1(Table 2). (10.98) (14.61) (16.68) (14.12) (17.19) Field emergence (%): Results in Table 3 indicated that there LSD 0.05 NS (2.77) were significant differences between the combinations of Dry seed 6.00 (8.80) study factors compared with the control. Coated seeds with LSD 0.05 1.19 (3.74) gypsum and treated with 400 ml-1 of Ag-NPs treatment *Speed of field emergence (seedling day-1) in parentheses significantly gave the highest average of field emergence Table 3. Effect of seed coated with gypsum and Ag-NPs on (88.75%) compared with 51.5 % for dry seeds. Seeds coated the field emergence (day) with local gypsum was significantly superior in giving the Seeds Ag-NPs (mg l-1) Average highest average (77.75%) of field emergence compared with treated 0 100 200 300 400 72.45% for un coated seeds. Seeds treated with 400 mg l-1 of Ag-NPs gave the highest average (85.62%) of field Coated 63.25 72.50 81.75 82.50 88.75 77.75 (5.50) (0.00) (0.00) 0.00) (0.00) (1.10) emergence compared with 60.38% for seeds treated with Uncoated 57.50 71.25 76.25 74.75 82.50 72.45 only distilled water (Table 3). (85.0) (14.0) (7.75) (0.00) (0.00) (21.35) Infection with fungus F. verticillioides (%): There were LSD 0.05 NS (5.95) 2.23 with significant differences between all combination (2.66) compared with the control (Table 3). All combinations were Average 60.38 71.88 79.00 78.62 85.62 (42.25) (7.00) (3.88) (0.00) (0.00) superior over dry seeds which gave the highest percentage LSD 0.05 3.52 (4.20) of infection (93.5%). However, coated seeds with gypsum Dry seed 51.50 (93.5) recorded the lowest percentage of infection (1.10%) LSD 0.05 4.77 (5.93) compared with 21.35% for uncoated seeds. Also, seeds *Infection percentage (%) in parentheses treated with Ag-NPs levels were significantly superior over seeds treated with only distilled water which gave the highest dry seeds. This may be due to the physical retardation caused average of infection 45.25% (Table 3). For interaction, there by local gypsum at the beginning of emergence process. were significant differences between study factors. All Whereas, seeds coated with gypsum and treated with 400 mg combinations of coated seeds with gypsum, all levels of Ag- l-1 of Ag-NPs showed highest average of emergence speed NPs and uncoated seeds treated with 300 and 400 mg l-1of and field emergence percentage. All levels of Ag-NPs, in Ag-NPs were superior without significant differences particular, 400 mg l-1 gave the best results of studied between them and seeds treated with only distilled water characters compared with seeds treated with only distilled which recorded the highest average 85% of infection. water. This may be due to the lime present in high percentage Field emergence is considered the key factor of (40%) in the gypsum, which stimulates seed germination. profitable agricultural production which can be realized when This result agrees with the findings of Scott (1975), who seedling vigour and its resistance to the biotic and abiotic achieved significant increase in field emergence percentage stresses have been improved. Among, biotic stresses due to the seed coated with different materials among them seedlings death which caused by F. verticillioides. This fungus lime, besides the role of Ag-NPs in controlling fungi, rots and is the most common disease especially in maize (Tesso et al other plant diseases and enhancing the plant growth (Sharon 2009, Choi and Xu 2010). Therefore, providing protection et al 2010). Many researches referred that using Ag-NPs measures for seeds at this stage is of vital importance. helped in increasing germination percentages and seedling Results showed that un coated seeds with local gypsum and growth via their enhancement to absorbs water (Zheng et al treated with 400 mg l-1 of Ag-NPs gave the lowest average of 2005), increasing nitrate reduction in the seeds, and days to reach the first day of field emergence compared with increasing the activity of oxidized enzymes (Lu et al 2002), 112 Saddam H. Cheyed, A. Ch. Al-Fahad, Eman A. Al-Qaisy and Aalaa K. Hassan

which increases the speed of physiological activities inside 2009. Genetic characteristics of F. verticillioides from corn in the Philippines. Journal of General Plant Pathology 75(6): 405 -412. the seed, speeding up germination percentage, and reducing Garg S, Chandra A, Mazumder A and Mazumder R 2014. Analgeic time required to emerge and completion of emergence. It is potential of hydrogels of silver nano-particales using aqueous clear from results of Table 3 the clear effect of combination of extract of Saracaindica bark. International Journal of Pharma Sciences and Research 5(1): 240-245. coated with local gypsum and treatment with all Ag-NPs Gutiérrez F, Mussons M, Gatón P and Rojo R 2012. Nanotechnology concentrations in preventing the infection with F. and food industry. Scientific, Health and Social Aspects of the verticillioides. It is worth to mention that seed coated Food Industry, Nanotechnology-and-Food-Industry, pp. 96- 128. treatment with gypsum only without Ag-NPs provided a Halmer P 2006. Seed technology and seed enhancement. In: XXVII protection to the seeds against the fungus infection which International Horticultural Congress-IHC2006: International reached 85%. This may be due to the positive role of coating Symposium on Seed Enhancement and Seedling Production 771. pp. 17–26. material in increasing the activity of seed coat to protect its Juber KS, Hassan AK and Alhamiri YN 2014. Evaluation of biocontrol internal contents against fungi and insects infection and agents and chemical inducers for managing vascular wilt of guaranteed the successful field emergence which, in turn, tomato caused by Fusarium oxysporum f sp. Lycopersici. Journal of Biology, Agriculture and Healthcare 4(27):335-343. allows a good adhesion with seeds and better protection of Kader MA 2005. A comparison of seed germination calculation compacted compounds with coating materials like fungicides formulae and the associated interpretation of resulting data. and insecticides (Pereira et al 2005 and Kunkur et al 2007). Journal and Proceeding of the Royal Society of New South Wales 138: 65-75. Also, the positive role of Ag-NPs which approved a good Kunkur V, Hunje NKB, Patil and Vyakarnhal BS 2007. Effect of seed protection against diseases. coating with polymer, fungicide and insecticide on seed quality in cotton during storage. Karnataka Journal of Agricultural Sciences 20(1): 137-139. CONCLUSION Lu CM, Zhang CY, Wen JQ, Wu GR and Tao MX 2002. Research of It can be local gypsum and Ag-NPs can improve the field the effect of nanometer materials on germination and growth enhancement of Glycine max and its mechanism. Soybean emergence properties, speed, percentage and providing the Science 21: 168-172. full protection of seeds against F. verticillioides with all Ag- Moretti A, Mulé G, Ritieni A, Láday M, Stubnya V, Hornok L and NPs concentration. However, Ag-NPs cannot provide such Logrieco A 2008. Cryptic subspecies and beauvericin production by Fusarium subglutinans from Europe. protection without coating with local gypsum especially at International Journal of Food Microbiology 127: 312-315. -1 higher concentration 300 and 400 mg l . This approves the Ni B 1997. Seed coating, film coating and pelleting. Seed Industry activity of local gypsum in increase the activity of Ag-NPs, and Agricultural Development. pp. 737-747. reducing its lost due its protection from washing off to be Rashid MU, Bhuiyan MKH and Quayum ME 2013. Synthesis of silver nanoparticles (Ag-NPs) and their uses for quantitative analysis available inside the seed bed for a longer period. of vitamin C Tablets. Dhaka University Journal of Pharmaceutical Sciences 12(1): 29-33. REFERENCES Safavi K, Mortazaeinezahad F, Esfahanizadeh M and Dastjerd H 2011. The study of nano silver (NS) antimicrobial activity and Abdullah A and Hamid Z 2013. Antimicrobial activity of silver evaluation of using NS in tissue culture media. International nanoparticles from Capsicum sp. against Staphylococcus sp., Conference on Life Science and Technology, IPCBEE 3: 159- Bacillus sp., Pseudomonas sp. and Escherichia coli. Journal of 161. Biology, Agriculture and Healthcare 3(7):67-72. Scott D 1975. Effects of seed coating on establishment. Journal of Al-Juboory HH 2011. Effect the variation of some Fusarium section Agricultural Research 18: 59-67. Liseola species on its virulence and mycotoxin production on maize and sorghum crops. Ph.D. Thesis College of Agriculture Steel R and Torrie J 1980. Principles and Procedures of Statistics a University of Baghdad. pp:125. Biometrical Approach, P: 633. New York. USA. Cheyed SH and Elsahookie MM 2011. Relationship between seed Szike CT, Bonisi AP and Szecs A 2009. Fusarium stalk rot: A biotic position on the cob, N level and harvesting data in maize seed stress factor decisive for maize stalk strength. VIII. Alps-Adria quality. The Iraqi Journal of Agricultural Sciences 42(5): 1-14. Scientific Workshop 37(2): 337-340 Tesso TT, Ochandaa N, Choi YE and Xu JR 2010. The Signaling pathway in Fusarium Little CR, Claflin L and Tuinstra MR 2010. Analysis of host plant verticillioides is important for condition, plant infection and resistance to multiple Fusarium species associated with stalk stress responses but not fumonisins production. rot disease in sorghum Sorghum bicolor L. (Moench). Field Phytopathology 23(4): 522-533. Crops Research 118: 177–182. Clements MJ, Kleinschmidt CE, Maragos CM, Pataky JK and White Vyn TJ and Marua M 2001. Polymer seed coatings: Sufficient Risk DG 2003. Evaluation of inoculation techniques for fusarium ear Reduction for early plant corn. Traditional risks of early planting rot and fumonisin contamination of corn. Plant Disease 87(2): of uncoated seeds. 56th Annual Corn and Sorghum Research 147-153. Conference, 1-11, Purdue University. Crocetti G and Miller G 2011. Nano silver policy failure puts public Yuan-Yuan S, Yonglian S, Tian WM, Yi LX, Xiang G, Rong H and Jun hea lthat risk. University of Adelaide and current president of the M 2010. Effects of seed priming on germination and seedling Australian Society for Microbiology, pp.1-23. growth of rice under water stress. Acta Agronomica Sinica 36(11): 1931-1940. Cumagun CJR, Ramos JS, Dimaano AO, Munaut F and Van Hove F

Received 22 September, 2019; Accepted 04 November, 2019 Manuscript Number: S-264 Indian Journal of Ecology (2019) 46 Special Issue (8): 113-118 NAAS Rating: 4.96

Effect of Glutathione Foliar Spray on Some Growth Traits in Maize Varieties

Ruqaya H. Mahmood and Najat H. Zeboon1 Ministry of Agriculture, Iraq 1College of Agricultural Engineering Sciences University of Baghdad, Baghdad, Iraq E-mail: [email protected]

Abstract: A field experiment was conducted to investigate the response of some growth traits of four maize varieties (Fajer, Baghdad, 5018, and Al-Maha) to the foliar Glutathione spray concentrations (0, 100, 200, and 300 mg l-1). The Baghdad variety showed superiority in the traits of plant height, leaf area, and number of leaves traits, while Fajer var. recorded the least average of days till75%tasseling and silking75% (75% of plants)i.e., 57.08 and 68.58 days, respectively and Al-Maha gave the highest number of days till tasseling and silking 75% of plants and the lowest values of leaf number, chlorophyll content, and plant dry weight. The treatment of spraying of Glutathione300mg l-1 was superior to the other concentrations in the most studied traits except for the number of leaves. This concentration increased the plant height, leaf area, plant dry weight, and total chlorophyll content by 4.26%, 5.532%, and 23.95%, respectively compared to the treatment of no-spray, which recording the lowest average of the studied traits. The interaction between two factors was significant in all the growth traits.

Keywords: Glutathione, Varieties, Maize, Growth traits

The cultivation of maize in Iraq faces a significant metals and herbicides. Moreover, it forms a defensive means reduction in cultivated area and low productivity compared to against pathogens. It has a role in the process of flower global production, although this crop has a wide development in plants (Noctor et al 2011). In addition to its environmental range and adapts to different environmental role in the release of salicylic acid (Rouhier et al 2008), which conditions. It's cultivation extends from latitudes 50oN and works to stimulate the pigments responsible for 40oS and in areas with cold climates to high temperature photosynthesis and thus increase the rate of carbon areas (Hallauer 1994). Maize yield production in Iraq (1133.8 representation and the accumulation of dry matter ton in the areas of 558 dunams according to the Ministry of (Rajasekaran and Blake 1990) Planning 2018) still low compared to the world production, for The important functions performed by Glutathione many reasons involving the failure to follow the modern mentioned above has a significant impact in increasing agricultural practices including the use of safe and growth and yield (Abdellatif 2017, Ebtihal et al 2018). environmentally friendly materials, whereas, recently the The main object of growing maize is the grain yield, world has trended towards the use of these materials as a which is greatly affected by the environment, so it's the result of the large problems caused by the excessive use of outcome of the interaction among a group of major and minor chemical fertilizers affecting adversely the animal and human components governed by genetics. Thus, it is necessary to health. One of the safe approach is using Glutathione, which concentrate efforts in order to improve this crop. is a tripeptide composed of three amino acids (Glutamic acid, Understanding the behavior, and performance of each Cysteine and Glycine). It is a powerful antioxidant variety in addition to their response to the growth factors, the responsible for the balance between oxidation and field practices are essential to the yield increment. Due to the antioxidants and protects against free radicals and regulates previously mentioned and the absence of a study showing many cell functions such as repair and formation of DNA and the effect of Glutathione on maize plants, this study was proteins as well as regulate the work of plant enzymes conducted aimed to knowing the response of the growth traits (Pompella et al 2003). The main ingredient in the of the genotypes to Glutathione spraying. Glutathione–ascorbatecycle, which reduces hydrogen peroxide, then preserves cells when exposed to oxidative MATERIAL AND METHODS stress (Noctor and Foyer 1998). Glutathione plays an A factorial experiment was conducted during 2018 in a important role in the metabolism processes and in cell field experiment at College of Agricultural Engineering division and differentiation, as well as it acts on gathering and Sciences, University of Baghdad Jadrya. The experiment receiving the optical signals in addition to resisting heavy was laid out in Randomized Complete Block Design (RCBD) 114 Ruqaya H. Mahmood and Najat H. Zeboon to investigate the response of four maize varieties to days, respectively, while the Al-Maha variety differed Glutathione spraying. significantly from the others and recorded the longest period The soil of the experiment field was ploughed using the to tasseling 75% of plant (60.75 days). This difference may flattening plough and the soil was prepared with rotovator be due to the genetic variances among varieties in the period plough and then leveled and divided into experimental units length preceding tasseling or to the extent they are with dimensions (3 m × 3 m). The number of experimental influenced by environmental conditions. This stage of growth units reached 48. The experimental unit included five lines of is most affected by the Light intensity and temperature and 3 m length and the distance between the lines was 75 cm and their interaction with the genetic structure of the plant. the distance between plants was 25 cm to obtain a plant Table 1 refers to the different behavior of the varieties density of 53000 plants ha-1 related to the different Glutathione spray concentrations. The The experiment contains two factors, the first factor number of days until reaching 75% tasseling significantly included four maize varieties (Fajer, Baghdad, 5018 and Al- decreased when the concentration of glutathione sprayed Maha). It is represented by symbols (V1, V2 , V3 and V4 ) increased from 0-100 at the variety 5018 and without sequentially, the second included four concentrations of significant difference between concentrations 100, 200 and Glutathione (0, 100, 200, and 300 mg l-1) and it's represented 300 mgl-1. The behavior of Al- Maha var. varied by the effect of by symbols (C0, C100, C200and C300) Sequentially that increasing concentrations of spraying. The number of days were sprayed during two growth stages: first, at six-leaves till75% tasseling increased by increasing the spray stage (75% of plants) and second, at twelve-leaves stage concentrations with a difference of 9.66 days when (75% plants). The Glutathione was sprayed during the increasing the spray concentration to 300 mg l-1 of evening on the plant vegetative parts using a pressure glutathione. sprayer until full wetness and a diffuser (dish detergent) was Number of days till silking 75% of plants: Data in Table 1 added to reduce the water surface tension. As a precaution, illustrated significant differences among the study factors the stem borer insect was controlled using 6kg ha-1 of (varieties and glutathione concentrations) in addition to the granular diazinon (10% effective material). The land of the significant interaction between them in the number of days till experiment was planted in July 27, 2018. Urea fertilizer of 75%silking. The data shows the significant superiority of 400 kg ha-1 was added in two batches, the first one was added 5018 var. giving the least number of days (68.58 days) with 20 days after emergence and the second was added at the no significant difference from Baghdad (68.83 days), while beginning of the flowering (Ministry of Agriculture 2011). Crop Al-Maha var. differed significantly from the other varieties and service operations were conducted manually whenever recorded highest number of days for 75% silking. This may needed. The parameters number of days from planting till be due to the difference of these varieties to reach 75% tasseling 75% of plants, number of days from planting till tasseling (Table 1), where female inflorescences appear silking (75% of plants), plant height (m2), number of leaves between 7-3 days after the emergence of male per plant (leaves plant-1), leaf area, cm2 (El-sahookie 1985), dry weight at the physiological maturity stage and chlorophyll Table 1. Effect of varieties, Glutathione concentrations and content (Goodwin 1976) were recorded. All data were their interaction on the number of days till the analyzed using the gestate software. Analysis of variance tasseling 75% of plants* and mean comparison was carried out with 5% probability. Varieties Glutathione concentrations (mg l-1) Plants were harvested when they reached full matured. C 0 C 100 C 200 C 300 Mean V 1 55.33 59.00 58.33 55.67 57.08 RESULTS AND DISCUSSION (64.67) (66.00) (74.33) (69.33) (68.58) Number of days till tasseling (75 % of plants): The maize V 2 56.00 54.67 63.00 58.33 58.00 (66.67) (64.67) (75.67) (68.33) (68.33) varieties and their interaction with the Glutathione V 3 62.67 55.33 55.67 55.67 57.33 concentrations on the number of days till the tasseling (75% (64.00) (65.00) (74.67) (70.67) (68.58) of plants) showed significant impact while the difference in V 4 55.67 61.67 60.33 65.33 60.75 the glutathione concentrations did not affect this trait (76.33) (65.00) (71.33) (76.33) (72.25) significantly. L.S.D 3.358 1.679 (4.052) (2.026) The table shows the superiority of Fajer variety. Mean 58.33 57.67 59.33 58.75 recording the least number of days (57.08 days) reaching (67.92) (65.17) (74.00) (71.17) tasseling 75% of plants. However, it did not differ significantly L.S.D NS (2.026) from Bagdad and 5018var. that recorded 58.00 and 58.25 *Number of days till silking 75% of plants Glutathione Spray on Growth Trait in Maize 115 inflorescences. Results showed the superiority of the cm that differed significantly from 0 mgl-1 giving 181.46 cm concentration 100 mg l-1 giving the lowest average of the trait plant height, as well as from the concentrations 100, and 200 (65.17 days) that differed significantly from the other mg l-1 that gave plant height averaged, 184.77 and 185.80 cm, concentrations (0, 200, and 300 mg l-1) that gave 6.92, 74.00, respectively. However, the difference between these two and 71.17 days, respectively (Table 1). This difference may concentrations was not significant. The difference among the be due to the glutathione role in developing the flowering Glutathione concentrations in this trait may be due to the process in plants (Rouhier et al 2008). It plays an important glutathione role in the processes of the cell division, and essential role in the nucleus for the transition of cells from elongation, and differentiation (Potters et al 2004, Henmi et al vegetative growth stage G1 to reproduction growth(S) (Diaz- 2005). It is a natural antioxidant in the plant that prevents or Vivancos et al 2010) regulates this process during growth and development from The interaction between maize varieties and glutathione cell division and elongation to aging and death (Sharma et al concentrations was significant. Fajer and 5018var. behaved 2012 and Kasote et al 2015). These results are consistent similarly in this trait when the Glutathione concentration with those of Ghoname (2010) and El-Awadiet al (2014). The increased from 0 to 200 mg l-1. The trait maximized at the same table demonstrates the differences between the concentration of 200 mg-1 and declined at the concentration responses of the varieties to Glutathione concentrations. 300 mg l-1, while Bagdad and A-Maha variety behaved Plant height of the Fajr variety increased positively by differently. The trait decreased at the concentration 100 mg l-1 increasing the Glutathione concentration to 300 mgl-1 and then recorded a significant increment at the Glutathione gave the highest plant height with a increasing concentration 200 mg l-1 compare to the no-spray treatment. ratio of 10.18% compared to non-spray treatment, while the Plant height: Table 2 refers to the significant different effect two varieties, Baghdad and Al-Maha varieties, gave the of maize varieties and spraying glutathione and the highest plant height averaged 196.27 and 188.67 cm, interaction between them on the plant height. Baghdad var. respectively when sprayed by 200 mg l-1 of glutathione. was superior in this trait giving the highest average of the Afterwards the plant height declined insignificantly. On the plant height (190.07 cm) followed by 5018 (187.97 cm) and contrary, the plant height of 5018 variety plants decreased AL-Maha variety (185.34 cm), while Fajer variety gave significantly when they were sprayed by 200 mgl-1 and minimum average of the plant height (177.88 cm).However, increased significantly when they were sprayed by 300 mg l-1 the difference between Baghdad and 5018 var. was not producing the highest plant height averaged 196.67 cm. significant. The differences among varieties may be due to Number of leaves per plant: Baghdad var. produced the their genetic differences in responding to the environmental highest leaf area averaged 14.683 leaves plant-1 that did not conditions, especially light and temperature. There was differ significantly from the variety 5018 producing 14.283 significant difference among the treatments spraying leaves plant-1 on average, while Al-Mahavar. produced the Glutathione in the plant height (Table 2). Spraying 300 mg l-1 lowest value averaged 13.733 leaves plant-1 (Table 2). The produced the highest values of plant height averaged 189.32 reason for this variance may be due to their variance in the plant height that may make plants have more leaves. The table also referred that the effect of different Table 2. Effect of varieties, Glutathione concentrations and their interaction on plant height (cm)* glutathione spray concentrations on the plant leaf number -1 was not significant, in contrast to the results obtained by Varieties Glutathione concentrations (mg l ) Bekheta and Talaat (2009) and El-Awadi and AbdElwahed C 0 C 100 C 200 C 300 Mean (2012) and Hussein et al (2014) who reported that spraying V 1 169.83 176.07 178.47 187.13 177.88 (13.220) (13.800) (14.200) (14.400) (13.905) plants of mung bean, scallion and cotton by different V 2 185.13 187.33 196.27 191.53 190.07 glutathione concentrations increased the number of leaves (14.200) (14.800) (15.733) (14.000) (14.683) plant-1 significantly. V 3 186.95 188.47 179.80 196.67 187.97 The results show a significant effect of the interaction (14.533) (14.867) (13.133) (14.600) (14.283) between the varieties and glutathione spray concentrations. V 4 183.91 187.20 188.67 181.60 185.34 (13.533) (14.067) (13.133) (14.200) (13.733) The variety Fajer, behaved differently from the other varieties L.S.D 6.031 (1.052) 3.015 where the number of leaves in this variety increased (0.526) positively when the glutathione spray concentration Mean 181.46 184.77 185.80 189.32 increased, whereas spraying 300mgl-1glutathione on (13.872) (14.383) (14.050) (14.300) Baghdad plants decreased the number of leaves, and the Al- L.S.D 3.015 (NS) Maha and 5018 behaviors were similar as the number of *Number of leaves plant-1 116 Ruqaya H. Mahmood and Najat H. Zeboon leaves decreased to 13.1333 leaves plant-1 when the area of the varieties Fajer and Al-Maha firstly increased when glutathione concentration was 200 mg l-1. the glutathione concentration increased, but it decreased Leaf area (m2): Baghdad proved to the superior variety insignificantly at the concentration 200 mg l-1. On the other producing the highest leaf area averaged 0.6696 m2 hand, the leaf area reached its maximum mean for Baghdad exceeding the varieties Fajer, 5018 and Al-Maha that variety at concentration of 200 mg l-1 of glutathione 0.6949 m2 produced leaf area averaged 0.6478, 0.6452, and 0.6623 m2 and then decreased significantly decrease at concentration by 3.365, 3.781 and 1.10%, respectively, yet Baghdad did not 300 mg l-1. differ significantly from Al-Maha. The reason behind this Plant dry weight (g plant-1): The maize varieties and the difference may be due to the genetic variances among Glutathione spray concentration with reference to plant dry varieties in this trait as well as the relationship between this weight showed significant difference in addition to the trait and number of leaves in plant (Table 3), which increased interaction between them (Table 3). The variety 5018 was the plant leaf area. superior to the others and recorded the highest plant dry In addition, there was a significant difference among weight averaged 346.2 g plant-1 that did not differ significantly glutathione spray concentration in this trait as the plants from Baghdad, while the lowest average of the plant dry sprayed by 300 mg l-1 were superior and produced the highest weight (316.7 g plant-1) was recorded by Al-Maha that did not plant leaf area averaged 0.6695m2 that did not differ differ significantly from Fajer. Perhaps the reason for the significantly from the two concentrations 100 and 200 mg l-1, superiority of the variety 5018 in this capacity is that it did not which in turn did not differ from each other. This may be due to differ significantly from the variety Baghdad in some growth the role of glutathione in the plant, as it is found in the traits (plant height and the number of leaves (Table 2). The Photosynthesis tissues and other tissues and protects the table illustrates the superiority of spraying 300 mg l-1 of cell and its structures from oxidative damage as a result of the glutathione giving the highest plant dry weight (367.9 g plant- biological activities in which it controls the division of cells as 1) compared to the other concentrations 0, 100, 200 mg l-1 with well as its role in the removal of toxins and its role in the a increase ratio 17.69, 16.75 and 9.13%, respectively. The pathways of biological representation and increase the difference between the first two concentrations, 0 and 100 activity of many enzymes (Gillbert et al 1990, Sanchez- mg l-1, was not significant. The reason may be that glutathione Fernandaze et al 1997, Noctor et al 2012). These results are is an antioxidant acting as a defensive barrier in a plant, consistent with those reached by Sadak et al (2014). protects it against the antioxidant damage as well as its role in The interaction effect between the varieties and increasing dry weight (indirect) through the release of glutathione concentrations was significant on leaf area. The salicylic acid (Rouhier et al 2008) which has a role in plant leaf area of 5018 increased positively with the increase catalyzing the pigments responsible for photosynthesis and of Glutathione concentration where the highest leaf area of thus increasing the rate of this process and the accumulation 2 this variety averaged 0.6708 m was produced when the of dry matter (Rajasekaran and Blake 1999). Or due to its -1 plants were sprayed by 300 mg l of Glutathione while the leaf direct role in increasing this trait through increasing some of the growth traits studied (plant height and leaf area Table 2). These results are consistent with those obtained by Table 3. Effect of varieties, Glutathione concentrations and their interaction on leaf area (m2) Ghoname et al (2010), Zaki and Mohamed (2017), and Eid et Varieties Glutathione concentrations (mg l-1) al (2011). Regarding the interaction between the maize C 0 C 100 C 200 C 300 Mean varieties and the Glutathione concentrations, Fajer, Baghdad V 1 0.6157 0.6584 0.6533 0.6635 0.6478 and 5018 were similar in increasing the plant dry weight (308.6) (316.3) (322.5) (358.1) (326.4) positively when the glutathione concentration increased, V 2 0.6620 0.6797 0.6949 0.6418 0.6696 where the variety 5018 plants gave the highest average of the (314.7) (328.5) (356.8) (373.8) (343.5) plant dry weight (401.3 g plant-1) when they were sprayed by V 3 0.6164 0.6357 0.6577 0.6708 0.6452 -1 (315.7) (324.8) (343.1) (401.3) (346.2) 300 mg l glutathione. Al-Maha showed a fluctuated response V 4 0.6434 0.6583 0.6457 0.7017 0.6623 to the spray concentration. It recorded a significant increment (311.4) (390.9) (326.0) (338.5) (316.7) in the plant dry weight when the plants were sprayed by 100 L.S.D 0.02631 (19.81) 0.01316 mg l-1 compare to 0 mg l-1 and significant decrease when they (9.90) were sprayed by 200 mg l-1 and finally an insignificant Mean 0.6344 0.6580 0.6629 0.6695 -1 (312.6) (315.1) (337.1) (367.9) increment when the plants were sprayed by 300 mg l . -1 L.S.D 0.01316 (9.90) Total chlorophyll content (mg g fresh weight) *Plant dry weight (g plant-1) The data in Table 4 showed significant differences Glutathione Spray on Growth Trait in Maize 117

Table 4. Effect of varieties, Glutathione concentrations and Abdellatif YMR 2017. Effect of glutathione, 24 epibrassinolide and their interaction on total chlorophyll content (mg g-1 proline on wheat growth and antioxidant enzymes activity under fresh weight) salt stress. Ame-Eur. Journal of Agriculture and Environmental Sciences 17(5): 410-421. Varieties Glutathione concentrations (mg l-1) Akladious SA and Abbas SM 2013. Alleviation of seawater stress on C 0 C 100 C 200 C 300 Mean tomato by foliar application of aspartic acid and glutathione. Journal of Stress Physiology and Biochemistry 9(3): 282-298. V 1 37.39 38.94 40.82 43.39 40.13 Bekheta MA and Talaat IM 2009. Physiological of mungbean Vigna V 2 34.88 36.84 42.16 45.18 39.76 radiate plants to some bio regulators. Journal of Applied Botany and Food Quality 83: 76-84. V 3 36.84 39.91 43.40 44.79 41.24 Cai Y, Cao F, Cheng W, Zhang G and Wu F 2010. Modulation and V 4 30.16 30.91 34.12 39.29 33.62 exogenous glutathione and photosynthetic performance against Cd stress in the two rice genotypes different in Cd L.S.D 1.997 0.998 tolerance. Biological Trace Element Research 43(2): 1159- Mean 34.82 36.65 40.13 43.16 1173. Diaz-Vivancos P, Wolff T, MarkovicJ, Pallard OFV and Foyer CH L.S.D 0.998 2010b. A nuclear glutathione cycle within the cell cycle. Biochemisty Journal 431: 169-178. Diaz-Vivancos P, Dong YP, Ziegler K, Markovic J, Pallardo F, Pellny among varieties and Glutathione spray concentrations in TK, Verrier P and Foyer CH 2010a. Recruitment of glutathione into the nucleus during cell proliferation adjusts whole-cell addition to the interaction between them in the chlorophyll redox homeostasis in Arabidopsis thaliana and lowers the content. The leaves of 5018 variety contained the highest oxidative defence shield. The Plant Journal 64: 825-838. -1 Ebtihal ME, Mervat SS and Tawfik MM 2018. Glutathione treatment total chlorophyll content of 41.24 mg gm fresh weight alleviates salinity adverse effects on growth, some biochemical indicating superiority to this variety. The presence of aspects, yield quantity and nutritional value of chickpea plant. significant difference between the varieties in the content of SFJ Global Warming 2: 2. Eid RA, Taha LS and Ibrahiem SMM 2011. Alleviation of adverse chlorophyll in their leaves may be mainly due to the different effect of salinity on growth and chemical constituents of genetic susceptibility. The results of the same table indicated marigold plants by using glutathione and ascorbate. Journal of Applied Sciences Research 7(5): 714-721. the superiority of spray treatment with a concentration of 300 -1 El-Awadi MEl, El-Lethy SR and El-Rokiek KG 2014. Effect of the two mg l of glutathione in the content of the total chlorophyll of antioxidants, glutathione and ascorbic acid on vegetative leaves of plants, which has an average value of 43.16 mg g-1 growth, yield and some biochemical changes in two wheat cultivars. Journal of Plant Sciences 2(5): 215-221. fresh weight, superior to the rest of the treatments (0, 100 and -1 El-Awadi ME and AbdElwahed M 2012. Improvement the growth and 200) mg l with an increase of 23.95%, 17.76% and 7.36%, quality of green onion allium cepa.plant by some bio regulators respectively and the reason for this effect of glutathioneis that in the new reclaimed area at nob arid region,Egypt. New York Science Journal 5(9):114-120. representation photosynthesis possesses organs have EL-Sahookie MM 1985. A short cut method for estimating plant leaf powerful enzyme antioxidants and the antioxidant systems area in maize. Journal of Agronomy and Crop Science 15(4): and small molecules (glutathione) plays an important role in 157-160. the regulation of biological processes during photosynthesis Ghoname AA, Dawood MG, ShSadak M and Hegazi AMA 2010. Improving nutritional quality of hot pepper Capsicum annuum L. process. Glutathione enhances the effectiveness of this plant via foliar application with arginine or tryptophan or process and the biosynthesis of chlorophyll and delays its glutathione. Journal of Biological Chemistry and Environmental Sciences 5(1): 409-429. degradation (Kattab 2007, Latifi et al 2009, Noctor and Foyer Gilbert HF, Mclean V and Mclean M 1990. Molecular and culluar 2009). These results are in agreement with those of Cai et al aspects of thioldisulphate exchange Advanced Enzyme 63: (2010), Son et al (2013), AbdElwahed and Abouziena (2014) 169-172. Goodwin TW 1976. Chemistry and Biochemistry of Plant Pigment. and Ebtihal et al (2018). It is noted from the same table that 2nd Academic. Press. London, New York. San Francisco: 373. the four varieties showed a positive response to all spraying Hallauer AR and Sears JH 1994. Integrating exotic germplasm into treatments with glutathione, but the response of the Al Maha corn belt maize breeding programs. Crop Science 12: 203-206. variety to increase the concentration of 300-0 mg l-1 of HenmiK, Demura T, Tsuboi S, Fukuda H, Iwabuchi M and Ogawa K 2005. Change in the redox state of glutathione requlates glutathione was more than the rest varieties in this capacity. differentiation of tracheary elements in zinnia cells and Baghdad's variety response came close to it, with an arabidopsis roots. Plant and Cell Physiology 46: 1757-1765. Hussein MM, Okasha EM and Mehanna HM 2014. Response of increase of 29.52% compared to 16.04%, 21.57% for Fajer cotton plants to glutathione rates under saline conditions. and 5018, respectively. Middle East Journal of Applied Sciences 4(1): 47-53. Kasote DM, Katyare SS, Hegde MV and Bae H 2015. Significance of antioxidant potential of plants and its relevance to therapeutic REFERENCES applications. International Journal of Biological Sciences 11(8): AbdElwahed MSA and Abouziena HF 2014. Efficacy comparison of 982–991. stearic acid, glutathione and salicylic acid on wheat Trticum Kattab H 2007. Role of glutathione and polyadenylic acid on the aestivum L. cultivars productivity in sandy soil. International oxidative defense systems of two different cultivars of canola Journal of Plant Soil Science 3(6): 554-574. seedlings grown under saline condition. Australian Journal of 118 Ruqaya H. Mahmood and Najat H. Zeboon

Basic and Applied Sciences 1: 323-334. 175-181. Latifi A, Ruiz M and Zhang CC 2009. Oxidative stress in Rouhier N, Lemaire SD and Jacquot JP 2008. The role of glutathione cyanbacteria-F E M S. Microbiological Research 33: 258-278. photosynthetic organisms: emerging function for glutaredoxins Ministry of Agriculture 2011. Guidelines for cultivation and production and glutathionylation. Annual Review of Plant Biology 59: 143- of maize. General authority for agricultural extension and 166. cooperation. Iraq. pp16. Sadak MSH, El-Lethy SR, Ahmed MA and El-Rokiek KJ 2014. Response of two cultivars of wheat plant foliar treatment of Noctor G and Foyer CH 1998. Ascorbate and glutathione: keeping glutathione. Middle East Journal of Agriculture Research 3(4): active oxygen under control. Annual Review of Plant 732-737. Physiology and Plant Molecular Biology 49: 249-279. Sanchez-Fernandez R, Fricker M, Corben LB, Nick SW, Nicki S, Noctor G, Mhamdi A, Chaouch S, Han Y, Neukermans J, Christopher JL, Marc VM, Dirk I and Mike JM 1997. Cell Marquez–Garcia B, Queval G and Foyer CH 2012. proliferation and hair tip growth in the arabidopsis root are Glutathaione in plants: An integrated overview. Plant, Cell and under mechanistically different forms of redox control. Environment 35(2): 454-484. Proceedings of the National Academy of Sciences of the United Noctor G, Queval G, Mhamdi A, Chaouch S and Foyer CH 2011. States of America 94: 2745-2750. Glutathione the Araidopsis Book 9: 1-32. Sharma P, Jha AB, Rama SD and Mohammad P 2012. Reactive Pompella A, Visvikis A, Paolicchi A, Tata V and Casini AF 2003. The oxygen species, oxidative damage, and antioxidative defense changing faces of glutathione, acelluar protagonist. Bioc. Phar mechanism in plants under stressful conditions. Journal of 66(8): 503-1499. Botany 26 pages. https://doi.org/10.1155/2012/217037 Potters G, Horemans N, Bellone S, Caubergs J, Trost P, Guisez Y Son JA, Narayanankutty DP and Roh KS 2013. Influence of and Asard H 2004. Dehydroascorbate influences the plant cell exogenous application of glutathione on rubisco and cycle through a glutathione–independent reduction rubiscoactivase in heavy metal-stressed tobacco plant grown in mechanism. Plant Physiology 134(4): 1479-1487. vitro. Biological Sciences 21: 89-97. Rajasekaran LR and Blake TJ 1999. New plant growth regulators Zaki SS and Mohamed GF 2017. Alleviating effects of ascorbic protect photosynthesis and enhance growth under drought of aciwed and glutathione for faba bean plants irrigated with saline jack pine seedlings. Journal of Plant Growth Regulation 18: water. SDRP Journal of Plant Science 2(2): 87-95.

Received 30 September, 2019; Accepted 21 November, 2019 Manuscript Number: S-265 Indian Journal of Ecology (2019) 46 Special Issue (8): 119-122 NAAS Rating: 4.96

Effectiveness of Oxidation Enzymes in the Ratio of Gluten to Wheat Bread Via Different treatments of Weeds Control

Alaa A.J. AL-Behadili and Ziyad A. Abed Department of Field Crops, College of Agricultural Engineering Sciences, University of Baghdad, Baghdad, Iraq E-mail: [email protected]

Abstract: A field study was carried out to evaluate the efficacy of various extracts, with Atlantis herbicide and their interactions on controlling grassy weeds of wheat.. The 1/4 Atlantis herbicide sorghum and sunflower extracts increased catylase (8.92) unit absorption mg-1 protein, while weed free treatment gave the lowest value measured at 1.27 unit absorption mg-1 protein. In addition to this, the 1/3 Atlantis herbicides and Atlantis + sunflower + sorghum extracts gave highest value of superoxide dismutase and peroxidase measurement at affecting 0.67, 26.70 unit absorption mg-1 protein, respectively. Atlantis herbicide reduced weeds and their competition with wheat. In turn, the gluten has increased from 15.8 to 17.5 mgl-1. Thus, Atlantis herbicide can be used with sunflower and sorghum extracts to increase the peroxidase and catylase against reactive oxygen species (ROS), and for accumulation of high concentration of gluten in grains.

Keywords: Atlantis herbicide, Reactive oxygen species, Wheat

Wheat is one of the most important crops worldwide. It's weed interference through a change of anti-oxidant enzymes a major source of energy in human nutrition. It provides about activity (Asada 1999). 50 per cent of total of carbohydrates of the world population The functions of anti-oxidant enzymes are to restore (FAO 2010). Wheat is unique among cereal crops due to the cellular homeostasis. Unlikely, weeds elevated the level of 1 - fact that the wheat flour has complex protein called glutin, O2 , H2O2 and OH , that will be required for generation of that can be formed into dough with rubbery properties to catylase and superoxide dismutase to scavenge their toxicity make leavened bread. The biotic stress such as sufficient of (Foyer and Nocoter 2005). The study aimed to measure a nutrient element, weeds, herbicides, that can limit of growth concentration of anti-oxidant enzymes under treatment of of crop and its productivity. Herbicides are often Atlantis herbicides and plant extracts. accompanied with hamper of metabolite process in plant cell.

Most of metabolites are synthesized of CO2 by Shikimate MATERIAL AND METHODS and mevalonate (Ahrens et al 2010). Atlantis herbicides have A field study was carried out at College of Science and induced the production of oxidative stress to produce Agricultural engineering Station – University of Baghdad, antioxidant and oxidant enzymes against reactive of oxygen Iraq during winter 2019, to investigate the effect of Atlantis - species (ROS) in plant tissues like O2, superoxide radical (O herbicide, sunflower and sorghum extracts with their 2 - ), hydrogen peroxide (H2O2) and hydrogen radical (OH ).The interactions of grassy weeds on wheat and their effects on symptoms of ROS caused damage of chlorophyll molecules, anti-oxidant enzymes, like catylase, peroxidase and proteins and DNA molecule that would cause affects to the superoxide dismutase. formatting fatty acids and plant growth (Asada 1999). The The experiments were laid out in randomized complete generated antioxidants enzymes such as catylase, block design with three replications. The net plot size of 2 2 m peroxidase and superoxide dismutase play a key role in was maintained for each treatment. Wheat cultivar (IPA 99) removing effects of ROS, and maintenance of plant live was sown on Nov. 20, 2018 with seed rate of 100 kg ha-1. The (Foyer and Nocoter 2005). While glutathione peroxides, NPK fertilizer dose of 70 kgha-1 was applied in the form of ascorbate and glutathione reductase showed high activities urea, whole of P was applied in form (P2O5) as basal, while N under different levels of herbicides (Scandalios 2002). fertilizer was applied at three time through out wheat life Herbicides caused breakdown of cytokinin content (CC) cycle. and kernel filling (Ramos and Pitelli 1998), however some of Experiments was comprised of 12 treatments namely plants could endure detoxify herbicides by scavenging of sorghum extract (T1), sunflower extract (T2), sorghum + reactive oxygen species (ROS) via antioxidant enzymes sunflower extract (T3), 1/2 Atlantis herbicide + sunflower

(Ciniglia et al 2015). Several studies have demonstrated extract (T4), 1/3 Atlantis herbicide + sorghum and sunflower change of ROS production of wheat varieties as response to extract (T5), 1/4 Atlantis herbicide sorghum and sunflower 120 Alaa A.J. AL-Behadili and Ziyad A. Abed

35 extracts (T6), 1/2 atlantis (T7), 1/3 atlantis (T8), 1/4 atlantis (T9), 30 28.76 full recommended of atlantis (T10), Free control weed (T11) and otiein weedy treatment (T12). Anti-oxidants enzymes have been 25 1 pr - extracted from plants during bloating stage. 20 .mg Catylase (CAT): The activity of CAT was measured by using 15 13.06

spectrophotometer according method (Mitter 2002): tylase U 8.92 8.96 10 8.3 D Readings 4.59 5 3.14 -1 2.01 2.3 Activity of CAT (U mg protein) = Dt 1.47 1.62 1.27 0.62 0.01 ´ 0.1´ concentration of protein mg ml-1

Activity of ca 0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 L.S.D -5 Superoxide dismutase (SOD): The activity of SOD was 0.05 Treatments measured according to method (Mitter 2002): Fig. 1. Effect of some plant extracts and Atlantis herbicide on SOD inhibition % D.F Activity of SOD (U mg-1) = ´ catylase activity of IPA wheat cultivar

50 % Vs (mL) Activity of peroxidase (POD): The activity of POD was 0.6 measured according to method given by Mitter (2002): 0.48 0.5 D Readings 1 -1 Dt - POD Activity (U mg protein)= ase unit of 0.4 0.36 0.01 ´ 0.1´ concentration of protein mg ml-1 0.35 0.32 0.32 0.31 0.32 0.28 otien ml 0.3 0.24 0.25 Proteins = Total proteins were measured according to the 0.22 xide dismut method given by Wrigley (1996): o 0.2 0.091 Gluten = Gluten was measured according to method given 0.1 by AA CC (2000). absorption of pr 0.021 0

Activity of super T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 L.S.D RESULTS AND DISCUSSION -0.1 0.05 Treatments Catylase: Catylase is one of antioxidants enzymes (Pen et al Fig. 2. Effect of some plant extracts and Atlantis herbicide on 2007). This enzyme consist of polypeptides approximately the activity of superoxide dismutase of IPA wheat 50-70 KDa in cell of plant, that provide insight to sufficient cultivar

H2O2 for chloroplast to synthesized proteins (Cakmak and Horst, 1991). The catylase is very rapidly reduced through act as thylakoid membrane of chloroplast, as well as it - - two electrons that come from molecule of water (H2O2) and transferred O2 through O2 formation (Dodge 1994). involves successive NADPH and caused loss of glucose-6, Atlantis herbicide consist of 1, 1-dimethyl–2,4- phosphate dehydrogenase (McCunn 1997). The results bipyridylium dichloride, and it is widely used to mediate showed that there were significant differences between damage of ROS in plants (Dodge 1999),while the substrate treatments and control (Fig. 1). The treatment T5 gave effects to chloroplast by increased SOD, with removal of highest level of catylase concentration (28.76) unit of ROS (Dodge 1994). The treatment of sorghum extract (T1) -1 absorption gm protein when compared with T12 that gave a caused control of weeds in very early age of weed life, lowest level of catylase (1.27 unit of absorption gm-1 of because of allopathy and not accumulation of antioxidant protein). The combination between 1/2 Atlantis, sorghum and enzymes: superoxide dismutase and peroxidase to sunflower extracts are enhanced by catylase activity via scavenging of reactive oxygen species (Altieri and Dall 2002) reduction to ascorbate by using NADPH-independent Peroxidase: The results (Fig. 3) showed that there were enzyme to protect cell membranes against oxidative damage significant differences between treatments under study. The to reactive oxygen species (Yang and Poovalah 2002). treatment T10 appear highest value 26.70 unit of ansorption -1 While, the treatment (control) caused decrease of enzymatic mg protein. Compared with T1, which gave the lowest value and non–enzymatic antioxidant function such as superoxide (10.01 units) of absorption. The sorghum extract did not have dismutase, caylase, ascorbate peroxidase, peroxidase and capacity to control of weeds. In addition to increase of flavoinds (Willekens 1993). reactive oxygen species ROS, and elevate of peroxidase to Superoxide dismutase (SOD): The results (Fig. 2) showed be factor scavenge (ROS) that there were significant differences between treatments Wheat proteins: Wheat protein consist of four fractions: and control. The treatment T10 gave the highest level of SOD albumins, globulins, gliadins and glutens. In addition to this, (0.88 unit of absorption ml-1). Atlantis herbicide has ability to gliadins and glutens comprise from 58.17 to 65.27 and 56.25 Weeds Control by Oxidation Enzymes 121 to 64.4%, respectively while albumins and globulins are Glutin: The (Fig. 5) showed that there were significant present 20 to 25 of whole quantity of protein (Verik- Ken et al differences between treatments, whereas, the treatment T10 -1 2000). The results (Fig. 4) showed that there were significant gave the highest value 15.8 mgl . Compared with T1. The differences between treatments. The treatment T2 showed increasing of gluten value in T10, is due to Atlantis herbicide, the highest quantity of protein (21.87%) compared to control which controlled the whole weeds and leads to increase the treatment (14.75%). The increasing of proteins are due to synthesis of glutin in mature wheat grain (Weegels et al elevate of salicylic (SA) levels, that have promoted Betaine- 1996). While, T1showed reduced glutin concentration biosynthesis (Woston 1996). Also, enhanced of long period trended to in format of anti-oxidant: enzymes, catylase and of stay green until the maturity of grains (Slooten et al 1995). peroxidase (Miflin and Habash 2002).

CONCLUSION

100 The antioxidant enzymes in plant cells protect cell 90 organelles against reactive oxygen species (ROS). It is often 81.41 80 considered that concentrations of factors of this study, like 1 - 70 atlantis herbicide, sorghum and sunflower extracts, as well .mg 60 as their interactions are determinate of the free- radical of 50 oxygen. Therefore, reactive oxygen species (ROS) are more xidase U o 40 useful conceptual paradigm to indentify the damage involved 30.3 26.77 26.77 30 22.63 23.67 in intracellular and perturbation of redox and high activity of 19.89 21.22 20 13.48 14.02 11.43 9.74 antioxidant enzymes such as catylase. Which increases Activity of per 10 2.4 recommended limit 1/3 of herbicide and sorghum extract. 0 Also with superoxide dismutase, whereas, T2 appears to be -10 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 L.S.D high level. While, gluten increased in treatments, which did Treatments 0.05 not contain weeds, due to control via Atlantis herbicide, which Fig. 3. Effect of some plant extracts and Atlantis herbicide on peroxidase activity on IPA wheat cultivar lead not subjected wheat plants to stress and accumulation of ROS and eventually increased of glutens.

25 21.87 REFERENCES 20 17.5 17.3 17.38 Apel K and Hirt H 2004. Reactive oxygen species: Metabolism, ein 15.31 15.31 15 15.19 ot 14.75 oxidative stress and signal transduction. Annual Review of Plant 13.56 13.45 13.2 15 Biology 55: 373-399.

tity of pr 10 Asada K 1999. The water-water cycle in chloroplast: Scavenging of active oxygen and dissipation of excess photons. Annual

Quan 5 Review of Plant Physiology and Plant Molecular Biology 50: 1.73 601-639. 0 Cakmak L and Horst J 1991. Effect of aluminum on lipid peroxidation T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12L.S.D 0.05 superoxide dismutase, catylase and peroxidase activities in root Treatments tips of soybean. Physiology Plant 83: 463-468. Fig. 4. Effect of some plant extracts and Atlantis herbicide on Ciniglia C, Mastrobuoni F, Scortichini M and Petriccione M 2015. the ratio of protein of IPA wheat cultivar Oxidative damage and cell programmed death induced in Zeamay l. By allelochemical stress. Ecotoxicology 24: 926-937. 25 Dodge A 1994. Herbicide action and effects on detoxification process. In: Mullineax D and Foyer C (eds) Causes of Photo – 19.2 Oxidative Stress and Amelloration of Defense Systems in 1 - 20 18 Plants. BOCA Raton : CRC press, pp. 219-236. 15.6 15.8 14.6 Food and Agriculture Organization (FAO) 2010. FAO STAT en mg l 15 12.3 12 Database. Available Online http://www.faostat.fao. Accessed 10.8 9.6 10 on 10 December 2010. 10 9 9.1

tity of glut Foyer CH and Nocotor G 2005. Redox homeostasis and antioxidants signaling: A metabolic interface between stress perception and

Quan 5 physiological responses. Plant Cell 17: 1866-1875. 0.75 McCunn CR 1997. Regulation of catylases in arabidopsis free radical 0 Biology and Medicine 23: 489-496. T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 Mitter R 2002. Oxidative stress, anti oxidants and stress tolerance. -5 Treatments Trends in Plant Science 7: 405-410. Fig. 5. Effect of some plant extracts and Atlantis herbicide on Ramos LRM and Pitelli RA 1998. Effect of different weed control period on productivity of corn crop. Resq. Agropecy. Bras. 29: the quantity of gluten of IPA wheat cultivar 1523-1531. 122 Alaa A.J. AL-Behadili and Ziyad A. Abed

Scandalios JG 2002. The rise of ROS trends. Biochemical Science of wheat glutenin. Journal of Cereal Science 23: 1-18. 27: 483-486. Wrigley CW 1996. Giant proteins with flour power. Nature 381: 738- Verik- Ken JM, Klosterman VLC, Beckers FHR, Spek King WTJ and 739. Graveland A 2000. Invective variation in proportion of wheat Yang T and Poovalah BW 2002. Hydrogen peroxidase protein. Fractions and relation to mixing behavior. Journal of homeostasis: Activation of plant catylases by calcium / Cereal Science 32: 159-167. modulin. Processing of the National Academy of Science, USA Weegels PL, Hamer RJ and Schofied JD 1996. Functional properties 99: 4097-4102.

Received 24 September, 2019; Accepted 21 November, 2019 Manuscript Number: S-266 Indian Journal of Ecology (2019) 46 Special Issue (8): 123-127 NAAS Rating: 4.96

Effect of Ratooning of Sorghum Cultivars on Growth and Yield Characters

Adawiya S. Mustafa Al-rawi, Sadam H. Chiad* and Arige M. Saleh1 College of Agriculture Engineering Science, University of Baghdad, Baghdad, Iraq 1Ministry of Water Resources, Iraq *E-mail: [email protected]

Abstract: A field experiment was carried out in the spring and autum seasons to study the response of five varieties of sorghum (Inkath, lilu, Ishtar, Buhooth 70 and caviar) to ratooning agriculture. Results showed that the pattern of agriculture has a significant effect on most studied traits. Ratoon plants were superior to plants grown from seeds in headm-2 (15.33 m-2), The plants grown from the seed were significantly higher than ratooning plants by giving the highest average number of grains, 200 grain weight, grain yield, plant and harvest index, and lowest mean number of empty heads 191.36 cm, 4910 cm2, 1999 grain head-1, 5.77 g, 57.03 gm plant-1, 639.6 g mm-2, 32.26% and 4.87%, respectively. The plants of the Inkath variety were superior to the weight of 200 grains, grain and plant yield, and harvest index 5.46 g, 57.56 g-1, 673.4 gm-2 and 30.40%, respectively.

Keywords: Sorghum ratoon, Planting pattern, Cultivars

The adoption of traditional methods in the cultivation yield traits that differ according to the genotype of the of sorghum crop leads to the acquisition of similar and cultivated cultivar which determines the phenotypic and comparable production and revenue in each season or anatomical characteristics of the species and determines the agricultural year, which is reflected negatively in the failure to extent of the response to the agricultural processes. Serhan increase the total profits in terms of production quantities and et al (2016) showed a significant difference between total revenue. The change in the pattern of production of a sorghum cultivars in the characters of weight of 500 grains, given crop achieves results that differ from the results the number of grains in the head and grain yield. obtained when repeating the same cultivation method. In the respect of the above, we can invest these Therefore, the development of modern agricultural methods methods in the cultivation of sorghum with the selection of to ensure increased revenues and the lowest cost may improved varieties for high productivity and the appropriate encourage farmers to grow the crop. The sorghum crop has compatibility between them to overcome the problems of the potential to stimulate its expansion of its cultivation, cultivation and to achieve an increase in the amount yield and where it can be grown from seeds and harvested more than therefore this study to evaluate the response of varieties of once for grain varieties (Chiad et al 2014). As the living roots sorghum for the ratoon cultivation. of the crop and basal buds develop in to tillers, stem and flowers and grain yield (Gerik et al 2003). As the date of MATERIAL AND METHODS cultivation of sorghum in the autumn season is the date of A field experiment was carried out in the spring and harvesting of the spring season, making conditions suitable autumn seasons of 2017 in the field of Field Crops for the application of the pattern of ratooning under the Iraqi Department – College of Agriculture to study the response of environment. Ratoon agriculture has many advantages as five varieties of sorghum (Inkath, lilu, Ishtar, Buhooth 70 and well as an economic method that does not need to be re- Caviar) to ratooning agriculture. The treatments were cultivated and requires land preparation, ploughing, and allocated to the experiment units according to randomized seeding (Ramburan et al 2013). Several studies have shown complete block design (RCBD) in split plot arrangement with that there is a significant superiority of the plants of the ratoon three replications. The main-plot consists of ratoon on the plants resulting from the cultivation of seeds directly to cultivation (Spring-Autumn ratoon) for sorghum cultivars, the crop of sorghum. As the number of heads, the weight of whereas, the sub-plots included sorghum varieties. Seed of the grains, the grain yield and harvest index were superior for varieties were planted in both spring and autumn seasons ratoon plants compared to plants grown from seed (Chiad with a plant density of 133300 plants ha-1. The experimental and Saleh 2018). Several studies have indicated that there is land was prepared with ploughing, smoothing and settling. It a wide variation between the varieties in most vegetative and was then divided into experimental units with dimensions of 3 124 Adawiya S. Mustafa Al-rawi, Sadam H. Chiad and Arige M. Saleh

x 2.5 m to give six lines. The distance between lines was 50 analysis of variance using the statistical program GenStat cm and the distance between plants was 15 cm. Three seeds v.12.1. Means were compared using the least significant were planted and then thinned to one plant in two stages, the difference at the probability level of 0.05 (Steel and Torrie first after the emergence and the second after ten days. Urea 1980). fertilizer (N% 46) was added to the soil as a source of nitrogen fertilizer at rate of 200 kg N ha-1, which was added at three RESULTS AND DISCUSSION stages, first at the end of field emergence, the second stage Number of Heads (head m-2): The results in Table 3 showed 4-6 leaves and the third at the beginning of the flowering a significant effect on the cultivation method. The ratoon stage. Phosphate fertilizer was added to the soil before cultivation method had significantly highest average number planting at the rate of 100 kg P ha-1 in the form of super- of heads count of 15.33 head m-2, while the normal seeding

phosphate (45% P2O5) in one dose. The soil of the method gave the lowest average number of heads, which experiment was irrigated immediately after planting, quietly, reached 11.25 head m-2. The increase in the number of heads to ensure that the seeds remained in the furrows in the of ratoon plants is due to the increase in the number of tillers planting lines. The experimental units were planted in the after the first harvest, which is reflected in the increase in the spring season on 2/4/2017 and plants were cut at a height of number of heads and grains in the unit area, which is 7-10 cm at maturity to protect the basal bud and new shoots positively reflected in the increase grain yield (Gerik and and left for the subsequent autumn season to obtain sorghum Neely 1987). These results are in agreement with what ratoon. The seeds of the sorghum cultivars were planted after Chiadand Saleh (2018) who found the superiority of the the completion of the cuttings of the ratoon plants and ratoon plants in the number of heads over plants produced irrigated with ratoon plants on same day of 22/7/2017. from seed cultivation. Weeding was done manually whenever needed. The results indicated that there were significant Characters studied: The number of heads per square meter differences between the cultivars in the study (Table 1). The (head m-2) and the number of empty heads (%) was plants of the variety Buhooth 70 were significantly higher by calculated. The number of grains per head and the weight of giving the highest average number of heads (16.79 head m-2). 200 grains (g) were taken from a grain sample. 200 grains While the plants of the variety Inkath and Lilohad the were calculated manually and weighed by a sensitive lowest average of this trait and without a significant difference balance, grain yield per plant (gm plant-1) and grain yield (t ha- between them (11.83 and 11.00 head m-2 respectively). The 1) were calculated from the 1 m2 harvested plants and then superiority of the plants of the Buhooth 70 in this trait is due to converted to ton ha-1 and dry matter yield (t ha-1), it was its superiority in the number of tillers, which was reflected in calculated from the harvesting 1 m2 at the maturity and then the increase in the number of heads. converted into units of tonsha-1dried naturally until the weight The results also showed a significant effect of the is stable.The harvest index was calculated as(total grain yield interaction between the two factors of the study. The ratoon / biological yield) *100. treatment of the Buhooth 70 was significantly higher with the Statistical analysis: The data obtained for the studied traits highest mean of 19.67 head m-2. While the plants of the were statistically analyzed according to the method of variety Inkath and Liloin the normal planting of seeds

Table 1. Effect of cultivars and cultivation pattern on the number of heads (head m-2), the number of empty heads (%) and the number of grains (grains head-1) Number of heads (head m-2) Empty head (%) Number of grains (grain head-1) Cultivar Cultivation pattern Mean Cultivation pattern Mean Cultivation pattern Mean Ratoon Seed Ratoon Seed Ratoon Seed Inkath 13.67 10 11.83 17 5 11 2243 2006 2124 Lilo 12.33 9.67 11 20 7.67 13.83 2303 2233 2268 Ishtar 14 10.67 12.33 23 5.33 14.17 1847 1797 1822 Buhoot 70 19.67 13.92 16.79 37.36 2.67 20.01 1188 1874 1531 Caviar 17 12 14.5 42.45 3.67 23.06 1223 2083 1653 LSD 0.05 1.34 1.01 1.71 1.19 157 Mean 15.33 11.25 27.96 4.87 1761 1999 LSD 0.05 0.92 1.63 66 Ratooning of Sorghum Cultivars on Yield 125 recorded the lowest average of this character and with no were significant differences between the varieties of the significant difference between them (10.00 and 9.67 head study. The plants of the Lilo cultivar were significantly higher m-2). than other varieties with 2268 grains head-1. While the Number of empty heads (%): The results of the statistical cultivars Buhooth70 and Caviar plants recorded the lowest analysis showed a significant effect among the cultivation average of grain per head and without a significant difference methods of agriculture. The normal cultivation pattern of between them (1531 and 1653 grains head-1). Differences in seeds was lower in the rate of empty heads with the lowest this characteristic may be due to the influence of genetic and average 4.87% (Table 1), while the highest rate of empty environmental factors and their interaction, which is due to heads was recorded by ratoon cultivation method the difference in the number of florets formed in (27.96%).This is due to the superiority of the ratoon plants in inflorescences, which turns in to grains later. The results the number of tillers and the absence of apical dominance showed significant interaction between the two factors. after the process of cutting plants in the first harvest. The ratoon pattern cultivation of cultivar Lilo gave the The results of Table 1 showed significant differences highest grains per head (2303 grains head-1) which did not between varieties in the study. The variety Inkath was differ significantly from same cultivar in normally seed superior to the other varieties with the lowest rate of number cultivation treatment and ratoon cultivation of the cultivar of empty heads of 11.00%, whereas, Caviar plants recorded Inkath (2233, and 2243 grains head-1). Whereas, the cultivar the highest average number of empty heads (23.06%). This Buhooth 70 and Caviar for the normal seed cultivation, is due to the variation in genetic material between the recorded the lowest grains (1188 and 1223 grains head-1, cultivars, which resulted in a difference in the resulting traits respectively). of each cultivars. Weight of 200 grains (gm): The results of the statistical The results also showed the significant of the analysis showed a significant effect among the cultivation interaction between the two factors of the study (Table 1). The methods of agriculture. The normal cultivation pattern of cultivar Buhooth 70 and Caviar of the normal planting pattern seeds was significantly higher in average weight of 200 of the seeds were superior to other cultivars with no grains (5.77 gm), while the ratoon cultivation method gave significant different between them, while the treatment of the the lowest average weight of 200 grains with 4.02 gm (Table Caviar ratoon plants gave the highest average of 42.45%. 2). The superiority of normal cultivation may be due to the Number of grains (grains head-1): The results of Table 1 lowest number of heads, which is reflected in increase of gain showed a significant effect of the two cultivation patterns. The weight (Table 1). Due to the lack of competition of grain for normal cultivation pattern of seeds was significantly higher nutrients, which reflected in the increase of grain size and with the highest number of grains per head (1999 grain head- weight, unlike ratoon plants, which there is a great 1). While, the ratoon pattern of cultivation recorded the lowest competition for the products of photosynthesis and light with 1761 grains head-1. This is due to the presence of greater because of increased branching, which reflected negatively competition between the ratoon plants because of the in the weight of grains. The results also showed that there increase of the tillers, which reflected negatively on the were significant differences between the cultivars in the number of grains in the head. The results showed that there study, where the plants of Inkath and Ishtar cultivars were

Table 2. Effect of cultivars and cultivation pattern on weight of 200 grains (g), grain yield (g plant-1) and plant yield (gm-2) Weight of 200 grain (g) Grain Yield (gplant-1) Plant Yield (gm-2) Cultivar Cultivation pattern Mean Cultivation pattern Mean Cultivation pattern Mean Ratoon Seed Ratoon Seed Ratoon Seed Inkath 4.75 6.17 5.46 53.25 61.86 57.56 728.1 618.6 673.4 Lilo 4.55 5.25 4.9 52.41 58.55 55.48 646 565.4 605.7 Ishtar 4.27 6.55 5.41 39.4 58.23 48.81 551.2 620.4 585.8 Buhoot70 3.3 6.46 4.88 19.57 60.5 40.04 384.8 841.9 613.4 Caviar 3.23 4.42 3.83 19.75 46.03 32.89 335.6 551.9 443.7 LSD 0.05 0.47 0.36 1.83 1.44 1.44 Mean 4.02 5.77 36.88 57.03 529.1 639.6 LSD 0.05 0.27 0.42 39.8 126 Adawiya S. Mustafa Al-rawi, Sadam H. Chiad and Arige M. Saleh

significantly higher in 200 gm with 5.46 and 5.41 gm, (639.6 gm-2), while the ratoon cultivation method gave the respectively, while Caviar cultivars recorded the lowest mean lowest average yield 529.1 gm-2 (Table 2). Also, the results (3.83 gm). The superiority of Ishtar and Inkath plants in this showed that there were significant differences between trait may be attributed to the lowest number of heads, which cultivars in the study. Inkath cultivar was significantly superior was reflected in the gain weight (Table 1). The results also by giving the highest average plant yield (673.4 gm-2), showed a significant effect of interaction between the two whereas, Caviar plants recorded the lowest mean for this factors of study. The treatment of Ishtar cultivars for the character (443.7 gm-2). The results also showed a significant normal cultivation pattern gave the highest mean of 6.55 gm, effect of interaction between the two factors of study. The which did not differ significantly with Buhooth 70 and Inkath treatment of Buhooth 70 cultivars for the normal cultivation plants (6.46 and 6.17 gm), respectively. While the cultivars of pattern gave the highest mean of 841.9 gm-2, while the Buhooth and Caviar plants in ratoon cultivation recorded the cultivars Buhooth and Caviar in ratoon cultivation pattern lowest mean of this trait, with no significant differences recorded the lowest mean of this trait (384.8 and 335.6 gm-2, between them (3.30 and 3.23 gm, respectively). respectively). Grain yield (g plant-1): The results showed a significant Grain yield (t ha-1): The results of the statistical analysis effect of the cultivation method, where the normal seed showed a significant effect among the cultivation methods. cultivation pattern was significantly superior by giving it the The normal cultivation pattern of seeds was significantly highest average grain yield of 57.03 gm plant-1, while the higher in grain yield (6.397 t ha-1), while the ratoon cultivation ratoon method gave the lowest average grain yield of 36.88 method gave the lowest average of grain yield 5.291 t ha-1 gm plant-1 (Table 2). This superiority is due to the increase in (Table 3). Also, the results showed significant differences the number of grains of plants from directly seed cultivation between the cultivars in the study. Inkath cultivar was pattern (Table 1). Also, the results of Table 2 showed that significantly superior by giving the highest average grain there were significant differences between the cultivars in the yield (6.734 t ha-1), whereas, Caviar plants recorded the study, Inkath cultivar was significantly superior by giving the lowest mean for this character (4.437 t ha-1). The results also highest average grain yield (57.56 g plant-1), whereas, Caviar showed a significant effect of interaction between the two plants recorded the lowest mean for this character (32.89 g factors of study. The treatment of Buhooth 70 cultivars for the plant-1). The results also showed a significant effect of normal cultivation pattern gave the highest mean of 8.420 t interaction between the two factors of study. The treatment of ha-1. While the cultivars Caviar in ratoon cultivation recorded Inkath and Buhooth 70 cultivars for the normal cultivation the lowest mean of this trait 3.355 t ha-1. pattern gave the highest mean of 61.86 and 60.50 g plant-1, Dry matter yield (t ha-1): The results in Table 3 showed a respectively which did not differ significantly between them. significant effect on the cultivation method. The ratoon While the cultivars of Buhooth 70 and Caviar plants in ratoon cultivation method were significantly higher with the highest cultivation recorded the lowest mean of this trait, with no average dry matter yield of 42.57 t ha-1 while the normal significant differences between them (19.57 and 19.75 g seeding method gave the lowest average dry matter yield plant-1, respectively). 20.26 t ha-1. This is due to superiority of ratoon plants in the Plant Yield (g m-2): The normal cultivation pattern of seeds yield of green fodder and the number of branches, this result was significantly superior by giving the highest average yield is consistent with the findings of Wiseman et al (2010) with

Table 3. Effect of cultivars and cultivation pattern on grain yield (t ha-1), dry matter yield (t ha-1), and harvest index (%) Grain yield (t ha-1) Dry matter yield (t ha-1) Harvest Index % Cultivar Cultivation pattern Mean Cultivation pattern Mean Cultivation pattern Mean Ratoon Seed Ratoon Seed Ratoon Seed Inkath 7.281 6.186 6.734 28.2 17.7 22.95 25.85 34.95 30.4 Lilo 6.46 5.654 6.057 26.33 16.83 21.58 24.57 33.59 29.08 Ishtar 5.512 6.204 5.858 30.57 18.07 24.32 18.03 34.35 26.19 Buhoot70 3.848 8.42 6.134 84.33 25.87 55.1 9.75 28.12 18.93 Caviar 3.355 5.519 4.437 43.4 22.83 33.12 10.64 30.29 20.47 LSD 0.05 0.46 0.33 3.57 3.57 2.11 1.62 Mean 5.291 6.397 42.57 20.26 17.77 32.26 LSD 0.05 0.39 3.61 1.17 Ratooning of Sorghum Cultivars on Yield 127 the superiority of ratoon plants by giving it the highest as being an economic method does not need to replant and average of the biological yield compared with normal seeding the resulting soil service and crop compared to the normal method. Also, the results showed significant differences pattern of seed cultivation. The varieties also differed between the cultivars in the study. Buhooth 70 cultivar was significantly among them and superiority in the variety of significantly superior by giving the highest average dry matter Inkath in most of the yield characters. yield (55.10 t ha-1), whereas, Inkath and Lilo cultivars recorded the lowest mean for this character (22.95 and 21.58 REFERENCES t ha-1, respectively). The results also showed a significant Chalabi, Fayeq Tawfiq and Alaa Abdul Hussein Bahadli 2010. Competitiveness of some sorghum varieties of the associated effect of interaction between the two factors of study. The weeds. Journal of Iraqi Agricultural Sciences 41(3): 63-78. treatment of Buhooth 70 cultivars for the ratoon cultivation Chiad, Saddam Hakim and Areej Mohammed Saleh 2018. Seed pattern gave the highest mean of 84.33 t ha-1. While the yield and quality of sorghum ratoon by nitrogen fertilization effect. Journal of Engineering and Technology 36(3): 247-252. cultivars Inkath, Lilo, and Ishtar in normal cultivation recorded Chiad, Saddam Hakim, Mujahid Ismail, Al-Kubaisi and Naim -1 the lowest mean of this trait 17.70, 16.83, and 18.07 t ha , Abdullah Mutlaq 2014. Effect of nitrogen and potassium respectively. fertilizer on seed yield and components in sorghum ratoon. Journal of Iraqi Agriculture (special Issue) 19(6): 111-120. Harvest Index: The results showed a significant effect of the Duncan RR, Bockholt AJ and Miller FR 1981. Descriptive cultivation method, where the normal seed cultivation pattern comparison of senescent and non-senescent sorghum was significantly superior by giving it the highest average genotypes. Agronomy Journal 73: 849-853. harvest index of 32.26 %, while the ratoon method gave the Enserink HJ 1995. Sorghum Agronomy in West Kenya: investigation from a farming systems perspective. Royal Tropical Institute, lowest average harvest index of 17.77% (Table 3). This is due Amsterdam. The Netherlands ISBN 90-6832-100-5. to the superiority of plants produced in the normal seed Gerik TJ and Neely CI 1987. Crop ecology, production, and management: plant density effects on main culm and tiller cultivation method in grain yield (Table 2). Also, the results development of grain sorghum. Crop Science 27: 1225-1230. showed that there were significant differences between Gerik TJ, Bean B and Vanderlip R 2003. Sorghum growth and cultivars in the study. The two cultivars Inkath and Lilo were development. Texas Cooperative Extension and The Texas superior, with no significant differences between them by A&M Univ. System, Temple. KARI 1992. Kenya Agricultural Research Institute, 1992 Annual giving them the highest average of harvesting index (30.40 Report. pp: 54-56. and 29.08%) respectively. While the cultivars Buhooth 70 Kirda C, Kanber R and Tulucu K 1996. Yield response of cotton, and Caviar recorded the lowest mean for this trait, with no maize, soybean, sugar beet, sunflower and wheat to deficit irrigation in nuclear techniques to assess irrigation schedules significant differences between them 18.93 and 20.47%, for field crops. IAEA TECDOC 888: 131-138. respectively, this is due to the superiority of the Inkath and Oizumi H 1977. Role of organic reserves upon the re-growth of Lilo cultivars in plant yield (Table 2), which is directly forage plant: I. Elongation of tiller buds of sorghum (Sorghum bicolor Monarch) plant after cutting as influenced by levels of proportional to the harvesting index. The results also showed organic reserves. Technical Bulletin of Faculty of Horticulture, a significant effect of interaction between the two factors of Chiba University, Japan 25: 55-62. study. The treatment of Inkath cultivars for the normal Ramburana S. Wettergreena T, Berrya SD and Shongwe B 2013. Genetic, environmental and management contributions to cultivation pattern gave the highest mean of harvest index ratoon decline in sugarcane. Field Crops Research 34.95%. While the cultivars Caviar in ratoon cultivation International Journal 146: 105–112. recorded the lowest mean of this trait 3.355 t ha-1, which did Sarhan, Ismail Ahmed, Ziad Abdul Jabbar Abdul Hamid and Sinan Abdullah Abbas 2016. Performance evaluation of three not significantly differ with the Lilo and Ishtar cultivars for the varieties of sorghum under different concentrations of zinc same cultivation pattern 33.59 and 34.35%, respectively. spraying. Journal of Tikrit University for Agricultural Sciences 16(4): 220-231. While the cultivar Buhooth 70 in ratoon cultivation pattern Steel RGD and Torrie JH 1980. Principles and Procedures of recorded the lowest average of this characteristic was Statistics. 2nd Edition. McGraw Hill Book Company Inc., New 9.75%. York. pp. 507. Wilson KSL 2011. Sorghum rationing as anapproach to manage covered kernel smut and the stem borer Chilopartellus. Ph.D. CONCLUSIONS Thesis. Univ. of Greenwich. p: 284. We conclude from the foregoing that the cultivation of Wiseman AJ, Kaminski RM, Riffell SK, ReineckeKJ and Larson EJ 2010. Ratoongrain sorghum and other seeds for waterfowl in sorghum plants by the ratoon method is preferable because sorghum croplands. Annual Conference SEAFWA. pp:106- they significantly outperformed in the yield of the crop as well 111.

Received 08 October, 2019; Accepted 20 November, 2019 Manuscript Number: S-267 Indian Journal of Ecology (2019) 46 Special Issue (8): 128-138 NAAS Rating: 4.96

Genetic Parameters and Hybrid Vigor of Single and Double Hybrids of Maize (Zea mays L.) in Two Plant Densities

Banan Hassan Hadi and Ahmad Nama Abdel-Amir1* Department of Field Crops, University of Baghdad, Iraq 1Directorate of Agriculture, Karbala, Iraq *E-mail: [email protected]

Abstract: A field experiment was carried out at the fields of field crops Department. Collage of Agriculture. University of Baghdad in fall season 2017 in order to estimate the some genetic parameters, including genetic, phenotypic and environmental variations, coefficient of genetic and phenotypic variation and heritability in the broad sense of double, Single hybrids, and their inbred lines of maize of each plant density. As well as estimating the hybrid vigor ratio for single and double hybrids of maize in each plant density for all studied traits (agronomic and yield components). The experiment was conducted according to the randomized complete block design (RCBD) with three replications using the split plot arrangement. Where the two densities (60000 and 8000) plant ha-1 were represented in the main plots and 30 genotypes (five inbred lines, ten single hybrids, and fifteen double hybrids) in the subplots. The results showed genetic variation was higher than environmental variation in the traits of the day number to silking, leaf area, leaf area index, days number to physiological maturity, ear length, grain number per row, grain number per plant, total dry matter, plant growth rate, yield (gm), yield (t ha-1) for the first density (60 thousand plants per hectare); also for the second density (80 thousand plants per hectare), it was higher in the traits of the days number to silking, plant height, ear height, leaf area, leaf area index, physiological maturity, grain weight, individual plant yield, total yield, dry matter, and crop growth rate. The values of phenotypic coefficient of variation P.C.V were higher than those of genotypic coefficient of variation G.C.V for all studied traits, for both densities indicating that the existing variation in addition to the presence of the environmental components. The highest was essentially genetic proportion of heritability in broad sense of the traits of the numbers of days to silking, dry matter, leaf area index and plant growth rate were 0.767, 0.717, 0.710 and 0.705 yield respectively in the low density; whereas, for the traits of plant height, ear height, yield and grain weight were 0.971, 0.829, 0.842 and 0.785, respectively in the high density. Most of the single and double hybrids showed significant hybrid vigor in most of the phenotypic traits under effect of the two plant densities, referred to altered control of over dominance and partial dominance genes of the high parent in traits inheritance.

Keywords: Maize, Plant densities, Over dominance, Hybrid vigor, Partial dominance

The progress and success of any plant breeding and study of genetic features and heritability of the crop and its improvement program depends primarily on the size of the components are necessary for plant breeders to develop the genetic variations found in the plant population under crop for inbred lines and hybrids. Shakoor et al (2007) studied consideration, so the first step in any plant breeding program thirty double hybrids compared with Hicorn984 and Hicornllt is to study genetic variations, as phenotypic changes in any hybrids for plant height attributes, tassling, silking, yield and environment can be measured, but in fact Not only is the its components. Wannows et al (2010). Coefficient impact of genetic variations, but also environmental and phenotypic variation was higher than the coefficient of growth factors, as well as the interaction between them and genetic variation of the studied traits except for the weight of genetic variations, the outward appearance of the plant is a ears as the coefficient of phenotypic variation and the picture of the genetic and environmental impact and their coefficient of genetic variation were close to 27.30 and 27.76 interaction (Bello et al 2007). The study of genetic features respectively. Heritability in the broad sense ranged between gives an idea of ​​the extent of heterogeneity in population and 36.18% for the number of rows and the weight of the ear the efficiency of the selection of genotypes based on the 93.81 followed by 95.88 for the yield. Al-Dulami and Al-Draji external appearance in a population with high genetic (2011) showed in his study of eight pure inbred lines of maize heterogeneity. Hybrids for commercial production have a to produce sixteen single hybrids that the heritability in the narrow genetic base, largely due to commercial pressures broad sense is high for all traits, the highest was for leaf area that force breeders to produce pure genetic material with a 93.6% followed by plant grain yield 82% followed by silking narrow range of variance (Hadi and Wuhaib 2015). 80.3%, this is due to the high value of genetic variation and Moreover, the development of pure lines in breeding program the low value of environmental variation, which makes the is frequent or recurrent cycles to bind or collect favorite transfer of the character to the first generation more likely. alleles in new inbred lines or hybrids (Cunha et al 2012). The Lahmode (2012) showed in his study of eight pure inbred of Some Genetic Parameters of Maize 129 maize introduced in partial diallel in the fall season and then variation coefficient, with the highest PCV for the individual introduced twelve single hybrids for comparison, that the rate plant yield 16.288 followed by the weight of 300 grains 12.028 of heritability in the broad sense was high for all traits and the followed by the height of the ear 10.423 and then the length of highest 99.37% for the yield of grain and 99.15 The number of the ear 8.482. Higher GCV traits 15.619, 11.436, 9.910, and grains, the weight of 1000 grains and the number of rows per 8.116 sequentially, higher values ​​of the genetic variation ear, respectively, is 98.68% and 97.86%. Bello et al (2012) coefficient reflect the extent to which traits respond to the found that the highest phenotypic variance was 2348.42, selection process, while higher values ​​for phenotypic followed by plant height 632.58, grain yield 181.48 and then parameters indicate the significant influence of weight of ears 149.45 and highest genetic variance for traits environmental factors on the phenotypic expression of the 2245.32, 472.54, 114.52 and 136.32 sequentially. The results trait to some degree (Mohanty 2010). A smpile increase in the of Dhannoon and Al-Jumaily (2014) showed that the phenotypic variation coefficient on the genetic variation heritability in the broad sense was high for all traits, the coefficient indicated a slight effect of the environment on the highest number of rows was 99.49%, the plant height heritability of traits, similar results obtained by Rafiq et al 99.06%, the height of ear 98.91%, the length of ear 98.24% (2010), and the values ​​of heritability ratio in the broad sense, and the weight of the seed 96.90%. Kannosh and Al-Dulami which were high in most traits, ranged from 72% for the length (2014) observed in their study six pure inbred lines of maize of the ear to 89% for a grain weight of 100 grain. Abed et al and fifteen hybrids for comparison, they found the heritability (2017) noted that the heritability rate in the broad sense is ratio in the broad sense high for some traits and low for high for all traits, the highest of which was 99.94% followed by others. It was high for individual plant yield 99.08, yield the weight of 300 grains 98.84% followed by the number of growth rate 89.49 and number of grains in row 73.71 due to grains 92.54% and the number of rows of ear 91.11% and high genetic variance and low environmental variance, low then the number of ears per plant 89.72%. The aim of this weight 100 grain 57.92 and female flowering 48.64 due to low study was to estimate the some genetic parameters, genetic variance and high environmental variance. The including genetic, phenotypic and environmental variations, results of Al-Jumaily and Al-Zubaidy (2014) indicated in their coefficient of genetic and phenotypic variation and heritability study of ten pure inbred lines of maize in a half-diallel in the broad sense of each plant density. As well as estimating compared with the commercial (Drachma) variety. In their the hybrid vigor ratio for single and double hybrids in each study, Bandar and Abed (2015) showed that the heritability plant density rate in the broad sense was high for all traits and ranged from 89.30 and 94.32% for the tassling in the spring and fall MATERIAL AND METHODS seasons in succession to 76.69 and 92.56% as the leaf area A field experiment was conducted in the fields of Field for the spring and fall seasons in succession. Mani et al. Crop Department, College of Agriculture, University of (2016), in a study of ninety-eight and four inbred lines for Baghdad, to estimate some genetic parameters and hybrid comparison, confirmed that phenotypic variability is higher vigor by comparing the performance of five parents than genetic variance, and noted the environmental (ZM43WIZE, ZM60, ZM49W3E, ZM19,CDCN5) with their variability of the studied traits. Riberiro et al (2016) estimated single and double hybrids using 30genotypes By conducting the genetic parameters of twenty hybrid maize hybrids for the all the Diallel Crosses between the inbred lines to produce number of row grains, weight of 100 grains and grain yield. Al- ten single hybrids according to the method of Griffing (1956) Roumi (2016) obtained in his study of eight strains and the second fixed model. At the same time, selfing for parental twenty-nine hybrids to compare the inheritance ratio in the inbred lines was conducted to propagation their seeds, broad sense, high, 85.18%, 90%, 90% and 98.75% for the double hybrids were also produced in a later season (fall of character of the paper area and the number of rows in ear and 2016) by crossing between single hybrids to produce 15 the number of grains in the row and the total product double hybrids. At the same time, parents were propagated in sequentially, this confirms the size Genetic variation in the above manner and a Diallel Crosses was conducted transmission and heritability of trait. Nasser et al (2016) in his between parents to produce single hybrids. study of six pure inbred lines of maize found that the Agricultural practices: All soil service operations were heritability ratio in the broad sense is high for all traits, ranging conducted by plowing, smoothing, settling and dividing as from 85,546 for the height of the ear to 99.39 for the weight of recommended. Calcium superphosphate fertilizer was -1 300 grains. Al-Rawi et al (2016), when studying six pure added to 46% P2O5 by (200 kg ha ) P2O5 when soil inbreds and their single crosses, confirmed that the preparation. Urea fertilizer was added to 46% with amount of phenotypic variation coefficient is higher than the genetic (300 kg Nha-1) in three doses at cultivating and one month 130 Banan Hassan Hadi and Ahmad Nama Abdel-Amir after adding the first dose and at flowering. The thicket control Variance. in all seasons was conducted with the use of Atrazine MSg: Mean squares of genotypes substance (85%) (effective substance at 3.25 kg ha-1), MSE: Mean squares for experimental error (Sesamia criteco) was treated using 10% Diazinon, with rate R: Number of replications of (4 kg ha-1) on the two doses, the first one when the plant h2b.s: Broad sense heritability reached the four leaves stage and the second one after The genetic coefficient of variation (GCV) and Phenotypic twenty days from the first control. Crop service operations Coefficient of variation were also applied from irrigation and weeding when ever (PCV) were estimated as in the following equations: needed. PCV= (√σ2 p/x) x 100 In this season, a evaluating experiment was conducted GCV = (√σ2 g/x) x 100 to evaluate the performance of single and double hybrids Estimation of Hybrid vigor (%) plants with their parents (inbred lines) on 21-7-2017. Their The hybrid vigor H.V% was calculated on the basis of the seeds were cultivated (5 inbred lines, 10 single hybrids and first generation deviation from the mean of the highest 15 double hybrids) and with two densities (60, 80 thousand parents as a percentage. plants ha-1), the cultivation was conducted on 21/7/2017 on According to the following equation: the furrows with length of (5 m), the distance between furrow H.V% = (F1 – HP/HP) x 100 and another is 0.75 m, the distance between plant and the HV% = Hybrid vigor other (0.222, 0.166 m) for the two densities mentioned, F1 = The first generation respectively. The randomized complete block design HP = Mean of the highest parent (RCBD) was used with three replications, using the spilt plots arrangement, where the two densities (60000 and 80000 RESULTS AND DISCUSSION plant ha-1) were represented in the main plots while the Genetic parameters: Genetic variability is a key factor for genotypes were sub-plots. Soil and crop service operations plant breeders as it is the basis for selection. As a result, were conducted as mentioned above. A random sample estimation of phenotypic, genetic, environmental, phenotypic consisting of five intermediate plants was collected from each and genetic variability factors and the degree of heritability in experimental unit to study the field traits which included the a broad sense provide important information from which the following traits. The statistical analysis of each trait was most appropriate methods can be derived to improve crops. analyzed by using the RCBD design, using the spilt plots Genetic analyzes were carried out to estimate these arrangement, a significant was tested by using F test. The parameters for each plant density individually. Variance means were compared using the least significant difference analysis Table in 1 and 2 shows that there are highly (LSD), with a significant level of 0.05 and for all the traits significant differences between the genotypes in all studied using Excel program 2014 and statistical program Gestate traits and both plant densities. 2014. Genetic parameters in plant density 60000 plant ha-1: The Statistical standards and genetic parameters: The results of Table 3 show that all the standard error values ​​of the coefficient of variance (CV%) was calculated to estimate the measured traits are low except for the number of plant grains homogeneity of the samples, as well as the calculation of the whose value is high. The low standard error values ​​indicate standard error (SE) to estimate the homogeneity between the the similarity of the attribute data and its proximity to the data for each trait and for each plant density and for each of arithmetic mean. The value of the CV difference of the the investigated trait. The differences in phenotypic, genetic, studied traits was less than 20% and indicated the environmental variations and broad sense heritability were acceptance of homogeneity of the samples. The genetic estimated as mentioned by Singh and Chuadhary (1985) variation values ​​were greater than the values ​​of using SPAR2.0 software according to the following environmental variability in most studied traits, indicating that equations: the heterogeneity of the genotypes was mainly genetic, with σ2 G = (Msg – MSE)/r little environmental impact and therefore inherited. However, σ2 E = MSE some traits such astassling, plant height, ear height, leaves σ2 P = σ2 G + σ2 e number and ears number per plant The values ​​of genetic h2b.s = (δ2 g / δ2 p) x 100 variation were low for environmental differences (64.14%, PCV = (√σ2 p/x) x 100 5.48%, 21.68%, 68.8%, 80% and 88.18%, respectively), GCV = (√σ2 g/x) x 100 indicating that the environment contribution was greater in σ2 P, σ2 g, σ2: Phenotypic, Geneotypic, and Environmental showing variability in these characteristics. Some Genetic Parameters of Maize 131

The genetic variation values ​​were greater than the ​​were lower than the environmental variance coefficient and values ​​of environmental variability in most studied traits, the lowest proportion of phenotypic variability. In such indicating that the heterogeneity of the genotypes was mainly characteristics, environmental conditions have a significant genetic, with little environmental impact and therefore role in controlling and inheriting traits in such highly inherited. However, some traits such as male flowering, plant influenced traits Environmental conditions, the election shall height, The values ​​of genetic variation were low for be slow and take a long process of optimization. This is environmental differences (64.14%, 5.48%, 21.68%, 68.8%, consistent with the results of Al-Rawi et al (2016), Mani et al 80%, and 88.18%, respectively), indicating that the (2017), Hadi et al (2017) and Wuhaib et al 2017. Because the environment contribution was greater in showing variability in phenotypic effect of the effects of genetic factors and these characteristics. These traits also showed a decrease in environmental conditions and the overlap between them, the the ratio of genetic variation to environmental heterogeneity, inheritance of these attributes is different depending on the where it was less than 1. Of the environmental variability of difference of these factors and their degree of impact in female flowering leaf area ,leaf area index ,The number of character. From Table 23, the heritability ratio varied plant grains, the weight of the dry matter, the maturation of significantly, ranging from 0.106 to the number of rows of ear the plant, the rate of plant growth, the plant yield, and the unit to 0.767 for female masturbation, the heritability ratio was yield. The percentage of the soil was higher than one, and its high (0.767, 0.717, 0.710, 0.705 and 0.701) silking, dry percentage ranged from 55.6% to 76.7%. The values ​​of the matter weight, leaf area, plant growth rate and leaf area genetic variation coefficient (GCV) for traits with high genetic sequentially are the same traits with high genetic variance variability and a significant proportion of the phenotypic values ​​and high phenotypic variance values. The days to variance of the PCV were indicative of the extent to which the maturity of the number of grains of the row and the length of genetic variance of phenotypic variance was measured and the ear, the average grain weight, which was also inherited controlled by the traits and its greatest responsibility in the high, ranged from 0.510 per grain weight to 0.688 per unit transmission of traits. The genetic variance coefficient values area and these characteristics had high genetic variation and

Table 1. Mean squares analysis of maize genotypes in the density of 60 thousand plants ha-1 S.O.V Df Tassling Silking Plant Ear Leaves Leaf area Leaf area Ear length Number of height height number index rows ear-1

Rep 2 7.077 5.133 924.4 154.2 1.894 0.019 0.69 0.853 3.743 Genotypes 29 5.124** 38.16** 2061.1** 473.6** 1.360* 0.019** 0.694** 15.41** 1.216* Error 58 2.468 3.502 537.6 141.3 0.696 0.002 0.086 3.235 0.897 S.O.V Df Number of Number of Number of Grain Total dry Days to Plant Yield (gm) Yield (t ha-1) grains row-1 grains plant-1 ears plant-1 weight matter physiological growth rate maturity Rep 2 11.2 38371.5 0.059 0.0001 78.23 31.74 0.066 1588.8 5.475 Genotypes 29 112.1** 50316.3** 0.026* 0.003** 7982.2** 27.53** 0.930** 4394.9** 15.99** Error 58 22.61 8639.7 0.015 0.0009 927.8 5.123 0.113 586.32 2.099

Table 2. Mean squares analysis of maize genotypes in the density of 80 thousand plants ha-1 S.O.V Df Tassling Silking Plant height Ear height Leaves Leaf area Leaf area Ear length Number of number index rows ear-1

Rep 2 28.43 8.044 24.4 74.84 2.285 0.001 0.183 0.707 7.473 Genotypes 29 6.229** 9.096** 1931.8** 558.5** 1.272** 0.012** 0.805** 8.441** 2.207* Error 58 2.628 2.101 16.51 35.71 0.429 0.001 0.134 2.271 1.188 S.O.V Df Number of Number of Number of Grain Total dry Days to Plant growth Yield (gm) Yield (t ha-1) grains row-1 grains plant-1 ears plant-1 weight matter physiological rate maturity

Rep 2 14.06 31970.5 0.014 0.0006 78.87 29.47 0.271 785.1 0.204 Genotypes 29 47.88** 25576.0** 0.009** 0.005** 5101.0** 44.01** 0.709** 1969.7** 13.31** Error 58 16.76 8834.1 0.003 0.0004 664.4 4.523 0.115 292.2 0.782 132 Banan Hassan Hadi and Ahmad Nama Abdel-Amir the ratio of phenotypic variation from 1 to 2.205. The height of row, number of grains per plant, number of ears, weight of dry the plant and the height of the ear had an average heritability matter, plant growth rate, individual plant yield, area unit ratio of 0.485 and 0.439. As for the other traits, their yield, the average weight of the grain and physiological heritability rate was low and ranged from 0.106 for the maturity. Only five traits increased the tassling, leaf area number of rows for ear to 0.263 for tassling. This is because index, number of rows, grain weight rate and physiological they are affected by environmental conditions, which led to maturity. high environmental variation and its contribution to a large Although the values ​​of phenotypic and genetic variation proportion of phenotypic variation and reduced genetic in this density were lower than the low density, the genetic variation. The amount of heritability refers to the degree of variation formed a large percentage of the phenotypic confidence that the genotype can be distinguished by its variability of plant height, ear height, leaf area and leaf area phenotypic expression. High heritability rates for these traits index, dry matter weight, physiological maturity and unit area may be due to the effects of the additive gene act and these yield, respectively (38.66, 4.878, 3, 2.225, 1.913 and 5.341). traits are likely to respond to direct selection. The genetic variation is close to the environmental variation Genetic parameters in plant density 80000 plants ha-1: of the characteristics of the tassling, the height of the plant The SE values ​​and the CV values ​​for the studied traits and the growth rate of the plant. The characteristics of differed in the density of 80 thousand plants ha-1 compared tassling, the number of leaves, the number of rows of ear, the with the density of 60 thousand plantsha-1, whether increased number of grains of row, the number of grains of plant, and or decreased, however, all values ​​are below 20% and are the weight of the grain was the effect of environmental statistically acceptable (Table 4). The values ​​of phenotypic conditions (increase plant density) is evident from the variance and environmental variation in this density differed increase of environmental variation from genetic variation from that of the low density of micro environment due to the and the formation of a high proportion of phenotypic contrast increase of plants per unit area that leads to increased and the contrast ratio was genetic Similar results obtained by competition among plants for growth inputs. This means that Rafiq et al (2010), Wuhaib et al (2017), Hadi et al (2017), plants have become more homogeneous (genetically and Abed et al (2017). phenotypically) under this density. Phenotypic variation in The values ​​of the phenotypic and environmental this density decreased for silking, plant height, plant height, variation coefficient in this density varied from the low density number of leaves, leaf area, ear length, number of grains per depending on the phenotypic, genetic and environmental

Table 3. Some genetic parameters of the studied traits in maize in the density of 60 thousand plants ha-1 Traits S.E C.V S2ɡ S2e S2p S2e / S2ɡ P.C.V G.C.V S.b2 h Day to tassling 0.907 2.707 0.885 2.468 3.354 0.358 3.156 1.621 0.263 Days to silking 1.08 3.17 11.55 3.502 15.05 3.298 6.573 5.758 0.767 Plant height 13.38 10.51 508.09 537.6 1045.7 0.945 14.65 10.21 0.485 Ear height 6.865 12.54 110.73 141.39 252.13 0.783 16.75 11.1 0.439 Leaves Number 0.481 5.203 0.221 0.696 0.917 0.317 5.973 2.934 0.241 Leaf Area 0.027 8.844 0.005 0.002 0.007 2.5 16.43 13.85 0.71 Leaf Area Index 0.169 9.039 0.202 0.086 0.289 2.348 16.54 13.85 0.701 Ear length 1.038 9.586 4.059 3.235 7.295 1.254 14.39 10.73 0.556 Number of row ear-1 0.546 6.3 0.106 0.897 1.003 0.118 6.663 2.17 0.106 Number of grains row-1 2.745 12.94 29.84 22.61 52.46 1.319 19.7 14.86 0.568 Number of grains plant -1 53.66 15.23 13892.1 8639.7 22531.9 1.607 24.6 19.31 0.616 Number of ears plant-1 0.071 11.11 0.003 0.015 0.019 0.2 12.43 5.57 0.2 Grain weight 0.017 9.633 0.0009 0.0009 0.0019 1 13.77 9.841 0.51 Total dry matter 17.58 9.483 2351.4 927.88 3279.3 2.534 17.82 15.09 0.717 Days to physiological maturity 1.306 2.256 7.471 5.123 12.59 1.458 3.537 2.724 0.593 Crop growth rate 0.194 10.47 0.272 0.113 0.386 2.407 19.29 16.2 0.705 Plant yield (gm) 13.98 13.58 1269.5 586.32 1855.8 2.165 27.72 22.93 0.684 Yield t ha-1 0.836 8.78 4.63 2.099 6.73 2.205 27.93 23.16 0.688 Some Genetic Parameters of Maize 133 variations as well as the performance of the genotypes. It was 0.222.The change in heritability values by increasing plant 23.7 for the individual plant yield, 23.03 for the number of density is due to environmental genetic interaction and its plant grains and 22.10 for the unit area. The genetic variation impact on the genetic action and its impact on environmental coefficient increased in this density from the low density of conditions. the characteristics of tassling, plant height, ear height, Days to tassling and days to silking: The results in Tables number of leaves, number of rows and physiological maturity 5 and 6 indicate significant differences in the hybrid vigor (15%, 11%, 22%, 14%, 83% and 31%, respectively), as a values ​​of the studied traits of ten single hybrids and fifteen result of changing plant conditions surrounding it. While the double hybrids in plant densities of 60 and 80 thousand values ​​of genetic variance coefficient for the characteristics plants per hectare. Single hybrids showed negative and of the silking, ear height, leaf area and its index, ear length, significant hybrid vigor in the desired direction in both tassling the number of grains per row, the number of grains per plant, and silkingtraits. The lowest for the hybrid (1 × 4) reached (- grain weight, the weight of dry matter, the rate of plant growth, 10.07%, -19.05%) and (-9.68, -10.00%) respectively in low the individual plant yield and the unit area yield by 59%, 17%, and high densities. Single hybrids exhibited similar behavior 18%, 22%, 35% and 26%. It is clear how the general by giving a negative hybrid vigor earlier than earliest parents environmental conditions (Macro environment and plant at the both densities except the hybrid (2 × 5) and (3× 4) conditions) affect the performance of genotypes as a result of hybrids, They gave positive hybrid vigor to delay tassling and changing variations in values ​​and their interactions. The most silking when increasing plant density to 80,000 plants per closely related characteristics of plant height (11.30 and hectare. The positive hybrid vigor values in these two hybrids 11.44, respectively), and the yield unit of area (22.10 and at high plant density indicates the complete dominance of the 20.28), followed by the characteristics of the height of ear, the dominant genes for parents 4 and 5.While the negative physiological maturity (14.88 and 13.55) and (4.127 and values ​​of the hybrid vigor in the low density indicate the 3.561). This shows that despite the impact of environmental control of the over dominance genes. conditions However, the most influential is genetic variation The double hybrids did not differ much in the ratios of the and these traits are genetically controlled, so the heritability hybrid vigor of the two traits mentioned in the low density has rate was high. The plant growth rate is 0.631, the plant growth been most of them were early in arriving at the silking of their rate is 0.631, and the leaf area index is 0.623, tassling 0.525 parents (single hybrid), While in high density, the double and ear length 0.475. The lowest heritability ratio was hybrids are delayed by their silking than their parents, except

Table 4. Some genetic parameters of the studied traits in maize in the density of 80 thousand plants ha-1 Traits S.E C.V S2ɡ S2e S2p S2e / S2ɡ P.C.V G.C.V S.b 2 h Day to tassling 0.936 2.748 1.2 2.628 3.829 0.456 3.316 1.856 0.313 Days to silking 0.837 2.25 2.331 2.101 4.433 1.109 3.368 2.37 0.525 Plant height 2.346 1.817 638.4 16.51 654.9 38.66 11.44 11.3 0.971 Ear height 3.45 6.137 174.2 35.71 209.9 4.878 14.88 13.55 0.829 Leaves number 0.378 4.121 0.281 0.429 0.71 0.655 5.302 3.336 0.395 Leaf area 0.02 6.688 0.003 0.001 0.004 3 13.27 11.47 0.746 Leaf area index 0.212 8.779 0.223 0.135 0.358 1.651 14.3 11.29 0.623 Ear length 0.87 8.747 2.056 2.271 4.327 0.905 12.07 8.323 0.475 Number of row ear-1 0.629 7.418 0.339 1.188 1.528 0.285 8.411 3.965 0.222 Number of grains row-1 2.364 12.28 10.37 16.76 27.14 0.618 15.63 9.665 0.382 Number of grains plant -1 54.26 18.03 5580.6 8834.1 14414.7 0.631 23.03 14.33 0.387 Number of ears plant-1 0.033 5.435 0.001 0.003 0.005 0.333 6.823 4.125 0.365 Grain weight 0.033 5.435 0.001 0.003 0.005 0.333 6.823 4.125 0.365 Total dry matter 14.88 8.367 1478.8 664.4 2143.3 2.225 15.02 12.48 0.689 Days to physiological maturity 1.227 2.087 13.16 4.523 17.68 2.909 4.127 3.561 0.744 Crop growth rate 0.196 11.22 0.197 0.115 0.313 1.713 18.48 14.68 0.631 Plant yield (gm) 9.87 13.58 559.1 292.2 851.4 1.913 23.17 18.78 0.656 Yield t ha-1 0.51 8.78 4.178 5.342 4.961 5.342 22.1 20.28 0.842 134 Banan Hassan Hadi and Ahmad Nama Abdel-Amir for the hybrids (1 × 2) (3 × 4) and (2 × 5) (3 × 4). The reason showed a positive hybrid vigor at low density and gave the may be due to the control of over dominance genes of the hybrid (3 × 4) the highest value of 25.9%. While at high single hybrid (4 × 3) and its stability in changing densities. density, the single hybrids exhibited a similar behavior to the Plant height and ear height: Hybrid vigor values ​​of plant low density except (3 × 4) which gave a negative hybrid vigor height and ear height varied significantly under plant density of 1.20%.While twelve double hybrids gave negative hybrid (Tables 5 and 6). It was noted that eight single hybrids gave vigor and the rest gave a positive hybrid vigor in the low positive and significant hybrid vigor for plant height as the density while ten even hybrids gave Negative hybrid vigor hybrids (3 × 1),(3 × 2)and (4 × 2) (25.73, 25.64 and 22.61) and five hybrids gave the highest positive hybrid vigor for the sequentially, the hybrids itself also excelled by giving highest double hybrid (2 × 5) (3 × 4) reached 11.03% for hybrid. hybrid vigor of the ear high trait was the highest 29.15% for Single hybrids in low density showed a positive and hybrid (1×3). As for the high plant density, we find that the significant hybrid vigor for eight hybrids reached 6.35% for majority of the hybrids gave a negative hybrid vigor for the hybrids (2 × 4) for the number of rows per ear while seven plant height and ear height it reached the highest -40.48% hybrids gave negative hybrid vigor. At high density, five of the hybrid (3×4).The remaining hybrids gave a hybrids gave positive hybrid vigor, (1 x 5) hybrid gave 0% . ٪and٤١٫٨ positive hybrid vigor above 12.71% for the hybrid (1 × 4 ) for hybrid vigor and the other hybrid gave negative hybrid vigor. the height of the plant and 2.82% for the hybrid (1 × 3) for the While the double hybrid gave a positive hybrid vigor of five of height of the ear. The double hybrids showed a different which the highest hybrid (2 × 3) (1 × 5) as it reached 5.16% in response from their parents than the single hybrids in the low density. While ten double hybrids gave a positive hybrid plant densities. The hybrids in the low plant density gave a vigor, the highest was 12.05% for hybrid (4 × 5) (2 × 3) at high negative and significant hybrid vigor for eleven hybrids, the density. highest of which was(1x2 )(3x5) reached -20%. remaining Grain number per row and grains number per plant: The double hybrids Positively hybrid vigor above the hybrid (4 × significant difference between the genotypes in the average 1) (3 × 5) amounted to 13.37%.In the high density, nine of number of grain number row-1 and grain number plant-1was a double hybrids showed a positive and significant hybrid vigor, significant effect on the variance of the values ​​of the hybrid the highest hybrid (4 × 3) (2 × 5) was 35.13%, while the vigor shown by the single hybrids. was characterized by a remaining six hybrids gave negative hybrid vigor, the highest significant positive hybrid vigor in the desired direction under hybrid was (1 × 3) (4 × 5) reached 16.41%. Five double the low and high densities except the hybrid 1 × 4 in low hybrids gave a positive and significant hybrid vigor for the ear density and hybrid 2 × 3 in high density gave negative values height in both densities. The highest hybrid (1 × 5) (3 × 4) was ​​for the hybrid vigor, while the double hybrids were 16.39% in the low density and 15.03 for the hybrid (2 × 4) (3 × characterized by the majority of negative hybrid vigor of eight 5) in the high density. This may be attributed to the fact that hybrids under low density and twelve hybrids under high the increase in the number of plants per unit area leads to the density. Single hybrids exhibited similar behavior by giving a negative hybrid vigor shown by the genetic or epigenetic high positive and significant hybrid vigor at low and high effect of plant density due to the effect of the gene action. density of the total number of grains of the plant. While the These results are consistent with Wuhaib et al (2016) and double hybrids gave negative hybrid vigor for nine hybrids in Elchneiter et al (2018). low density and eleven hybrids in high density. These results Leaf area and leaf area index: Table 5 and 6 show the indicate that the number of grains and the total number of values ​​of hybrid vigor values ​​of leaf area and its index in the grains were controlled by over dominance at one time and 10 individual and 15 double hybrids in two plant densities. partial dominance at other times. The genetic action was The results indicated a clear control of the over dominance clearly influenced by the growth environment of plant density, genes of the best parent in the heritability of leaf area and its which is consistent with the results Dhoot et al (2017) and index which gave a positive hybrid vigor at low density for all Shenawa (2018). single hybrids. It has a negative hybrid vigor except for (1 × Grain weight: The data of Tables 5 and 6 of the hybrid vigor 4), (1 × 5) and (4 × 5) hybrids which gave a positive hybrid indicate that the weight of the grain was under almost vigor at high density. As for the double hybrid, it gave negative absolute control of the effects of partial dominance. The hybrid vigor to twelve hybrids the hybrid (1 × 4) (2 × 5) gave majority of single and double hybrids gave negative hybrid highest value amounted to -20.55%, while in the high density vigor under both densities. Six single hybrids showed gave seven hybrids positive hybrid vigor and the rest hybrid negative hybrid vigor at low density and seven at high gave negative hybrid vigor. density, while ten double hybrids gave negative hybrid vigor Ear length and number of rows per ear: Single hybrids at low density and twelve hybrids at high density Table 5. The hybrid vigor values of single and double hybrids in maize at a density of 60 thousand plants ha-1 Traits Single hybrid Y(T.H-1) PY(gm) CGR DTM TDM GW NEP NGP NGR NRE EL LAI LA LN EH PH DTS DTT 84.81 18.02 42.19 -8.41 35.44 -14.91 -8.41 -1.9 -0.29 -5.87 5.22 10.6 11.7 5.44 6.68 2.06 -15.71 -3.28 1×2 67.43 59.57 56.76 -7.17 47.31 -12.28 -7.17 44.5 40.84 2.56 21.85 38.29 40 29.15 2.89 25.64 -18.57 -6.56 1×3 75 54.55 65.57 -9.35 49.08 -18.42 -9.35 52.86 45.01 -2.61 30.71 51.96 51.47 27.32 7.32 20.24 -19.05 -7.1 1×4 60.89 39.95 36.92 -6.79 27 -21.05 -6.79 47.13 52.91 -4.91 20.53 18.51 17.96 -26.01 -6.01 -15.17 -14.29 -1.64 1×5 73.14 38.09 32.43 -8.81 20.77 9.68 -8.81 12.55 12.16 -4.7 11.85 12.87 12.77 17.37 7.37 25.59 -18.36 -6.11 2×3 80.38 80 60.94 -9.35 53.67 18.95 -9.35 20.13 11.76 6.35 22.49 9.82 9.57 27 2 25.73 -7.89 -2.78 2×4 64.8 38.02 47.69 7.10- 37.86 -11.46 7.10- 29.5 12.16 3.85 25.9 16 15.96 -3.28 3.28 8.1 -11.62 -2.79 2×5 78.29 39.57 36.49 -7.79 27.82 13.68 -7.79 12.96 14.61 -1.07 5.74 25.24 25.88 19.39 9.39 22.61 -16.91 -4.44 3×4

116.2 40.43 29.73 -6.48 23.72 -10.42 -6.48 18.62 21.3 -2.56 9.26 28.72 29.41 8.33 6.33 11.96 -16.43 -3.89 3×5 Some GeneticParametersofMaize 62.57 79.62 53.85 -9.57 39.29 -4.17 -9.57 62.98 52.16 -2.99 41.9 16.87 17.95 -24.46 4.46 -15.04 -12.12 -3.33 4×5 5.092 6.222 3.975 0.368 3.584 4.471 0.368 6.628 6.163 1.298 3.713 4.278 4.297 6.449 2.449 5.074 1.141 0.577 SE Double hybrids 0.32 0.58 -0.99 2.36 1 -25.93 12.5 9.53 6.64 1.94 7.71 -3.74 -2.83 1.05 -0.27 -9.12 -1.53 -1.69 1X2 3X4 -14.47 -14.37 -1.04 -1.32 -1.45 -17.53 -23.08 -5.33 1.44 -5.7 7.12 -3.64 -3.81 -7.41 -22.15 -20.04 -3.22 -3.39 1X2 3X5 -22.18 -22.04 -7.76 0 -7.05 -14.16 -14.29 -20.52 -12.49 -8.96 -8.36 -15.97 -15.07 -7.56 -1.29 -1.37 -3 -4 1X3 2X4 9.49 9.74 -6.9 -1.33 -7.48 -15 0 -11.8 -8.47 -4.53 -9.12 -7.56 -7.11 -3.87 -2.88 -9.59 -2 -2.29 1X3 2X5 20.48 20.2 -15.52 -1.68 -17.06 -24 -2.7 -33.95 -24.11 -13.75 -17.78 -21.85 -21.52 -3.66 -21 -10.48 -0.49 -0.57 1X3 4X5 6.6 6.69 12.87 4.47 18.56 9.8 8.82 4.97 -1.45 6.7 -6.95 -3.77 -4.55 2.05 3.94 -2.61 2.44 2.94 1X4 2X3 36.61 34.69 8.91 1 13.74 -12.9 9.38 -5.2 -5.02 -3.29 -11.16 -21.1 -20.55 -3.87 -15.49 -17.18 -2.14 -1.14 1X4 2X5 -38.76 -38.64 2.97 -2.64 4.92 1.08 -17.95 -13.47 -10.52 -3.51 -11.69 -11.82 -12.54 2.4 -18.55 -13.46 -2.77 -2.89 1X4 3X5 -13.53 -13.58 0 0 4.99 -19.61 -5.88 4.46 -0.44 5.16 -7.28 -17.92 -17.73 0.41 -5.76 -11.17 -1.45 -1.11 1X5 2X3 2.43 2.28 6.8 -1.99 8.26 -13.27 8.82 6.71 4.56 1.77 -2.95 -11.65 -12.29 4.2 -9.63 -16.99 -3.78 -3.89 1X5 2X4 0.69 0.2 6 -2.65 5.85 0 0 -7.16 0.08 -7.05 -2.35 15.22 14.62 4.07 16.39 11.07 3 -2.78 1X5 3X4 30.69 30.31 16.33 2.41 19.64 3.92 0 10 7.95 2.69 7.45 5.66 4.75 -0.21 3.45 -5.56 2.3 2.37 2X3 4X5 -8.07 -8.39 3.88 2.41 5.47 -25.66 -11.76 5.21 8.42 -3.54 6.89 7.77 7.18 6.93 15.03 11.25 -1.78 -0.57 2X4 3X5 3.05 3.45 27.08 0.66 28.19 27.06 3.13 -1.22 10.14 -10.29 -2.07 -8.26 -8.15 2.04 13.57 3.01 -2 -2.29 2X5 3X4 -1.19 -1.18 -7.92 1.72 -6.26 -5.38 19.35 -16.35 -15.05 -1.34 -6.38 -0.97 -4.55 9.58 3.98 13.37 -0.56 -0.59 4X1 3X5 5.02 4.94 2.79 0.56 3.08 3.83 3.24 2.56 1.5 2.06 2.76 2.66 1.28 3.28 2.79 0.49 0.52 SE Traits*:PH: Plant height, EH: Ear Height, LN; Leaf Number, LA; Leaf Area, LAI: Leaf Area Index, EL: Ear Length, NRE: Number of Rows Ear-1, NGR:Number Grain row-1, NGP: Number Grain Plant-1,NEP: Number ear -1 -1 -1

plant ,GW:Grain Weight, TDM: Total Dry Matter, DTM: Days to Maturity, CGR: Crop Growth Rate, PY: Plant yield, Y(TH ): Yield in area unit (t ha ) 135 136

Table 6. The hybrid vigor values of single and double hybrids in maize at a density of 80 thousand plants ha-1 Traits* Single hybrid Y(T.H-1) PY(gm) CGR DTM TDM GW NEP NGP NGR NRE EL LAI LA LN EH PH DTS DTT 44.17 170.61 96.3 -10 81.31 39.19 6.67 85.43 47.21 6.48 27.86 -3.7 -2.45 0.85 0.47 11.92 -7.62 -5.38 1×2 38.39 9.13 36.36 -9.09 24.79 -20 0 36.21 21.09 12.63 11.48 -11.33 -1.64 3.63 2.82 -13.31 -8.57 -9.14 1×3 89.22 46.38 52.24 -12.73 33.2 6.25 6.68 29.6 17.83 9.76 13.86 16.22 -4.7 -0.95 -0.85 12.71 -11.9 -9.68 1×4 45.29 92.41 35.82 -9.01 22.58 21.62 10 34.24 34.27 0 30.63 10.81 -8 0.896 -18.84 -8.31 -10 -6.99 1×5 17.5 -14.56 12.99 -8.48 3.68 -16.92 0 2.7 -3.11 -3.94 0.62 -20.57 3.38 2.67 -7.14 -16.99 4.3 -0.56 2×3 25.83 17.96 32.84 -9.7 21.17 -4.17 3.33 17.71 9.57 2.08 4.02 -2.96 -8.44 3.58 -1.58 0.68 -0.53 -2.22 2×4 Banan HassanHadiand 41.25 135.17 43.28 -6.91 33.6 50 20 20.07 26.64 -5.18 13.33 -20.74 -10.29 5.77 -31.77 -23.81 1.04 1.11 2×5 46.45 -2.14 10.39 -7.88 1.78 -21.54 3.34 14.18 8.91 4.15 -1.2 -45.33 7.65 4.54 -41.88 -40.48 9.57 0 3×4 63.68 11.46 24.68 -9.01 13.88 -17.69 10 17.87 4.99 5.63 7.29 -12.67 -14.86 7.66 -17.66 -9.82 -1.58 -5 3×5 29.15 21.45 31.34 -8.11 19.89 -7.29 3.35 27.78 19.67 -1.13 1.81 7.62 -10.67 5.09 -15.09 -5.21 1.04 -2.78 4×5 6.439 19.856 7.653 0.494 7.05 8.158 1.889 7.076 4.677 1.827 3.475 5.751 2.17 4.703 4.703 5.067 2.185 1.198 SE Double hybrids -19.36 -19.35 -20.75 -0.66 -18.09 -15.53 0 -16.38 -13.42 -1.96 -4.86 -3.08 -3.11 -3.72 -11.69 -6.78 -2.58 -2.22 1X2 3X4 Ahmad Nama 11.51 11.35 -8.49 0 -5.45 -9.35 -3.03 -12.16 -9.46 -4.48 2.72 -12.21 -11.57 4.61 -0.78 3.38 1.03 1.7 1X2 3X5 16.23 16.36 1.9 0.33 2.19 0 9.68 4.42 1.92 1.79 -4.11 4.51 4.36 -2.08 -9.9 6.24 3.65 3.98 1X3 2X4 -4.72 -4.48 1.9 -5.48 -1.71 -12.61 -11.11 10.28 8.71 1.12 10.48 -2.26 -2.67 -1.25 -11.91 -3.85 -2.06 -2.2 1X3 2X5 51.03 37.6 -9.52 -0.65 -18.63 0 12.9 -7.26 -8.9 0.45 2.24 -11.28 -11.28 3.33 -25.9 -16.41 4.64 4 1X3 4X5 Abdel-Amir 13.92 13.89 0 1.32 7.69 -21.3 3.12 2.99 2.31 0.44 -4.94 7.75 8.22 -1.25 15.83 14.52 1.03 -0.56 1X4 2X3 -15.04 -14.86 0 -4.19 2.21 -2.7 -16.67 -7.33 -8.03 0.89 -14.81 11.63 11.93 -5.73 -9.19 -0.33 1.03 -1.65 1X4 2X5 -26.85 -26.86 -9.8 -1.65 -8.9 -24.92 -6.06 -4.55 -6.18 -3.2 -13.23 -3.82 -3.51 2.58 6.1 2.9 3.17 -1.75 1X4 3X5 -0.31 -0.49 16.48 -0.33 16.12 -3.7 -6.06 -0.17 -9 9.46 -8.77 13.82 11 -3.31 0.83 0.87 0 -1.12 1X5 2X3 -7.1 -6.88 0 -1.32 2.3 -4.35 -3.03 -7.88 -5.24 -2.48 -15.95 -3.82 -1.9 0.62 -14.47 -7.9 0.53 -1.7 1X5 2X4 -6.48 -6.63 3.3 -1.96 4.93 13.33 -9.09 -16.36 -11.45 -5.63 -14.19 2.44 1.87 1.24 -4.18 -5.08 1.55 0 1X5 3X4 28.37 27.97 14.94 -0.99 14.3 -12.96 10 40.19 26.46 12.05 9.73 18.49 19.19 2.92 -7.52 8.13 0.52 -2.79 2X3 4X5 2.32 -7.65 -3.37 3.69 -0.68 -9.78 6.45 -9.03 -13.29 4.99 -4.25 11.45 10.62 16.22 16.57 13.77 5.35 1.14 2X4 3X12 -2.95 6.37 10.42 -3.55 7.85 -8.11 -13.89 -10.44 -15.01 6.65 11.03 25.23 24.8 0.85 32.03 35.13 -0.52 -2.75 2X5 3X4 -10.44 -10.61 -7.84 4.86 -3.59 -10.78 0 -0.04 -2.31 2 -12.87 -1.55 -0.87 5.36 3.63 -4.29 5.41 7.14 4X1 3X5 5.12 4.58 2.56 0.7 2.6 2.41 2.32 3.61 2.78 1.3 2.45 2.78 2.7 1.34 3.77 3.15 0.64 0.76 SE Traits*:PH: Plant height, EH: Ear Height, LN; Leaf Number, LA; Leaf Area, LAI: Leaf Area Index, EL: Ear Length, NRE: Number of Rows Ear-1, NGR:Number Grain row-1, NGP: Number Grain Plant-1,NEP: Number ear plant-1,GW:Grain Weight, TDM: Total Dry Matter, DTM: Days to Maturity, CGR: Crop Growth Rate, PY: Plant yield, Y(TH-1): Yield in area unit (t ha-1) Some Genetic Parameters of Maize 137

Total dry matter, days to maturity and crop growth rate: grain yield and some of iIts components in maize (Zea mays L.) in diallel cross among ten pure lines. Tikrit University of The results indicated that the hybrid vigor was positive and Agricultural Sciences (Special Issue): 248-255. significant for the total dry matter of all single hybrids under AL-Rawi WMH, Anees AHA and Al-Dawoode SAM 2017. Genetic both densities. The hybrid 1 × 4 gave highest hybrid vigor was analysis of half diallel crosses of inbred line of maize (Zea mays L.). Tikrit University of Agricultural Sciences 17(1): 65-78. 49.08% in low density. Hybrid vigor values ​​are also Al-Roumi AH 2016. Estimation of some genetic parameters for decreased by increasing the plant density of all single inbrids be accustomed to poaceae and it's hybrids of (Zea mays hybrids, indicating the environmental impact of increased L.). University of Karbala Scientific 14(4): 87-100. plant density. Ten double hybrids gave positive hybrid vigor at Bandar SJ and Abed RT 2015. Heterosis and gene action of diallel cross among inbred lines of maize. The Iraqi Journal of low density and eight at high density. It is evident from the Agricultural Sciences 46(2): 169-176. hybrid vigor values ​​for the number of days to the Bello D, Kadams AM, Simon SY and Mashi DS 2007. Studies on physiological maturity that the values ​​were negative in the genetic variability in cultivated sorghum (Sorghum bicolor L. Moench) cultivars of adamawa state Nigeria. American- desired direction towards the early maturity of all single Eurasian Journal of Agricultural & Environmental Sciences 2(3): hybrids in both densities and the lowest for hybrids 4 × 5 and 297-302. Bello OB, Ige MS, Afolabi SY, Abdulmaliq S and Mahamood J 2012. 1 × 4 was (-9.57% and -12.73) respectively in both densities. Heritability and genetic advance for grain yield and its While six double hybrids gave negative hybrid vigor in low component characters in maize (Zea mays L.). International Journal ofPlant Breeding Research 2(5): 138-145. density and nine in high density. The lowest for hybrids (3 × 4) Cunha KS, Pereira MG, Goncalves LSA, Berilli AP and Amaral AT (1 × 5) and (1 × 3) (2 × 5) was (-2.65 and -5.48%) in both 2012. Full-sib reciprocal recurrent selection in the maize densities, respectively. populations cimmyt and piranao. Genetic and Molecular Research 11(3): 3398-3408. Single hybrids showed positive and significant hybrid Dhannoon OM and Al-Jumaily AM 2014. Estimation of gene action vigor in the desired direction by increasing plant growth rate and some genetic parameters in maize using triple test cross. in both densities. The highest values ​​for hybrids (1 × 4) and (1 University of Al-Anbar-College of Agriculture –Dept. of Crops 12(2): 182-190. × 2) were 65.57% and 96.50% in both densities respectively. Dhoot M, Dubey A, Ameta RB, Dhoot KD, Kumar R andBadaya VK While ten double hybrids gave a positive hybrid vigor 27.08% 2017.Estimation ohhetrosis for grain yield and architectural for hybrid (3 × 4) (5 × 2) in low density, while in high density traits in yellow seed maize (Zea mays L.). International Journal of Current Microbiology and Applied Sciences 6(7): 4536-4542. nine double hybrids showed a positive hybrid vigor The Elschneiter ZTH 2018.The Molecular Effect of SelfingIn the Genetic highest was 16.48% for hybrid( 2 × 3) (1 × 5). and Eipegentic Diversity and Their Relationship With Hybrid Individual plant yield(gm) and yield in area unit (t ha-1): It Vigor in Maize. Thesis Master. College of Agriculture -University of Anbar, PP.120. is evident from the observation of the hybrid vigor values ​​of Griffing BA 1956. Concept of general and specific combining ability in the grain yield of the individual plant and the total yield that relation to diallel crossing systems. Australian Journal of Biological Sciences 9: 463-493. the hybrid vigor was positive and highly significant for single Hadi BH and Wuhaib KM 2015. Estimation of genetic parameters of hybrids except the hybrid (2 × 3) which gave negative hybrid growth and yield characters of yellow maize (Zea mays L.) vigor at high density and the highest hybrid vigor values under tow levels of nitrogen and plant density. Egyptian Journal of Applied Sciences 30(2): 108-129. ​​were79.62% for hybrid (4 × 5) in low density and for hybrid (1 Kannosh OA and Al-Dulami HJH 2014. Genetic analysis for some × 2) at high density reached 170.61%. The influence of the physiological characters, yield and its components in maize by over dominance genes of the highest parent was significant. using half–diallel. Al- Anbar Journal of Agricultural Sciences 12(4): 227-240. Also, the hybrid vigor values ​​of the unit area yield decreased Lahmode AM, Kathem SH and Al-Rome AH 2012. Estimation of significantly increasing the plant density and this shows the some genetic features in corn using partial mutual crossing Zea mays L. Al-Taqani Journal of Agricultural Science 25(4): 62-75. environmental effect in showing the genetic action of over Mohanty BK 2010. Genetic variability inter-relationship and path dominance. While the double hybrids differed in their hybrid analysis in onion. Journal of Tropical Agriculture 39: 17-20. vigor values ​​ of the total yield between the negative and the Nasser NM, Latheeth HR and Ahmed AM 2016. Genetic analysis for positive in both densities. We find five hybrids gave a combining ability and estimation some genetic parameters for inbred lines and single cross hybrids of maize. Al-Anbar Journal negative hybrid vigor in low density and nine in high density. of Agricultural Science 14(1): 155-162. Rafiq MA, Ali A, Malik MA and Hussain M 2010. Effect of fertilizer and REFERENCES plant densities of yield and protein of autumn planted maize. Pakistan Journal of Agricultural Sciences 47(3): 201-208. Abed NY,Hadi BH, Hassan WA, Wuhaib KM 2017. Growth trait's and Riberiro LP, Teodoro PE, Correa CC, Oliverita EF, Silva FA and yield evaluation of Italian maize inbred lines by full diallel cross. Torres FE 2016. Correlation and genetic parameters in maize The Iraqi Journal of Agricultural Sciences 48(3): 773-781. hybrids. Pakistan Journal of Agricultural Sciences 44(2): 213- Al-Dulami HJH, Al-Draji ZAA 2011. The genetic analysis of 216. combining ability and some parameters in maize (Zea mays L.) Shakoor MS, Muhammad A and Hussain A 2007. Correlation and By using factorial hybridization. The Iraqi Journal of Agricultural path Coefficient studies of some morpho physiological traits in Sciences1 (3): 24-30. maize double crosses. Pakistan Journal of Agricultural Al-Jumaily ARA and Al-Zubaidy KMD 2014. Genetic analysis for Sciences 44(2): 213-216. 138 Banan Hassan Hadi and Ahmad Nama Abdel-Amir

Shenawa MHA 2018.Molecular effect of plant density in DNA Wannows AA, Azzam HK and Ahmad AA 2010. Genetic variances, methylation level, gene expression and some inbred lines and heritability, Correlation and path coefficient analysis in yellow hybrids of maize (Zea mays L.). M.Sc. Thesis (Field Crops maize crosses (Zea mays L.). Agriculture and Biology Journal of Dept.) Al-Anbar university, pp:130. North America 1(4): 630-637. Singh RK and Chaudary BD 1985. Biometrical Methods in Wuhaib KM, Hadi BH and Hassan WA 2016. Hybrid vigor, heterosis, Quantitative Genetic Analysis. Rev. Ed., Kalyani Publishers and genetic parameters in maize by diallel cross analysis. Ludhiana, India, pp. 318. International Journal of Applied Agricultural Sciences 2(1):1-11.

Received 08 October, 2019; Accepted 21 November, 2019 Manuscript Number: S-268 Indian Journal of Ecology (2019) 46 Special Issue (8): 139-144 NAAS Rating: 4.96

Effect of Water Stress on Growth and Green Forage Yield of Ratoon Sorghum

Waleed A.T. El.Fahdawi, Adwea M. Al-Rawi1, Saddam H. Cheyed1, Areej M.S. AL-Mohammedi2 and Adwiya S.M. Al-Rawi2 Department of Field Crop, University of Anbar.,1 Department of Field Crop, University of Baghdad, Baghdad, Iraq 2Ministry of Water Rescuers, Iraq E-mail: [email protected]

Abstract: A field trial was conducted to investigate the effect of three levels of intermittent irrigation (R1 : Irrigation at 50-55% of field capacity, R2 and R3 : withholding one and two irrigations, respectively when control treatment reach 50-55% of field capacity and continue to the end of season), and cultivation pattern (ratoon and by using seeds) on some growth characters and green forage yard of three sorghum cultivars

(Inqath, Lilo and Caffier). Results revealed significant effect of Irrigation treatment on all studied characters where control treatment R1 gave the highest averages of all characters. Also, cultivation pattern significantly affected most characters, where ratoon plants were superior over cultivation by seeds on number of day, from sowing to 75% flowering, tillers number, plant height and leaf area. Cultivars significantly affected all characters where Inqath was superior in the leaf area. However, Caffier was superior in the plant height, number of tillers and green forage yield. It also gave the lowest concentrating of HCN. The interactions at the first and second levels were different in its effects according to the different characters. However, they showed that plants of Caffier were distinguished when sown by ratoon in the characters of green forage yield and HCN concentration, while plants of Inqath sown by seeds were distinguished when sown by direct drilling of seeds in the leaf area less affected by water stress treatments.

Keywords: Ratoon, Hydro cyanic acid, Green forage, Field capacity.

Sorghum (Sorghum bicolor L.) is considered one of the (Cheyed et al 2014). This ratoon does not need land important summer crops as food for human being in some preparation and replanting which makes at economic method developing countries and animal feed as green forages or dry (Ramburan et al 2013,Jassem etal 2016). Ratoon plants are or silage. This crop is characterized by giving plenty yield in characterized by high growth rate, early has vesting and the seasons where forages are scarce. Also, it tolerates competition to the weed plants compared with the plants of drought and grown easily in most soil types. As this crop is direct seeding (KARI 1992, Cheyed et al 2014). Plants of both efficient in utilizing environmental factors, which benefit cultivation patterns are identical in terms of performance and photosynthesis, it is grown successfully under climatic stress tillering (Gerk et al 1990). Many studies recorded that there and relatively high water stress (Mutlaq et al 2015). Water were significant differences between ratoon plants and plays a Key role in the manipulation of all plant activities, direct- seeding in the growth characters i.e., number of days which reflected on growth and yield. Due to the recent from sowing to the flowering stage (AL- Mohammedi 2015), shortage of water resources Iraq, researchers pay attention plant height and green forage (Wiseman et al 2010, Botelho to this factor via using intermittent irrigation as good et al 2010, Cheyed et al 2014 and AL-Mohammedi 2015). technique to save water and increasing its use efficiency The productivity of sorghum is the reflection of (Kirda et al 1996). By controlling water use in irrigation of interaction between genotype, agronomy practices and crops (in terms of magnitude and suitable timing), which environment. Sany et al (1991) indicated that sorghum depend upon the nature and plant growth stages, climatic cultivars were different in reaching 50% flowering stage, conditions, application of chemical fertilizers and the plant height and leaf area (Ahmed and Abood 2016), green capability of soil to capture water (Nielson 2002). AL-Edlabi et forage (Hussein and Dhahir 2006, AL-Jubouri and AL- al (2015) indicated that there were significant differences Zubaidy 2013), harvest index (AL-Edlabi et al 2015), HCN between full and intermittent (deficit) irrigation of Sorghum (AL-Sadi and AL-Rubaie 2014). Eric et al (1995) referred that where full irrigation was superior in the growth and green late mature sorghum cultivars gave highest forage yield forage character. compared with early mature ones. In the light of above, that Sorghum crop also characterized by possibility of it's study was conducted to evaluate the ability of sorghum cutting many times from one sowing. Then, after has cutting, cultivars to tolerate drought via deficit Irrigation under ratoon crop can regrowth from the basal buds, this known as ratoon pattern of cultivation through some growth criteria. 140 Waleed A.T. El.Fahdawi et al

MATERIAL AND METHODS Table 1. Some chemical and physical properties of soil A field trial was conducted during the spring and fall before sowing* Character Value Dissolved positive ions seasons of 2018 at the experimental farm of Department of -1 and negative (Meg l ) Field Crop Sciences- College of Agriculture-AL-Anbar Organic matter 0.50 Meg l-1 Na 11.7 University (alternative site, Abu-Ghraib), to investigate the Carbonate 20.2 Meg l-1 Chloride 38.8 response of three sorghum cultivars (Inqath, Lilo and Caffier) -1 to the ratoon cultivation (spring ratoon- full ratoon) compared Gypsum 0.31 Meg l Bicarbonate 4.10 with the convention method by seeds (control) under the Available nitrogen 0.02% Sulphar 12.1 influence of deficit irrigation. Available phosphorus 41.3% Magnesium 17.5

Three treatments followed were; R1 -full irrigation at Available potassium 73.6% Calcium 23.0

-1 -1/2 100% depletion of available water (control treatment); R2 - SAR 2.52 (Meg l ) Soil texture (g -1 withholding one irrigation after reaching 50-55% of field m kg ) PH 7.13 ds m-1 sand Silt Clay capacity and continue to the end of season and R3 - withholding two irrigation after reaching 50-55% of field ECe 4.31ds m-1 280 498 222 capacity and continue to the end of season. Texture : silty Irrigation treatment were chosen in this method due to its Volumetric moisture content Volumetric moisture simplicity in the field water-deficit treatment were applied content (33 K Bascal) -3 -3 3 -3 when plants reached six leaf stage. Treatments were 0.15 cm cm 0.32 cm cm distributed according to the Randomized Complete Block *Soil analysis was preferment at the Central / Lab.; Coll. of Agric. Engineering Sci. Univ. of Baghdad Design in the arrangement of split - split plots design with three replicates. Water - deficit treatments occupied the mine RESULTS AND DISCUSSION plots. Cultivation patterns occupied the sub- plots while Number of days from sowing to 75% flowering (day): It is cultivars occupied the sub- sub plots. Seeds of all cultivars clear from Table 2 that there was significant effect of irrigation were sown in both seasons with 133300 plant ha-1. Soil and cultivation treatments, cultivars and bi- and triple samples at (0 -30 cm) depth were taken before sowing for interactions on this character. For irrigation treatments, R1 physical and chemical analysis (Table 1). After seed bed treatment gave significantly less average of number of days preparation, soil divided to the experimental units with from sowing to 75% flowering (74.45) compared with 82.67 dimension 3*205 m to give 6 (six) rows 50 cm a part and 15 day for R3 treatment. Plants of ratoon cultivation followed cm between main plots was left. Three (3) seeds were sown similar trend as irrigation treatment (82.46), which gave on 2/4/2018, and then they were thinned to one plant per hill 74.21 day compared with 82.46 day for direct seeding. These at two stages, the first after emergence and the second after results agree with the finding of AL-Mohammedi (2015), -1 ten days from the first one. Nitrogen fertilizer 200 kg ha as where plants of ratoon gave 56.7 days compared with 81.0 urea (40% N) was applied at three stage, the first at the day for direct seeding. In fact, KARI(1992) and AL- completion of field emergence, at 4-6 leaves and at beginning Mohammedi (2015) mentioned that ratoon cultivation of flowering. Super phosphate fertilizer 45% P2O5 (100 kg p method were characterized by their earliness in growth and -1 ha ) was added with the soil before sowing. All plots were harvesting with 10-25 day compared with the direct seeding irrigated immediately after sowing. Plants were cut at 7-10 cm method of sorghum. In respect to the cultivars, table2 shows height at maturity to protect new buds and left to the next fall that Lilo cultivar recorded the lowest average of this season to obtain the ratoon. Seeds of sorghum were sown at character, 77.46 day compared with 78.88 and 78.67 day for the completion of cutting ratoon and watered at the same time Inqath and Caffire, respectively with no significant on July 22, 2018. Hand weeding were done as necessary. differences between them. These varietal differences may be Number of days from sowing to 75% flowering; plant due to their genetical differences i.e. early or late in maturity. height was measured after full flowering for 5 plants taken Concerning interaction, the control and ratoon pattern randomly from middle rows for each treatment from soil recorded the lowest average 66.07 day, while the interaction surface up to the tip of head; number of tillers and green between irrigation treatment R3 and direct seeding gave the forage yield were counted for full row and HCN concentration highest value (85.22 day). For interaction between irrigation plant-1 was estimated according to the Haskins et al (1978). and cultivar, R1 treatment and Caffier recorded the lowest Statistical analysis: All data were analysed for ANOVA by value (69.00 day) compared with highest 86 day for R3 using statistical programme Genstat and least significant treatment and Caffier cultivar. differences (LSD) at 0.05 probability level. For interaction between cultivation pattern and varieties, Water Stress on Yield of Ratoon Sorghum 141

Table 2. Effect of irrigation treatments and cultivation pattern Table 3. Effect of irrigation treatments and cultivation pattern on 75% flowering (day) of sorghum cultivars on 75% flowering (day) of sorghum cultivars Irrigation Cultivatio Cultivars (C) A × R Irrigation Cultivation Cultivars (C) A × R treatment (R) n pattern treatment (R) pattern (A) Caffier Lilo Inqath (A) Caffier Lilo Inqath

R1 Ratoon 69.28 67.11 61.83 66.07 R1 Ratoon 129.33 130.33 173.20 144.29 Seed 86.33 86.00 76.17 82.83 Seed 156.67 148.33 202.57 169.19 R2 Ratoon 76.00 74.67 78.67 76.44 R2 Ratoon 125.31 127.87 166.51 139.90 Seed 81.33 74.33 82.33 79.33 Seed 147.67 146.40 185.75 159.94 R3 Ratoon 76.33 79.33 84.67 80.11 R3 Ratoon 109.66 122.07 156.48 129.40 Seed 84.00 83.33 88.33 85.22 Seed 129.57 124.89 168.47 141.11 Cultivar Average 78.88 77.46 78.67 Cultivar Average 133.03 133.32 175.56 LSD 0.05 C 0.90 LSD 0.05 C 2.78 A × R 1.24 A × R 3.71 A × R×C 2.24 A × R×C NS Irrigation Cultivars Average Irrigation Cultivars (C) Average Treatment (R) (C) (R) Treatment (R) (R). Caffier Lilo Inqath Caffier Lilo Inqath

R1 77.81 76.55 69.00 74.45 R1 143.00 139.33 187.88 156.74 R2 78.67 74.50 80.50 77.89 R2 136.49 137.14 176.13 149.92 R3 80.17 81.33 86.50 82.67 R3 119.61 123.48 162.68 135.25 LSD 0.05 R 0.77 LSD 0.05 R 3.70 R×C 1.49 R×C 4.82 cultivation Cultivars Average cultivation Cultivars (C) Average pattern (A) (C) (A) pattern (A) (A) Caffier Lilo Inqath Caffier Lilo Inqath

Ratoon 73.87 73.70 75.06 74.21 Ratoon 121.43 126.76 165.40 137.86 Seed 83.89 81.22 82.28 82.46 Seed 144.63 139.87 185.73 156.75 LSD 0.05 A 0.94 LSD 0.05 A 1.64 A×C 1.38 A×C 3.46 it was noticed that ratoon plants of Inqath and Lilo cultivars This may be due to the earliness of ratoon plants in growth gave the lowest value of interaction 73.87 and 73.70 days, and maturity to reach 75% flowering stage (Table 3) as a respectively with no significant difference between them result to the shortening of life cycle of these plants. These compared with 83.89 day for Inqath sown by direct seeding. results are in agreement of AL-Mohammedi (2015). Caffire For triple interaction, control irrigation treatment of ratoon cultivar gave the highest average of plant height of 178.37 cm cultivation for Caffier recorded the lowest value of 61.83 day compared with Inqath and Lilo cultivars (133.03 and 133.32 compared with 86.33 day for R3 of Caffier sown by direct cm, respectively). These varietal differences were due to seeding with no significant differences between R1 treatment their differences in genetic-make up and number of days from of Inqath sown by direct seeding. sowing to the 75% flowering stage (Table 3). This results Plant height (cm): Table 3 indicated that there was were in accordance with the findings of Hussein and Thahir significant effect of irrigation treatments, cultivation pattern, (2006), AL-Jubory and AL-Zubaidy (2013), Ahmed and cultivars and bi interaction on plant height. For irrigation Abood (2016) who reported varietaldifferences between treatment, R1 gave the highest average of plant height of sorghum cultivars in plant height. Concerning interaction, two

156.74 cm compared with 135.25cm for R3 treatment. These factor interaction (irrigation treatment × cultivation pattern) result were in agreement with the finding of Hadif and Lafta showed significant effect where R1 and direct seeding gave (2013) that full irrigation increased plat height. Direct seeding the highest average 169.19 cm compared with 129.40 cm for increased plant height by giving the highest average of R3 and ratoon cultivation. For bi interaction (irrigation and

156.75 cm compared with 137.86 cm for ratoon cultivation. cultivars), R1 and caffier gave the highest value 187.88 cm 142 Waleed A.T. El.Fahdawi et al

compared with 119.61 cm for R3 and Inqath cultivar. different genetic make- up of these cultivars (Hussein and Concerning cultivation pattern and cultivars, plants of direct Thahir 2006, AL-Jubory and AL-Zubaidy 2013). seeding of Caffier recorded the highest value 185.73 cm Number of tillers m-2: Table 5 shows significant effect of compared with 121.43 cm for ratoon cultivation of Inqath. irrigation treatments, cultivation pattern, cultivars and bi Leaf area (cm2): Table 4 showed significant effect of interaction except bi interaction of irrigation × cultivars and irrigation treatments, cultivation method, cultivars, two factor triple interaction. The number of tillers m-2 declined with the -2 interaction and three factor interaction on this character. For increase of irrigation treatments i.e., 12.14 tiller m at R1 2 -2 irrigation treatment, R1 gave the highest value 422.3 cm treatment compared with 17.94 tiller m at R3. 2 compared with 3574.2 cm for R3 treatment. This may be due This reduction in tiller number may be due to the water that water stress adversely affects the growth and size of stress which adversely affects growth and development of cells especially the leaves which in turn reduced the plant leaf lateral bud, of sorghum AL-Muhammadi (2015) reported that area this was confirmed by the fin clings of Hadif and Lafta bud formation in sorghum depends up on the availability of (2013). Concerning, cultivation method, direct seeding growth pastors like water and nutritional elements. recorded the highest average 4000.6 cm2 compared with Table 5 indicates that ratoon cultivation recorded the 3682.0 cm2 for ratoon cultivation. This could be due to the highest average 25.71 tiller m-2 compared with 15.56 tiller m-2 increase of tiller number and fertile tiller number of ratoon for plant resulted from direct seeding. The superiority of plants compared with direct seeding method which in turn ratoon plants may be due to the limited apical dominance and reflected in the total leaf area of plant (Hammer et al 1987). promotion of lateral buds and then increased tiller number For cultivars, plants of Inqath gave the highest value of compared with direct seeding. This result confirms the 3999.9 cm2 compared with the lowest value 3673.6 cm2 for findings of Cheyed and AL-Mohammedi (2016). Who found Caffier plants. This varietal difference may be due to the that ratoon plants of sorghum were superior in their tiller

Table 4. Effect of irrigation treatments and cultivation pattern Table 5. Effect of irrigation treatments and cultivation on on leaf area (cm2) of sorghum cultivars number of tillers m2 of sorghum cultivars Irrigation Cultivation Cultivars (C) A × R Irrigation Cultivation Cultivars (C) A × R treatment pattern (A) treatment pattern (A) (R) Caffier Lilo Inqath (R) Caffier Lilo Inqath

R1 Ratoon 4034.0 3892.3 3655.0 3860.4 R1 Ratoon 28.20 26.33 31.54 3860.4 Seed 4595.3 4540.0 4605.0 4580.1 Seed 17.70 16.83 18.21 4580.1

R2 Ratoon 3787.3 3599.0 3400.3 3595.6 R2 Ratoon 25.89 23.11 28.43 3595.6 Seed 4026.0 3930.3 3633.3 3863.2 Seed 16.51 14.55 16.46 3863.2

R3 Ratoon 3830.0 3606.3 3333.3 3589.9 R3 Ratoon 22.56 20.06 25.22 3589.9 Seed 3723.3 3537.7 3414.3 3558.4 Seed 13.79 11.71 14.31 3558.4 Cultivar Average 3999.3 3850.9 3673.6 Cultivar Average 20.77 18.77 22.36 LSD 0.05 C 42.1 LSD 0.05 C 0.39 A × R 68.2 A × R 0.44 A × R×C 103.1 A × R×C NS Irrigation Treatment Cultivars (C) Average Irrigation Treatment Cultivars (C) Average (R) (R). (R) (R). Caffier Lilo Inqath Caffier Lilo Inqath

R1 4314.7 4216.2 4130.0 4220.3 R1 22.95 21.58 24.87 23.14

R2 3906.7 3764.7 3516.8 3729.4 R2 21.20 18.83 22.45 20.83

R3 3776.7 3572.0 3373.8 3574.2 R3 18.18 15.88 19.76 17.94 LSD 0.05 R 65.1 LSD 0.05 R 0.31 R×C 78.0 R×C NS cultivation pattern (A) Cultivars (C) Average cultivation pattern (A) Cultivars (C) Average (A) (A) Caffier Lilo Inqath Caffier Lilo Inqath Ratoon 3883.8 3699.2 3462.9 3682.0 Ratoon 25.55 23.17 28.40 25.71 Seed 4114.9 4002.7 3884.2 4000.6 Seed 16.00 14.36 16.33 15.56 LSD 0.05 A 36.6 LSD 0.05 A 0.33 A×C 57.17 A×C 0.52 Water Stress on Yield of Ratoon Sorghum 143 number compared with direct seeding plants. Concerning Table 6. Effect of irrigation treatments and cultivation pattern -1 cultivars, Caffier recorded the highest average of tiller on green forage yield (t ha ) of sorghum cultivars number m-2 22.36 compared with 18.77 of Lilo cultivar. This Irrigation Cultivation Cultivars (C) A × R treatment pattern (A) may be due to the differences in the genetic-make up which (R) Caffier Lilo Inqath reflected in the growth characters. Also, Caffier was R Ratoon 20.07 19.07 26.38 21.84 characterized on forage crop with vigorous vegetative 1 growth compared with Inqath and Lilo. Seed 12.99 12.83 21.31 15.71 -1 R Ratoon 19.17 18.35 23.96 20.49 Green forage yield (t ha ): It is clear from Table 6that there 2 was significant effect of irrigation treatments, cultivation Seed 12.33 12.28 18.91 14.51 pattern, cultivars bi interaction of irrigation ×cultivar, cultivation R3 Ratoon 17.34 15.86 19.76 17.65 pattern × cultivar and the triple interaction on that character. Seed 9.65 9.26 16.37 11.76

For irrigation treatments, R1 gave the highest average of green Cultivar Average 15.26 14.61 21.11 -1 -1 forage yield 18.77 t ha compared with 14.71 t ha for R3 LSD 0.05 C 0.27 irrigation treatment. This reduction in the green forage yield A × R N.S with the increase of water stress was attributed to the A × R×C 0.67 reduction in plant height (Table 3), leaf area (Table 4) and tiller -2 Irrigation Treatment Cultivars (C) Average number m (Table 5) which in tour, reduced the size of (R) (R) vegetative growth. This results confirmed the findings of AL- Caffier Lilo Inqath Edlabe et al (2015). It is also clear from Table 6 that ratoon R1 16.53 15.95 23.84 18.77 pattern was superior in giving highestgreen forage yield 20 t R 15.75 15.32 21.44 17.50 ha-1 compared with 13.99 t ha-1 for plants resulted from direct 2 R 13.50 12.56 18.06 14.71 seeding. This may be due to the superiority of ratoon plants in 3 -2 LSD 0.05 R 0.17 the number of tiller m (Table 5) where 39.48% increase in this character over the plants of direct seeding. This result was in R×C 0.40 agreement with the finding of AL-Mohammedi (2015) who cultivation pattern (A) Cultivars (C) Average (A) found significant increase in the green forage yield of ratoon Caffier Lilo Inqath plants compared with direct seeding. Ratoon 18.86 17.76 23.37 20.00 Concerning cultivars, Table 6 shows that Caffier recorded the highest average of green forage yield 21.11t ha- Seed 11.66 11.46 18.86 13.99 1 compared with 14.61 t ha-1 for Lilo cultivar. This may be due LSD 0.05 A 0.37 to superiority of Caffier in plant height (Table3), number of A×C `0.45 tiller m-2 (Table 5), which, in turn, reflected in the increase of vegetative growth weight compared with other cultivars. This compared with 59.13 mg kg-1 for ratoon method. This may be result was in agreement with findings of Hussein and Thahir due to the reduction of nitrogen accumulation in the plant (2006) and AL-Edlabi et al (2015). tissues of ratoon method as a result of increased this size of -2 For interaction, R1 irrigation treatment × Caffier cultivar vegetative growth (number of tiller m ), which in turn reduced recorded the highest average of 23.84 t ha-1 compared with its HCN content. These results were in agreement with the -1 lowest value of 12.56t ha for R3 ×Lilo cultivar. Ratoon plants findings of Cheyed and AL-Mohammedi (2016) who found ofCaffier gave the height value of 23.37 t ha-1 compared with significant reduction in the HCN content of ratoon plants 11.46 and 11.66t ha-1 for direct seeding of Lilo and Inqath, compared with the plants resulted from direct seeding. respectively with no significant differences between them. Concerning cultivars, Lilo gave the highest average 75.12 mg -1 -1 For triple interaction, R1 treatment × ratoon pattern ×Caffier kg compared with 61.73mg kg for Caffier. This result was in recorded the highest value 26.38 t ha-1 compared with 9.26 t agreement with the findings of AL-Sady and AL-Rubaie -1 ha for R3 treatment ×Lilo× direct seeding. (2014) where significant differences were found in the HCN HCN (mg kg-1): Results of Table 7 shows only significant concentration in three sorghum cultivars. Bi interaction effect of irrigation treatments, cultivation pattern, cultivars between cultivation pattern ×cultivars had significant effect on and bi interaction between cultivation pattern and cultivars on HCN as direct seeding of Lilo recorded the highest value -1 -1 the character. R1 treatment gave the lowest average HCN 84.97 mg kg compared with 45.88 mg kg for ratoon method -1 -1 67.06 mg kg compared with 72.73 mg kg for R3 treatment. of Caffier. Also, direct seeding gave the highest value 80.73mg kg-1 It is concluded that the irrigation system had significant 144 Waleed A.T. El.Fahdawi et al

Table 7. Effect of irrigation treatments and cultivation pattern AL-Jubory RK and Al-Zubaidy SA 2013. Effects of plant densities on on HCN (mg kg-1) of sorghum cultivars growth and fodder yield of two cultivars of sorghum (Sorghum bicolor (L.) Moench). EL-Phrates Journal of Agricultural Irrigation Cultivation Cultivars (C) A × R Science 5(2): 111-120. treatment (R) pattern (A) Caffier Lilo Inqath Al-Mohammedi AMS 2015. Effect of production style and nitrogen fertilization on growth and yield of sorghum. M.Sc. Thesis. R1 Ratoon 63.71 63.38 42.39 56.49 Faculty of Agriculture, Baghdad University, 78p. Seed 75.38 82.54 74.94 77.62 AL-Sadi ILR and AL-Rubaie MMA 2014. Effect of cutting numbers in R Ratoon 65.52 64.52 45.65 58.56 the yield and quality of green yield to several feed cultivers of 2 Sorgum bicolor L. Moench. EL-Phrates Journal of Agricultural Seed 81.27 84.61 78.39 81.42 Science 6(4): 337-347.

R3 Ratoon 69.45 67.95 49.59 62.33 Botelho PR, Pires OD and Jayme DG 2010. Evaluation of genotypes Seed 82.24 87.75 79.42 83.13 of sorghum in first cutting and regrowth for silage production. Revista Brasileira de Milhoe Sorgo 9(3): 287-297. Cultivar Average 72.93 75.12 61.73 Cheyed SH and AL-Mohammedi AMS 2016. Same growth LSD 0.05 C 1.02 parameters of sorghum ratoons under nitrogen fertilizer. The A × R N.S International Journal of the Environment and Water 5(1): 83-89. Cheyed SH, Al-kubaisy MI and Motlag NA 2014. Effect of nitrogen A × R×C N.S and potassium fertilizers on seed yield and its components of Irrigation Treatment (R) Cultivars (C) Average sorghum ratoon. Iraqi Journal of Agric. Ministry of Agriculture (R) 19(6): 111-120. Caffier Lilo Inqath Eric PB, Cupina V and Pataki I 1995. Effect of sowing and cutting term on the. Yield and quality of varieties of sudangrass. R1 69.54 72.96 58.66 67.06 Savremena Poljoprivreda (yugo –Slavia) 43(1): 53-59.

R2 73.39 74.56 62.02 69.99 Gerik TJ, Rosenthal WD and Seavey WF 1990. Phenology, early growth, and yield of planted ratoon grain sorghum. Field Crops R3 75.84 77.85 64.50 72.73 Research 23: 205-219. LSD 0.05 R 2.02 Hadif WM and Lafta LR 2013. Effect of planting dates and levels of moisture stress on some vegetative and yield characteristics of R×C N.S sorghum. Journal of University of Thi-Qar 8(2):18-35. cultivation pattern (A) Cultivars (C) Average Hammer GL, Hill K and Schrodter G 1987. Leaf area production and (A) senescence of diverse grain sorghum hybrids. Field Crops Caffier Lilo Inqath Research 17: 305-317. Haskins FA, Gorz HJ and Johnson BE 1987. Seasonal variation in Ratoon 66.23 65.28 45.88 59.13 leaf hydrocyanic acid potential of low high dhurrin sorghum. Seed 79.63 84.97 77.58 80.73 Crop Science 27: 903–906. Hussein AS and Thahir AT 2006. Response of two varieties of LSD 0.05 A 1.13 sorghum (Sorghum bicolar L) to the sowing distance and A×C 1.51 regions. Technical Journal 19(3): 55-63. Jassam KT, Ali IH, Cheyed SH and Naif SH 2016. Financial analysis of sorghum crop planted by ratoon method. Association of th effect on sorghum growth with the highest negative effect of Genetic and Environmental Resources. The 4 International Scientific Congress. Cairo, Egypt 4(2): 11-18. R irrigation method compared with the R method, which 3 2 KARI 1992. Kenya Agricultural Research Institute, Annual Report. showed less negative effect. For cultivation pattern, recorded pp: 54-56. superiority in the characters of number of days from sowing to Kirda C, Kanber R and Tulucu K 1996. Yield response of cotton, 75% flowering, number of tillers, green forage yield and HCN. maize, soybean, sugarbeet, sunflower and wheat to deficit irrigation In nuclear techniques to assess irrigation schedules However, direct seeding was superior in the plant height and for field crops. IAEA. TECDOC 888: 131-138. leaf area. There were clear differences between cultivars Mutlag A, Saleem QA and Kadhum FA 2015. Effect of deficit irrigation where Inqath was superior in the leaf area, while Lilo was and potassium fertilizer on water consumption and crop coefficient of sorghum. The Iraqi Journal of Agricultural Science superior in the number of days from sowing to the 75% 46(5): 937-957. flowering. However, Caffier was superior in the plant height, Nielsen RL 2002. Drought and heat stress effects on corn pollination. number of tillers, green forage yield and HCN concentration. Purdue Coop. Ext. Ser. URL: http://www.agry.purdue.edu/ext/ corn/pubs/corn-07.htm. Ramburana S, Wettergreena T, Berrya SD and Shongw B 2013. REFERENCES Genetic, environmental and management contributions to Ahmed YA and Abood NM 2015. Response of two varieties of ratoon decline in sugarcane. Field Crops Research, sorghum (Sorghum bicolor L. Moench) to plant density. AL- International Journal 146: 105–112. Anbar Journal of Agricultural Science1 4(2): 188-203. Sang TG, Sak SL and Jun HB 1991. Forage productivity of silage AL Edelbi R, Sabouh M and Al Lahham 2015. Evaluation of some corn and – sorghum at different planting dates in rice black – productivity and quality traits of some sorghum (Sorghum streak dwarf virus pre – valemt area. Journal of Korean Grass bicolor L. Moench) genotypes under water stress conditions Science 11(2): 129-136. during grain filling stage. Journal of Jorden in Agricultural Wiseman AJ, Kaminski RM, Riffell SK, Reinecke KJ and Larson EJ Science 4(2):11-18. 2010. Ratoongrain sorghum and other seeds for waterfowl in sorghum croplands. Annual Conf. SEAFWA.pp:106-111. Received 19 September, 2019; Accepted 20 November, 2019 Manuscript Number: S-269 Indian Journal of Ecology (2019) 46 Special Issue (8): 145-150 NAAS Rating: 4.96

Response of Growth and Yield Characters of Three Sorghum Cultivars to the Biological Catalyst (Amino triplex)

Ahmed H. Ahmed and Sinan A. Abbas Field Crop Department, College of Agriculture,University of Anbar, Anbar, Iraq E-mail: [email protected]

Abstract: Two field experiments were conducted to study response of growth and yield characters of three sorghum cultivars to the biocatalyst (amino triplex) using different concentrations. The main plots included the bio-catalyst concentrations (0, 2, 4 and 6 gm L-1), while the sorghum cultivars (Giza 113, Rabeh and Warkaa). The results showed that the Giza cultivar was significantly superior in plant height as it gave the highest value of plant height in both seasons, also it gave the highest values of leaf area in the spring season, the highest number of grains per panicle that in fall season, the highest weight of 500 grain reached in both seasons and grain yield in fall season, while Warkaa cultivar was superior by giving the highest number of grains per panicle and highest yield grains in spring season. It also had the highest protein percentage in both seasons. In terms of spraying concentrations, the concentration of 6 gml-1 was superior and gave the highest values of plant height, leaf area, grain yield, protein percent, the same concentration was superior also in number of grain per panicle in the spring season and the weight of 500 grains in fall season. The effect of interaction between cultivars and biocatalyst concentrations was significantly in the character of leaf area in both seasons, the number of grains per panicle, the weight of 500 grains in the spring season and the percentage of protein in fall season only.

Keywords: Grain crop, Fertilizer, Bio-catalyst, Seeds protein

The sorghum crop Sorghum bicolor L. Moench is the fifth lack of arable land and environmental fluctuations. The grain crop in terms of economic importance and globally increase in agricultural land, coupled with the problems of production. It is a major economic source for many of the providing arable land and providing water, has become a world's population, especially in developing countries in arid major global crisis. The use of plant nutrients in terms of their and semi-arid regions in Asia and Africa. It is also a staple type and concentrations and the use of foliar feeding on the food, energy source, protein, vitamins and minerals for plant is one of the alternative scientific methods for the use of people living in poor areas. It is useful for people with fertilizers that require increasing the number of irrigation diabetes as well as for those with allergies to gluten and is times as well as its negative effects on the environment and considered as antioxidants. sorghum grains contain 11.3% agriculture. Therefore, bio-fertilizers are a modern method of protein, 3.3% fat and 56-70% starch. It is relatively rich in iron, increasing and safety of the food product, improving its zinc, phosphorus and vitamins (Ratnavathi 2016). Its grains quality and increasing the plant's ability to resist are also used in poultry diets as well as green fodder in times environmental stresses by regulating plant content of plant of forage scarcity during the summer to fill the shortage in the growth regulators (PuGuxin et al 2008). production of forage in the livestock sector, which contributed In light of the new climate challenges of rising significantly to the solution of this problem, as it is temperatures and lack of rain. The search for crops that resist distinguished from the rest of the summer crops tolerance to these conditions has become necessary, including sorghum drought and lack of water (crop adaptability in dry land crop. It is expected that this crop will increase significantly as conditions). Despite the importance of this crop, the a source of human and animal nutrition and the production of cultivated areas decreased by 0.15 million hectares annually ethanol. also, the selection of the most suitable variety for the from the mid-1980s until recently. Subsequently, the prevailing environmental conditions and the most responsive producing countries began to improve cultivars of sorghum to the addition of nutrients within those conditions is an species due to the recent environmental fluctuations important means that will increased the rate of production of (Hariprasanna and Rakshit 2017). The productivity of this this crop per area unit and improve the chemical composition crop was 1.57 t ha-1 in 2015 (USDA 2016). of seeds (quality). In Iraq, this crop has received attention only recently by The objective of this study is to determine the suitable some workers for research. Vertical expansion is one of the cultivars FOR best productivity and quality, and test the effect solutions that need to be studied to solve the problem of food of the biological catalyst (Amino triplex) on the growth, yield shortage in the world due to the increase in population and and quality of different cultivars. 146 Ahmed H. Ahmed and Sinan A. Abbas

MATERIAL AND METHODS Table 1. Some chemical and physical properties of Two field experiments were conducted in the spring and experiments soil before planting in spring and fall seasons of 2018 fall seasons of 2018 in one of the agricultural fields in Indicators Values of both seasons of 2018 Units Habbaniyah district-Anbar province. in order to study Spring season Fall season response of growth and yield characters of three sorghum (PH) 7.21 7.21 -- cultivars to the growth regulator (amino triplex) using Electrical Conductivity EC 1.72 1.72 ds/m different concentrations as well as control treatment. The layout of the experiment was RCBD design using split plot Available N 26.13 26.13 Ppm pattern with three replicates. The main plots included the bio- Available P 18.72 18.72 Ppm catalyst concentrations (0, 2, 4 and 6 gm l-1), while the Available K 212 212 Ppm sorghum cultivars (Giza 113, Rabeh and Warkaa) occupy Organic matter OM 0.95 0.95 % the sub-plots. The number of experimental units in each Soil structure ------replicate reached 12 experimental units resulting from the Sand 32 32 % study factors. The dimensions of the experimental unit was Silt 57 57 % 3×3 m. All field operations such as plowing, smoothing and Clay 11 11 % leveling were conducted and then a random sample of field Texture Silt loam Silt loam -- soil was taken in both seasons from different locations of the experiment field at a depth of 0-30 cm before planting to determine their chemical and physical properties as shown RESULTS AND DISCUSSION in Table 1. The soil sample was analyzed in the laboratories Plant height (cm): The results of Table 2 indicate that there of the department of soil affiliate of the agricultural research was significant difference between the sorghum cultivars and department - Abu Ghraib. After the completion of the field the concentrations of bio catalyst (Amino triplex) in plant operations, the field was divided into experimental units height in both seasons, while the interaction between the two each unit included six lines with distance reached 50cm factors had no significant effect in both seasons. The cultivar between them and 25cm between holes. The length of each of Giza 113 was superior significantly by giving the highest line is 3m. A distance of 1.5 m was left between the main plant height amounted to 343.4 and 248.0 cm in both plots to prevent transmission of the spray solution when seasons respectively, while the Rabeh cultivar gave the spraying and 50 cm between the subplots and 2 m between lowest value reached 155.1 and 180 cm in both seasons the replicates. Planting was done on 10/4/2018 in the spring respectively, while the height of the Warkaa cultivar was in season and on 1/8/2018 in the fall season by placing 2-3 middle between Giza 113 and Rabah which gave 179.2 and seeds in the hole and then thinned to one plant in the stage of 209.0 cm in both seasons respectively. This difference 5 leaves. between species may be due to their difference in genetic The first irrigation was conducted slowly to prevent origin as well as their responsiveness to environmental drifting of seeds from planting lines, and then irrigation was conditions, these results are in agreement with results of repeated as needed. Weeding was conducted as needed (Rana et al 2014) and (Ahmed et al 2017). Who found a also. As a precautionary measure, the liquid diazinon was difference between sorghum cultivars in plantheight. The sprayed to control the corn borer stalk at a concentration of results of the same table indicate that the plant height 400 cm3 ha-1. Nitrogen fertilizer was added at a rate of 200 kg increased gradually with the increase of the biocatalyst N ha-1 with the form of urea (46% N) and in three batches concentrations to reach the maximum height at the level of 6 -1 phosphate fertilizer was added at the rate of 100 kg P2O5 gml by giving (241.0 and 242.0 cm) in both seasons (46% P) with one batch before planting. The data were respectively and differed significantly compared to other statistically analyzed using the Genstat programme by concentrations except the concentration of plant height gm L- analysis of variance and for all studied characters using the 1 in the spring season. While the control treatment gave the least significant difference test (LSD) to compare statistical lowest plant height (211.1 and 191.0 cm) in both seasons. averages at a significance level of 0.05 (Al-Rawi and Khala Leaf area (cm2): The results of Table 3 showed that the Fallah 1980). varieties were significantly different in leaf area in both Parameters recorded: Plant height (cm), leaf area (cm2), seasons respectively. The cultivar of Giza 113 was superior in number of grain per panicle (grain panicle-1), weight of 500 this character and gave the highest average leaf area grains (gm), grains yield (t ha-1) and the percentage of seeds reached 4542 cm2 in spring season, while the Rabeh cultivar protein (%). gave the highest leaf area in the fall season reached 6128 Growth and Yield Characters of Sorghum Cultivars 147 cm2 which did not differ significantly from Warkaa cultivar. period. Finally this reflected on the increase of leaf area, The difference in the leaf area may be due to the cultivars these results are in agreement with the results of (Ogundare different in their genetic origin and their response to et al 2015) and (Obid et al 2016). The interaction between environmental conditions. This was observed from the cultivars and biocatalyst showed significant effect in both cultivars different during the two seasons. It was noted that seasons in this character, where the cultivars of Giza 113 and cultivar of Giza 113 recorded the highest value of this Warkaa which treated with concentration of 6 gml-1 gave the character in the spring season while the lowest average highest values reached 6540 and 4762 cm2, while the two obtained from the same cultivar in fall season. This was also cultivars of rabeh and Giza 113 gave the lowest values of leaf observed with the Rabeh cultivar which reinforces our results area reached 4628 and 3550 cm2 in both seasons, that cultivars vary depending on the growing season. These respectively. results are in agreement with results of (Abood and Salh Number of grains per panicle (grainpanicle-1): The results 2018). The increase of the used biocatalyst concentrations of Table 4 showed that the varieties were significantly on the sorghum plant resulted in a significant increase in the different among them in the number of grains per panicle. leaf area where the concentration of 6 gm l-1 gave the highest The cultivar of Warkaa gave the highest value of this value of leaf area reached 6092 and 4624 cm2 in both character reached 1586 grain panicle-1, which did not differ seasons, respectively, while the control treatment gave the significantly compared to the Rabeh cultivar which gave 1570 lowest value reached 3989 and 5417 cm2 in both seasons. grain panicle-1, while the cultivar of Giza 113 recorded the The superiority of the biocatalyst treatments may be due lowest average reached 1313 grain panicle-1. The reason for to the positive role of the biocatalyst components of nutrients the cultivars different in this character is due to their and amino acids which sprayed on the plant at the time of the differences in their genotypes. The results of the same table plant need, which contributed to increase the biological also indicate a significant effect of the concentrations of processes in the leaves and increase cell division, which Amino triplex in this character. The concentration of 6 gm l-1 increased the leaves area and stay effective for a longer recorded the highest value of this character reached 1727

Table 2. Effect of sorghum cultivars, bio-catalyst (Amino triplex) and their interaction on plant height in spring and fall seasons of 2018 Cultivars Spring season Cultivars Fall season Cultivars mean mean Bio catalyst concentration gm.L-1 Bio catalyst concentration gm.L-1 0 2 4 6 0 2 4 6 Rabeh 141.6 156.2 160.5 161.9 155.1 156 160 179 223 180 Warkaa 160.9 169.2 184.4 202.1 179.2 197 208 209 221 209 Giza 113 330.7 340.7 343.1 359 343.4 220 232 261 282 248 L.S.D 0.05 N.S L.S.D 0.05 N.S L.S.D 0.05 Conc. mean 211.1 222 229.3 241 24.25 191 200 216 242 20.3 L.S.D 0.05 20.25 29.3

Table 3. The effect of sorghum cultivars, bio-catalyst (amino triplex) and their interaction on leaf area in spring and fall seasons of 2018 Cultivars Spring season Cultivars Fall season Cultivars mean mean Bio catalyst concentration gm.L-1 Bio catalyst concentration gm.L-1 0 2 4 6 0 2 4 6 Rabeh 3550 4439 4511 4639 4285 5716 6489 6131 6182 6128 Warkaa 4047 4124 4557 4472 4300 5908 5688 5902 6540 6010 Giza 113 4371 4457 4577 4762 4542 4628 4601 5094 5555 4969 L.S.D 0.05 626.8 L.S.D 0.05 602.8 L.S.D 0.05 Conc. mean 3989 4340 4548 4624 313.4 5417 5591 5709 6092 301.4 L.S.D 0.05 215 NS 148 Ahmed H. Ahmed and Sinan A. Abbas and 2018 grainpanicle-1, while the control treatment gave the relative increase of 17.01 and 15.09% in both seasons lowest number of grain per panicle reached 1150 and 1483 respectively. The superiority of Giza 113 cultivar in this grainpanicle-1, this may be due to the role of these nutrients, character may be due to its superiority in the leaf area (Table which contributed to increasing the leaf area for both seasons 3). It may also be due to the fact that the cultivars differ in their (Table 3). This has been positively reflected in the increase of genotypes, the response of each cultivar to environmental plant efficiencies in the conversion of photosynthesis conditions and the duration during which the leaves remain products and their delivery to developing and emerging effective longer to carry out photosynthesis and creation the grains. These results are consistent with the results of (Salah carbohydrates and transport them to grains, this result is 2017), which showed the role of this substance in increasing consistent with the results of (Muflahi and Basuaid 2017). the number of grains per panicle. The interaction between the Who found a significant difference between the cultivars in two study factors had a significant effect in this character in the mean of this character. Also, there was a significant spring season only, the interaction treatment of Warkaa difference between the mean of this character by increasing cultivar with the concentration of 6 gm l-1 gave the highest the concentrations of biocatalyst in both seasons (Table 5). value reached 1852 grain panicle-1 while the interaction The treated plants with concentrations of 6 gm l-1 gave the treatment of Giza113 cultivar with control treatment gave the highest values reached 26.92 and 32.43 gm, while the control lowest value reached 892 grain panicle-1. treatment recorded the lowest values reached 20.92 and Weight of 500 grains (gm): The results of Table 5 indicate 26.47 gm in both seasons respectively. The superiority of this that there was significant effect of cultivars and biocatalyst concentration may be due to its superiority in the leaf area concentrations between them in spring and fall seasons, and (Table 3). This enhances the role of the leaf area in increasing interaction between the two factors in the first season only. the carbohydrates creation processes in the leaves and their The cultivar of Giza 113 was significantly superior transfer to the grain in the panicles. These results are in compared to other cultivars by giving 27.16 and 32.80 gm in agreement with results of other researchers who found a both seasons respectively, with a relative increase by 13.60 significant effect of foliar fertilizers and biocatalysts in weight and 9.93% compared to cultivars of Warkaa and rabeh with a of 500 grains (Liamelo et al 2016). The results of the same

Table 4. Effect of sorghum cultivars, bio-catalyst (amino triplex) and their interaction on number of grains per panicle in spring and fall seasons of 2018 Cultivars Spring season Cultivars Fall season Cultivars mean mean Bio catalyst concentration gm.L-1 Bio catalyst concentration gm.L-1 0 2 4 6 0 2 4 6 Rabeh 1383 1852 1700 1646 1570 1340 1823 1970 2090 1805 Warkaa 1175 1634 1684 1852 1586 1492 1670 1154 1960 1819 Giza 113 892 1232 1445 1684 1313 1618 1921 1942 2004 1871 L.S.D 0.05 347 L.S.D 0.05 NS L.S.D 0.05 Conc. mean 1150 1472 1609 1727 173 1483 1804 2022 2018 NS L.S.D 0.05 284 291.3

Table 5. The effect of sorghum cultivars, bio-catalyst (amino triplex) and their interaction on weight of 500 grains in spring and fall seasons of 2018 Cultivars Spring season Cultivars Fall season Cultivars mean mean Bio catalyst concentration gm.L-1 Bio catalyst concentration gm.L-1 0 2 4 6 0 2 4 6 Rabeh 20.45 23.27 23.13 26 23.21 31 30 26.7 26.3 28.5 Warkaa 19.89 22.3 28.54 24.89 23.9 23.7 27 30.3 37.7 29.9 Giza 113 22.41 27.2 29.16 29.87 27.16 24.7 34 39.3 33.3 32.8 L.S.D 0.05 6.02 L.S.D 0.05 NS L.S.D 0.05 Conc. mean 20.92 24.25 26.94 26.92 3.01 26.47 30.67 32.1 32.43 3.69 L.S.D 0.05 2.17 2.92 Growth and Yield Characters of Sorghum Cultivars 149 table also showed a significant effect of the interaction table also indicate the significance of the biocatalyst in this between the two study factors in this character in the spring trait. 4.88 and 6.59 tons E-1 for both seasons in succession. season only. The interaction treatment of Giza 113 cultivar This is in line with what Sadak et al (2012) and Hassanein et with concentrations of 6 gm l-1 gave the highest value reached al (2017) found. The results of the same table also showed 29.87 gm. While the interaction treatment of warkaa cultivar significant differences between the concentrations of the with control treatment gave the lowest value reached biocatalyst in this character. The highest concentration of the 19.89gm. biocatalyst (6gm l-1) gave the highest value of grain yield Grains yield (t ha-1): The results of Table 6 indicate that there reached 6.73 and 10.49 t ha-1 in both seasons respectively, were significant differences between both of cultivars and while the control treatment gave the lowest value reached biocatalyst concentrations in both seasons, while there was 4.88 and 6.59 tha-1 in both seasons, respectively. This is in no effect from the interaction between the two factors in this agreement with results of (Sadak et al 2012, Hassanein et al characters in both seasons. The cultivar of warkaa was 2017). superior and gave the highest grain yield reached 6.02 t ha-1 The percentage of seeds protein (%): The results of Table in the spring season. The lowest yield was 5.51 and 8.10 t ha- 7 indicate that there was a significant effect of the cultivars 1 in both seasons respectively. The reason for the superiority and the biocatalyst and their interaction in this character in of the two cultivars in the grain yield is due to the superiority of both seasons except the interaction between the two factors the warkaa cultivar with the highest number of grain per of study in the spring season. panicle in the spring season (Table 4). While in the fall The cultivar and warkaa was significantly superior season, the cultivar of Giza 113 may be superior in this compared to the other two cultivars (Rabeh and Giza 113) character due to its superiority in weight of 500 grains and which gave the highest averages of this character reached number of grains per panicle (Tables 4 and 5). These results 10.50 and 10.42% in both seasons, respectively, while the are in agreement with results of (Abood and Abduilhmed cultivar of Giza 113 gave the lowest protein percentage in 2017, Abood and Salh 2018) who found significant both seasons reached (9.21 and 9.13%, respectively). The differences between the cultivars. The results of the same variance among cultivars in this character may be due to the

Table 6. The effect of sorghum cultivars, bio-catalyst (amino triplex) and their interaction on grains yield in spring and fall seasons of 2018 Cultivars Spring season Cultivars Fall season Cultivars mean mean Bio catalyst concentration gm.L-1 Bio catalyst concentration gm.L-1 0 2 4 6 0 2 4 6 Rabeh 4.57 4.74 6.34 6.4 5.51 5.67 8.14 8.37 10.22 8.1 Warkaa 5.01 5.82 5.91 7.38 6.02 6.18 7.55 8.57 10.5 8.2 Giza 113 5.07 5.96 6.13 6.43 5.89 7.94 8.88 10.25 10.76 9.45 L.S.D 0.05 NS L.S.D 0.05 NS L.S.D 0.05 Conc. mean 4.88 5.51 6.13 6.73 0.89 6.59 8.19 9.06 10.49 1.06 L.S.D 0.05 1.11 2.06

Table 7. The effect of sorghum cultivars, bio-catalyst (amino triplex) and their interaction on the percentage of seeds protein in spring and fall seasons of 2018 Cultivars Spring season Cultivars Fall season Cultivars mean mean Bio catalyst concentration gm.L-1 Bio catalyst concentration gm.L-1 0 2 4 6 0 2 4 6 Rabeh 8.57 9.48 10.08 11.54 9.92 7.67 9.48 10.31 12.21 9.84 Warkaa 9.95 10.93 9.72 11.13 10.5 8.95 10.26 10.39 12.16 10.42 Giza 113 7.91 8.97 9.88 10.1 9.21 7.57 8.64 10.22 10.46 9.13 L.S.D 0.05 NS L.S.D 0.05 1.05 L.S.D 0.05 Conc. mean 8.81 9.79 9.9 11.01 0.7 8.03 9.46 10.23 11.46 0.52 L.S.D 0.05 1.99 1.29 150 Ahmed H. Ahmed and Sinan A. Abbas

difference of their genotype, this results is in agreement with performance of forage sorghum genotypes as affected by watering interval in semi-arid environment. World Journal of results of (Salh 2018), who found a significant difference Agricultural Research 5(1): 1-4. between the cultivars in percentage of protein. The results of Al-Rawi K and Khalafallah MAA 1980. Design and Analysi the same table also showed that the concentration of 6 gm l-1 Agricultural AOAC. 1965. Official method of Analysis of the Association of the Official Agricultural Chemists. Washington. gave the highest values reached 11.01 and 11.46% in both Experiments. Ministry of Higher Education and Scientific seasons respectively. Where the increase in the percentage Research, University of Mosul - College of Agriculture and Forestry, pp.488. of protein was associated with the increase of used Hariprasanna K and Rakshit S 2017. Economic of sorghum (Chap.): concentrations, which reached the maximum at the highest In The Sorghum Genome, pp. 1-25. concentration, while the control treatment gave the lowest Hassaneni MS, Ahamed AG and Zaki NM 2017.Grain sorghum as percentage of protein reached 8.81 and 8.03%, respectively influenced by foliar fertilizer stimufol amino. Journal by Innovative Scientific Information 14(2): 362-369. in both seasons. The increase of protein percent when Jabereldar AA, EL-Naim AM, Abdalla AA and Dagash YM 2017. increasing the concentrations of bio-catalyst may be Effect of water stress on yield and water use efficiency of attributed to the role of this substance in increasing the leaf sorghum (Sorghum bicolor (L.) Moench) in semi- arid environment. International Journal of Agriculture and Forestry area (Table 3) and then increase the physiological and 7(1): 1-6. carbohydrates creation processes inside the plant, This Liamelo N, Bulalin SP and Pattung A 2016. Effect of different bio- fertilizers applied as supplemental foliarspray on the growth contributed to the transport of these carbohydrates to the and yield of corn (Zea mays L.). Asia pacific Journal of grain and increased the weight of the grain (Table 5). These Multidisciplinary Research 4(4): 119-125. results are in agreement with results of Ahmed et al (2015) Muflahi AAM and Basuaid AS 2017. Effect of nitrogen fertilizer and its application efficiency on two local sorghum cultivars (Sorghum and Hassaneine t al (2017). It is evident from the results of the bicolor L. Moench). Journal of Agric. Scieence 9(4): 1916-9760. interaction between the two factors in the same table that Obid SA, Idris AE and Abdelgadir BA 2016. Effect of bio-fertilizer on there was an increase in the percentage of protein in the growth and yield of tow maize (Zea mays L.) cultivars at Shambat, Sudan. Scholars Journal of Agriculture 3(4): 313- grains in the fall season with increasing the concentrations of 317. biocatalyst with all cultivars. The interaction treatment of Ogundare SK, Aduloju MO, Ayodele FG and Olorunfemi SD 2015. TM Rabeh cultivar with concentration of 6 gm l-1 gave the highest Effect of tillage methods and foliar fertilization (Boost Extra ) on soil physical properties, weed dry matter and grain yield of mean of protein percent reached 12.21% and did not differ sorghum in Ejiba, kogi state Nigeria. Natural Science 7(10): significantly with cultivar of Warkaa with the same 338-345. concentration; while the interaction treatment of Giza 113 PuGuixin B, Barry M, Bell G and Want MP 2008. Fate of applied bio solids nitrogen in a cut and remove forage system on alluvial with the control gave the lowest protein percent reached clay loam soil. Australian Journal of Soil Research 46(8): 703- 7.57%. 709. Rana AS, Ahmad AH, Saleem N, Nawaz A, Hussian T andSaad M 2014. Differential response of sorghum cultivars for fodder yield REFERENCES and quality. Journal of Global Innovations in Agricultural and Abood NM and Salh MA 2018. Response of several sorghum Social Sciences 2(1): 6 -10. varieties to potassium foliar. Iraqi Journal of Agricultural Ratnavanthi CV 2016. Sorghum processing and utilization (Chap.6): Sciences 49(6): 973-981. 1 Sorghum Biochemistry, pp. 311-327. Abood NM and Abdulhameed ZA 2017.Response of some sorghum Salah BH 2017. The effect of genotype and spry with bio-stimulator in (Sorghum bicolor L. Moench) cultivars to foliar spraying of some anatomical indicators, yield and components of sorghum. riboflavin growth, grain yield and proline content. Journal of Thesis-College of Agriculture – Anbar University-Field Plant Production, Mansoura Univrsity 8(11): 1093-1101. Crops,106 p. Ahmed SO, AdelWahad HA, Magdi OA, Tomoe I, Ping A and Elfadil Shafeec MR, Helmy YI, Shalaby MAF and Omer NM 2012. BE 2015. Effect of different levels of micronutrients fertilizer on Response of onion plant to foliar application of sources and protein and basic and acidic amino acids contents of grains of levels of some amino acid under sandy soil conditions. Journal sorghum cultivars. Journal of Agricultural Science 21: 159- of Applied Science Research 8(11): 5521-5527. 166. USAD 2016. World Agricultural production. Circular series, foreign Ahmed SE, El-Naim AM and Dagash YM 2017. Agronomic agricultural services- Office of global analysis: WAP 20-16.

Received 28 September, 2019; Accepted 21 November, 2019 Manuscript Number: S-270 Indian Journal of Ecology (2019) 46 Special Issue (8): 151-156 NAAS Rating: 4.96

Effect of Zero Tilled Dibbled and Phosphorus Fertilizer on Growth and Yield of Sunflower (Helianthus annuus L.)

Edrees S. Najm, Malath A. Hamed1, Q. Snaa2 and S.H. Cheyed Field Crops Dept., Coll. of Agric. Eng. Sci, University of Baghdad, Baghdad, Iraq 1Field Crops Dept., Coll. of Agric., University of Anbar, Anbar, Iraq ٢Foundation of Technical Education, Technical Coll., Al-Msiaab, Iraq E-mail: [email protected]

Abstract: A field trial was carried out to investigate the effect of cultivation method (no tillage versus conventional ploughing) and phosphorus fertilization (100, 200 and 300 kg ha-1) on some growth characters of sunflower field variety Aqmar. The results showed that there were not significant differences between the two methods of cultivation on seed yield and plant height. However, ploughing method was superior in the rest of the parameters. Levels of phosphorus fertilizer had significant effect on all the characters with the exception of number of days from sowing to the 50% flowering where 200 and 300 kg ha-1 were superior over 100 kg ha-1 in all characters. The use of 200 kg ha-1 in improving growth characters and increased seed yield is appropriate. The interaction of ploughing and 300 kg ha-1 phosphor fertilizer gave the best values.

Keywords: Conservative cultivation, Industrial crops, Nutritive element, Ploughing

Sunflower (Helianthus annuus L.) is considered one of study was conducted to investigate the response of oily the major crops in the Iraq due to its various uses among sunflower (Aqmar oily cultivar) to the zero-tillage and them oil from oily cultivars and feeds for animals (Cheyed phosphorus levels. 2014). It is the best fourth oily crop in the world (Nisar 2011). This crop is characterized by its high capability of adaption in MATERIAL AND METHODS a wide range of environmental conditions; therefore it is A field trial was conducted at the alternate site (site of grown widely around the world (Agele 2007). Iraqi college of agriculture- university of Baghdad) Abu-Ghraib environment is suitable to produce sunflower, yet its during the spring and fall seasons of 2015. The objective was productivity still low due to the bad application of husbandry to investigate the effect of zero-tillage compared with the practices and not using modern agricultural systems, which conventional method and phosphorus levels (100, 200, and -1 conserve the soil texture and improve the plant growth in the 300) kg P2O5 ha which triple super phosphate was the main field, in turn increases the seed yield of unit area. Zero (no) source of phosphorus on some growth characters and seed tillage method improve the physical chemical and biological yield of Aqmar oily cultivar. A piece of land previously grown characteristic of soil, which has led to more stability of with sunflower in the spring season was used to be the same productivity (Alrijbo and Hussan 2011, Chauhan et al 2015), for cultivation in the fall season. This land was divided to six besides it is economical method reduces the costs of stripes, three of the were ploughed after the removal of plant production (Piggin et al 2002) via lessening of energy waste residues random and the rest of land left without ploughing manpower and reduces the required time for seed bed with the plant residual been left, each stripe was divided to preparation. Also, it increases the rain water harvesting in the experimental units 3 × 4 m as the same dimension of dry and semi dry regions compared with the conventional unploughed plots to assure the homogenous experimental method of cultivation (Anaerson et al 2000), especially in the units. Sowing was performed on rows 75 cm between them region of very low water table(Humphreys et al 2004). and 25 cm between plants in rows to achieve plant population Nutritional element in seed play a key role in agricultural density 53000. A distance of 75 cm between experimental production, as Iraqi sails contain high levels of calcium units was left. Three-five seeds were placed in each hill, then carbonates which makes the pH soil close to the alkalinity, a thinning was done after the completion of full field there for, soils suffer from the reduction of essential aliments emergence and the formation of first pair of true leaves. The availability for plant growth among them phosphorus design was split-block design where the main factor was no- (Zeboon 2006). The phosphorus is the second important tillage and conventional ploughing and the sub plots for element after nitrogen, which affects all growth stage of phosphorus fertilizer with three replication. All phosphorus sunflower, seed yield and quality (Salih 2013), there for, this fertilizer was applied at cultivation with the cultivation rows. 152 Edrees S. Najm, Malath A. Hamed, Q. Snaa and S.H. Cheyed

Nitrogen fertilizer (urea 40% N) with the rate 280 Kg ha-1 was except the effect of cultivation method with no significant applied twice the first after field emergence and the second differences between them. Plants of conventional method one at the beginning of floral bud formation. where superior in the plant height 178.03 cm compared with Five plants were randomly chosen from the middle 170.80 cm for no-tillage method in the fall season. This may rows to record the period from sowing to the 50% flowering, be that ploughing helps in the increase of root system via plant height (cm), leaf area (cm2), chlorophyll content of breaking down sub-down layers of soil. The increase of leaves, Phosphorus ratio of leaves and seed yield plant-1. phosphorus fertilizer from 100 to 300 kg ha-1 increased plant Data were analyzed according to the used design height from 161 and 106 cm and 174.83 and 119.25 cm, average using LSD at 0.05 for comparison. respectively with no significant difference with 200 kg ha-1 in the increasing root system and plant growth which, in turn, RESULTS AND DISCUSSION increases plant height. This result confirms the findings of Number of days from sowing to the 50% flowering: Table Zeboon (2006). 1 show the significant effect of phosphorus fertilizer only in Concerning interaction, plants of cultivated area and the spring season with no significant effect of cultivation fertilized with 300 kg ha-1 gave the highest value in both method and interaction in both seasons. Increased seasons (189.41 and 127.66 cm) compared with 158.29 and phosphorus fertilizer from 100 to 300 kg ha-1 increased the 106.1 cm during both seasons for no tillage × 100 kg ha-1 of number of days from sowing to the 50% flowering by 1.5 day phosphorus. in spring season. Leaf area (m2 plant-1): It is clear from Table 3 that both study Plant height (cm): Table 2 indicates that there was factors and their interactions had significant effect on leaf significant effect of both study factors and their interaction area. Plants of tilled cultivation gave the highest average

Table 1. Effect of zero tilled dibbled and phosphorus fertilizer on number of days from sowing to the 50% flowering in spring and full season Phosphorus Spring season Fall season fertilizer (kg h-1) (P) Cultivation system (C) Mean (P) Cultivation system Mean (P)

Zero- tillage Ploughed Zero- tillage Ploughed

100 73.33 72.67 73 54.93 55.44 55.18 200 73.67 73.33 73.5 55.78 54.93 55.35 300 75 74.5 74.5 55.44 56.59 56.02 Mean (C) 74 73.33 55.38 55.65 LSD 0.05 (P) 1.3 N.S LSD 0.05 (C) N.S* N.S LSD 0.05 (P×C) N.S N.S NS: Non significant

Table 2. Effect of zero tilled dibbled and phosphorus fertilizer on plant height (cm)in spring and full season Phosphorus Spring season Fall season fertilizer (kg h-1) (P) Cultivation system (C) Mean (P) Cultivation system Mean (P)

Zero- tillage Ploughed Zero- tillage Ploughed

100 158.29 164.63 161.46 106.01 106.62 106.31 200 179.27 180.07 179.67 116.29 120.01 118.15 300 174.83 189.41 178.83 110.85 127.66 119.25 Mean (C) 170.80 178.03 111.05 118.09 LSD 0.05 (P) 2.9 5.17 LSD 0.05 (C) 4.01 N.S* LSD 0.05 (P×C) 3.57 6.96 NS: Non significant Sunflower Eeffectiveness by Tilled Dibbled and phosphorus Fertilizer 153

0.777 and 0.366 m2 plant-1 in both seasons compared with for plants resulted from un-tilled land and fertilized with 100 0.690 and 0.333 m2 plant-1 for no-tilled cultivation, kg ha-1 in both seasons. The increase of leaf area with the respectively. This may be due that ploughing (tilling) increases of phosphorus level in the tilled area may be increases root system of plants as a result of breaking down attributed to the fact that excited soil because of tilling allow sub-surface layers of soil, which ultimately improve the easy movement and distribution of phosphorus around the vegetative growth characters of plants among them root system which increase its absorption by plants and then increased plant height (Table 2). increases plant growth characters among them plant height It is also clear from Table 3 that phosphorus fertilizer had (Table 2) and leaf area (Table 3). significant effect on this character. The highest level 200 and Plant dry weight (gm):A significant effect of both factors and 300 kg ha-1 recorded the highest average 0.707 and 0.775 m2 most their interaction was obtained on plant dry weight (Table and 0.383 and 0.388 m-2 in both spring and fall seasons, 4). Plants of tilled method gave the highest value of dry respectively with no significant differences between them. weight 138.9 gm plant-1 compared with 130.4 gm of un-tilled This may be due to the role of phosphorus in formation land in the season. This may be due to the high plant height cell membranes and transportation of sugar (saccharides) (Table 2) and leaf area (Table 3) which in turn, increased plant from their sites of formation in leaves to the other parts of dry weight. It is also clear from Table 4. That high level of plants of increase of photosynthesis which increases plant phosphorus 300 and 200 kg ha-1 recorded the highest activities among them leaf area. This result was in agreement averages (277.7, 272.5, 147.3 and 148.1 gm plant-1) with no with the findings of AL-Tammimi (2010) and Alias et al (2003). significant differences between these two levels in both Concerning interaction, plants of tilled land and fertilized season, respectively, compared with 207.1 and 108.4 gm with the 300 kg ha-1 of phosphorus gave the highest value in plant-1 for 100 kg ha-1 level in both season, respectively. This both seasons (0.836 and 0.423 m2 plant-1), respectively was due to the increase of both plant height and leaf area. compared with the lowest value (0.606 and 0.262 m2 plant-1) This result confirmed the finding, of Salih (2013) who

Table 3. Effect of zero tilled dibbled and phosphorus fertilizer on leaf area (m2 plant-1) in spring and full season Phosphorus Spring season Fall season fertilizer (kg h-1) (P) Cultivation system (C) Mean (P) Cultivation system Mean (P)

Zero- tillage Ploughed Zero- tillage Ploughed

100 0.606 0.71 0.658 0.262 0.296 0.279 200 0.765 0.785 0.775 0.384 0.381 0.383 300 0.699 0.836 0.767 0.352 0.423 0.388 Mean (C) 0.69 0.777 0.333 0.366 LSD 0.05 (P) 0.036 0.012 LSD 0.05 (C) 0.035 0.032 LSD 0.05 (P×C) 0.038 0.026

Table 4. Effect of zero tilled dibbled and phosphorus fertilizer on plant dry weight (gm) in spring and full season Phosphorus Spring season Fall season fertilizer (kg h-1) (P) Cultivation system (C) Mean (P) Cultivation system Mean (P)

Zero- tillage Ploughed Zero- tillage Ploughed

100 198.2 216 207.1 106.5 110.4 108.4 200 274.4 270.6 272.5 146.3 150.0 148.1 300 265.1 290.3 277.7 138.3 156.2 147.3 Mean (C) 245.9 259 130.4 138.9 LSD 0.05 (P) 5.5 7.9 LSD 0.05 (C) NS* 2.8 LSD 0.05 (P×C) 10.2 7.7 NS: Non significant 154 Edrees S. Najm, Malath A. Hamed, Q. Snaa and S.H. Cheyed reported that phosphorus increases growth characters at all 11.22 for the 100 kg ha-1 in both season, respectively. This plant growth stages. may be attributed to the role of phosphorus in enhancement Concerning interactions, highest level of phosphorus of many vital biochemical reaction in the plants especially 300 kg ha-1 and tilled treatment gave the highest value (290.3 improvement of root system (Griffith 2010). and 156.2 gm plant-1) in both seasons with no significant Concerning interactions, the highest level of differences with the 200 kg ha-1× tilled treatment in the fall phosphorus 300 kg ha-1 in the tilled area gave the highest seasons compared with 100 kg ha-1 × un-tilled treatment value in both season. Respectively 30.23 and 17.61 gm with which gave the lowest values 198.2 and 106.5 gm plant-1 no significant differences with the 200 kg ha-1 in the tilled area respectively in both season with no significant differences in fall season compared with 19.82 and 10.65 gm for 100 kg with the same level × tilled treatment in the fall season. ha-1 in tilled areas in both seasons, respectively. Dry weight of root system (gm): Table 5 illustrates the Chlorophyll content of leaves (mg gm-1 fresh weight): significant effect of both factors and their interaction except Table 6 shows significant effect of both study factors and their the effect of tilled treatment in the spring season. Plants of interaction in both seasons. Plants of tilled areas recorded tilled area gave the highest value 15.28 gm compared with the highest average of chlorophyll content 38.67 and 43.79 13.15 gm for plants growth in the no-tilled area. This may be mg gm-1 fresh weight in both seasons compared with 36.43 due to the high plant height (Table 2) and leaf area (Table 3), and 40.92 mg gm-1 fresh weight for plants resulted from un hence, increased dry weight of plant including root dry tilled areas. Tis may be due the effect of tilling on root dry weight. Table 5 also shows that high level of phosphorus 300 weight (Table 5) which in turn improving vegetative growth * 200 kg ha-1 gave the highest averages 28.07 and 27.38 gm (Table 2 and 3), hence improving photosynthesis efficiency with no significant differences between for spring season and and plant chlorophyll contest. Also, phosphorus fertilizer level 16.00 and 15.96 gm for fall season compared with 22.50 and significantly increased this character where 200 and 300 kg

Table 5. Effect of zero tilled dibbled and phosphorus fertilizer on dry weight of root system (gm) in spring and full season Phosphorus Spring season Fall season fertilizer (kg h-1) (P) Cultivation system (C) Mean (P) Cultivation system Mean (P)

Zero- tillage Ploughed Zero- tillage Ploughed

100 21.02 23.77 22.5 10.27 12.17 11.22 200 27.90 26.86 27.38 15.86 16.06 15.96 300 25.92 30.23 28.07 14.40 17.61 16.00 Mean (C) 25.02 26.95 13.51 15.28 LSD 0.05 (P) 1.85 1.52 LSD 0.05 (C) NS* 1.56 LSD 0.05 (P×C) 2.05 1.61 NS: Non significant

Table 6. Effect of zero tilled dibbled and phosphorusus fertilizer on chlorophyll content of leaves (mg gm-1 fresh weight)in spring and full season Phosphorus Spring season Fall season fertilizer (kg h-1) (P) Cultivation system (C) Mean (P) Cultivation system Mean (P)

Zero- tillage Ploughed Zero- tillage Ploughed

100 32.63 34.02 33.33 38.83 39.89 38.86 200 40.08 39.37 39.73 43.64 44.12 43.88 300 36.58 42.61 39.59 41.29 47.37 44.33 Mean (C) 36.43 38.67 40.92 43.79 LSD 0.05 (P) 1.12 1.43 LSD 0.05 (C) 2.13 1.43 LSD 0.05 (P×C) 1.79 1.52 Sunflower Eeffectiveness by Tilled Dibbled and phosphorus Fertilizer 155

Table 7. Effect of zero tilled dibbled and phosphorus fertilizer on phosphorus ratio of leaves (%) in spring and full season Phosphorus Spring season Fall season fertilizer (kg h-1) (P) Cultivation system (C) Mean (P) Cultivation system Mean (P)

Zero- tillage Ploughed Zero- tillage Ploughed

100 0.417 0.446 0.432 0.274 0.295 0.284 200 0.61 0.63 0.62 0.413 0.428 0.42 300 0.526 0.65 0.588 0.376 0.454 0.415 Mean (C) 0.518 0.576 0.354 0.392 LSD 0.05 (P) 0.029 0.015 LSD 0.05 (C) 0.055 0.015 LSD 0.05 (P×C) NS* 0.016 NS: Non significant

Table 8. Effect of zero tilled dibbled and phosphorus fertilizer on plant seed yield (gm plant-1) in spring and full season Phosphorus Spring season Fall season fertilizer (kg h-1) (P) Cultivation system (C) Mean (P) Cultivation system Mean (P)

Zero- tillage Ploughed Zero- tillage Ploughed

100 43.77 45.31 44.54 52.88 52.95 52.92 200 51.09 51.13 51.11 57.41 58.61 58.01 300 53.71 54.71 54.20 60.17 60.14 60.15 Mean (C) 49.52 50.38 56.82 57.23 LSD 0.05 (P) 2.63 1.82 LSD 0.05 (C) NS* NS LSD 0.05 (P×C) NS NS NS: Non significant ha-1 level recorded highest average 39.73, 39.59 and 43.88, phosphorus gave the highest averages 0.620 and 0.42% in 44.33 mg gm-1 fresh weight in spring and seasons, both seasons with no significant differences with 300 kg ha-1 respectively with no significant differences between them. in the fall season. This may be due to the effect of increasing This may be due to the effect of phosphorus on root system application phosphorus in the soil which increased the and plants growth which, in turn, reflected in the increased of available phosphorus in the soil and the absorption via root plants highest (Table 2) and leaf area (Table 3), which system. This result was in agreement with the finding of AL- increased photosynthesis. This result was in agreement with Dilaimi (2006). Plants growth in the tilled area fertilized with the findings of Salih (2013). 300 kg ha-1 gave the highest value 0.454% in the fall season Concerning interaction the highest level of phosphorus compared with untilled area fertilized with 100 kg ha-1 which 300 kg ha-1 in tilled area gave the highest values in both gave 0.274% in the fall season. seasons (42.61 and 47.37 mg gm-1 fresh weight) compared Plant seed yield (gm plant-1): Results of Table 8 show with 32.63 and 38.83 mg gm-1 fresh weight for the 100 kg ha-1 significant effect of phosphorus fertilizer on seed yield of in both seasons, respectively. plants in both season with no significant effect of interaction. Phosphorus ratio of leaves (%): Table 7 illustrates the Increased phosphorus level from 100 to 300 kg ha-1 increased significant effect of both factors and their interaction except seed yield by 21.69 to 15.55% in both season, respectively. the interaction in the spring season. Plants of tilled areas This was due to the role of phosphorus in improving growth recorded the highest average 0.576 and 0.392% in both characters and increasing leaf content of chlorophyll which in seasons compared with the plant of untilled areas. This may turn, increased plant seed yield. be due to the effect of tilling in the improving of plant root system as a result of breaking down the sub layers of soil CONCLUSION which improves the absorption of nutritive element by plants. There was no significant differences between tilled and It is also clear from Table 7 that 200 kg ha-1 level of untilled soil in respect of number of days from sowing to the 156 Edrees S. Najm, Malath A. Hamed, Q. Snaa and S.H. Cheyed

50% flowering in both seasons, plant height in the in the fall N, Shackley BJ, Amjad M and Zaicou- Kunesch C 2005. The role management in yield improvement of the wheat crop-a season, plant dry weight and root dry weight in the spring review with special emphasis on western Australia. season and seed yield of plant in both seasons. However, Chauhan SK, Saini KS, Saralch HS, Rani S and Verma A 2015. tilled treatment was significantly superior in the other Wheat and barley crop performance under different sowing methods under poplar based agroforestry system. Indian characters. Despite that the differences in both tilled and Journal of Ecology 42(2): 528-530. untilled treatment were significant in those characters we Cheyed SH 2014. The relation of sunflower seed qualities and suggest that further studies may be needed to find the capiltlum diameter and seed position. AL-Anbar Journal of Agricultural Sciences 12(2): 217-226. economical feasibility. Results also showed the positive role Elsahookie MM and Eldabas EE 1982. One leaf dimension to of phosphorus in both high level, compared with the lowest estimate leaf area in sunflower. Journal of Agronomy and Crop one, which suggest the possibility of using 200 kg ha-1 in Science 151: 199-204. improving plant growth and increasing seed yield. Griffith B 2010. Efficient Fertilizer Use-Phosphorusus, Pp. 1-7. Humphreys E, Meisner C, Gupta RK, Tinsina J, Beecher HG, Tang YL, Singh Y, Gill MA, Masih I, Guo ZI and Thompson JA 2004. REFERENCES Water savings in rice-wheat systems. Proceedings of the 4th International Crop Science Congress, Brisbane, Australia. Al-Dilaimi HY 2006. Effect of phosphorus applied foliar and mixed in soil on the availability and uptake of p, Zn and cu by maize. The Nisar M, Hussain S, Nausheen A, Khan N and Siddique MF Iraqi Journal of Agricultural Sciences 37(2): 15-22. 2011.Chemical composition of open pollinated and hybrid population of sunflower (Helianthus annus L.).Pakistan Journal Agele SO, Maraiyesa IO and Adeniji IA 2007. Effects of variety seed of Botany 43(1): 157-163. set efficiency in late season sunflower (Helianthus annus L.) in a humid zone of Nigeria. Academic Journals 2: 80-88. Piggin CM, Gracia CO and Janiya JD 2002. Establishment of irrigated rice under zero and conventional tillage system in the Alias A, Usman M, Ullah E and Warraich EA 2003. Effect of different Philippines in ProcInt workshop on herbicide resistance phosphorusus levels on the growth and yield of two cultivars of management and zero tillage in rice-wheat system. March 4-6, maize (Zea mays L.). International Journal of Agriculture and 2002, Hisar, India: 190-195. Biology 5(4): 632-634. Salih MNT 2013. Response of sunflower (Helianthus annuus L.) to Alrijabo AA and Hassan HH 2011. Effect of seed grading, seed rate phosphorusus and nitrogen fertilization under rainfed and zero tillage planting method on growth, yield and its conditions, Blue Nile State-Sudan. HELIA, 36(58): 101-110. components of durum wheat (Triticum durum DESF.) under Zeboon NH 2006. Effect of sulphur and phosphorusus fertilizer on rainfed area. Mesopotamia Journal of Agriculture 39(1): 177- growth and yield of sunflower (Helianthus annus L). M.Sc. 190. Thesis, Dept. Field Crop Sci., College of Agriculture, Univ. of Anderson WK, Hamza MA, Sharma DL, Antuono MFD, Hoyle F, Hill Baghdad, pp. 142. Received 28September, 2019; Accepted 21 November, 2019 Manuscript Number: S-271 Indian Journal of Ecology (2019) 46 Special Issue (8): 157-160 NAAS Rating: 4.96

Management of Wheat Weeds using Sorghum bicolor (L.) Leaves Extract and Effect on Yield Components

Watheq Falhi Hammood and Suhad Mathkoor A. Safi* College of Agricultural Engineering Sciences, University of Baghdad, Baghdad, Iraq *E-mail: [email protected]

Abstract: An experiment was carried out to take check samples of Sorghum bicolor (L.) leaves for their potential for allelopathy. The experiment was also carried out to knowing the effect of Sorghum bicolor (L.) leaves extract and Chevalier herbicide on the weed associated with the wheat crop cultivar IPA-99 and its effect on the yield. The results showed that the S. bicolor (L.) leaves extract at 8 leaves stage gave good results in completely eliminating the spread of weed. The other treatments reduced the number of weeds by 94.2% and 97.5%. The superiority of sorghum -2 -2 leaf extract treatment was also reflected in the yield and its components, which gave the highest number of spikem (472.3 spikesm ) the same treatment also gave the highest number of spikes amounted to56.23 grain The spraying of the wheat crop weeds with the Sorghum bicolor (L.) resulted in complete elimination.

Keywords: Wheat, Weeds, Sorghum bicolor, Yield

Wheat comes at the forefront of the grain that humans of spikes plant-1 to 20.32%, and the number of grains spike-1 need, and represent the main source of food for more than with a percentage of 20.18%, in addition to the weight of grain 35% of the world population because of the gluten that spike-1 by 26.88%, and the weight of 1000 grains with a rate of produces the best bread types (Jamali et al 2000). The total 29.64%. As well as, the increase in grain yield plant-1 by quantity of wheat production in Iraq for 2018 was two million 57.45% compared to the weedy treatment. In spite of the and 178 thousand tons, while the total cultivated area efficiency of chemical herbicides in eliminating weeds and it amounted to three million compared to the production is reflected in the yield and its components, but the damage amount of two million and 974 thousand and 136 tons during caused by chemical herbicides such as groundwater 2017 (Central Statistical Organization, 2018).Thus, it does pollution, the emergence of health problems as a result of not meet the local need for local consumption and therefore a their widely and excessive use, and the long-term remaining balance must be created between the gross domestic of the herbicide residues in the soil, which affected many vital product and demand and this can be achieved only through ecosystems. In order to invest Allelopathic phenomena in good management and right service operations, such as the controlling the weeds, using of plant extracts was widely controlling of weeds. The wheat crop is considered as a weak increased, since it act as natural herbicides. It was observed competitor to the weeds compared to other types of crops, that spraying the Sorghum bicolor (L.) extract, sunflower, and where the growth of both broad and narrow- leaves weeds in Brassica napus L. reduced the dry weight of the weeds by wheat fields are one of the most important problems that 57%, and its numbers by 55% and increased the yield of reduce crop productivity and deteriorates its quality (Al- wheat by 13-21% (Arif et al 2015). The Sorghum bicolor (L.) Chalabi 2003). The reduction in the wheat yield was 50-80%, extract inhibited the dry weed weight at a percentage of 22- (Gianessi 2009) and it was increased by 37% after 46%, and the yield of wheat grains increased by 21% eliminating the weeds with chemical herbicides (Hammood (Cheema et al 2000). Sorghum bicolor (L.) extracts spraying and Safi 2018). Furthermore, one of the most commonly on the wheat gave numbers of fertile tiller and the highest used herbicides at present is the Chevalier herbicide, which numbers of grains in spike to (Abdul Razzaq et al has been proven to be effectively used in eliminating the 2012).These extracts contain multiple phenolic acids, weeds, where as using of herbicide at the spraying rate of including p- syringic and vanilic, Gallic, Caffeic, P-Coumaric, 300 gha-1led to reduction in number of broad-leaves weeds Ferulic, Catechol, Benzoic, Hydroxyl, Protocatechuic from 200 plant m-2 to 5 plant m-2 and the number of narrow leaf (Farooq et al 2013, Alsaadawi et al 2007) and this was the weeds from 132 plant m-2 to 4.3 plant m-2. At the same main reason for its effectiveness in controlling the weeds spraying rate it led to an increase in the wheat grain yield to (Hammood 2018). Finally, for the purpose of investing these 4.98 t ha-1 compared to the weedy treatment that gave 3.1 t allelopathic compounds in control the weed, the experiment ha-1 (Habib et al 2003), which led to an increase in the number was carried out to determine the wheat responded to 158 Watheq Falhi Hammood and Suhad Mathkoor A. Safi

Sorghum bicolor (L.) extracts and compared to the Chevalier Table 1. Types and amount of allelopathic compounds (ppm) herbicide and its effect on the yield and weeds. Leaves age Benzoic acid Cimannic acid Coumarrin 6 leaves 229.86 18.02 9.87 MATERIAL AND METHODS 8 leaves 486.08 38.81 24.53 Two experiments were carried out in the field of experiments of the Field Crops Department, College of iv. Weedy treatment: leaving the weed to grow till the Agricultural Engineering Sciences, University of Baghdad end of the season (T4) –Jadiriya during the summer 2018 and winter seasons 2018- The field was divided into three replicates, where the 2019. wheat seeds cultivar IPA-99 were planted on 16/12/2018 with The first experiment was carried out during summer a seeding rate of 120 kg ha-1 in lines, and the distance season of 2018 for obtaining sample from Sorghum bicolor between lines were 20 cm. The experimental land was

(L.) leaves, where the field was divided into replicates and the fertilized with triple phosphate fertilizer (P2O5, 46%) at a rate S. bicolor (L.) seeds Inkath class planted at 1/8/2018 within a of 100 kg ha-1 before planting. Urea (N 46%) was also added -1 line by maintaining of a distance between each line. The soil at a rate of 200 kg N ha in three batches, where, the first of the field experiment has been fertilized according to the batch was added at the beginning of the tillering stage, the recommendations and irrigated whenever needed, while second at the beginning of the elongation stage, and the third samples were randomly taken from S. bicolor (L.) at 6 leaves at the booting stage. The plant extract of S. bicolor (L.) leaves and 8 leaves. and Chevalier herbicide was sprayed after 45 days of Extraction of allelopathic compounds: After the sampling planting on the vegetative growth of the crop and the weeds, process was finished, the leaves were dried, milled and especially at the beginning of the tillering stage with a specialized knapsack sprayer according to the analyzed in the laboratories of the Ministry of Science and concentrations under study by using 100 l ha-1 of water. Technology by the German HPLC device (SYKAM) (Mradu et To determine the degree of weeds spread in each al 2012), where a S. bicolor (L.) leaves extract has been experimental unit, the types and density of the weeds were prepared at a concentration of 5%, and the results of the recorded after 30 days of spraying by squares method using analysis were shown in the Table 1. a square with dimensions 50x50 cm, for an area of ​​1 m from The second field experiment was carried out during the each experimental unit (Al-Chalabi and Al-Majdi 2001). The winter season of 2018-2019 in order to compare the effect of types of weeds are summarized as shown in Table 2. weed herbicide Chevalier and plant extract of the S. bicolor -2 Weeds dry weight (gm m ) and percentage of weed (L.) leaves in the weeds associated with wheat crop yield and control was recorded inhibition of weed dry weight was its components. The experiment was laid out in Randomized calculated according to the following equation, % inhibition= Complete Block Design. The treatments included in the 100 - (A / B × 100) research were: Where: -1 i. Spray Chevalier at the rate of 300 g ha (as A = dry weight of the weeds in the control treatment recommended by the manufacturer) (T1) B = dry weight of the weeds in the comparison treatment ii. Spray the S. bicolor (L.) leaves extract at 6 leaves The effect of different treatments on the yield and its stage (T2) components of the wheat crop was recorded in terms of iii. Spray the S. bicolor (L.) leaves extract at 8 leaves Number of spikes m-2, Number of grains spike, Weight of stage (T3) 1,000 grain and Grains yield (t ha-1).

Table 2. Types of weed spread in the experiment land Local name Scientific name Family name Type Life cycle Lesser canary Pharlaris minor L. Gramineae Narrow leaves Annual Dwarf mallow Malvarolundi folia L. Malvaceae Broad leaves Annual Prickly lettuce Lactuca serriola L. Compositeae Broad leaves Annual Wild carrot Daucus carota L. Umbelifereae Broad leaves Annual Saw bane Chenopodium album L. Chenopodiaceae Broad leaves Annual Milk thistle Silybum marianum L. Campositeae Broad leaves Annual Prostatelant Polygoumum aviculare L. Plantaginaceae Broad leaves Annual Sybmbrium septulatum Sisymbrium irio L. Cruciferae Broad leaves Annual Management of Wheat Weeds 159

Statistical analysis: After collecting and sampling the data Table 3. Effect of different treatments on the number of under study, it was statistically analyzed according to weeds, its dry weights, % control and % inhibition in dry weight of weeds Randomized Complete Block Design and the arithmetic Treatments* No. of No. of Control Dry Inhibition mean compared using the least significant difference at a weeds weeds (%) weight of (%) level of 5% (Steel and Torri 1980). before after weeds spraying spraying (gm m-2) (plant m-2) (plant m-2) RESULTS AND DISCUSSION T1 51.33 3 95.29 1.8 98.22 Number of weeds, dry weights, control and inhibition 2T 52.33 1.3 97.91 1.4 99.07 percentages: After spraying a significant difference was T3 55.33 0 100 0 100 observed on the number of weeds, control (%), dry weight of weeds and percentage inhibition (Table 3). The treatment T4 53.67 62.7 0,0 151.8 0,0 (T3) exceeded the treatment, which gave the highest number LSD (0.05) NS 7.06 1.09 18.7 1.11 Note: T1* Chevalier herbicide spray at 300 g ha-1, T2 Sorghum bicolor (L.) of weeds 62.7 by completely eliminating the weeds and leaves extract at age of 6 leaves, T3 spray the Sorghum bicolor (L.) giving the highest control percentages that reached 100% leaves extract at age of 8 leaves, T4 weedy treatment compared. Treatment T1 and T2 did not differ significantly from treatment T3, where both treatments reduced the Table 4. Effect of different treatments on the yield and its number of weeds by 94.2% and 97.5%, respectively. It was components of wheat crop Treatments* No. of No. of grains Weight of 1000 Grains yield noted that the Sorghum leaves extract has given the best -2 -1 -1 spikes m spike (grain gm) (t ha ) results in eliminating the weeds found the good control T1 445 52.17 43.22 1.85 percentage, that may be due to the fact that the extract 2T 381.7 55.6 46.14 2.63 contains the Allelopathic compounds represented by T3 472.3 56.23 59.7 3.04 phenolic acids (Lahmoud 2012, ALsaadawi 2007). This component in high concentrations cell division, the T4 326 42.17 41.31 1.28 biosynthesis of hormones, the absorption of elements (Riziv L.S.D 40.1 6.28 5.76 0.08 et al 1992), the permeability of cell membranes (Harper and *As in table 3 Balke 1981), photosynthesis, opening and closing the stomata mechanism (Einhellig and Rasmussen 1979), which was resulted from achieving of the highest control respiration and protein synthesis (Kruse et al 2000), which percentage of 100%, and completely eliminating the weeds negatively affect the growth of the weeds and consequently that reduce its dry weights (Table 4). This has led to a lack of the reduction of its numbers. The reduction in the number of competition from the weeds for basic growth requirements, weeds was also reflected in their dry weights, where allowing the crop to grow, exploit all the essential elements, treatment T3 gave the lowest dry weight (0 gm-2), which did and give the best yield components. The study results were not significantly differ from the treatments T1 and T2, T4 consistent with the findings of Arif et al (2015) and Abdul -2 Razzaq et al (2012). gave the highest dry weight of 151.8 g m , leading to superior of the treatments by giving the highest inhibition percentages CONCLUSION for dry weight reached 100.0%, 99.0% and 98.2% for T3, T2 The spraying of wheat crop weeds with the Sorghum and T1 respectively. The results were consistent with the bicolor (L.) leaves extract at 8 leaves stage gave an excellent study of (Habib et al 2003). It was also agreed with the results in terms of eliminating the weeds and increasing the Lahmoud (2012), Al- Saadawi (2007), Hammood (2018). yield of wheat grains due to the presence of phenolic Yield and its components of wheat: Treatment T3 was compounds worked as allelopathic compounds. superior in most traits and achieved the highest number of spikes m-2 measured at 472.3 spike m-2 compared with the REFERENCES comparison treatment (T4) that gave the least number of -2 Abdul Razzaq, Cheema ZA, Jabron K, Hussain M, Farooq M and spikes (326.0 spikes m ). The same treatment gave the Zafar M 2012. Reduced herbicide doses used together with highest number of grains in spike (56.23 grin spikes-1), which allelopathic sorghum and sunflower water extracts for weed control in wheat. Journal of Plant Protection Research 50(2): was significantly not different from T1 and T2. In addition to 24-31. this, T3 gave the highest weight of 1000 grains gm measured Al-Chalabi FT 2003. Biological response of wheat to weeds control at 59.7 gm and the highest grain yield (3.04 t ha-1) compared with Diclofop-methyl herbicide sequentially with 2,4-D and its effect on grain yield. Journal of Iraqi Agricultural Science 34(1): to T4 (Weedy) treatment. The reason for the highest grain 89-100. yield giving treatment (T3) was highest yield components, Al-Chalabi FT and Al-Majdi LE 2001. Weed plants scattered on the 160 Watheq Falhi Hammood and Suhad Mathkoor A. Safi

Iraqi railways. Journal of Iraqi Agricultural Science 32(4) :123-130. and Applied Sciences 9(1): 37-44. ALsaadawiI S, AL-Ekeelie MHS and AL-Hamzawi MK 2007. Hammood WF and Safi SMA 2018. Effect of weed competition in the Differential allelopathic potential of grain sorghum genotypes to characteristics of growth and yield and its components of wheat weeds. Allelopathy Journal 19: 135-160. crop Triticum aestivum L.: A mini review. Journal of Research in Arif M, Cheema ZA, Khaliq A and Hassan A 2015. Organic weed Ecology 6(1):1637-1646. management in wheat through Allelopathy. International Harper JR and Balke NE 1984. Characterization of the inhibition of Journal of Agriculture & Biology 17(1): 127-134. K+ absorption in oat roots by salicylic acid. Plant Physiology 68: Central Statistical Organization 2018. Annual report of wheat 1349-1353. production. Ministry of Planning and Development Jamali KD, Arain MA and Mhamd M 2000. Comparative performance Cooperation. Iraq. of semi-dwarf Wheat (Triticum aestivum L.) genotypes. Wheat Cheema ZA, Khaliq MN and Tariq M 2002. Evaluation of Information Service 90: 45-46. concentrated sorghum alone in combination with three pre- Kruse M, Strandberg M and Strandberg B 2000. Ecological effects of emergence herbicides for weed control in cotton (Gossypum Allelopathic plant-A Review. pp: 66. National Environmental hirsutum L.). International Journal of Agriculture & Biology 4: Research Institute. NERI, Technical report NO. 315, Silkeborg, 549-552. available online at: http//www2.dmu.dk/1_viden/ Einhellig FA and Rasmussen JA 1979. Effects of three phenolic acids 2_publikationer/3…/rapporter/fr315.pdf. on chlorophyll content and growth of soybean and grain sorghum seedlings. Journal of Chemical Ecology 5: 815-824. Lahmoud NR 2012. Allopathic effects of Sorghum bicolor L. in the weeds associated with subsequent crop. Ph.D. thesis. College Farooq M, Bajwa AA. Cheema SA and Cheema ZA 2013. Application of Agriculture, University of Baghdad, Baghdad, Iraq, 207. pp of allelopathy in crop production. International Journal of Agriculture and Biology 6:1367-1378. Mradu G, Saumykanti S, Sohini M and Arup M 2012. HPLC profiles of standard phenolic compounds present in medicinal plants. Gianessi L 2009. Solving Africa's Weed Problem: Increasing Crop International Journal of Pharmacognosy and Phytochemical Production and Improving the Lives of Women. Crop Protection Research 4(3): 162-167. Research Institute. pp.35. www.croplifefoundation.org. Rizvi SJH, Haque H, Singh VK and Rizvi V 1992. A discipline called Habib SA, Ali M, Hussian W, AliI and Alani A 2003. Selectivity of Chevalier in major Wheat varieties in Iraq. Documented, allelopathy. In: Allelopathy basic and applied aspects. pp:1-8. Report, Technical Bureau. Bayer Crop Science. Rizvi SJH and Rizvi V (eds.). Chapman & Hall, London. Hammood WF 2018. Effect of Sorghum ratoon plants on Steel RGD and Torrie JH 1980. Principles of Statistics. McGraw-Hill accompanied weeds of crops. International Journal of Natural Book Co. Inc. New York. USA. pp. 485.

Received 30 September, 2019; Accepted 21 November, 2019 Manuscript Number: S-272 Indian Journal of Ecology (2019) 46 Special Issue (8): 161-164 NAAS Rating: 4.96

Effect of Foliar Application of Urea and Boric Acid on Growth Parameters and Yield of Eggplant Solanum Melongena L

Aqeel Kareem Hasan Al-Tufaili, Jamal Ahmed Abbass, Mansoor Abed Aboohanah and Ali Razzak Jasim Mohammed Hraen 1Department of Horticulture and Landscape Gardening, Faculty of Agriculture, University of Kufa, Kufa, Iraq E-mail: [email protected].

Abstract: A field experiment was conducted to study the effect of three foliar application with Urea (CO (NH2)2) and Boric acid (H3BO3). The spraying urea at a concentration of 0.5 gm l-1 had a significant superiority in all vegetative growth indicators, where it gave a highest plant height, leaves number, number of main branches, fruits number, main stem diameter, total yield and total chlorophyll of leaves compared with the control treatment. Boric acid spraying effected growth parameters at a concentration 30 mg l-1 that gave a highest plant height, leaves number, number of main branches, the main stem diameter, total yield and total chlorophyll of leaves. The best interaction treatment was 0.5 g urea l-1 x 30 mg Boric acid l-1), which significantly affected the vegetative parameters, except the number of branches, where (0.5 g urea l-1 x 15 mg B l-1) treatment, had significantly impact.

Keywords: Urea, Boric acid, Foliar application, Eggplant

Eggplant/eliminate, Solanum melongena L. belongs to fruit weight and total chlorophyll content of leaf. The aim of Solanaceae family and considers as an important vegetables this study was to improve growth and yield parameters by crops in the world and Iraq. Consumers are interested in foliar application with urea and boric acid. eggplant fruit due to its content from bio-compounds, which have an important role in the human life safety like phenols MATERIAL AND METHODS and other phytochemicals (Gorinstein et al 2009), in addition An experiment was conducted during 2017-2018 in Al- to Potassium, Magnesium, Calcium and Iron (Zenia and Heera, Najaf province, Iraq. The experimental field was Halina 2008). prepared by tillage and leveling and sterilizing by pentanol at Nitrogen is one of the main nutrients, which an important a concentration (1ml l-1) and then left for 10 days after roles in the photosynthesis of, plant growth and development sterilizing. Then field was divided to three blocks, every block (Singh et al, 2003). Urea is the best source of nitrogen contain one row (36 m) and nine experimental units (every element favorable for foliar application because its experimental units was 4m). Experimental units included ten absorption and quick transition. Aujla et al (2007) mentioned plants, with distances between plants is 40 cm, and among that the eggplant fertilization by nitrogen increased yield, rows 75 cm. Eggplant seedlings (Syrian cultivar) were number of fruits, the fruit weight average and its size transplanted on May 10, 2018. significantly. Similarly, Tanko and Yusuf (2015) observed that This experiment was in randomized complete block application of nitrogen fertilizer to eggplant increased plant design with three replicates in two factors spraying with urea -1 height, number of leaves, number of nodes, number of (CO(NH2)2) at three concentrations (0, 0.25 and 0.5) gm l , branches, number of fruits plant-1 and fruit weight. Boron had spraying three times, 30,40 and 50 days from planting, The several physiological roles inplant like cell wall strength and second parameter include spraying with boric acid (H3BO3) development, RNA metabolism, sugar transfer, hormone (17%B) at three concentrations (0, 15 and 30) mg l-1, applied development, respiration, cell division, growth regulator thrice, 31,41 and 51 days from planting. metabolism (IAA) and as a part of cell membranes (Marchner Plant growth parameters plant height, leaves number, 1995). Abd El-Gawad and Osman (2014) showed that foliar number of main branches, stem diameter, number of fruits, application of boric acid on eggplant significant increased total yield and total chlorophyll of leaves were recorded. plant height, leaves number, leaf area and shoot dry weight compared with control treatment, Hussein and Muhammed RESULTS AND DISCUSSION (2017) found that spraying white eggplant with boric acid The foliar application of urea at (0.5g l-1) showed increase significantly number of leaves, leaf area, average significantly effect on the plant height, with highest plant 162 Aqeel Kareem Hasan Al-Tufaili et al height (78.41cm), compared with control. Foliar application experiment factors, where the (0.5 gm. l-1) of urea gave a of boric acid (30 mg l-1) significantly increased plant height, highest average (4.77 branch plant-1), reverse control (75.01cm), as compared with control. The best interaction treatment, which gave lowest average (2.51 branches plant- treatment (0.5 g urea l-1 x 30 mg B l-1) which significantly 1). Treatment (30 mg l-1) of Boric acid gave a highest average increased plant height (84.95 cm). (4.41 branches plant-1), control without using of Boric acid In a Table 1, results showed that the 0.5 gm l-1 (3.25 branches plant-1). The interaction treatment (0.5 g urea concentration of urea gave a highest result (76.66 leaf plant- l-1 x 15 mg B l-1) significantly superiority, where it gave the 1), compared with control treatment (41.59 leaf plant-1). While highest rate reached, (5.44 branch plant-1), compared with the 30 mg l-1concentration of Boric acid gave a highest interaction treatment (control of urea x 15 mg B l-1), (1.67 average (59.65 leaf plant-1), but the control treatment gave a branch plant-1). least number of leaves (51.27 leaf plant-1). While, the An experiment treatments significant influence on fruits interaction between two factors, the highest rate of leaf number, where results of the Table 1 that the Urea number was (85.08 leaf plant-1) at the interaction treatment concentration of 0.5 gm l-1 gave a highest average (35.77 (0.5 g urea l-1 x 30 mg B l-1) compared with control treatment fruit plant-1), compared with the control treatment (21.44 fruit for both factors (39.00 leaf plant-1) plant -1). While the Boric acid spraying didnot had significant The number of main branches affected by both effect on fruits number. Fruits number have been affected by

Table 1. Effect foliar application of urea and boric acid on plant growth and yield Boric acid (mg l-1) Plant height (cm) Urea (gm l-1) 0 0.25 0.5 Boric acid average 0 67.18 70.67 74.66 70.83 15 67.01 73.63 75.64 72.09 30 68.14 71.94 84.95 75.01 Urea average 67.44 72.08 78.41 LSD (P≤0.05) Urea=2.20 Boric acid =2.20 Interaction= 3.81 Boric acid (mg l-1) Leaves number (leaf plant-1) 0 0.25 0.5 Boric acid average 0 39.00 48.22 66.60 51.27 15 41.34 49.82 78.31 56.49 30 44.45 49.42 85.08 59.65 Urea average 41.59 49.15 76.66 LSD (P≤0.05) Urea= 1.68 Boric acid = 1.68 Interaction= 2.92 Boric acid (mg l-1) Number of main branches (branch plant-1) 0 0.25 0.5 Boric acid average 0 2.41 3.77 3.59 3.25 15 1.67 3.72 5.44 3.61 30 3.47 4.50 5.28 4.41 Urea average 2.51 3.99 4.77 LSD(P≤0.05) Urea=0.247 Boric acid = 0.247 Interaction= 0.428 Boric acid (mg l-1) Fruit number (fruit plant-1) 0 0.25 0.5 Boric acid average 0 20.33 26.33 34.33 26.99 15 19.00 27.66 34.33 26.99 30 25.00 26.00 38.66 29.88 Urea average 21.44 26.66 35.77 LSD (P≤0.05) Urea= 4.09 Boric acid = NS Interaction=7.09 Foliar Application Urea and Boric Acid on Growth of Egg Plant 163 the interaction between experiment factors, where the reached (22.08 t ha-1), compared with control treatment interaction treatment (0.5 g urea l-1 x 30 mg B l-1) gave highest (17.53 t ha-1). While, the interaction between two factors rate (38.66 fruit plant-1), compared with interaction treatment resulted in the highest total yield of 31.63 t ha-1) at the (control of urea x 15 mg B l-1) i.e., 19.00 fruit plant-1. interaction treatment of 0.5 g urea l-1 x 30 mg B l-1 compared The data in Table 2 depicted that the foliar application of with the control treatment of urea and 15 mg l-1 of Boric acid urea had effect on the main stem diameter, where the i.e., 11.89 t ha-1). average highest was at the concentration (0.5gm l-1), and The total chlorophyll content of leaves in Table 2 was reached (1.80 cm), compared with the control treatment influenced by both the factors, where the concentration of 0.5 (1.31 cm). Boric acid spraying also had significant impact on gm l-1 of urea gave a highest result (56.61 mg 100gm-1 fresh the main stem diameter, where the 30 mg l-1concentration weight), compared with control treatment which gave a gave a highest result (1.70 cm) compared with control lowest average (33.63 mg. 100gm-1. fresh weight). While the treatment (1.44 cm). Interaction between two factors has a spraying of Boric acid, the concentration (30 mg l-1) gave a significant influence on the main stem diameter rate, the highest average (55.35 mg 100 gm-1 fresh weight), compared interaction treatment (0.5 g urea l-1 x 30 mg B l-1) gave a with (15 mg l-1), which gave a lowest average (36.46 mg 100 highest rate (1.95 cm),compared with interaction treatment gm-1 fresh weight). The interaction between urea and boric (control treatment for both factors), which gave a lowest acid have effected on the total chlorophyll content of leaves, average (1.17 cm). where the interaction treatment (0.5 g Urea l-1 x 30 mg B l-1) Results of a Table 2 refers that the concentration of 0.5 gave a highest average of 66.97 mg 100 gm-1 fresh weight, gm l-1 of urea gave highest rate of total yield (25.65 t ha-1), but but the interaction treatment (control of urea x 15 mg B l-1) the control treatment gave a lowest average, reached (14.36 gave a lowest average of 24.90 mg 100 gm-1 fresh weight. t ha-1). While the second factor, where the concentration (30 The foliar application of urea had significant influence mg l-1) of Boric acid gave a highest result of total yield, on all vegetative traits (Tables 1, 2) due to the role of nitrogen

Table 2. Effect foliar application of Urea and Boric acid on main stem diameter (cm), total yield (t h-1) and total chlorophyll content of leaves (mg 100 gm-1 fresh weight) Boric acid (mg l-1) Urea (gm l-1) Main stem diameter (cm) 0 0.25 0.5 Boric acid average 0 1.17 1.47 1.69 1.44 15 1.31 1.58 1.78 1.55 30 1.46 1.71 1.95 1.70 Urea average 1.31 1.58 1.80 LSD (P≤0.05) Urea=0.112 Boric acid =0.112 Interaction= 0.195 Boric acid (mg l-1) Total yield (t ha-1) 0 0.25 0.5 Boric acid average 0 13.67 16.84 22.10 17.53 15 11.89 18.22 23.22 17.77 30 17.53 17.09 31.63 22.08 Urea average 14.36 17.38 25.65 LSD (P≤0.05) Urea= 3.10 Boric acid = 3.10 Interaction= 5.37 Boric acid (mg l-1) Total chlorophyll content of leaves (mg 100gm-1 fresh weight) 0 0.25 0.5 Boric acid average 0 28.26 50.91 56.20 45.12 15 24.90 37.80 46.68 36.46 30 47.73 51.36 66.97 55.35 Urea average 33.63 46.69 56.61 LSD (P≤0.05) Urea=12.22 Boric acid = 12.22 Interaction= 21.17 164 Aqeel Kareem Hasan Al-Tufaili et al

element, where it is consider as a one of the important growth European Agriculture 11(4): 453-458. elements in controlling yield and quality of most vegetable Aujla MS, Thind HS and Buttar GS 2007. Fruit yield and water use efficiency of eggplant (Solanum melongena L.) as influenced by crops (Sorensen et al 1995). Also Ullah et al (2010) different quantities of nitrogenand water applied through drip mentioned that nitrogen play an important role in the and furrow irrigation. Journal of Scientific Horticulture 112(2): 142-148. processes activation in several of enzymes and co-enzymes Broadley M, Brown P, Cakmak I, Rengel Z and Zhao F 2012. Chapter which is characterized by contributing to many of the bio- 7 - Function of Nutrients: Micronutrients. (In Marschner P ed, processes like photosynthesis, and this also leads to an Marschner's Mineral Nutrition of Higher Plants (Third Edition).Academic Press, San Diego, pp.191-248. increase in cells division and elongation, which forms Brown PH, Bellaloui N, Wimmer MA, Bassil ES, Ruiz J, Hu H, Pfeffer meristematic tissues and increase the size of leaf cells H, Dannel F and Römheld V 2002. Boron in plant biology. Plant (Walch et al 2000). This agreement with Aminifard et al Biology 4(2): 205-223. (2010) found that increasing nitrogen fertilizers influenced Elwan MWM and Elhamahmy MAM 2015. Boron improved growth and fruit productivity of eggplant (Solanum melongana L.). plant vegetative growth parameters (plant height, lateral Zagazig Journal of Agricultural Research 42(5):1101-1112. stem number, and leaf chlorophyll content) in eggplant. Gorinstein S, Jastrzebski Z, Leontowicz H, Leontowicz M, Namiesnik J, Najman K, Park YS, Heo BG, Cho JY and Bae JH 2009. Boron also effect on vegetative growth parameters of Comparative control of the inactivity of some frequently eggplant, due to its role in plant growth, cell wall composition, consumed vegetables subjected to different processing conditions. Food Control 20(4): 407-413. membrane stability, sugar transportation, carbohydrate Hussein WA and Muhammed MM 2017. The response of white metabolism and phenol metabolism (Brown et al 2002, eggplant plants to foliar application with boron and potassium Broadley et al 2012). Several recent reviews propose that silicate. Assiut Journal of Agricultural Science 48(1): 394-401. boron is implicated in supporting metabolic activities. Finally Marchner H 1995. Mineral Nutrition of Higher Plants. 2nd ed. Academic Press, London ,England. there was a positive relation between boric acid application Ouda BA and Mahadeen AY 2008. Effect of fertilizers on growth, and enhancing of the growth and yield of some crops like yield, yield components, quality and certain nutrient contents in eggplant (Suganiya and Harris 2015), and these findings broccoli (Brassica oleracea L.). Journal of Agricultural Biology 10(6): 627–632. agreement with Elwan and Elhamahmy (2015), where foliar Singh SS, Gupta P and Gupta AK 2003. Handbook of Agricultural application of boric acid at three concentrations (0, 5 and 10 Sciences. Kalyani Publishers, New Delhi, India. 184-185. mg l-1) on eggplant produced good results. Suganiya S and Harris KD 2015. Effect of boron on flower and fruit set and yield of ratoon Brinjal crop. International Journal of Sci. Res. and Inn. Tech. 2(1):135-141. CONCLUSION Sorensen JN, Johansen AS and Kaack K 1995. Marketable and Foliar application of urea and boric acid had significantly nutritional quality of leeks as affected by water and nitrogen supply and plant age at harvest. Journal of Science of Food and effect on plant growth parameters. Interaction treatments Agriculture 68: 367-373. between urea and boric acid also achieved good results Tanko UM and Yusuf MJ 2015. Influence of N-Fertilizer application about all studied indicators. Foliar application of fertilizers onthe yield and yield components of two varieties of eggplant (Solanum melongena L.) In Anyigba, Kogi State. Journal of was observed one of the best systems of plant nutrition Biology, Agriculture and Healthcare 5(17): 201-208. compared with traditional nutrition system. Ullah MA, Anwar M and Rana AS 2010. Effect of nitrogen fertilization and harvesting intervals on the yield and forage quality of elephant grass (Pennisetum purpureum L.) under mesic REFERENCES climate of Pothowar plateau. Pakistan Journal of Agricultural Abd El-Gawad HG and Osman HS 2014. Effect of exogenous Science 47(3): 231-234. application of boric acid and seaweed extract on growth, Walch LP, Neumann G, Bangerth F and Engels G 2000. Rapid biochemical content and yield of eggplant. Journal of effects of nitrogen form on leaf morphogenesis. Journal of Horticulture Science and Ornamental Plants 6(3): 133-143. Experimental Botany 51(343): 227-237. Aminifard MH, Aroiee H, Fatemi H, Ameri A and Karimpour A 2010. Zenia M and Halina B 2008. Content of microelements in eggplant Responses of eggplant (Solanum melongens L.) to different fruits depending on nitrogen fertilization and plant training rates of nitrogen under field conditions. Journal of Central method. Journal of Elementology 13(2): 269-274.

Received 29 August, 2019; Accepted 04 September, 2019 Manuscript Number: S-273 Indian Journal of Ecology (2019) 46 Special Issue (8): 165-167 NAAS Rating: 4.96

EFficacy of Rigenal P and Liqhumis 18 in Rooting and Vegetative Growth of Olive, Olea Europaea Cuttings

G.B.A. Al-Abbasi, F.S.S. Alabbasi1 and M.A.A. Abdulhussein Department of Horticulture and Landscaping, College of Agriculture, University of Kufa, Najaf, Iraq 1Department of Biology, College of Education, University of Kufa, Najaf, Iraq *E-mail: [email protected]

Abstract: The research was carried out to evaluate the response of three types of cuttings (limb basal, medial and lateral) to individual or combined treatments of rooting agent Rigenal P and LIQHUMUS *18 in pots under greenhouse conditions. There were significant differences between the three olive cutting types. The basal cuttings significantly had higher responses for rooting than the two other types showing the highest 30% rooting, 6.25cm root length, and 1.67cm shoots length. The highest percentage of rooting was in the combined treatment (Rigenal P+LIQHUMUS *18) in basal and medial cuttings followed by basal/Rigenal P and basal/LIQHUMUS *18. Number of roots/cutting was the highest in lateral/LIQHUMUS *18, which did not differ from that of basal/Rigenal P+LIQHUMUS *18. Root length and number of vegetative buds were the highest in basal/Rigenal P+LIQHUMUS *18 followed by the medial in same treatment. Basal cuttings also had the highest vegetative growth and number of leaves/cutting especially in the Rigenal P treatment followed by the combined treatment with medial cuttings.

Keywords: Olive cuttings, LIQHUMUS * 18, Organic fertilizer, Rooting, Rigenal P

Olive tree is an evergreen fruit tree of warm temperate MATERIAL AND METHODS regions, which is rich in high quality oil and contains a low The research was carried out at Ibn Al-Bitar in the percentage of protein. The vegetative propagation of olive province Karbala during the 2016 infactorial design) with two varieties is the best method to produce seedlings (young factors, three types of cuttings (basal, medial and lateral) and trees) using the stem cuttings of various types, seeds four soaking treatments each replicated thrice with 15 germinating and grafting. The semi-wooden cutting is one of cuttings per replicate. Each selected olive branch was cut the most common types of cuttings used for propagation on a into three 13 cm cuttings (basal, median and lateral) with four commercial scale. This type of olive propagation is low cost, leaves on the lateral end of each cutting and were subjected does not require special skills and economically reasonable. to treatments of Rigenal P (alpha naphtallenacetic acid ANA) The success of rooting of the cuttings depends on the content and/ or LIQHUMUS *18(2ml l-1). Bases of the cuttings (3 cm) of internal hormones and nutrients in the cutting, growth were dipped in distilled water for 10 min left to dry and then stimulants and other factors that help in rooting (Al-Tai 2004). dipped in the Rigenal P powder. The dipping the cuttings in Type of cutting in terms of location on a branch (basal, medial LIQHUMUS *18 for 10 min while the combined treatment was or lateral cutting) is of great importance that affecting roots dipping the cuttings in the LIQHUMUS *18 for 10 min and formation. Fontanazza et al (2001) found that semi-woody then dipping in the Rigenal P. Cuttings that dipped only in olive cuttings produce significantly much higher roots than distilled water served as control. The treated cuttings were those produced from leafy lateral cuttings. Plant auxins play planted in 2 polythen bags filled with river sand and peat an important role in increasing the successful propagation in moss 1:1 (v/v) and d cuttings were irrigated as needed and plants that are difficult to root (Al-Imam 2011). Treating basal maintained under the green house conditions. At the end of part of the cutting with humic acid increased the number of experiment, data were recorded, including percent of rooted transverse roots and reduced the time required to rooting cuttings, number of roots/cutting, root length, number of leafy (Hmaleki et al 2012). Similar results were found in studies by buds/cutting, length of new grown shoots and number of Al-Ali (2007) and Riffae and Dogogic (2013), they showed leaves/cutting. that auxins played an important role in roots producing on olive cuttings used in their studies. Therefore, the study was RESULTS AND DISCUSSION conducted to evaluate the effect of the type of cutting and The rooted cuttings, basal cuttings showed significantly basal treatments (dipping) of Rigenal P and LIQHUMUS *18 (P≤0.05) higher rates of rooted cuttings than the other two individually or in combination in successful rooting and types of cuttings (Table 1). Number of rooted cuttings was seedling growth parameters. also affected by treatments. LIQHUMUS *18 and the highest 166 G.B.A. Al-Abbasi, F.S.S. Alabbasi and M.A.A. Abdulhussein percentage of rooted cuttings which did not differ from those regardless the type of treatment, had always higher numbers resulted from the combined treatment (LIQHUMUS of grown buds than those of medial and lateral, respectively. *18+Rigenal P.) followed by Rigenal P (42.22%) and Within treatments, LIQHUMUS *18+Rigenal P did not differ similarly, the highest number of rooted cuttings from the from Rigenal P alone, all the treatments resulted in interaction treatment of LIQHUMUS *18+Rigenal P (Table1). significantly higher numbers of green buds compared to the The number of roots/cutting) showed that basal cuttings control (Table 2). Although treatment of LIQHUMUS *18 had almost always higher numbers of roots/cutting). Over its +Rigenal P in basal cuttings resulted in the highest number of corresponding control, treatment of LIQHUMUS vegetative buds (6) among all the experimental units, the *18+Rigenal P significantly (P≤0.05) increased rooting same treatment with lateral cuttings had the lowest (1.2) percentage upto86 and 40% in medial and basal cuttings among all treatments including the control. Similar to number followed by LIQHUMUS *18 alone in case of basal cuttings of vegetative grown buds, numbers of leaves/cutting were and Rigenal P alone in case of medial cuttings, respectively. also higher in basal cuttings and LIQHUMUS *18 +Rigenal P. Results also showed (Table 1) that lateral cuttings within the treatments (Table 2). The highest number ofleaves/cutting control had significantly (P≤0.05) higher numbers of was recorded in the interacted treatment of basal/Rigenal P. roots/cutting than the other two types of cuttings. Average of (16) with slight difference (P≤0.05) from same type of cutting/ root length also differed among treatments and types of LIQHUMUS *18 +Rigenal P. (15) while the same treatment cuttings (Table 1). The root length was significantly higher in with lateral cuttings had the lowest number (2.7) of basal cuttings. The combined LIQHUMUS *18+Rigenal P leaves/cutting (Table 2). treatment resulted in highest average of root length The findings showed that basal cuttings produced higher compared to other treatments. In case of interactions, the number of vegetative grown buds with higher length and highest root length was in LIQHUMUS *18+Rigenal P/medial higher number of leaves/cutting over the other types of followed by LIQHUMUS *18/Lateral (Table 1). Relative to cuttings. This may due to the interrelationship between the number of grown vegetative buds/cutting, basal cuttings, number and length of roots and the vegetative growth

Table1. Effect of different treatments with rooting powder Rigenal P. (Rig.) and LIQHUMUS*18 (LIQ.) on root parameters of olive cuttings Cutting' s Rooting percentage Number of rootscutting-1 Root length (cm) type Cont. Rig. Rig.+ LIQ. Average Cont. Rig. Rig.+ LIQ. Average Cont. Rig. Rig.+ LIQ. Average LIQ. LIQ. LIQ. Lateral 43.33 30.00 23.33 46.67 35.38 17.00 9.00 3.00 48.00 19.25 4.25 1.90 1.50 9.40 4.26 Medial 30.00 43.33 56.67 36.67 41.67 4.00 21.00 36.00 5.00 16.50 2.30 5.50 10.50 3.10 5.35 Basal 40.00 53.33 56.67 50.00 50.00 7.00 14.00 47.00 22.00 22.50 3.50 7.25 8.25 6.00 6.25 Average 37.78 42.22 45.56 44.44 9.33 14.67 28.67 25.00 3.35 4.88 6.75 6.17 CD Cuttings Treatments Interaction Rootstocks LIQ. Concent. Interaction Rootstocks LIQ. Concent. Interaction (p=0.05) 1.85 2.13 3.70 2.73 3.16 5.47 0.51 0.59 1.02 Values are means of three replicates (15 olive cuttings per replicate)

Table 2. Effect of different treatments with rooting powder Rigenal P. (Rig.) and LIQHUMUS*18 (LIQ.) on number of vegetative buds, length of green buds and number of leaves/cutting in three types of olive cuttings Cutting's Number of vegetative buds Length of green buds Number of leaves/cutting type Cont. Rig. Rig.+ LIQ. Average Cont. Rig. Rig.+ LIQ. Average Cont. Rig. Rig.+ LIQ. Average LIQ. LIQ. LIQ. Lateral 2.65 2.00 1.20 3.00 2.21 0.60 0.45 0.20 1.90 0.78 6.30 4.20 2.70 12.00 6.30 Medial 1.80 3.00 3.20 2.00 2.50 0.30 1.10 2.10 0.60 1.2 3.00 6.80 13.00 4.00 6.70 Basal 4.00 5.00 6.00 4.50 4.87 0.70 2.30 0.50 1.20 1.67 7.60 16.00 15.00 10.00 12.15 Average 2.81 3.33 3.46 3.16 0.53 1.28 1.60 1.23 5.63 9.00 10.23 8.67 CD (p=0.05) Cuttings Treatments Interaction Cuttings Treatments Interaction Cuttings Treatments Interaction 0.24 0.28 0.40 0.14 0.16 1.28 0.79 0.92 1.59 Values are means of three replicates (15 olive cuttings per replicate) EFficacy of Rigenal P and Liqhumis 18 on Vegetative Growth of Olive 167 indicators (Al-Tai 2004). It is well known thathumic acid REFERENCES participates as a supplementary source polyphenol in the Al-Ali HH 2007. Effect of indole-butyric acid and the agricultural center and certain factors on the rooting of the olive cutting Olea early stages of the plant growth, which acts as a chemical europaea L. Anbar Journal of Agricultural Sciences 5(1): 15-19. respiratory agent leading to increase the biological efficiency Al-Imam NMA 2011. Effect of some factors on rooting percentage of the plant by increasing plant enzymatic system efficiency, and subsequent growth of manzanillo olive cuttings (Olea europaea L.). Mesopotamia Journal of Agriculture 39(2): 540. cell division, root system development and the production of Al-Rifai FAR and Dugji EHA 2013. Effect of organic manure and IBA dry matter. It was also found by Turkmen et al 2004a and b) on rooting of two types of Ola. Journal of the University of that spraying with humic acid increases nutrients absorption Karbala Scientific 11(4): 55-62 and thus increases the plant growth rate. Thehumic acid can Al-Tai DKA 2004. Effect of IBA and IBA in rooting four varieties of olives (Olea europaea L.). M.Sc. Thesis, Faculty of Agriculture - increase the size of a plant by increasing water University of Kufa, Iraq. exchange(effectiveness of the water system) within the roots Atiyeh RM, Lee S and Edwards CA 2002. The influence of humic acids derived from earth worm processed organic wastes on and thus facilitate the absorption process of nutrients and plant growth. Bioresearch Technology 84: 7- 14. minerals, which reflects positively on the overall growth rate Ferrara G and Brunetti G 2010. Effects of the times of application of a of plants (Ozdamarullu et al 2011). Ferrara and Brunetti soil humic acid on berry quality of table grape (Vitis vinifera L.) Cv. Italia. Spanish Journal of Agricultural Research 8(3): 817-822. (2010) and Pouzeshi et al (2011) indicated that the increase Fontanazza G, Bartolozzi F and Cipriani M 2001. New technique to in quantitative and qualitative of plant traits using thehumic grow mother plants for continuous olive propagation. Olivae 89: acid may be due to the complex composition between the 42-46 humic acid and the mineral elements by the aid of plant Hmaleki S, Peivast G and Ghasemnejad M 2012. Effect of humic acid on vegetative traits and yield of tomato c. vlzabela. Journal enzymes. This may explain the increase in LIQHUMUS * 18 of Agricultural Sciences and lndustries 26(4): 358-363. effects when used in combination with Rigenal P in this study. Howard BH 1973. Factors affecting the rooting response of plants to Our findings confirmed that humic acid is affecting plant growth regulator application.Acta Horticulture 34: 93-106. Ozdamarullu HU,Nlu H, Kkarakurt Y and Padem H 2011. Changes in respiration and photosynthesis, stimulating the metabolism fruit yield and quality in response to foliar and soil humic acid of nucleic acids and increasing the effectiveness of plant application in cucumber. Scientific Re Scientific Research and Essrch and Essays 6(13): 2800 -2803. hormones, and thus positively affects plant growth indicators Pears HL 1946. Rooting of nine plum cutting as affected by nutrition (Turkmen et al 2004a). of the parent plantand treatment with phytohormons. Sci Bul. Dept of Agriculture. Tech. Ser No. (61): 1–13. Plants for continuous olive propagation.Olivae 89: 42–46. U.S.A. CONCLUSION Pouzeshi R, Zabihi MR, Ramezani–Moghadam MR, Rajabzadeh M It can be concluded from this study that ability of and Rajabzadeh AM 2011. Effect of foliar application of zinc rooting is different in different type of cutting. The basal ,humic acid and acetic acid on yield , yield com ponentes and mineral concentration in Grape c. vpeikani. Horticultural cuttings had always higher responses than the other types Sciences Journal (Agricultural Sciences and Industries) 25(3): of cuttings in terms of percentage of rooting, root length, 351-360. number of vegetative buds, length of new shoots and Turkmen OM, Bozkurt A, Yilddiz M and Mcimrin K 2004. Effect of nitrogen and humicacid and applications on head weight, number of leaves/cutting. We also showed that a nutrient and nitrate contents in lettuce. Advance in Food combination of more than one humic acid derivatives, Science 26: 1-6. Rigenal P+LIQHUMUS *18 in this study, were always more Turkmen O, Dursun A, Turan M and Erdinc C 2004a. Calcium and humic acid affect seed germination. Growth and nutrient effective than individual treatment in increasing all the content of tomato (Lycopersiconesculentum L.) seedlings studied growth parameters. under saline soil conditions. Acta Agriculturae Scandinavica. Section B – Soil and plant Science 54: 168-174. Received 29 September, 2019; Accepted 15 November, 2019 Manuscript Number: S-274 Indian Journal of Ecology (2019) 46 Special Issue (8): 168-172 NAAS Rating: 4.96

Effect of Genotype and Nano-Fertilizers on Some Traits and Yield of Potato

Abbas K. Mijwel and Hawraa H. Muhsin College of Agriculture, University of AL-Qassim green, Babylon,51001, Iraq E-mail: [email protected]

Abstract: The experiment was conducted with five cultivars of potato (Solanum tuberosum L.) (Arnova, Arizona, Rivera, Sifra, and Burren) in main plots and the spraying with nano-fertilizer (macronutrients + micronutrients), with three concentrations (0, 1, 2 g l-1) in the sub-plots. The Arizona cultivar gave the best results in the leaf area (10924 cm2plant-1)and the total yield amounted to 23.22 tha-1. The percentage of starch in the tubers, the percentage of phosphorus in the leaves and the percentage of potassium in the leaves were maximum in Rivera cultivar. Burren cultivar gave the highest percentage of nitrogen in the leaves.The nano-fertilizer with a concentration of (2 g l-1) gave maximum starch in the tubers and the phosphorus in the leaves. The interaction between Arizona cultivar and spraying with nano-fertilizer at a concentration of 2 g l-1 has gave highest total yield. The interaction between Burren cultivar and spraying with nano-fertilizer at a concentration of 2 g l-1 recorded the percentage of nitrogen in the leaves.

Keywords: Nanotechnology, Genotype, Starch, Phosphorus

Potato (Solanum tuberosum L.) is considered an science that processes materials at the nanoscale. This important food crop belonging to the Solanaceae family, science occupies an important place because of its many which ranks fourth in the world among food crops, and the applications in the agricultural field at all stages from importance of this crop stand out as the staple food for many production, processing, storage and packaging to of the world population. The production of this crop is transporting agricultural products (Ali et al 2014). Mijwel and influenced by many factors including suitable cultivar, Abod (2018) studied to know the effect of nano-fertilizers on climatic factors, fertilization, seed size, planting date, treating potato plant (Solanum tuberosum L.). The results showed a with hormone, agricultural operations, cultivation distance, significant effect of total fertilizer combinations + nano- its depth, and irrigation. All these factors seem to influence fertilizer in most of the traits. It should be noted here that the the competition process on the necessary building products current agricultural system in many countries of the world is between different sink centres, including tubers, which effect characterized by low productivity over time. It is therefore on the yield both quantitatively and qualitatively (Al-Bahash recommended to use modern methods with chemical 2006). Genetic factors determine the degree of growth and fertilizers for the purpose of increasing the yield and development of the organism, and cultivated cultivar greatly improving its quality. affects the quantity and quality of the crop. Kumar et al(2000)mentioned that morphological traits and productivity MATERIAL AND METHODS indicators of three potato cultivars are Diamant, Lady Rosetta The experiment was conducted at Al-Dabla region, Spunta. There is a discrepancy in the performance of the which lies south of Babylon province within 32.38 degrees studied cultivars between spring and autumn seasons, north latitude and 44.53 degrees east longitude for a season. Spunta was significantly excelled in its production in the The samples were taken at a depth of (0.3 m) from five spring season, while its productivity decreased and delayed locations. All samples were mixed in a homogeneous germination in the autumn season, on the other hand, the manner. Chemical and physical analyses for soil samples Diamant cultivar was characterized by stable performance were conducted as per standard procedure. The physical and relative stability in both spring and autumn season and and chemical properties for soil samples are given in Table 1. gave the highest productivity in autumn season. With the The experiment involved the study of two factors: the main increase in population, climate change and depletion of factor include five cultivars (Arnova, Arizona, Rivera, Sifra, natural resources, traditional agriculture is almost incapable and Burren), and the second factor (sub-plots) included of meeting people's food requirements. Therefore, nano-fertilizer (macronutrients + micronutrients) spraying on alternative and modern methods have to be used. These the total vegetative at three concentrations of (0, 1, 2 g l-1). methods include the use of nanotechnology, which is modern The plants were sprayed thrice in the early morning. First, 45 Potato Effectiveness by Genotype, Nano-fertilizers 169 days after the emergence, the second spraying after 15 days be due to the genetic difference between the cultivars of the first spraying (the emergence of tubers) and the third resulting from the variation of genetic factors responsible for spraying after 15 days of the second spraying (the large size the traits of vegetative growth or perhaps to the different of the tubers) according to different treatments. The leaf area content of these cultivars from growth hormones (Saqr (cm2 plant-1), and total yield (t ha-1) were estimated. The 2009). A positive effect of nano-fertilizer in the studied traits percent nitrogen by Micro Kjeldahl device (Black 1965), P by was recorded. This is due to the nano-fertilizers entering the Olsen and Sommers (1982) by Spectrophotometer and the plant leaves through the stomata very quickly and easily. It is percentage of potassium in the leaves by the Flame converted into xylem and phloem texture more efficiently photometer (AOAC 1970). The specific density for tubers on (Wang et al 2013). Leaf surfaces are highly porous on a the basis of dry matter estimated by method of Hassan nanometer scale and form the main gates for nutrient entry (1999) and the percentage of starch in tubers as follows, into plants (Eichert and Goldbach 2008). Nano-fertilizers Starch in tubers (%) = 17.55 + 0.891 (percentage of dry have special characteristics that are different from traditional matter - 24.182) according to (AOAC 1980). fertilizers, where they have been shown to be effective when used with many agricultural crops in growth and productivity RESULTS AND DISCUSSION due to their small size and increasing their surface area that Leaf area (cm2 plant-1): There was significant effect of the leads to increase absorption surface. Thus the ability to cultivar in the leaf area where Arizona cultivar gave improve the efficiency of the used nutrients through certain significantly higher leaf area (10924 cm2 plant-1), than other mechanisms such as targeted delivery and slow or controlled cultivars. The interaction between cultivar and nano-fertilizer release leading to increase in photosynthesis process and had no significant difference in the leaf area. It is clear from increasing nutrient production in the plant (Siddiqi and Husen Table 2 that the cultivar had a significant effect on vegetative 2017, Sohair et al 2018). The chelated iron nanoparticles growth indicators. This may be due to the genetic factors for affect the activity of the enzymes involved in the each cultivar and their suitability with the environmental photosynthesis process (Phogat et al 2016). conditions for the region such as temperature and intensity of Total yield (t ha-1): The effect of the cultivar on the total yield lighting as well as soil properties. These results agree with for potato plant was significant, where Arizona cultivar (Geremew et al 2007), Arizona cultivar significantly excelled recorded the highest yield (23.22 t ha-1), while the Sifra gave in the trait of leaf area compared to other cultivars. This may the lowest yield. There was no significant effect of nano- fertilizer on total yield. However, significant differences were observed in interaction between cultivars and the spraying Table 1. The physical and chemical properties for soil samples PH EC ds.m-1 OM N % P % K % Sand % Silt % Clay % with nano-fertilizer, where the interaction between Arizona cultivar and spraying with nano-fertilizer at concentration of 7.6 3.3 1.54 0.34 0.13 1.02 22 54 24 (2 g l-1) gave the highest yield (25.57 t ha-1). Table 2 shows the superiority of the Arizona cultivar in the total yield (tha-1). Table 2. Effect of cultivar and nano-fertilizers and their Perhaps the reason is that the yield traits are controlled by the interaction on the leaf area for potato plant (cm2 -1 genetic traits for the cultivar (Mahmoud 2003). Or the reason plant )

-1 may be due to the suitability of environmental conditions such Cultivars Nano-fertilizers gl Mean as temperature and length of a lighting period in addition to 0 1 2 the type of soil for one cultivar rather than another. It may be Arnova 6005 8057 6697 6920 (25.52) (18.94) (18.43) (20.96) due to the superiority of Arizona cultivar significantly in the 2 -1 Arizona 11183 9955 11634 10924 trait of the leaf area (cm plant ), which reflected on (21.13) (22.96) (25.57) (23.22) increasing the total yield (t ha-1). This agrees with Zelelew et Rivera 5158 6400 8699 6752 al (2016) who found that there are differences between the (16.56) (22.35) (25.37) (21.42) cultivars of the potato plant in the total yield. It may be due to Sifra 5466 4977 4402 4948 (15.11) (16.33) (15.27) (15.57) increasing the percentage of nitrogen, phosphorus, and Burren 5832 6243 8685 6920 potassium in the leaves where it is the main component for (16.40) (17.72) (17.30) (17.14) proteins and protoplasm, and plays a vital role in increasing Mean 6729 7126 8023 biomass and reproduction in plants (Meharg and Marschner (18.94) (19.66) (20.39) 2012). LSD ≤ 0.05 V=1716 (2.208) N= NS (NS) V*N = NS (2.983) LSD ≤ 0.01 V=2498 (3.213) N= NS (NS) V*N = NS (4.037) Percentage of nitrogen in leaves: The effect of the cultivar * Yield for potato plant (t ha -1) on the percentage of nitrogen in leaves for potato plant was 170 Abbas K. Mijwel and Hawraa H. Muhsin significant where Burren cultivar gave the highest differences between the cultivar and the nano-fertilizer on the percentage (4.58 %), while the Rivera cultivar recorded the percentage of potassium in the leaves for potato plant, where lowest percentage. The effect of the nano-fertilizer in the the interaction between Arnova cultivar and spraying with percentage of nitrogen in the leaves, where the spraying with nano-fertilizer at a concentration of (2 g l-1) gave the highest nano-fertilizer at concentration of (1 g l-1) gave the highest percentage of nitrogen in the leaves and there was no significant effect of from spraying with nano-fertilizer at Table 3. Effect of cultivar and nano-fertilizers and their -1 interaction on the percentage of nitrogen in concentration of (2 g l ), compared to the control treatment leaves* for potato plant (%) which gave the lowest percentage. There were also Cultivars Nano-fertilizers g l-1 Mean significant differences in the interaction between study 0 1 2 factors, where the Burren cultivar and spraying with Nano- fertilizer at a concentration of (2 g l-1) gave the highest Arnova 3.36 (0.18) 4.15 (0.30) 3.48 (0.33) 3.66 (0.27) percentage of nitrogen 5.13%, excelling on all treatments Arizona 3.44 (0.21) 3.84 (0.21) 3.89 (0.32) 3.72 (0.25) where increasing the percentage of nitrogen in the leaves as Rivera 3.61 (0.21) 3.45 (0.29) 3.61 (0.30) 3.55 (0.27) shown in Table 3 is due to the role of the elements in different Sifra 3.84 (0.13) 3.87 (0.19) 3.98 (0.16) 3.90 (0.16) biological processes, its effect on increasing the plant's Burren 3.69 (0.17) 4.93 (0.19) 5.13 (0.25) 4.58 (0.20) efficiency, its ability to absorb nitrogen and then increase the Mean 3.59 (0.18) 4.05 (0.24) 4.02 (0.27) percentage of nitrogen in the leaves as well as the effective contribution for potassium in increasing the plant's ability to LSD ≤ 0.05 V= 0.187(0.022) N= 0.092 (0.013) V x N = 0.237 (0.031) LSD ≤ 0.01 V=0.272 (0.032) N= 0.126 (0.018) V x N = 0.322 (0.042) utilize nitrogen and increase its absorption rate (Chaudhary *Percentage of phosphorus in leaves et al 2002) The Percentage of phosphorus in the leaves: The effect of Table 4. Effect of cultivar and nano-fertilizers and their the cultivar on the percentage of phosphorus in leaves for interaction on the percentage of potassium*in potato plant was significant where Arnova cultivar and Rivera leaves for potato plant (%) cultivar excelled on the other cultivars by giving it the highest Cultivars Nano-fertilizers gl-1 Effect of the cultivar percentage of phosphorus (0.27%), while Sifra cultivar gave 0 1 2 the lowest percentage. The nano-fertilizer at a concentration Arnova 1.55 (1.044) 1.33 (1.010) 1.62 (1.051) 1.50 (1.048) of (2 g l-1) gave the highest percentage of phosphorus in the Arizona 1.24 (1.052) 1.54 (1.054) 1.43 (1.051) 1.40 (1.048) leaves (0.27%), while the lowest percentage of phosphorus Rivera 1.49 (1.054) 1.46 (1.059) 1.46 (1.056) 1.74 (1.056) in the leaves at the control treatment. There were also Sifra 1.27 (1.052) 1.49 (1.056) 1.27 (1.059) 1.35 (1.056) significant differences in the interaction between study Burren 1.28 (1.05) 1.46 (1.049) 1.33 (1.051) 1.36 (1.050) factors, where Arnova cultivar and spraying with nano- fertilizer at a concentration of (2 g l-1) gave the highest Effect of the 1.37 (1.05) 1.46 (1.053) 1.42 (1.054) nano- percentage of phosphorus in the leaves (0.33%). The fertilizer N increase in the percentage of phosphorus in the leaves as LSD ≤ 0.05 V=0.029 (0.0037) N= 0.031 (0.0020) V x N= 0.062 (0.0050) LSD ≤ 0.01 V= 0.043 (0.0054) N= 0.042 (0.0028) V x N= 0.083 (0.0067) shown in Table 3 may be due to the direct addition of this *Specific density of tubers element, which increases its absorption by plant tissues, as well as the role of micronutrients present in the fertilizer, Table 5. Effect of cultivar and nano-fertilizers and their especially iron (Fe), which helps increase the absorption of interaction on the percentage of starch in tubers phosphorus in the tissues of the plant (Abdul Rasul et al for potato plant (%) 2010). Cultivars Nano-fertilizers gl-1 Mean Percentage of potassium in the leaves: The effect of the 0 1 2 cultivar in the percentage of potassium in the leaves for Arnova 8.24 8.28 8.57 8.36 potato plant was significant, where the Rivera excelled on all Arizona 8.83 9.16 8.55 8.85 cultivars in the leaves by giving it the highest percentage of Rivera 9.06 10.1 9.58 9.58 potassium (1.74%), while the Sifra cultivar had the lowest Sifra 8.71 9.43 10.09 9.41 percentage of potassium in the leaves (Table 4). The Burren 8.36 8.12 8.51 8.33 spraying with nano-fertilizer at a concentration of (1 g l-1) gave Mean 8.64 9.02 9.06 the highest value (1.46%) compared to the control treatment, which recorded the lowest value. There were significant LSD ≤ 0.05 V=0.509 N= 0.219 VxN = 0.611 LSD ≤ 0.01 V= 0.741 N= 0.300 VxN = 0.833 Potato Effectiveness by Genotype, Nano-fertilizers 171 percentage (1.62%), excelling on all interaction treatments. this cultivar can be adopted for cultivation in Babylon The specific density of tubers: The effect of the cultivar on province. The spraying with nano-fertilizer at a concentration the specific density of tubers was significant, where the two of (2 g l-1) recorded the best results. The interaction between cultivars (Rivera and Sifra) gave the highest value (1.056) Arnova cultivar and spraying with nano-fertilizer at a while Arnova cultivar gave the lowest value. The effect of the concentration of (2 g l-1) gave the best total yield (t ha-1). nano-fertilizer in the percentage of nitrogen in the leaves, where the spraying with Nano-fertilizer at a concentration of REFERENCES (2 g l-1) gave the highest value amounted to (1.054), AOAC 1970. Official Methods of Analysis. 11th. Ed. Washington D.C. Association of the Official Analytical Chemist.1015 P. compared to the control treatment which recorded the lowest AOAC 1980. Official Methods of Analysis Association of Official value. However, the effect between the cultivar and the Analytical Chemists, 13thed, Washington. USA. Nano-fertilizers had no significant difference in the specific Abdul R, Iman J, Kadhim DH and Fadel HA 2010. Effect of foliar density for the tubers of the potato plant. application of ungreen and solu potash on yield and quality of potato tuber. Jordan Journal of Agricultural Sciences 6(1): 111- Percentage of starch in tubers: The effect of the cultivar on 119. the percentage of starch in tubers was significant, where Al-Sahuki MM, Bashir A and Francis G 2004. Nitrogen relationship to Rivera cultivar gave the highest percentage (9.58%), while soybean seed protein. Journal of Iraqi Agricultural Sciences 35(2): 53-58. Burren cultivar gave the lowest percentage. The effect of Al-Shehawi FA 2009. Studying the correlation between morpho- nano-fertilizer in the percentage of starch in tubers, where the physiological traits and productivity for several potato cultivars -1 (Solanum tuberosum L) at different levels of moisture. Ph.D. spraying with a concentration of (2 g l ) gave the highest Thesis, College of Agriculture.ALBaath University. Damascus. percentage of starch in tubers (9.06%), compared to the Syria. control treatment, which gave the lowest value (Table 4). Ali MA, Rehman I, Iqbal A, Din S, Rao AQ, Latif A, Samiullah TR, Azam S and Husnain T 2014. Nanotechnology a new frontier in There were also significant differences in the effect of agriculture. International Journal of Advances in Life Science cultivars and nano-fertilizers on the percentage of starch in 1(3): 129-138. potato tubers, where the interaction between Rivera cultivar Bahash NA 2006. Guidelines for potato production. Ministry of Agriculture - General Authority for Agricultural Extension and -1 and spraying with nano-fertilizer at a concentration of (1 g l ) Cooperation - Extension Bulletin. gave the highest percentage (10.10%), which did not differ Black A 1965. Methods of soil analysis part 2. Amererican Science of from the Sifra cultivar that sprayed with Nano-fertilizer at a Agronomy Inc. U.S.A. -1 Chaudhary AN, Ali S and Hassan I 2002. Effect of different color concentration of (2 g l ) amounted to (10.09%), while the plastic mulches on the yield and nutrient contents of the tomato interaction between Burren cultivar and spraying with Nano- plant.,Asian Journal of Plant Science 1(4): 388-389. fertilizer at a concentration of (1 g l-1) gave the lowest Eichert T and Goldbach HE 2008 .Equivalent pore radii of hydrophilic foliar uptake routes in sto-matous and astomatous leaf percentage of starch in tubers. The significant effect of nano- surfaces: further evidence for a stomatal pathway. Physiology fertilizer on the studied traits is attributed to the fact that its Plant 132(4): 491-502. use increases the rate of photosynthesis process and Geremew EB, Steyn JM and Annandale J .2007. Evaluation of growth performance and dry matter partitioning offour produces more dry matter (Qureshi 2018). The increase in processing potato (Solanum tuberosum L.) cultivars. New the density of tubers and the percentage of starch in the Zealand Journal of Crop and Horticultural Science 35(3): 385- 393. tubers due to the addition of nano-fertilizers and their role in Hassan A 1999. Potato Production, Vegetable Crops Series: stimulating the speed of biochemical reactions leading to the Production Technology and Advanced Agricultural Practices, production of growth substances and stimulating enzymatic 1st ed., Arab Publishing House, Egypt, p. 446. Jabour MA, Nizar H, Abdullah H and Nael A 2009. Studying activity, which in turn leads to increased production of performance of three potato cultivars (Solanum tuberosum L.) biomass and carbohydrates and increase the dry matter (Jan in spring and autumn seasons in Daraa province. General Commission for Scientific Agricultural Research. Horticulture Mohammadi et al 2016). It can also be attributed to the role of Research Department. Damascus. p.113. the boron element important in the life of the plant through its Janmohammadi M, Pornour N, Javanmard A and Sabaghnia N 2016. great importance in many vital processes, including the Effects of bio-organic, conventional and nano-fertilizers on growth, yield and quality of potato in cold steppe. Botanica transfer of saccharides from the places of its formation to Lithuanica 22(2):133-144. growth areas and storage in tubers, as well as its role in the Kumar A, Dahiya MS and Bhutani RD 2000. Performance of brinjal activation of many enzymatic reactions, thus increase growth (Solanum melongena L.) genotypes in different environments of the spring summer season. Haryana Journal of Horticulture expressed by increasing dry matter and starch. 11: 63-67. Mahmood SAW 2003. Studying some traits of vegetative growth and CONCLUSIONS yield for five potato cultivars (Solanum tuberosum L.) under the conditions of spring cultivation for the central region of Iraq. Cultivars varied in its response, where Arizona cultivar Tikrit Journal of Agricultural Sciences 3(5): 105-113. gave the best results in most of the studied indicators so that Meharg A and Marschner P 2012. Marschner's mineral nutrition of 172 Abbas K. Mijwel and Hawraa H. Muhsin

higher plants. Experimental Agriculture 48(2): 305-310. Saqr MT 2009. Growth and Flowering regulators. Mansoura Mijwel AK and Abboud AK 2018. Growth and yield of potato University, College of Agriculture, Department of Plant (Solanum tuberosum L.) as influenced by nano-fertilizers and Physiology, Egypt. different planting dates. Research on Crops 19(4): 649-654. Siddiqi KS and Husen A 2017. Plant response to engineered metal Mosli HA 2000. Potatoes: Cultivation, pests, storage and processing oxide nanoparticles. Nano Research Letter 12:92. of their products, Dar Aladdin Publications, Damascus, Syria. Sohair EED, Abdalla AA, Amany AM and Houda RA 2018. Effect of Olsen SK and Sommers LE 1982. Phosphorus. In Page AL et al nitrogen, phosphorus and potassium nano fertilizers with (Eds) Methods of Soil Analysis. Am. Agro. Inc. Madison, different application times, methods and rates on some Wisconsin, New York. growthparameters of Egyptian cotton (Gossypium barbadense L.). Bioscience Research 15(2): 549-564. Phogat N, Khan SA, Shankar S, Ansary AA and Uddin I 2016. The fate of inorganic nanoparticles in agriculture., Adv. Mater. Wang WN, Tarafdar JC and Biswas P 2013. Nano-particle synthesis Letters 7(1): 3-12. and delivery by an aerosol routefor watermelon plant foliar uptake. Journal of Nano Particle Research 15(1): 1–13. Qureshi A, Singh DK and Dwivedi S 2018. Nano-fertilizers:Anovel way for enhancing nutrient use efficiency and crop productivity. Zelelew DZ, Lal S, Kiddane TT and Ghebeslassie BM 2016. Effect of International Journal of Current Microbiology and Applied potassium levels on growth and productivity of potato varieties. Science 7(2): 3325-3335. American Journal of Plant Science7(12):1629-1638.

Received 22 September, 2019; Accepted 20 November, 2019 Manuscript Number: S-275 Indian Journal of Ecology (2019) 46 Special Issue (8): 173-178 NAAS Rating: 4.96

Effect of Niacin (Nicotinamide) and Humic Acid on Growth and Chemical Traits of Pelargonium hortorum L.

Thaer Y. Khudair, Fakhria A. Abdul Albbas and Khansaa A. Abdul Kreem Department of Horticulture and Landscape, College of Agriculture, University of Basrah, Basrah, Iraq E-mail: [email protected]

Abstract: This experiment was conducted in the lath house (covered with Saran cloth)at the College of Agriculture, University of Basra to study the response of the plant Pelargonium hortorum L. for the spraying with humic acid organic fertilizer at a concentration of 0, 50, 100 gm-1 and the nicotine amide at a concentration 0, 40, 80 gm-1 in the vegetative and flowering growth and chemical traits, with three spraying. Spraying led to the improvement of all of vegetative and flowering growth indicators and chemical traits, which were included plant height, number of leaves, the number of lateral branches, leaf area, flowering date, flowering duration, number of the inflorescence, number of florets, leaf content of total chlorophyll, anthocyanin pigment, nitrogen, phosphorus and potassium.

Keywords: Geranium, Humic acid, Nicotine amide, Growth

Pelargonium hortorum belongs to the Geraniaceae in many plants (Bearder 1980 and Arteca 1996). Vitamin B3 family, which includes more than 11 genus (Saad 1994). It (niacin) is one of the vitamins, also called Nicotineamide. It is originated in South Africa, where it is found naturally growing a type of vitamin B group that has an effect in many biological in hot and dry environments in sandy, rocky and sandy hills. processes in plants because it is a component of the enzyme (Vanderwalt 1977). Geranium has been propagated, hybrids NADP + and NAD + (Bearder 1980, San and Ota 1977). This and Improved qualitative traits since 904 until its species vitamin is also called nicotinic acid. For the purpose of not reached in 2004 more than 500 commercial species (Erilk mixing between nicotinic acid and nicotine tobacco, this is 2006). The demand increased for them significantly in recent called niacin, a white substance that dissolves with water, years. It has become one of the plant groups of economic basal substances, and alcohol, but does not dissolve in ether value important for the beauty of its leaves and flowers and (AlKilani and Abdul Hassan 1986). Organic fertilizers are the multiplicity of colors and sizes and the less of infection represented by humic acid is one of the humus compounds with pests and the length of the flowering period (Larson and resulting from the decomposition of organic matter (Al-

Fonterno 1992). It increased sales in the United States, Nuaimi 1999). It chemically symbolizes 2CO3(COOH) especially California, to reach 160 million dollars in the years C75H33O17N3. It contains carbon, hydrogen, nitrogen, and 1979-1999 (Wazir 1999). Foliar fertilization is one of the oxygen in its composition and in varying proportions, important factors in increasing the growth rate, completing resulting in the formation of compounds of varying molecular the formation of flowering and its quality. This is done by weights (Senesi 1992). The effect of humic acid on plant following proper and balanced fertilization programs that growth depends on the source of organic fertilizer and provides the plant all its nutrient needs (Abu Dahi and Al concentration and molecular weight of the humic molecule. Younis 1988). The reduction of the necessary nutrient has Low molecular weights easily reach to the cell membrane of negative effects on the biological processes that take place plant cells and enter the cell, while the high molecular weights inside the plant. Therefore, it became necessary to provide reaction with the cell wall and are not carried into the cell these elements by spraying them on the vegetative total to be (Nardi et al 2002). The studies indicate the possibility of using absorbed directly by the plant to avoid being exposed to humic acid as a growth regulator by regulating the level of washing and fixing when added to the soil (AlSahaf 1994). hormones in the plant, in addition to increasing plant Foliar mineral nutrition has been shown to be an effective resistance to water stress and stimulating plants method for better nutrient transport within the plant, followed Pelargonium to increase the percentage of flowers (Piccolo by its contribution to the natural growth of plants (Al-Jawari et al 1992, Nardi et al 2002, Morard et al 2011). This study 2002). Vitamins play a big role in their effects on vegetative was conducted due to the importance of geranium plant and and flowering growth. Many endogenous organic the need to expand the cultivation of its aesthetic and substances, such as plant growth regulators and vitamins, coordination value, whether cultivated pots or in the garden have been used to promote growth and improve productivity land. 174 Thaer Y. Khudair, Fakhria A. Abdul Albbas and Khansaa A. Abdul Kreem

MATERIAL AND METHODS increase of internal cytokinin and Auxin, which in turn This experiment was conducted in the lath house stimulate cell division in the plant (Ervin and Zhang 2004, covered with Saran cloth at the Department of Horticulture Schmidt and Zhang 2000). Humic acid treatment also and landscape gardening at the College of Agriculture, increased the accumulation of carbohydrates that stimulate University of Basra on the plant Geranium (Pelargonium lateral buds to grow (Turkmen et al 2004). The increase in the hortorum L) from Feb. to June 2019. Plants brought from one number of lateral branches leads to an increase in the of the nurseries in Baghdad on 10/2/2019 and at the age of a number of leaves, which reflected positively on the leaves month. Seedlings were transferred on 12/2/2019 to pots, area because humic acid contains micronutrient that has a where they were washed and sterilized with a 4% formalin positive role in the growth of the plant as it increases the cell solution. Before starting the agricultural operations, a division of the meristem (AbdEl_Monem et al 2011). random sample of soil was taken for analysis. Table 1 shows These results agree with Hafez and Mahmoud (2010) some chemical and physical traits of the river soil. The on the potato plant and Safana (2013) on the dahlia plant and experiment was conducted as a factorial experiment (3×5) Al- Aljubore (2014) on the geranium plant. This is due to the according to Randomized Complete Blocks Design with positive effect of being the main component of the three replicates and the averages were compared using the biosynthesis of NAD and NADP enzyme compounds as it is least difference test (LSD) below the 5% probability level needed by the plant in carbohydrate and amino acid (Alrawi and Khalaf Allah 1980). metabolism. It is also involved in the formation of the indole Plant height, number of leaves, number of lateral acetic acid (IAA) and is elongated by cells (Devlin and branches, leaf area, flowering date/duration, number of Witham 1985).It also enters the synthesis of proteins and inflorescence, number of florets, chlorophyll content, nucleic acids involved in the synthesis of organic bases and a n t h o c y a n i n c o n t e n t , N P K c o n t e n t s w e r e the growth hormones such as cytokinin (Robinson 1973). It measured/estimated. has a relationship with the tryptophan amino acid in plant tissues, which is considered the basis in the construction of RESULTS AND DISCUSSION Auxin(IAA) important in the expansion and elongation of cells Vegetative Growth (Abu Dahi and AlYounis 1988). The results in Table 3 showed that the spraying plants Flowering Growth with a humic acid organic fertilizer at a concentration of 100 The results in Table 4 showed that the spraying plants mg l-1 has a significantly in the effect as it recorded the highest with a humic acid organic fertilizer at a concentration of 100 average of plant height, number of leaves, leaf area, number mg l-1 was a significantly affect with the highest average of the of lateral branches, which amounted to 36.511, 42.76, flowering date, flowering duration, number of the 3116.0, 5.067, respectively compared with control plants inflorescence, number of florets which gave 69.641, 48.769, (30.256, 29.26, 2324.0, 2.589, respectively). The same table 5.833, 13.456, respective values compared with control also shows that the spraying with nicotine amide at a plants 91.708, 32.726, 3.467, 9.479, respectively. The same concentration of 80 mg was significant which gave the table also shows that the spraying with nicotine amide at a highest average of plant height, the number of leaves, concentration of 80 mg was significant which gave the number of lateral branches, leaf area of 34.467, 39.48, 2908.0, 4.556, respectively compared with control plants i.e., Table 1. Some chemical and physical traits of the study soil 31.389, 31.33, 2438.0, 2878, respectively. As for the bi- used in the experiment interaction between the organic fertilizer humic acid and Traits Unit Value nicotine amide, it was found to have a significant effect on Soil texture ____ Sandy loam improving the vegetative growth indicators, where the sand 1g-kg 861.8 treatment of spraying with organic fertilizer humic acid and silt 1g-kg 80.84 nicotine amide at a concentration of (100 + 40 mg)was clay 1g-kg 57.36 recorded the highest average of plant height, number of PH ____ 7.31 leaves, number of lateral branches and leaf area i.e., 39.113, E.C dsm-1 1.54 47.97, 3613.0 and 6.20, respectively compared to the lowest average amounted to 29.300, 27.30, 2099 and 1.767, Organic matter % 1.84 respectively for control. Zn++ ppm 1.37 The increase in plant height may be due to that the humic N ppm 171 acid contains cytokinin and that its addition results in an P ppm 2.47 Pelargonium effectiveness by Niacin and Humic Acid 175

Table 2. Chemical composition of POWHUMUS organic phosphorus, potassium and flower content of anthocyanin fertilizer according to Humintech GmbH pigment, which gave values of 78.03, 2.075, 0.609, 5.228, Potassium humate 85% 33.19, respectively compared to the control plants 66.14,

Water-soluble potassium (K2O) 11.00% 1.501, 0.451, 3769, 26.26, respectively for control. The same Nitrogen 0.80% table also shows that the spraying with nicotine amide at a Iron 1.00% concentration of 80 mg was significant which gave the Humidity 12-10% highest the leaf content of total chlorophyll ,Percentage of Dry matter 88-90% nitrogen, phosphorus, potassium and flower anthocyanin Degradation 99.80% pigment (75.59, 1.722, 0.583, 4.754, 31.04, respectively) compared with control plants (69.55, 1.704, 0.486, 4.166, Other materials 15% 27.77, respectively). highest average of the flowering date, flowering duration, As for the bi-interaction between the organic fertilizer number of the inflorescence, number of florets which gave humic acid and nicotine amide, it was found to have a 76.269, 45.413, 5.100, 12.289, respectively in comparison significant effect on improving the chemical traits, where the with control plants i.e., 84.510, 36.253, 3.900 and 10.558, treatment of spraying with organic fertilizer humic acid and respectively. As for the bi-interaction between the organic nicotine amide at a concentration of (100 + 40 mg)was fertilizer humic acid and nicotine amide, it was found to have recorded the highest average of the highest the leaf content a significant effect on improving the vegetative growth of total chlorophyll, percentage of nitrogen, phosphorus, indicators, where the treatment of spraying with organic potassium and flower content of anthocyanin pigment i.e., fertilizer humic acid and nicotine amide at a concentration of (100 + 40 mg)was recorded the highest average of the Table 3. Effect of spraying with organic fertilizer humicacid flowering date, flowering duration, number of the and niacin (nicotine amide) and their interactions on inflorescence, number of florets, which gave 68.400, 54.572, vegetative growth traits of geranium plant 6.733, 14.543 compared to the lowest average amounted to (Pelargonium hortorum L) 97.667, 30.201, 2.733, 8.637, respectively. The increase Traits/ Plant height Number of Leaves Number of Treatments (cm) leaves (leaf area (cm2) lateral may be due to the positive effect to organic fertilizer humic plant-1) branches acid, which provides natural stimulants such as Auxin and -1 Cytokinin and amino acids, which led to encouraging growth Humic acid levels (mg ) and therefore early flowering (Al-Ani 1987). The reason for 0 30.256 29.26 2324 2.589 the long vase life to the content of humic acid in the micro 50 33.467 36.48 2725 3.622 nutrients that stimulate the vegetative growth and reflected 100 36.511 42.76 3116 5.067 on improving the efficiency of photosynthesis and energy- LSD 0.42 1.664 53.3 0.27 rich compounds, thus improve plant growth and prolong Nicotine amide levels (mg-1) flowering duration (Adams and Winsor 1979). This allowed it 0 31.389 31.33 2438 2.878 to form the largest number of inflorescences and to increase 40 34.378 37.68 2818 3.844 the number of florets (Morard et al 2011). These results agree 80 34.467 39.48 2908 4.556 with Chang (2012) on Lilium and Ambreen et al (2014) on LSD 0.42 1.664 53.3 0.27 Petunia. While the reason for the increase in total flowering -1 when treated with nicotine amide due to a cycle in the Interaction levels between humic acid and nicotine amide (mg ) production of amino acids and nuclear which is reflected 0 29.3 27.3 2099 1.767 positively on the manufacture of carbohydrates and 40 29.9 29.1 2065 2.033 transported from the leaves to flower buds. This leads to a 80 31.657 31.37 2806 3.967 balance between carbohydrates and protein, which have a 0 50 31.2 30.3 2484 3.033 positive role in the flowering date, flowering period, number of 40 40 34.1 35.97 2777 3.3 inflorescences and florets (Florent 1986). 80 80 35.1 43.17 2914 4.533 Chemical Traits 0 100 33667 36.4 2731 3.833 The results in Table 5 showed that the spraying plants 40 40 39.113 47.97 3613 6.2 with a humic acid organic fertilizer at a concentration of 100 80 36.733 43.9 3003 5.167 mg l-1 was significantly affected with the highest average of 80 the leaf content of total chlorophyll, percentage of nitrogen, LSD 0.74 2.881 92.2 0.469 176 Thaer Y. Khudair, Fakhria A. Abdul Albbas and Khansaa A. Abdul Kreem

Table 4. Effect of spraying with organic fertilizer humicacid Table 5. Effect of spraying with organic fertilizer humicacid and niacin (nicotine amide) and their interactions on and niacin (nicotine amide) and their interactions on flowering growth traits of geranium plant chemical traits of geranium plant (Pelargonium (Pelargonium hortorum L) hortorum L) Traits Flowering Flowering Number of Number Trait/ Chlorophyll Anthocyanin Nitrogen Phosph Potassi Treatments date (day) duration the of florets Treatments (mg 100 g-1 (mg 100 g-1 % orus um (day) inflorescence fresh dry weight) % % weight) Humic acid levels (mg-1) Humic acid levels (mg-1) 0 91.708 32.726 3.467 9.479 0 66.14 26.26 1.501 0.451 3.769 50 75.856 43.165 4.622 11.36 50 75.46 29.06 1.72 0.507 4.526 100 69.641 48.769 5.833 13.456 100 78.03 33.19 2.075 0.609 5.228 LSD 0.304 0.133 0.303 0.274

-1 LSD 1.099 0.451 0.063 0.064 0.187 Nicotine amide levels (mg ) Nicotine amide levels (mg-1) 0 84.51 36.253 3.9 10.558 0 69.55 27.77 1.704 0.486 4.166 40 76.426 42.994 4.922 11.448 40 74.49 29.71 1.869 0.498 4.604 80 76.269 45.413 5.1 12.289 80 75.59 31.04 1.722 0.583 4.754 LSD 0.304 0.133 0.303 0.274

-1 LSD 1.099 0.451 0.063 0.064 0.187 Interaction levels between humic acid and nicotine amide (mg ) Interaction levels between humic acid and nicotine amide (mg-1) 0 0 97.667 30.201 2.733 8.637 0 0 63.1 24.38 1.508 0.433 3.624 40 90.51 31.092 3.167 8.6 40 66.18 24.79 1.434 0.426 3.375 80 86.947 36.884 4.5 11.2 80 69.13 29.62 1.562 0.495 4.308 0 50 85.38 35.472 4.067 10.713 0 50 70.86 27.49 1.618 0.496 4.082 40 70.367 43.317 4.867 11.2 40 75.58 28.66 1.776 0.458 4.602 80 71.82 50.707 4.933 12.167 80 79.94 31.04 1.765 0.565 4.895 0 100 70.483 43.087 4.9 12.323 0 100 74.68 31.44 1.987 0.528 4.792 40 68.4 54.572 6.733 14.543 40 81.72 35.7 2.397 0.611 5.835 80 70.04 48.649 5.867 13.5 80 77.7 32.45 1.841 0.689 5.058 LSD 0.74 0.527 0.23 0.525 LSD 1.904 0.781 0.11 0.111 0.324

81.72, 2.397, 0.611, 5.835, 35.70, respectively compared to positively on the photosynthesis products that increase the the lowest average amounted to 63.100, 1.508, 0.433, 3.624, nitrogen percentage (Minkel and Kirby 1984). The reason for 24.38, respectively. the increase in phosphorus is due to the role of iron in the The increase in leaf content of chlorophyll when treated organic fertilizer humic acid Table 2, which helps in with humic acid may be due to macro and micro nutrient and increasing plant growth as well as its effect in increasing the iron elements (Table 2). Nitrogen stimulates the activity of availability of the nutrients. The increase in the nutrients some enzymes in photosynthesis (Mohammed 1985). content (nitrogen, phosphorus, potassium) may be due to the Nitrogen and iron also enter the structure of the chlorophyll spraying plants with vitamin nicotine amide may lead to an molecule and stimulate them to build chlorophyll (AlSahaf increase in carbohydrates during photosynthesis, which 1989). While the increase of the anthocyanin pigment may be leads to increased synthesis of the enzyme nitrate reductase due to the increase of plant content of substances like auxin necessary to reduce nitrates and their representation, which and cytokinin and contains macro and micro nutrient and thus leads to an increase and accumulation of nitrogen in the increase the process of photosynthesis and carbohydrates leaves (Witham and Witham 1985). It is also involved in the that increase the construction of anthocyanin in flowers formation of both co-enzyme NAD + and NADP +, which are (Aljubore 2014). These results agree with Parandian et al composed of the reaction of nicotine amide and Adenosine (2012) and Aljubore (2014) on geranium and Abbas (2018) triphosphate (ATP), which work in the processes of oxidation on Matthiola. The positive effect of humic acid on leaf and reduction in carbohydrate transformation processes and nutrients for the geranium plant is due to the increase of thus increase phosphorus within the plant (Robinson 1973). nitrogen percentage through direct feeding of this nutrient, These results agree with Tarraf et al (1999) found in which increases the vegetative total. This is reflected Lemongrass and Al-Ali (2011) on dahlia. Pelargonium effectiveness by Niacin and Humic Acid 177

REFERENCES Consumption. Association of Floriculture Professianals Abbas Moayad Fadel and Mohsen Jalab Abbas 1992. Care and No.894. www.fao.org storage of fruit and vegetables (practical). Dar Al - Hikma Press, Florent J 1986. Vitamins in Biotechnology. In : Page , H and Basra University, Iraq Rehmeds, H.J., 4 : 115 – 158. VCH. Verlagsgesell-schaft MBH, Abbass JA and Bader KSM 2018. Effect of spraying activated dry D-6940 Weinheim, Fedral Republic of Germany . yeast extract and vitamin B3 on growth characteristics of stock Fuleki T and Francis FJ 1968. Quantitative methods for plant Mathiola incana L. Journal of Agricuttual Research 6(5): anthocyanins. 1. Extraction and determination of total 142-147 anthocyanins in cranberries. Journal of Food Science 33: 72- Abd El-Monem EAA, EL-Ashry SM and Mostafa EAM 2011. 77. Performance of coratina olive seedlings as affected by spraying Gharib A 2002. Effect of nicotin amide on growth and volatile oil humic acid and some micro elements. Journal of Applied composition of various parts of sweet fennel (Foeniculum Sciences Research 7(11): 1467-1471. vulgare var. dulce) plants. Journal of Biological Sciences 2(4): Abu Dahi Yousef Mohammed and Muayad Ahmad Al-Younis 1988. 250-253. Plant nutrition guide. Dar Al Hekma Press for Printing and Larson RA and Fonterno WC 1992. Geraniums introduction to Publishing. Ministry of Higher Education and Scientific floriculture. California. Academic press. Pp 451-475. U.S.A. Research. University of Baghdad. Iraq. Le C, Wu Y, Xu WW, Nikbakht A and Xia YP 2010. Effects of calcium Al-Ali Fakhria Abdullah Abdul Abbas 2011. Effect of pinching and and humic acid treatment on the growth and nutrient uptake of spraying with vitamin C and B3 and iron and zinc elements on oriental lily. African Journal of Biotechnology 11(9): 2218-2222. Dahlia variabilis L. growth and flowering. Doctoral dissertation, Mahmoud AR and Hafez MM 2010. Increasing productivity of potato College of Agriculture. University of Basra - Iraq. plants (Solanum tuberosum L.) by using potassium fertilizer Al-Ani TA 1987. Physiology of plant growth and composition. Ministry and humic acid application. International Journal of Academic of Higher Education and Scientific Research, University of Research 2(2): 83-88. Baghdad, Iraq. Minkel K and Kirby JA 2000. Principles of plant nutrition. Translated Al-Jawari Abdul Rahman Khamas Suhail 2002. Effect of spraying by Saadallah Najm Abdullah Al-Naimi, Directorate of the House with different nutrients on growth and yield of sweet pepper of Books for Printing and Publishing, University of Mosul _ Iraq. Capsicum annuum L. Master Thesis, College of Agriculture, Mohammed AAK and Abdul HR 1982. Plant Physiology. The second University of Baghdad, Iraq. part. Ministry of Higher Education and Scientific Research. Al-JubouriZeinaKhalif Mahmoud Khalif 2014. Effect of humic acid Morard P., Boris E, Marie M and Jerome S 2011. Direct effects of and cycocell on growth and flowering of geranium plant humic-like substance on growth, water, and mineral nutrition of Pelargonium x hortorum L. College of Agriculture, University of various species. Journal of Plant Nutrition 34: 46-59. Mosul. Iraq. Murphy T and Riley JR 1962. A modified single solution method for Al-Nuaimi, Saadallah Najm Abdullah 1999. Fertilizers and soil the determination of phosphate in natural water. Analytica fertility. Kutub House for Printing and Publishing, Ministry of Chemical Acta 27: 31-36. Higher Education and Scientific Research, Mosul University. Nardi S, Pizzeghello D, Muscolo A and Vianello A 2002. Alrawi Khasha Mahmoud and Abdul Aziz Mohammed Khalaf Allah Physiological effects of humic substances on higher plants. Soil 1980. Design and analysis of agricultural experiments. Dar Al Biology and Biochemistry 34: 1527-1536. Kutub Establishment for Printing and Publishing, University of Page AL, Miller RH and Keeney DR 1982. Method of Soil analysis Mosul / Iraq. Part 2, 2nd Ed. Madison Son, Wisconson, U.S.A.,1159 p. AlSahaf FH 1994. Effect of the number of times sprayed with liquid Parandian F and Samavat S2012. Effects of fulvic and humic acid on nutrient solution (Nahrain) on growth and yield of potatoes. anthocyanin, soluble Sugar, μ Amylase enzyme and some Journal of Iraqi Agricultural Sciences 25(1): 95-100. micro nutrient elements in Lilium. International Research Ambreen SM, Muhammed HB and Ahmed RB 2014. Influence of Journal of Applied and Basic Sciences 3(5): 924-929. humic acid and macronutrinents (mgso4+s) application on Piccolo A, Nardi S and Concheri G 1992. Structural characteristics of growth and yield of petumia (Petunia milliflora). International humus and biological activity. Soil Biology & Biochemistry 24: Journal of Agricultural Technology 10(6): 1493-1508 . 273-380. Amin ERM 2018. Effect of spraying with chelated iron and organic Robinson FA 1973. Vitamins in Phytochemistry, Vol. III,In : Lawrence fertilizer (ALGIDEX) and their interaction on the vegetative and P Miller (Ed.). pp. 195-220. Van-Nostrand Reinhold Co., New Flowering growth and the oil yield of Gardania jasminoides ellis York. L.. College of Agriculture. University of Basra. Saad SI 1994. Floral plants, its origins - evolution - classification. Dar Arteca EN 1996. Plant Growth Substances: Principles applications. Al-Fikr Al-Arabi for Printing and Publishing. Chapman and Hall, Now York. Safana HS 2013. Effect of humicacid spraying on some vegetative Atta JM 2010. Humic acids and plant nutrition. Arab Fertilizers growth characteristics of Dahlia hyprida. Master Thesis, Forum. College of Agriculture, Muthanna University, Iraq. Bearder IR 1980. Plant hormones and other growth substances. San JS and Ota Y 1977. Plant growth regulating activities of nicotin Their back ground , structure and occurrence in hormonal amide.ll. Effect of nicotin amide on growth of several crops. regulation and development. J. MacMillan (ed) Encyclopedia of Japan Journal of Crop Science 46:8-12. plant physiology, New Seties, Vol.9:112, Springer – Verlage, Senesi N 1992. Metal- Humic Substances Complexes In The Berlin. Environment Molecular and Mechanistic Aspects By Multiple Cresser MS and Parsons JW 1979. Sulphuricperchloric acid Spectroscopic Approach. Lewis Pub. Co., New York. digestion of plant material for the determination of nitrogen, Tarraf SA, El-Din KG and Balbaa LK 1999. The response of potassium, calicium and magnesium. Phosphorus chemical vegetative growth essential oil of lemongrass Cymbopogom Analytical Acta 109: 431- 436. citrates to foliar application of ascorbic acid, nicotin amide and Devlin RM and Witham FH 1985. Plant Physiology, Translated by some micro-nutrients . Arab Univ. Journal Agricultural Sciences Mohamed Fawzi Abdel Hamid, Mohamed Sharaqi, Abdel 7(1): 247-259. HadiKhedr, Nadia Kamel and Ali Saad Eddin Salama, Arab Turkmen O, Dursun A, Turan M and Erdinc C 2004. Calcium and Publishing and Distribution House, Arab Republic of Egypt. humic acid affect seed germination, growth, and nutrient Erilk R 2006. Temperature effects on floriculture crops and Energy content of tomato (Lycopersicon esculentum L.) seedlings 178 Thaer Y. Khudair, Fakhria A. Abdul Albbas and Khansaa A. Abdul Kreem

under saline soil conditions. Acta Agriculturae Scandinavica, Research, New Delhi. India). Section B- Plant Soil Science 54(3): 168-174. Youssef AA and Talaat IM 2003. Physiological response of rosmary Vanderwalt JJA 1977. Pelargonium of Southern Africa. Purnell and plant to some vitamins. Egypt Pharm Journal 1: 81-93. Sons, SA Limited, Cape Town. ISBN. 0-86843-006-4. Zhang X and Ervin EH 2004. Cytokinin- containing seaweed and Walsh LM and Beaton JD 1973. Soil testing and plant analysis. humic acid extracts associated with creepingbentgrass leaf Madison, Wis : Soil Science Society of America, Inc., 512 p. cytokinins and drought resistance. Crop Science 44: 1737- Wazir JS 1999. Performance studies in pelargoniums. Annual 1747 Report, RHRS, Mashobra,Shimla (Bhattacharjee SK Advances Zhang X and Schmidt RE 2000. Hormone-containing products in ornamental horticulture. Volume 2. Herbaceous perennials impact on antioxidant status of tall fescue and creeping bent and shade loving foliage plants Indian council of Agricultural grass subject to drought. Crop Science 40: 1344-1349.

Received 23 September, 2019; Accepted 20 November, 2019 Manuscript Number: S-276 Indian Journal of Ecology (2019) 46 Special Issue (8): 179-182 NAAS Rating: 4.96

Investigation of Intestinal Parasites in Fruits and Vegetables in Local Market of Tikrit, Iraq

Ashraf Jamal Mahmoud Zangana Department of Biology, College of Education for Women, University of Tikrit, Iraq E-mail: [email protected]

Abstract: The study aimed to identify the parasitic stages, which contaminants vegetables and fruits in local markets of Tikrit. Four hundred fifty samples of vegetables and fruits (six types of vegetables and four types of fruits i.e., Lactuca sativa, spinach, Solanum copersicum, Mentha piperita, Apium graveolens, Lepidium sativum, pyrusmalus, banana, clementia, Abutilon) taken randomly from local markets for the period October 2016 until the end of March 2017. The percentage of vegetables and fruit contamination was 47.4 and 36.66%, respectively. The percentage of contamination with parasitic infections was recorded with Entamoeba coli (3.77%), Entamoeba histolytica (9.33%), Idomoeba bustchlii (1.77%), Giardia lamblia (1.11%), Toxocara canis (6.88%) and Fasciolopsis buski (2.44%).

Keywords: Vegetables, Fruit, Parasites, Staining method

Fresh vegetables and fruits are an essential component (green onions, lettuce, radish, leeks) in Egypt, The highest of daily healthy food. In recent years, interest has increased contamination rate was 49% in summer and the lowest in in the study of registered cases due to contamination of fresh winter was 10.8%. These pathogens can contaminate vegetables and fruits (Drisdelle 2013). The outbreak of vegetables and fruits in a number of ways. They can be found intestinal parasitic infections has caused a decrease in the in the soil where these plants are grown or in animal feces health level in different age groups, especially in the used for fertilization or in water used for watering, They can underdeveloped societies, and the mixture of fruits and be found by the workers in these plants or in the water, Used vegetables can protect the human body from many diseases in washing or possible contamination also occurs during by providing it with many nutrients, minerals, vitamins, transport or storage of these plants (Drisdelle 2013). proteins, fiber and vice versa where possible can be caused Objective of the study was to identify the contagious Diseases by contamination of infectious pathogens of human contaminants of intestinal parasites contaminated to some pathogens (Abougrain et al 2016). We need to identify the fruits and vegetables in some areas Tikrit city and Study the source of this pollution, The World Health Organization percentage of the spread of these contaminations. (WHO) and the organizations concerned with health food programs emphasize fruit as vegetables as healthy food and MATERIAL AND METHODS must be clean these foods before eating them (Drisdelle Samples were collected from local markets (Alwa Tikrit, 2013). Registered percentage 9% in 544 samples of stored grocery stores in Al Arbaeen Street and Al Zohour Street) in vegetables were found in Canada (Damen et al 2007). Tikrit city, Salahadin Governorate for the period from October Cryptosporidium was the most contaminated vegetable, 2016 until March 2017. Total number of 450 specimens from followed by Giardia parasite. In south west of Iran (Jasem et Lactuca sativa, spinach, Solanum copersicum, Mentha al 2013) found three types of parasitic parasites (Giardia, piperita, Apium graveolens, Lepidium sativum, Pyrus malus, Entamoeba histolytica and Entamoeba coli) in 135 samples banana, clementia and Abutilon were randomly collected of nine species of vegetables with a contamination rate of from the markets in plastic bags. 15.5%. Giardia duodenalis; Cryptosporidium, Cyclospora Two hundred grams of each sample were wash with 500 cyetanensis are common parasitic infections causing ml of the normal saline solution to isolate the oval, larvae, contamination of 141 different vegetable samples by 9%, cyst, oocyt of parasites. After 24 hours of washing, 5 ml of 11% and 7%, respectively (Daryani et al 2008). Checkup sediment was taken. After a floatation method, wab was (Tefera et al 2014) in Ethiopia examined 360 samples of eight tested by loop and placed on the slides and stained by zeihl types of fruits and vegetables and recorded a contamination neelsen to distinguish the Cyclospora, Crypto sporidenm. rate of 57.8%, including the contagious pathways of five The luogle iodine was used to differentiate the cyst of species of parasitic worms and two parasitic protozoa. Eraky protozoa and helminthes eggs (John and Petri 2006). The et al (2014) recorded 29.6% of 530 samples of vegetables slides were maintained and scanned with a microscope 180 Ashraf Jamal Mahmoud Zangana under 100x and 40x magnification and parasitic phases were Toxocaracanis 6.88%, and the lowest of Giardia lamblia identified by (Yamaguti 1961, Soulsby 1982). (1.11%) (Table 2). There are many factors related to plant The results were statistically analyzed using a chi- pollution, such as the cultural level of workers, the square test, below the level of 0.05 ≤ p to determine the presence or absence of moral differences. Table 1. Percentage of distribution of intestinal parasitic stages contamination on fruits and vegetables in RESULTS AND DISCUSSION Tikrit city The number of positive samples was 191 (42.44%) and Samples type Samples Positive Per cent Number of examined samples stages the highest percentage of contamination was for Apium 1 2 3 graveolens vegetables (60%) and the lowest in Solanum copersicum (33.33%), while in fruit, highest infection rate was Lepidium sativum 45 18 40 13 3 2 in Clementia fruit (44.44%) and the lowest in apples Mentha piperita 45 21 46.66 12 5 4 (28.88%). The percentage of single infection with intestinal Spinach 45 22 51.11 15 6 1 parasites on vegetables and fruits was 67.53%, percentage Lactuca sativa 45 24 53.33 18 4 2 of double infection 24.08 and the tri infection with parasitic Solanum copersicum 45 15 33.33 12 3 -- infection reached 8.37% (Table 1). These intestinal parasites Apium graveolens 45 27 60 19 5 3 were diagnosed: Entamoeba coli, Entamoeba histolytica, Pyrus malus 45 13 28.88 8 4 1 Idomoeba bustchlii, Giardia lamblia, Toxocaracanis, Banana 45 12 31.11 9 3 -- Fasciolopsisbuski (Figs. 1-11). The high percentage Clementia 45 20 44.44 11 8 1 Entamoeba histolytica was 9.33%, followed by the Abutilon 45 19 42.22 12 5 2 Total 450 191 42.44 129 46 16 X2 6.06

Table 2. Percentage of parasitic contaminates which isolated from fruits and vegetables in Tikrit city during the study period Parasites Positive samples Per cent

Trophozoite of Entamoeba coli 17 3.77 Trophozoite of Idomoeb abustchlii 8 1.77 Trophozoite of Entamoeba histolytica 42 9.33 Cyst of Giardia lamblia 5 1.11 Ova of Toxocara canis 31 6.88 Fig. 1. Trophozoite of Idomoeba bustchli in spinachi Ova of Fasciolopsis buski 11 2.44 (Lugoliodin stain 1000x)

Fig. 2. Trophozoite of Entamoeba histolytica in Clementia Fig. 3. Ovum of Fasciolopsis buski in Apium graveolens (Lugoliodin stain 1000x) (Lugoliodin stain 1000x) Intestinal Parasites Stages in Fruits and Vegetables 181 educational level of the worker. Tefera et al (2014) confirmed result of human and animal products. This is an indication of that they collected 360 samples of vegetables and fruits from pollution of water and soil that cause pollution of plants. The local markets in Ethiopia, 57.8% attributed this contamination present study showed the percentage of plant contamination to intestinal parasitic pathways to vegetables and fruits as a of different species in Tikrit city was 42.44%. The results of

Fig. 4. Ovum of Fasciolopsis buski in spinach (Lugoliodin Fig. 7. Ovum of Toxocara canis in Lepidium sativum stain 1000x) (Lugoliodin stain 1000x)

Fig. 5. Ovum of Toxocara canis in Apium graveolens Fig. 8. Trophozoite of Entamoeba coliin in Apium graveolens (Lugoliodin stain 1000x) (Lugoliodin stain 1000x)

Fig. 6. Ovum of Toxocara canis in in Abutilon (Lugoliodin Fig. 9. Trophozoite of Entamoeba histolyticain Musa stain 1000x) (Lugoliodin stain 1000x) 182 Ashraf Jamal Mahmoud Zangana

thus constitute an important epidemiological factor for the transmission of diseases such as: Giardia as: Giardia lamblia, Entamoeba spp, Enterobius vermicularis, Hymenolepis nana, AscarisIum bricoides, Taniidae spp, Toxocara spp, Trichostorongylus spp.

REFERENCES Abougrain AK, Nahaisi MH, Madi NS, Msaied M and Ghenghesh KS 2010. Parasitological contamination in salad. Vegetables in Tripoli – Libqa. Food Control 21(5): 760-762. AL-Megrm WAI 2010. Drevalence intestinal parasites in leafy vegetables in Riyadh, Saudi Arabia. International Journal of Tropical Medicine 5(2): 20.23. Daryani A, Ettehad GH, Sharif M, Ghorbani L and Ziaei H 2008. Fig. 10. Trophozoite of Entamoeba histolytica in Apium Prevalence of intestinal parasites in vegetables consumed in graveolens (Lugoliodin stain 1000x) Ardabil, Iran. Food Control 19(8): 790-794. Damen JG, Banwat DZ, Egah DZ and Allanana JA 2007. Parasitic contamination of vegetables in jos, Nigeria. Annals of African Medicine 6(3): 115-118. Drisdelle R 2013. Parasitic protozoa on fruits and vegetables. Decoded Science 2(6): 7-11. Eraky MA, Rashed SM, Nasr ME, El-Hashary AM and El-Ghannam AS 2014. Parasitic contamination of commonly fresh leafy vegetables in Benha. Egyptian Journal of Parasitology Research. Jasem S, Raza A, Andshahram K 2013. Prevalence of intestiml parasites in Vegetables consumed in Ahvaz, south west of Iran. Journal of Medical Sciences 13: 488-492. John DT and Petri JW 2006. Medical parasitology. 9thed. Saunders Elseviver, Klapec T and Borecka A 2012. Contamination of vegetables, fruits and soil with geonelmints eggs on organic forms in Poland, Annals of Agricultural Medicine19 (3):421- 425. Fig. 11. Trophozoite of Giardia lambliain in Lactuca sativa) Myarango RM, Aloo PA, Kabiru EW and Nyanchongi BO 2008. The (Lugoliodin stain 1000x) risk of pathogenic intestinal parasite infections in kisii Municipality, Kenya BMC Public Health. 8. Ogbolu DO, Alli OA, Gunleye VFO, Olusoga–Ogbolu EF and Olaosnh I 2009. The presence of intestinonal parasites in earlier worker supported the present observations the study selected vegetables from open markets in south western (Tomas and Kidane 2012, Omagye 2010). The difference is Nageria. African Journal of Medicine and Medical 38(4): 319- 324. due to different geographical locations and the difference in Omagye OS and Audu PA 2012. Parasites contamination and temperature and relative humidity, the environmental distribution on fruits and vegetables in Kogi, Nigeria ccibtech. conditions surrounding the species and the different types of Journal of Bio-protocols 1(1): 44-47. specimens studied (Ogboul and Ali 2009, Said 2012, Tefera T, Birukesw A, Mekonnen Z and Eshetu T 2014. Parasitic contamination of fruits and vegetables collected from selected Mnyarango et al 2008). Toxocara eggs were the second in local markets of Jimma Town. Southwest Notices 2 (1):1-7. terms of plant contamination with parasitic infections, and Tomass Z and Kidona D 2012. Parasitological contamination of wastewater irrigated and Raw manure fertilized vegetables in perhaps because female worms produce 10,000 eggs per mekellecity and its suburb, Tigray Ethiopia. Mamona Ethiopian day and are resistant to environmental conditions. These Journal of Science 4(1): 77-89. plants are more coarse when they are more contaminated, Said GL, Mariaho CMR, Matti NSA and Famas GB 2012. Parasitic infestation of vegetables in selected markets in markets in where parasitic infections attach to them. The incidence of metro anole. Philippines Asian Pacific Journal of Tropical the infection of the parasite was strongyloid, as observed by Disease 2(1): 51-54. Said (2012). AL-megrm (2010) observed cases of illness due Soulsby EJL 1982. Helminthes, Arthropods and Protozoa of Domesticated animals. Bailliere Tindall, London. to food contamination by intestinal parasitic. Vegetables and Yamaguti S 1961. Systema Helminthum. Is ted., chancery Lane, fruits play a role in the transmission of these pathogens and London:67.

Received 21 August, 2019; Accepted 04 November, 2019 Manuscript Number: S-277 Indian Journal of Ecology (2019) 46 Special Issue (8): 183-187 NAAS Rating: 4.96

Effectiveness of Brassinolide and Dry Yeast Extract Spraying on Growth Parameters and the Chemical Content of the Grape Seedlings

Sabaa Jawad Abdulkadhim and Akram Abd-Alkadem Hadi Al-Musaib Technical College, University of Al- Furat Al-Awsat Technical, Iraq E-mail: [email protected]

Abstract: The study was conductedto identify the physiological effect of spraying with Brassinolide and addition methods of active dry yeast on growth parameters and the chemical content of the grape seedlings (Summer Royal CV).The factor is spraying with Brassinolide with three levels (0,4,8 gml-1), while the sub factor include four methods of spraying with dry yeast extracts is (spraying with water only, spraying on leaves, soil, leaves and soil).The spraying with Brassinolide showed a significant influence on all characters studied( seedling length, stem diameter, leaves number, leaf area, chlorophyll content in leaves, dry weight of shoot and root system and nutrient concentration(NPK) in leaves. The 8 gm l-1 gave a significant increase in all studied characters as compared to the control. However, the addition of dry yeast extract showed a significant influence on all characters spraying on leave and soil gave a significant increase. The interaction 8 gm l-1 with leave and soil spraying was effective combination.

Keywords: Grape, Summer royal, Brassinolide, Dry yeast extract

Grapes, Vitis vinifera L. is grown extensively in the tropical conducted to investigate the physiological effect of the growth to temperate area (AL-Saidi 2000). The world's cultivated area regulator of Brassinolide, addition method of dry yeast extracts is estimated at 8.8 million hectares, and the global production and its interaction with the vegetative and root growth is 6.89 million tons (FAO 2012). Grapes nutritional value parameters and the chemical content of the Summer Royal extracted from juice, containing vitamins, mineral, sugars, grape seedlings. organic acids, salts, proteins, fats, etc. in addition to its importance in medical uses in the treatment of many diseases MATERIAL AND METHODS (Chevalieh 2010). Samar Royal is a seedless seed early This research was carried out at Horticulture Station maturity needs 200 hours of coolness at a temperature of 7°C. and Forestry Ministry of Agriculture, Al-Mahaweel, Babylon in In order to increase the value of this product spraying plant season 2016 on the Summer Royal grape seedlings,180 growth regulators is of great importance in improving the seedlings were selected, one year old and homogeneous as growth parameters and increase the quantity and quality possible in size, vegetative growth and growing in sandy (Rajni and Deol 2015). Brassinosteroids a growth regulator loam soil, planted with black plastic bags made of known as a raw fatty extract extracted from Brassica nupus, as polyethylene with a capacity of 1.75 kg, it was transferred on detected in pollen, another, seeds, leaves, stems, roots and 10/2/2016 to bags measuring 30 * 25 cm and a capacity of flowers of other plants, are also available in plants on the free 5.25 kg and was filled with sandy loom soil (Table 1) .The form or linked with sugars or fatty acids were extracted about research was carried out as a factorial experiment with two 70 steroid compounds from plants. (Al-Hadethi 2015) found factors and three replicates, the first factor was the spraying that the spray of growth regulator Brassinolide had a of the growth regulator Brassinolide at three levels (0, 4, 8) significant effect in increasing the vegetative growth mg l-1 the second factor was four methods to add the dry yeast characteristics of apricot trees.(Al-Jumaily and Al-esawi 2016) extract (spraying the vegetative with water only, spraying on found that Brassinolide has a significant effect on vegetative the vegetative, spraying on the soil surface, spraying on the growth of apple trees. Biofertilizations is one of the topics that vegetative + the soil surface gl-1.The dry yeast extract had received great attention in recent years due to its similar produced by Turkish company Lesaffre was prepared by effect to organic fertilizers in reducing environmental pollution taking 8g dry yeast per liter of warm water with 1: 1 sugar and health damage to humans and animals (Dilpreet et al addition to activate and double dry yeast, kept for 24 hours 2016).The yeast (Saccharomyces cerevisiae) of the fungi and mixed for 5 minutes (EL-Tohamy et al 2009), and filtered (Ascomycola) of the family Saccharomycetacea, and the dry and use the leachate to spray the seedlings until the full wet yeast is a living organism, fungi (Lateifa 2012). This study was (Table 2). Brassinolide was four sprayed on April 1, 20 and 184 Sabaa Jawad Abdulkadhim and Akram Abd-Alkadem Hadi

May 10and 30/T The dry yeast extracts was sprayed after 5 The percentage of NPK was estimated in leaves using days from each date of spraying with Brassinolide using a 2 standard method. The N was estimated by method of liter hand spray and added The measurements of the straits Haynes (1980), Pby Page et al (1982) and K by flame were taken, and the data was analyzed using the Genestate photometer by method (Erwin and Houba 2004). statistical program. First vegetative characteristics include: High seedling, RESULTS AND DISCUSSION stem diameter, leaf area dsm2, chlorophyll content of the Vegetative characteristics: The Brassinolide at 8 mgl-1 leaves SPAD unit according to the method in (Felixloh and caused a significant increase in seedling height, stem Bassuk 2000), total leaves number seedling-1, dry weight of diameter, leaf area, leaves number, leaf content of vegetative and root groups were measured. chlorophyll and dry weight of shoot and root (Table 3) as compared to 0,4 mgl-1,which gave the lowest rate, the growth regulator Brassinolide has a similar effect to the auxins, Table 1. Physical and chemical property of soil Parameter Result analysis gibberellins and cytokinins this regulates many cellular and

-1 physiological processes that occur in plants such as cell Electrical conductivity EC (extract 1-1) 1.22 DSm elongation and division, resistance to stress and bio- Soil reaction pH (Extract 1-1) 6.9 processing of cell wall components, manufacture of nucleic Sand 870 g kg-1 acids DNA, RNA and various proteins, nitrogen fixation, -1 silt 43 g kg differentiation of vascular system, adventitious roots, etc.(Al- -1 Clay 87 g kg Khafaji 2014), which caused an increase in the Soil texture Sandy loam characteristics of vegetative growth above. The increase in Nitrogen 24.79 mg kg-1soil chlorophyll leaf content may be due to inhibited action of Phosphorus 0.47 mg kg-1soil chlorophyllase enzyme responsible for the degradation of Potassium 29.90 mg kg-1soil chlorophyll, which led to the accumulation of chlorophyll in

Table 2. Bread yeast components Sacchromyce cervisiae Seq. Amino Acids (mg. g-1) 0.2 K % 0.18 0.1 Glycine 0.103 0.3 Na % 0.12 0.2 Alanine 0.132 0.4 Mg % 0.1 0.3 Valine 0.312 0.5 Ca % 0.04 0.4 Leucine 0.067 0.6 µgg-1Mn 5.69 0.5 Isoleucine 0.421 0.7 µgg-1 Zn 69.5 0.6 Aspartic acid 0.274 0.8 µgg-1 Cu 12.78 0.7 Glutamic acid 0.367 0.9 µgg-1 Fe 30.5 0.8 Serine 0.523 Seq. Vitamins (mg g-1) 0.9 Threonine 0.206 0.1 Vit. B1 0.163 0.1 Tyrosine 0.031 0.2 Vit. B2 0.054 0.11 Phenyl alanine 0.116 0.3 Vit. B6 0.019 0.12 Proline 0.041 0.4 Pantothenic acid 0.058 0.13 Arginine 0.073 0.5 Biotin 0.091 0.14 Lysine 0.089 0.6 Niacin 0.112 0.15 Cysteine 0.025 7 Inositol 0.372 0.16 Methionine 0.012 Seq. Other ingredients (%) 17 Histidine 0.078 1 Total Nitrogen 7.69 0.18 Tryptophan 0.02 0.2 Carbohydrate 5.47 Seq. Metal structure 3 Ash 13.51 0.1 P % 0.4 Water 4.7 * The yeast components were analyzed at the Central Laboratory of the Bio Sciences University of Poland (Lublin) Brassinolide and Dry Yeast Extract Spraying 185 the leaves (Fariduddin et al 2003). The dry weight gain may the rest of the plant parts (El-Sherbeny et al 2007). also be due to the potential impact of BRs on the CO2 fixation Interaction treatments had a significant effect on the above -1 in photosynthesis, which increases the CO2 net represented characteristics, 8 mgl yeast extracts spraying on vegetative in the leaf, which increases the efficiency of photosynthesis group and soil surface caused a significant increase ​​for the (Sadeghi and Shekafandeh 2014). These results are same characteristics, the increase may be due to the impact consistent with Al-Jumaily and Alesawi (2016) on apples, Al- of combined factors. Hadethiand AL-Dujaili (2015)on apricots, and Al-Dulaimy Nutrients (NPK%) in leaves: Spray of Brassinolide had a on pomegranates that the spraying of the significant effect on the percentage of NPK nutrients in (٢٠١٥) and Alrawi Brassinosteroid positively effects in the vegetative leaves with 8 mgl-1 treatment caused a significant increase characteristics of the fruit trees. The method of adding dry (2.233%, 0.560%, 1.761%) respectively (Table 4). This may yeast extracts significantly affected the growth The treatment be due to the effect of Brasinolide, which is similar to auxins, of seedling spray with the yeast extracts on vegetative group gibberellins and cytokinins. This may also be due to increase and soil surface caused a significant increase in the above the water transport and necessary nutrients for plant growth characteristics were compared to the spraying the vegetative this in turn increases the content of leaves of nutrients. Ruiz with water only. This is due to the effect of the dry yeast et al (2000) found that there is a close relationship between extract on the metabolic and biological processes of the amount of chlorophyll in the leaves and the accumulation photosynthesis pigments (chlorophyll) and the effectiveness of N in the dry matter. The results were agreed with AL- on grapes, Abubakar et al (2013) on (٢٠١٦) of photosynthesis enzymes, which causes the conversion of Ahbaby large amounts of light energy into chemical energy, pomegranates and Al-Hadethi (2015) on apricots. The table producing larger amounts of carbohydrates and stimulating showed that the yeast extracts spraying on vegetative group plant organs, especially leaves that store and distributes to and soil surface was significantly affected in the above

Table 3. Effect of spraying with Brassinolide and method of adding dry yeast extracts invegetative characteristics Brassinolidemg l-1 Yeast Plant height Stem diameter Leaf area Leaves Leaves Dry weight of Dry weight of extract (cm) (mm) (dsm2) number chlorophyll vegetative root groups g l-1 content SPAD groups

B0 :0 Y0 81.12 8.7 30.33 11.28 28.12 21.22 10.42 Y1 95.3 10.14 56.35 19.85 31.88 25.63 11.62 Y2 86.22 9.13 45.31 16.41 30.45 23.61 12.53 Y3 101.55 10.35 59.37 25.71 33.18 29.14 13.24 B1 :4 Y0 90.56 8.81 48.3 15.21 33.53 26.13 11.33 Y1 110.11 10.25 60.36 22.68 35.77 29.52 12.49 Y2 98.32 9.96 62.33 19.86 34.67 28.45 14.62 Y3 118.23 11.43 90.05 28.6 36.8 31.47 15.42 B2 :8 Y0 120.22 9.55 50.39 18.37 35.16 30.42 13.46 Y1 134.44 11.71 76.47 27.93 38.61 35.64 15.59 Y2 127.65 10.74 69.33 24.6 37.32 33.69 17.79 Y3 141.65 12.17 88.46 33.15 39.25 37.75 19.65 LSD at 5% 8.1 1.95 5.13 8.26 1.98 1.36 1.2 Brassinolidemg l-1 B0 :0 91.05 9.58 47.84 18.31 30.91 24.9 11.95 B1 :4 104.31 10.11 65.26 21.59 35.19 28.89 13.47 B2 :8 130.99 11.04 71.16 26.01 37.59 34.38 16.62 LSD at 5% 3.12 0.14 2.16 2.49 0.5 0.59 0.49 Yeast extractg l-1 Y0 97.3 9.02 43.01 14.95 32.27 25.92 11.74 Y1 113.28 10.7 64.39 23.49 35.42 30.26 13.23 Y2 104.06 9.94 58.99 20.29 34.15 28.58 14.98 Y3 120.48 11.32 79.3 29.15 36.41 32.79 16.1 LSD at 5% 4.01 1.02 2.49 5.7 0.73 0.75 0.77 186 Sabaa Jawad Abdulkadhim and Akram Abd-Alkadem Hadi

Table 4. Effects of spraying with Brassinolide and method of fertilizers and its interactions with bread yeast Sacchromyce adding dry yeast extract in percentage of some Cervisiae. Tikreet University Journal of Agriculture Sciences nutrients in the leaves 15(4):73-84.

-1 -1 AL-Dulaimy AFZ 2012. Effect of foliar spray with yeast suspension, Brassinolide mg l Yeast extract g l N% P% K% Liquorice roots extract and amino Quelant-K on growth and B0 :0 Y0 1.624 0.302 1.262 yield of grapecv. Black Hamburg. Ph. D. Thesis .College of Agriculture, University of Baghdad, 119p. Y1 1.873 0.483 1.493 Al-Hadethi MEA and AL–Dujaili JA 2015. Effect of biofertilizers, algae Y2 1.885 0.461 1.461 extract and brassinolide on growth character of apricot trees. Euphrates Journal of Agriculture Science 7(3): 63-82. Y3 2.238 0.524 1.664 Al-Hadethi MEA 2015. Effect of different fertilization sources and the B1 :4 Y0 1.831 0.404 1.453 growth regulator (Brassinosteroids) on growth and yield of apricot trees. Ph.D. Dissertation, College of Agriculture, Y1 1.945 0.523 1.574 University of Baghdad, 153p. Y2 1.925 0.494 1.594 Al-Jumaily OJM and Alesawi SAA 2016. Effect of foliar application with Brassinolide and algae extract (tecamine) in vegetative Y3 2.383 0.564 1.753 and yield characteristics of apple tree (cv. Anna). The Iraqi B2 :8 Y0 1.981 0.544 1.483 Journal of Agricultural Sciences 47(5): 1225-1234. Al-Khafaji MA 2014. Plant Growth Regulators, Application and Y1 2.246 0.553 1.884 Utilization in Horticulture. Bookstore for Printing publishing and Y2 2.242 0.543 1.773 Translating, University of Baghdad, Iraq, pp.348. AL-Saidi IHM 2000. The Production of Grapes (Part I), The Ministry Y3 2.017 0.606 1.904 of Higher Education and Scientific Research, University of LSD at 5% 0.034 0.032 0.025 Mosul-Iraq, pp.1-200. Brassinolide mg l-1 B0 :0 1.905 0.442 1.47 Bajguz A and Tretyn A 2007. The chemical characteristic and distribution of Brassinosteristic in plant. Photochemistry 62: B1 :4 2.021 0.496 1.593 1027-1046. B2 :8 2.233 0.56 1.761 Chevalieh A 2010. Medicinal plant it's using and Handling with Ayoub translation – International Academy Beirut –Lebanon. LSD at 5% 0.017 0.016 0.012 Dilpreet, Kulbir Singh, Khurana DS and Variendar S 2016. Influence Yeast extract g l-1 Y0 1.812 0.416 1.399 of biofertilizers on growth, yield, quality and nutrient uptake in onion (Allium cepa L.). Indian Journal of Ecology 43(1): 124- Y1 2.021 0.52 1.65 127. Y2 2.017 0.499 1.609 Dinkha RF and Al Khazragji T 1990. Nutrition and Fungus Function Science, University of Salahaddin, Ministry of High Education, Y3 2.362 0.563 1.773 Iraq. LSD at 5% 0.019 0.018 0.014 El-Desouky SA, Wanas ZM, Khedr VA and Kandiannn K 2007. Utilization of parthasarathy horticulture, vegetable some natural plant extracts of garlic and yeastas seed soaked materials to characteristics (2.362%, 0.563% and 1.773%, respectively). squash (Cucurbita pepo L.) 1. Effect on growth, sex expression and fruit yield and quality. Journal of Agriculture Science .on Moshtohor, Zagazig University 35(2): 839-854 (٢٠١٥) The results were agreed with Al-Dulaimy and Alrawi pomegranate and with Hafez et al (2013) on olives and AL- El-Sherbeny SE, Khalil MY and Wahby MS 2007. Growth and -1 productivity of rue (Ruts graveolens) under different foliar Dulaimy (2012) on grapes. Interaction 8 mgl with yeast fertilizers application. International Journal of Applied Science extracts spraying on vegetative group and soil surface 3(5): 399-407. caused a significant increase of 2.466%, 0.606%, 1.904%, EL-Tohamy WA, EL-Abady HM and EL-Greadly NHM 2009. Studies on the effect of putrescine, yeast and vitamin C on growth, yield respectively. This may be due to the positive effect of and physiological responses of eggplant under sandy soil Brassinolide and yeast extract in the increased vegetative conditions. Australian Journal of Basic and Applied Sciences growth indicators shown above and reflected in the increased 2(2): 296-300. Erwin E and Houba V 2004. Plant Analysis procedures, Kluwer N, P and K content of the leaves. Academic publishers, The Netherlands, 179p. FAO, Food and Agriculture Organization 2012. The United Nation REFERENCES (UN) Bulletin of Statistic. Abubakar AR, Ashraf N and Ashraf M 2013. Effect of plant Fariduddin Q, Ahmad A and Hayat S 2003. Photosynthetic response biostimulants on growth, chlorophyll content, flower drop and of Vigna radiata to pre-sowing seed treatment with 28 - fruit set of pomegranate cv. Kandhari Kabuli. International homobrassinolide. Photosynthesis 41: 307-310. Journal of Agriculture, Environment and Biotechnology 6(2): Felixloh JG and Bassuk N 2000. Use of the Minolta SPAD-502 to 305-309. determine chlorophyll concentration in Ficus benjamina L. and AL-Ahbaby AJA 2016. The effect of foliar spraying with KT-30, Populus deltoides Marsh leaf tissue. Horticultural Science Brassinolide and seaweeds extract in some vegetative growth 35(3): 423. of seedlings vine varieties VitisviniferaL.Hallawani and Kamali. Hafez OM, Saleh MA and El-Lethy SR 2013. Response of some Al-Anbar Journal of Agricultural Sciences 41(special No.): 8e- seedlings olive cultivars to foliar spray of yeast and garlic 17e. extracts with or without vascular arbuscular mycorrhizal fungi. Al-Dulaimy AFZ and Alrawi MMO 2015. Response of pomegranate World Applied Sciences Journal 24(9): 1119-1129. trees (Punica granatum L.) Cv. Salimi and Wonderful to organic Haynes RJ 1980. A comparison of two modified kijeldahl digestion Brassinolide and Dry Yeast Extract Spraying 187

techniques for multielements plant analysis with conventional matter accumulation, partitioning and fruit retention of Btcotton. wet and dry washing methods. Communication, Soil Sciences Indian Journal of Ecology 42(1): 270-272. and Plant Analysis 11(5): 459-467. Ruiz JM, Castilla N and Romero L 2000. Nitrogen metabolism in Lateifa SA 2012. Biofertilizer and its role in reducing water pollution pepper plants applied with different Bioegulatos. Journal of problems with chemical fertilizers. Libyan Agriculture Research Agricultural and Food Chemistry 48: 2925-2929. Center Journal international 3(2): 1457-1466. Sadeghi F and Shekafandeh A 2014. Effects of 24-epibrassinolide on Page AL, Miller R and Keeny DR 1982. Method of soil and analysis nd growth, lipid peroxidation, protein andantioxidative enzyme part 2, 2 Ed, Argon 9 publisher Madison Wisconsin, 4SA, USA. activities in seedlings of loquat under salinity stress. Alban Rajniand Deol JS 2015. Effect of plant growth regulators on dry Journal of Agricultural Sciences 13(2): 116-124.

Received 22 August, 2019; Accepted 07 October, 2019 Manuscript Number: S-278 Indian Journal of Ecology (2019) 46 Special Issue (8): 188-191 NAAS Rating: 4.96

Effect of Salicylic Acid on Chlorophyll A, B and Carotene Content of Petunia sp.

Zahraa H. Ali and Faiq H. Ali Department of Biology, College of Science, University of Al-Muthanna, Samawa, Iraq E-mail: [email protected]

Abstract: The aimed current study to determine the effect of salicylic acid as a plant growth hormone to improve physiology parameters of plant, SA was used in four concentrations (0, 100, 200 and 300) ppm, in three different treatments namely soaked seeds before planting for 24 hours and spraying of small and young plants and soaked seed and spray plant with the above-mentioned concentrations. The results indicated the positive effect of salicylic acid on chlorophyll a, b and carotene at lower concentration at 100 ppm with soaked seed and spray plant treatment. The plant variety P. hybirda was superior in obtaining the highest concentrations and its values were for chlorophyll a, b and carotene (2.46 0, 1.083 and 9.63 mg g-1, respectively). While the lower values were recorded for other variety P. niglefera on chlorophyll a, b and carotene (0.886, 0.4 and 3.81 mg g-1, respectively) as control treatment.

Keywords: Petunia, Salicylic acid (SA), Chlorophyll a, b and carotene

Petunia belongs to the Solanaceae, a family with major and systemic plant defense responses against pathogens. It crops including tobacco (Nicotiana tabacum), tomato has also been found that SA plays a role during the plant (Solanum lycopersicon) and potato (Solanum tuberosum). response to abiotic stresses such as drought, chilling, heavy Petunia plant used in the summer for home gardens and metal toxicity, heat, and osmotic stress (Rivas-San Vicente commercial landscaping projects. The name of Petunia and Plasencia 2011). Zarghami et al (2014) obtaining the derives from ''petum'' or ''betum'', an original name for the highest corolla tube, chlorophyll content, plant height, leaf tobacco plant (Gautam 2012) South America, is origin where area, root and shoot dry weight and biomass, stomatal the various species have been documented from emollitions conductivity, primary and secondary shoots produced by made in Argentina, Brazil, Bolivia, Paraguay, and Iranian variety with salicyclic acid on petunia. Moreover, Uruguay(Drummond et al 2015) Petunias are heat-tolerant salicylic acid activated the synthesis of carotenoids, species and could adapt well to tropical climates(Hong et al xanthophylls and the rate of de-epoxidation, but decreased 2018). Petunia hybrida has been widely used as a model the level of chlorophyll pigments, both in wheat and moong system for molecular genetics providing a range of genetic plant and also the ratio of chlorophyll a/b, in wheat seedlings and molecular tools (Dell'Olivo et al 2011). The morphology (Moharekar et al 2003). Shinwari et al (2018) found of the beautiful Petunia describe that leaves shape are ovate. exogenously applied various levels of acetyl salicylic acid The trumpet-shaped flowers may be single or double and are (ASA) including ASA0 (Control), ASA1 (0.30mM), ASA2 across, calyx deeply 5-parted, corolla funnel form with 5 (0.60mM), ASA3 (0.90mM), ASA4 (1.20mM), ASA5 (1.50mM) rounded (white, red, yellow, pink, purple, or were investigated for induction of thermo-tolerance in tomato variegated).Seeds many subglobose, 0.5 mm across (Lim seedlings, indicated that chlorophyll contents was affected 2014). The petunia offers the potential for high economic significantly by both ASA treatments and growth conditions. returns and is an excellent source of additional income to the Therefore, this study was conducted to determine any flower industry (Zhang et al 2012). Salicylic acid (SA) is a application and any concentration of salicylic acid was phenolic derivative, distributed in a wide range of plant perfect and any variety of petunia was responded better to species. It is a natural product of phenylpropanoid this acid. metabolism (Shayat et al 2007). This acid was first discovered as a major component in the extracts from Salix MATERIAL AND METHODS (willow) and have a significant impact on plant growth and Preparation of salicylic acid: Salicylic acid (SA) prepared development, photosynthesis, transpiration, ion uptake and by dissolving 1 gm from SA in later distilled water to be it is transport and also induces specific changes in leaf anatomy concentration 1000 ppm, as stock solution. Along with this to and chloroplast structure(Hayat et al 2008). In recent years reduce mistake in chemical preparations is using titration law salicylic acid (SA) has been the focus of intensive research (N1×V1= N2×V2), to prepare the studied concentrations of due to its function as an endogenous signal mediating local 0,100,200 and 300 ppm. In addition with distilled water stirrer Chlorophyll A, B and Carotene Content of Petunia sp. 189

then quantities were placed separately in a 1 liter flask for volume of the diluted acetone is (80%). W= dry weight per

each treatment. The treatment namely: T1:100,200 and300 gram of extracted vegetable tissue.

ppm SA by soaking seed method. T2:100,200 and300 ppm

SA spraying on plant method and T3:100,200 and300 ppm SA RESULTS AND DISCUSSION soaking seeds and spraying plant method Effect of SA in Chlorophyll a: The results of Table 1 showed Growth condition: This experiment was conducted under the effects of salicylic acid in increasing the content of natural conditions in a Greenhouse of Muthanna University chlorophyll a, the best concentrations of SA at 100 ppm in the

by using two varieties of plant seeds soaking and non- third treatment T3 (seed soaking + spraying plants) and in soaking for P. hybirda and P. niglefera are putting in pots that plant P. hybrida, where it gave the highest value of chlorophyll weight 4 kg in three repeats for each treatment. Irrigation the A content at (2.46) mgg-1, compared with the other three pots with 75% field capacity. More than 20 seed/pot. When concentrations in the same coefficients or in the remaining the seedlings are begging in growth, then reduce its number coefficients, and for the same plant. On the other hand, the for 3 seedlings. At the age of around 75 days, the plants were lowest values ​​for chlorophyll content were 0.886 mgg-1 of P. sprayed in SA. Spraying in every week for 6 weeks. niglefera at 300 ppm concentration. Estimation of chlorophyll type and carotenoid content: In addition, all treatments with salicylic acid gave Collect of Petunia sp. leaves after end of treatment by SA and significant differences at the probability level of 5% and for all are washed by fresh water, then distal water to prepare for concentrations compared with treatment control for both measurement the pigments Chlorophyll and carotene plants at (0.66 and 0.55) mg g-1, respectively. contents was evaluated in fresh leaves by the method of Effect of SA in Chlorophyll b: The results of Table 2 showed (Makinny 1941, Arnon 1949 ). A total of 200 mg of fresh tissue the effects of salicylic acid in increasing the content of from inter venial leaf areas was crushed using a mortar and chlorophyll b The highest value at 100 ppm concentration -1 pestle containing 80% acetone. The optical density (OD) of approximately (1.083) mg g for the treatment T3 (soaking the solution was recorded at 645, 663nm for chlorophyll seeds + spraying plants), the values belong to a plant P. estimation and 452.5 nm for carotenoid estimation using a hyberada as for the P. niglefera. The highest value of spectrophotometer. The following relationships were used to concentration was 100 ppm and for same treatment (0.913) calculate the amount of chlorophyll: mg/g In addition to that we find the lowest value for the same

Chl. a = (12.7 (D663) – 2.69 (D645)) × V /(1000 × W). plant back to the concentration 100 ppm, but for T1 treatment Chl. b = (22.9 (D645) – 4.68 (D663)) × V /(1000 × W) . it was (0.4) mgg-1. Carotenoids=4.2×D452.5 –(0.0264 Chl.a+0.426 Chl. b). All concentrations with salicylic acid offered significant D= the optical density reads of chlorophyll extracted at differences at the probability level of 5%, in comparison to the wavelengths 663 and 645 nm, respectively. V= The final control.

Table 1. Effect of salicylic acid and treatments methods in chlorophyll a content (mg g-1) of Petunia sp Petunia sp. Treatment Con. Ppm Interference Effect of Effect of methods species× species treatment treatment 0 100 200 300

P. niglefera T1 0.55 c 0.84 c 1.06 bc 0.886 bc 0.861 b

T2 0.55 c 1.18 bc 1.086 bc 1.046 bc 0.993 b

T3 0.55 c 2.146 ab 1.45 abc 1.14 bc 1.349 ab

P. hyberada T1 0.66 c 1.326 abc 1.516 abc 1.346 abc 1.185 ab

T2 0.66 c 1.38 abc 1.82 abc 1.453 abc 1.295 ab

T3 0.66 c 2.46 a 1.82 abc 1.726 abc 1.639 a Interference P. niglefera 0.55 c 1.388 ab 1.198 abc 1.0244 bc 1.068 b plant × Con. P. hyberada 0.66 c 1.862 a 1.596 ab 1.484 ab 1.373 a

Interference T1 0.605 c 1.083 bc 1.288 bc 1.116 bc 1.023 b treatment × T 0.605 c 1.49 b 1.27 bc 1.213 bc 1.144 ab Con. 2

T3 0.605 c 2.303 a 1.635 ab 1.433 bc 1.494 a Effect of Con. 0.605 b 1.625 a 1.397 a 1.254 a *All numbers have the same letters are No significant difference at probability level 5% under Duncan polynomial test 190 Zahraa H. Ali and Faiq H. Ali

Effect of SA in Carotene concentration: The data in Table plant type and the mode of application of SA. The application 3 showed activity of salicylic acid increase in the carotene of SA (20 mg ml-1) to the foliage of the plants of Brassica content of the plant for the Petunia sp. It can be seen from the napus improved the chlorophyll contents (Ghaiet al 2002). values that the highest significant concentration is below the Similarly, soaking the grains of wheat in 10-5M of SA resulted -1 probability level 5% to the third treatment T3 was 9.63 mg g in the plants with higher pigment contents which declined as for the concentration of 100 ppm and it was share of the plant the concentration of SA was increased (Hayat et al 2005). P. hybridia, while the same concentration, for the same Moreover, 30 day old plants of Brassica juncea sprayed with treatment but for plant P. niglefera is 6.98 mg g-1. 10-5M of SA possessed chlorophyll higher than those The plant, the treatments and interference between sprayed by water only, however the maximum concentration plant and concentrations had significant differences from the (10-3M) decreased the chlorophyll contents and the values control concentration at the probability level of 5%. The were below that of water sprayed control, at 60 day stage metabolic aspects of plants, supplied with SA or its (Fariduddin et al 2003). However, the leaves of corn and derivatives, shifted to a varied degree, depending on the soybean applied with acetyl salicylic acid (ASA) or gentisic

Table 2. Effects of salicylic acid on Chlorophyll B concentration (mg g-1) of Petunia sp. Plant Petunia sp. Treatment Con. Ppm Interference Effect of Effect of methods species× species treatment 0 100 200 300 treatment

P. niglefera T1 0.55 bc 0.306 c 0.466 bc 0.4 bc 0.458 b

T2 0.55 bc 0.52 bc 0.486 bc 0.42 bc 0.521 b

T3 0.55 bc 0.913 ba 0.6 abc 0.5 bc 0.668 ab

P. hyberada T1 0.66 abc 0.54 bc 0.6 abc 0.546 abc 0.559 ab

T2 0.66 abc 0.76 abc 0.62 abc 0.586 abc 0.629 ab

T3 0.66 abc 1.083 a 0.78 abc 0.66 abc 0.768 a Interference P. niglefera 0.55 ab 0.58 ab 0.517 ab 0.44 b 0.549 a plant × Con. P. hyberada 0.66 ab 0.794 a 0.666 ab 0.597 ab 0.652 a

Interference T1 0.605 b 0.423 b 0.553 b 0.473 b 0.508 b treatment × T 0.605 b 0.64 b 0.553 b 0.503 b 0.575 ab Con. 2

T3 0.605 b 0.998 a 0.69 ab 0.58 b 0.718 a Effect of Con. 0.605 a 0.687 a 0.592 a 0.518 a *All numbers have the same letters are No significant difference at probability level 5 under Duncan polynomial test

Table 3. Effects of salicylic acid on carotene concentration (mg g-1) of Petunia sp. Petunia sp. Treatment Con. ppm Interference Effect of Effect of methods species×treatment species treatment 0 100 200 300

P. niglefera T1 2.55 b 6.033 ab 6.00 ab 3.81 b 4.01 a

T2 2.55 b 6.487 ab 6.033 ab 4.853 ab 5.236 a

T3 2.55 b 6.98 ab 6.05 ab 3.907 b 5.706 a

P. hyberada T1 3.47 b 6.807 ab 6.507 ab 5.887 ab 5.181 a

T2 3.47 b 7.42 ab 6.867 ab 5.187 ab 5.96 a

T3 3.47 b 9.63 a 7.003 ab 5.45 ab 6.15 a Interference P. niglefera 2.55 c 6.491 a 6.451 a 4.848 bc 5.113 a plant × Con. P. hyberada 3.47 bc 6.911 a 6.623 a 5.646 ab 5.633 a

Interference T1 3.01 b 6.42 ab 5.837 ab 4.953 ab 4.622 a treatment × T 3.01 b 6.507 ab 6.158 ab 5.68 ab 5.443 a Con. 2

T3 3.01 b 8.317 a 6.735 ab 4.848 ab 6.055 a Effect of Con. 3.01 b 6.557 a 6.278 a 5.649a *All numbers have the same letters are No significant difference at probability level 5% under Duncan polynomial test Chlorophyll A, B and Carotene Content of Petunia sp. 191 acid (GTA), exhibited no change in their chlorophyll contents salicylic acid on chlorophyll content, hill activity and yield components in Brassica napus L. (cv. GSL-1). (Khan et al 2003). The impact of exogenous application of Phytomorphology 52: 83-87. salicylic acid (SA 0.01 mM) in Arabidopsis (Borsani et al Hayat S, Fariduddin Q, Ali B and Ahmad A 2005. Effect of salicylic 2001) and in tomato plants (Kaydanet al 2007). acid on growth and enzyme activities of wheat seedlings. Acta Agronomica Hungarica 53(4): 433-437. Hong L, Hoang N and Kim WS 2018. Air temperature and humidity CONCLUSION affect petunia ornamental value. Korean Journal of Horticultural The increase of chlorophyll a, b and carotene Science and Technology 36(1): 10-19. concentration in Petunia sp. is treated by SA, perhaps was Kaydan D, Yagmur M and Okut N 2007. Effects of salicylic acid on the growth and some physiological characters in salt stressed because of its promotion of plant hormone to improvement in wheat (Triticum aestivum L.). Tarim Bilimleri Dergisi 13(2): 114- all physiological aspects. Pre-sowing soaking treatment of 119. seeds with SA, positively, affected the chlorophyll a, Khan W, Prithiviraj B and Smith DL 2003. Photosynthetic responses of corn and soybean to foliar application of salicylates. Journal chlorophyll b and carotenoids. of Plant Physiology 160(5): 485-492. Lim TK 2014. Edible medicinal and non-medicinal plants: flower. Springerpp.756-761. REFERENCES Mackinney G 1941. Absortion of light by chlorophyll solutions. The Arnon DI 1949. Copper enzymes in isolated chloroplasts. Journal of Biological Chemistry 140: 315-322. Polyphenol oxidase in Beta vulgaris. Plant Physiology 24(1): 1- 5. Manaa A, Gharbi E, Mimouni H, Wasti S, Aschi-Smiti S, Lutts S and Ahmed HB 2014. Simultaneous application of salicylic acid and Borsani O, Valpuesta V and Botella MA 2001. Evidence for a role of calcium improves salt tolerance in two contrasting tomato salicylic acid in the oxidative damage generated by NaCl and (Solanumly copersicum) cultivars. South African Journal of osmotic stress in Arabidopsis seedlings. Plant Physiology Botany 95: 32-39. 126(3): 1024-1030. Rivas-San Vicente M and Plasencia J 2011. Salicylic acid beyond Dell'Olivo, A, Hoballah ME, Gubitz T and Kuhlemeier C defense: its role in plant growth and development. Journal of 2011.Isolation barriers between Petunia axillaris and Petunia Experimental Botany 62(10): 3321-3338. integrifolia (solanaceae). Evolution 65(7): 1979-1991. Shinwari AU, Ahmad I, Khan I, Khattak H and Azimi AS 2018. Acetyl Drummond RS, Janssen BJ, Luo Z, Oplaat C, Ledger SE, Wohlers salicylic acid affects growth and productivity of tomato MW and Snowden K C 2015. Environmental control of (Solanumly copersicum L.) under different growth conditions. branching in petunia. Plant Physiology 168(2): 735-751. Annals of Horticulture 11(1): 55-61. Fariduddin Q, Hayat S and Ahmad A 2003. Salicylic acid influences Zarghami M, Shoor M, Ganjali A, Moshtaghi N and Tehranifar A 2104. net photosynthetic rate, carboxylation efficiency, nitrate Effect of salicylic acid on morphological and ornamental reductase activity, and seed yield in Brassica juncea. characteristics of Petunia hybrid at drought stress. Indian Photosynthetica 41(2): 281-284. Journal of Fundamental and Applied Life Sciences 4(3): 523- Gautam P 2012. Effect of light quality in the regulation of morphology 532. and flowering of petunia. M.Sc. Thesis, Department of Plant Zhang W, Li X, Cheng F and Lu J 2012. Accumulation and distribution and Environmental Sciences (IPM), Norwegian University of characteristics for nitrogen, phosphorus and potassium in Life Sciences, Norway 65 p. different cultivars of Petunia hybrid Vlim. Scientia Horticulturae Ghai N, Setia RC and Setia NE 2002. Effects of paclobutrazol and 141: 83-90.

Received 22 September, 2019; Accepted 20 November, 2019 Manuscript Number: S-279 Indian Journal of Ecology (2019) 46 Special Issue (8): 192-195 NAAS Rating: 4.96

Effect of Integrated Pest Management on Mortality of Whitefly Parasitoid Eretmocerus eremicus (Hymenoptera: Aphelinidae) on Egg Plant

S.A. Alani and Z.A.A. Al-Gadban Department of Plant Protection, College of Agricultural Engineering Science, University of Baghdad, Baghdad, Iraq E-mail:[email protected]

Abstract: The toxicity of some integrated pest management programs on the natural enemy of the whitefly Bemisia tabaci (Genn.) were evaluated for the parasitiod Eretmocerus eremicus using simple counting method on laboratory conditions. Mortalities from the two used IPM programs (Isaria fumosorosea + Applaud (a.i. Buprofezin) + Evisect (Thiocyclam hydrogen oxalate)) and (Applaud + encapsulated Evisect (nano formulation) were 100% for the adult parasitoid indicating that the parasitoid were seriously affected by these applications. The chemical used were seriously harmful to the parasitoid and no individual could survive the side effect of these applications. The results indicate that the entomopathogenic fungi, IGRs, classical pesticide and nanoformulation of pesticide used in this experiment are highlytoxic, and are not compatible with the use of parasitoids to support natural control of whiteflies under integrated pest management in greenhouse conditions.

Keywords: Eretmocerus eremicus, Bemisia tabaci, Natural enemy, side effect, Nano pesticide

Encarsia and Eretmocerus are the most commonly the pressure of the sprayer and used Plants were sprayed occurring parasitoids of B. tabaci (Shah et al 2015). bottom up to concentrate on the lower side of the leaves Generally, Eretmocerus are solitary, internal parasitoids of where all stages of the insect host B. tabaci are located. whitefly that complete larval development in the whitefly Application of classical Evisect and Nano Evisect: Both nymph, and pupate in the mummified fourth instar nymphs. WP and Nano Evisect were suspended in distilled water Adult Eretmocerus emerge from the mummified whitefly separately (according to insecticide label recommendations nymphs through an exit hole cut in the dorsum of the host for the classical Evisect and then sprayed as mentioned (Gould et al 2008). Eretmocerus species are tiny wasps. before. Eretmocerus is an ecto-endo parasitoid of females. Application of IPM programs: The IPM programs included two or three steps (Table 1) with seventy two hour time MATERIAL AND METHODS interval among treatments. The experiment was performed in a greenhouse (240 m2 Control: This was sprayed only with DW. on eggplant var. Barcelona). Each replicate consisted of 40 plants, only 10 plants were marked and numbered from 1-10 RESULTS AND DISCUSSION to be observed and monitored for the chosen time interval of Density of whitefly parasitoid Eretmocerus: The initial the experiment (winter season 2018). Plants were chosen densities shows low values in the beginning of the infestation, carefully to be homogenous in infection of the parasitoid host after few weeks the densities of the adult parasitoid Bemisia tabaci. increased as the population of their insect host (B. tabaci) Collection of the parasitoid E. eremicus: Three leaves start to increase (Table 2). were taken from each marked plant from base, middle and tip After application of the IPM programs no adult of E. of the vegetative parts. Leaves with no insects (pupa or eremicus survived in both samples taken after application exuvia) were excluded from the experiment while the infected (Table 1). The leaf samples taken from the two applied IPM ones were then taken into a squared plastic vials with wet programs did not contain any living individual of the cotton on the bottom and covered with a fine mesh under a parasitoid (100% mortality). However, the control treatment degree of 18 ± 2oC.The vials were observed for 4-6 hours showed that the adult parasitoid continue to exist and it could each day and all emerged parasitoid individuals were be due to the lack of toxic chemicals applied from one hand collected and preserved for the counting and classification. and availability of its insect host B. tabaci from the other. Application of Isaria fumosorosea: Five grams of I. Gerling and Sinai (1994) found that the young adult of the fumosorosea commercial product was diluted in aliter parasitoidE. eremicus are affected by the IGR (buprofezin) distilled water. Hand sprayer was calibrated manually to set used to control B. tabaci (Gennadius) while pupa of the Integrated Pest Management on Whitefly Parasitoid 193 parasitoid was not affected in laboratory trial. The harmful residues were highly repellant to parasitoids1.5 h after effect of buprofezin on young Eretmocerus sp. probably is drying. Following a 48-h drying period which significantly related to the ovipositional habits and immature reduced leaf abandonment rates for these two IGRs, E. development. Eretmocerus sp. lays its eggs under the host, eremicus females were not observed foraging on B. after that they hatch, larvae then penetrates the host. After argentifolii nymphs treated with pyriproxyfen and parasitoids penetration, the larva develops inside a capsule formed by spent significantly less time on kinoprene-treated leaf halves. the insect host that maintains contact with the surroundings Behavioral assays indicated that E. eremicus would readily (Gerling et al 1991).The contact with the external abandonleaf halves to forage on non-kinoprene and non- environment exhibited by eggs and young Eretmocerus sp. pyriproxyfen treated leaf surfaces (Hoddle et al 2000). In a larvae may expose them to buprofezin vapor. Buprofezin study on the natural enemies of B. tabaci, Naranjo et al applied to immature E. tejanus Rose and Zolnerowich (2004) found that insecticide regimes based on the initial use (Hymenoptera: Aphelinidae) reduced the longevity of of the IGR buprofezin or pyriproxyfen reduced densities of surviving adults but a similar effect on Eretmocerus mundus eight predator taxa out of 20 examined in at least one year, Mercet was not observed by Jones etal (1998). Hoddle et al including common species such as Geocoris punctipes (1997) assumed that If an IGR is to beused with E. eremicus (Say), Nabis alternatus Parshley, Chrysoperla carnea and in a reduced parasitoid release program then reduced the empidid fly Drapetis nr. divergens. Moreover, the parasitoid longevity and fecundity could have a significant researchers suggested that patterns of predator and pest effect on whitefly control, as host feeding and parasitization population change relative to IGR application dates suggest rates of parasitoid progeny may be reduced following IGR that factors such as reduction in prey availability, were likely applications tohosts within which parasitoids are developing. involved in the reduction of these species other than direct This effect could be significant, as parasitoid reproduction toxic effects of these compounds. within greenhouses can augment weekly releases and Other researchers showed different effects of IGRs on enhance whitefly control. some parasitoid species including E. eremicus, they found Foraging behaviors of female E. eremicus were affected out that mortalities recorded from the use of insect growth by dried IGR residues on poinsettia leaves when B. regulators (IGRs) (flufenoxuron and lufenuron) were <30% argentifolii patches were presented for host feeding and for both pupae and adults of all three species Eretmocerus oviposition. Kinoprene and pyriproxyfen (both liquid IGRs) mundus (Mercet), E. eremicus Rose and Encarsia formosa Gahan indicating that the parasitoids were not seriously affected by these compounds (Sugiyama et al 2011). Table 1. IPM programs used to control Bemisia tabaci on No previous results was study on the effect of the Evisect eggplants for the winter season 2018 under on the parasitoid E. eremicus. The present results showed protected culture that this broad spectrum pesticide was seriously harmful to Treatment 1st step 2nd step 3rd step the parasitoid both in classical and nano-formulation that it T1 IPM 1 Isaria Applaud (a.i. Evisect caused 100% mortality (Fig. 1). The used pesticide fumosorosea Buprofezin) (Thiocyclam hydrogen oxalate) decreased the pest population rapidly and this could be the T2 IPM 2 Applaud Evisect - reason of its lethal effect. The parasitoid of E. eremicus (encapsulated) nano-formulation T3 Control Distilled water DW DW 16.00 14.00 P1 P2 12.00 Table 1. Densities estimates of the whitefly adult parasitoid Control Eretmocerus eremicus (2018-2019) 10.00 Treatments No. of individuals/plant 8.00 6.00 Before application After application 4.00 1st 2nd 3rd 1st 2nd 2.00 sample sample sample sample sample 0.00 T1 5.33 8.7 14.0 0.00 0.00 1st sample 2nd sample 3rd sample 1st sample 2nd sample T2 6.00 8.0 14.3 0.00 0.00 Fig. 1. Effect of IPM programs on adult parasitoid Control 5.33 8.3 14.3 12.00 11.67 Eretmocerus eremicus for the winter season 2018- CD (p=0.05) N.S N.S N.S 4.61 2.40 2019 194 S.A. Alani and Z.A.A. Al-Gadban develop inside the host pupa, the death of the pupa could lead to the death of the parasitoid inside. Moreover, the lack of the prey due to the use of this pesticide could also contribute to expiration of the parasitoid other than the direct toxic effect of the pesticide itself (Naranjo et al 2004). Adult parasitoid of E. eremicus: The newly emerged adults are highly active, as soon as they rapture the exuvia dorsum then they start to push their head capsule to exit the hole (Fig. 2) and spread their wings and antennae after they exit successfully from the mummified pupa, few minutes after they start moving in the area around the mummy crust. All the

Fig. 4. The exuvia of the whitefly B. tabaci (Genn.) under magnification.a. The healthy exuvia with red eye- spots of B. tabaci.b. B. tabaci exuvia parasitized by E. eremicus upper view. c. E. eremicus in final developmental stage on prepared microscopic slide (lower light).

adults collected move to the lower side of the leaves where Fig. 2. Parasitized exuvia of the whitefly Bemisia tabaci their insect host were located and no individuals were ever (Genn.) under magnification showing the adult parasitoid Eretmocerus eremicus rapturing the observed and/or collected from the upper side of the leaves dorsum and emerging from host and this is a strong indication that this species is located mainly close to its insect host and do not fly or search their host away from their patches (Fig. 3). The development of this species parasitoid can be distinguished under microscope in the final stages when the color of the exuvia changes and dark spots of the eyes, wings and antennae start to be visible while in the healthy exuvia of B. tabaci, the color is creamy white to yellowish and the red eye spots of the host B. tabaci are seen shorty before emergence (Fig. 4).

REFERENCES De Cock A, Ishaaya I, Degheele D and Veierov D1990.Vapor toxicity and concentration-dependent persistence of buprofezin applied to cotton foliage for controlling the sweet potato whitefly (Homoptera: Aleyrodidae). Journal of Economic Entomology 83(4): 1254-1260. Gerling D 1990. Natural enemies of whiteflies: predators and parasitoids. In Whiteflies: their Bionomics. Pest Status and Management. Ed. D. Gerling, Intercept Ltd, Andover UK.147- 185. Gerling D and Sinai P 1994. Buprofezineffects on two parasitoid species of whitefly (Homoptera: Aleyrodidae). Journal of Economic Entomology 87(4): 842–846. Fig. 3. The adult of the whitefly B. tabaci (Genn.) parasitoid E. Gerling D, Alomar O and Arnó J 2001. Biological control of Bemisia eremicus under magnification. a. Adult parasitoid tabaci using predators and parasitoids. Crop Protection 20: wondering on lower leaf surface few minutes after 779–799. emergence. b. Dead adult parasitoid dorsal view. c. Gerling D, Tremblay E and Orion T 1991. Initial stages of the vital Dead adult parasitoid ventral view capsule formation in the Eretmocerus- Bemisia tabaci Integrated Pest Management on Whitefly Parasitoid 195

association. Redia 74(3): 411-415. sublethal effects of insecticides on two parasitoids attacking Gould J, Hoelmer K and Goolsby J 2008. Classical Biological Control Bemisia argentifolii (Homoptera: Aleyrodidae). Biological of Bemisia tabaci in the United States A Review of Interagency Control 11(1): 70-76. Research and Implementation. Vol.4. Springer. ISBN 978-1- Naranjo SE, Ellsworth PC and Hagler JR 2004. Conservation of 4020-6739-6. natural enemies in cotton: role of insect growth regulators in Hafez M, Tawfik MFS, Awadallah KT and Sarhan AA 1978. Natural management of Bemisia tabaci. Biological Control 30(1): 52- enemies of the cotton whitefly, Bemisia tabaci (Genn.), in the 72. world and in Egypt. Bulletin Society Entomology Egypte 62: 9- Qiu BL, Ren SX, Lin L and Wang XM 2004b. Effects of release 13. density of Eretmocerus sp. (Hymenoptera: Aphelinidae) on Hoddle MS, Van Driesche RG and Sanderson JP 1997. Biological the control effectiveness of Bemisia tabaci (Homoptera: control of Bemisia argentifolii (Homoptera: Aleyrodidae) on Aleyrodidae). Entomology Journal East China 13: 27-30. poinsettia with inundative releases of Encarsia Formosa Qiu YT, van Lenteren JC, Drost YC and Posthuma Doodeman CJAM Beltsville strain (Hymenoptera: Aphelinidae): Can parasitoid 2004a. Life history parameters of Encarsia formosa, reproduction augment inundative releases? Journal Economic Eretmocerus eremicus and E. mundus, aphelinid parasitoids of Entomology 90(4): 910-924. Bemisia argentifolii (Homoptera: Aleyrodidae). European Hoddle MS, Van Driesche RG, Lyon SM and Sanderson JP 2000a. Journal Entomology 101: 83-94. Compatibility of insect growth regulators with Eretmocerus Shah MR, Zhang SZ and Liu TX 2015. Whitefly, host plant and eremicus (Hymenoptera: Aphelinidae) for whitefly (Homoptera: parasitoid: A review on their interactions. Asian Journal of Aleyrodidae) control on poinsettias. I. laboratory assays. Applied Science and Engineering 4(10): 48-61. Biological Control 20: 122–131. Sugiyama K, Katayama H and Saito T 2011. Effect of insecticides on Hoddle MS, van Driesche RG and Sanderson JP 1998. Biology and the mortalities of three whitefly parasitoidspecies, Eretmocerus use of the whitefly parasitoid Encarsia formosa. Annual Review mundus, Eretmocerus eremicus and Encarsia Formosa Entomology 43: 645-669. (Hymenoptera: Aphelinidae). Applied Entomology Zoology Jones WA, Ciomperlik MA and Wolfenbarger DA 1998. Lethal and 46(3): 311-317.

Received 17 September, 2019; Accepted 04 November, 2019 Manuscript Number: S-280 Indian Journal of Ecology (2019) 46 Special Issue (8): 196-200 NAAS Rating: 4.96

Effect of Host Plant on a Amylase Activity in Cotton Aphid Aphis Gossypii (Glov.) (Hemiptera: Aphididae)

Hind Ibrahim AL-Khazraji Department of Plant Protection, College of Agriculture Engineering Sciences, University of Baghdad, Baghdad, Iraq E-mail: [email protected]

Abstract: The research was conducted to find out the seasonal abundance of cotton aphid on Cucumber (Cucumis sativus), zucchini (Cucurbita pepo) and pepper (Capsicum annum) cultivated during the spring season of 2017, the research also aimed to study the effect of host plant on α- amylase activity in cotton aphid. The results showed that the cotton aphid infestation started on the three crops with the beginning of growth and there was variation in the population density of cotton aphid on different crops and the highest population density was on cucumber crop which amounted to (21.3 individuals/disk leaf), while on zucchini and pepper crops amounted to (12.8 and 8 individuals/disk leaf), respectively. The results also showed that the food preference of cotton aphid on cucumber crop was due to reducing the leaves content of total phenols and carotenoids and increasing in the leaves content of carbohydrates compared to the leaves of both zucchini and pepper crops for the mentioned compounds. The highest activity of α- amylase was on aphid adults which feed on cucumber crop which amounted to (1.01 U ml-1) and decreased to (0.90 and 0.73 U ml-1) on aphid adults which feed on zucchini and pepper crops. Understanding the nutrition biology for insects and the responses of host plants, helps in developing the techniques for pest management.

Keywords: Seasonal abundance, Food preference, Polyphagous, insects, Cotton aphid, Host plant, α-Amylase, Solanaceae, Cucurbitaceae

The Solanaceous and Cucurbitaceous plants are behavior that enables them to feed on the different parts of economically important plants in Iraq and the world. The the plant. In addition to the difference in the nutritional habits crops of these families are grown in the fields and for the insects, some species were characterized as greenhouses, infested with several insect and non-insect monophagous or oligophagous, while others are pests such as Bemisia tabaci, Thrips tabaci, Aphis gossypii, polyphagous, insects have the ability to adapting plant spodoptera littoralis, Tetranychus urticae, etc., Many defenses through their ability to detoxify the plant's defensive methods were used to control pests in the field, greenhouse compounds by releasing antioxidant enzymes after feeding and laboratory (Ali and Ahmed 2009, Sataret al 2012, AL- to detoxify. Food preference may be depending the Khazrajiet al 2016, Khazrajiet al 2018). The cotton aphid secondary plant compounds, including phenolic compounds, Aphis gossypii (Glover) is one of insects that infested there are many studies conducted to know the role of the Solanaceous and Cucurbitaceous plants and the native of phenolic compounds in protection plants from insects the aphid is Southern Eastern Europe. The cotton aphid was attacked where these compounds have an inhibitory effect of diagnosed first time in 1877, which returned to the family growth and fertility, egg-laying and repellent insects, several Aphididae and the order Hemiptera. Aphis gossypii is previous studies have been conducted on the effect of important economically in many countries of the world phenolic compounds on Aphid, especially the effect on because of the economic damage that caused to the plants nutritional behavior, growth, and reproduction. Phenolic into two ways, firstly, the feeding on plant sap and secretion of compounds are considered one of the defenses for the plants the honeydew that causing several symptoms such as to influence the aphid due to its interaction with metabolism yellowing, leaves curl and growth fungi on the leaves as well and selecting the suitable nutritional host for feeding and as prevent of plant breath and photosynthesis because of the laying eggs, where the concentration of bioactive dust. Another damage can be caused by aphid, which compounds varies from plant to plant and even among plants transfer of plant viruses during feeding on infested plant by belonging to one plant family (Beinger et al 2004, viruses. Short life cycle of aphid and their feeding on multi- Czerniewicz et al 2016). Some researchers have indicated plant hosts in different plant families exceed 900 species of the role of carotenes resistant to insects infestation where it is host plants (Blackman and Eastop 2000, Dhaliwal et al considered one of the means of defense for the host plant 2010). Some of insects are characterized by a high nutritional against herbivores and pathogens. Carotenes are formed in preference where they prefer only a certain part of the plant chloroplasts, which are isopropyl compounds found in plants, and other insects characterized by a large nutritional algae and bacteria act as antioxidants and precursors of Amylase Activity in Cotton Aphid 197 phytohormones and attractants to pollinators (Egea et al of 15 pots per crop. It was divided into three replicates at the 2010, Nisar et al 2015, Cáceres et al 2016). Carbohydrates rate of five pots in each replicate for each of the three crops. are essential in feeding insects for optimal growth, increasing After several days, an artificial infection was conducted by fertility, and species survival. Insect requirements for transporting 10 nymphs and adults of cotton aphid for each carbohydrates differ depending on age and gender. There plant. The pots were then covered with organza cloth in a are many enzymes involved in the metabolism of controlled manner and each crop individually to ensure the carbohydrates in insects, especially Aphids, which feed on aphid was fed to the specified host plant. Adults were the plant sap, it is rich in sugar compared to nitrogen. collected after 25 days at a rate of 50 adults per replicates per Carbohydrates are absorbed as monosaccharaides, α- crop for the purpose of assessing the activity of the amylase hydrolyzes starch to maltose and glycogen to α–amylase enzyme according to the method described by glucose by decomposing the α- 1, 4- glucan chains present in Darvishzadeh et al (2014). Fifty individuals of cotton aphid disaccharide, Understanding the function of enzymes and were homogenizer and ground with distilled water. It was insects digestive physiology is essential for developing the placed in a tube with capacity of 1.5 ml in the centrifuge at 15 insect control methods (Pyati et al 2011, Douglas 2006). This 000 rpm for 15 min at a temperature of 4ºC, the experiment research aimed to study the effect of host plant and its was conducted at the rate of 3 replicates per crop. content of chemical compounds on the population density of α-amylase assay: The activity of the enzyme was measured aphid and the effect of host plant on the activity of α-amylase by dinitrosalicylic acid (DNS) based on the method described in cotton aphid. in (Bernfeld 1955) by taking 0.1 ml of the enzyme extract and adding 0.9 ml of the starch solution (1%) as substrate and MATERIAL AND METHODS incubated in a water bath at 35°C for 10 min then added 1 ml Green house plantation: The research was conducted in of DNSA solution. It was left in a water bath to boiling for 5 min the Department of Plant Protection, College of Agricultural and the absorbance was read using the Spectrophotometer Engineering Sciences, University of Baghdad in 2016- at 540 nm after being left in the ice for five min. use maltose to 2017.The greenhouses was cultivated with cucumber prepare standard curve (Fig. 1). The concentration of (Cucumis sativus) Baraka variety, zucchini (Cucurbita pepo) maltose released was estimated based on a standard curve Zyna Flvariety, and pepper (Capsicum annum) California of absorbance during the α-amylase assays. wonder variety that cultivated in overlapping form, the experiment was designed according to the randomized RESULTS AND DISCUSSIONS complete block design. The greenhouse was divided into four Seasonal abundance of cotton aphid on host plants: replicates for each crop and the population density of cotton Figure 2 shows that the first appearance of cotton aphid on aphid was calculated to study the seasonal abundance of the pepper crop at the beginning of March and the population -1 cotton aphid on different crops by taking ten leaves cut density amounted to (1 individuals disk leaf ), the highest randomly from each replicates and regularly weekly, the population density was in the second half of March and the leaves were placed in the labeled polyethylene bags and population densities started gradually decreased, then transferred to the laboratory, then examined leaves to began disappearance in the second half of the May with calculate the numbers of nymphs and adults of cotton aphid increasing the population density of the Aphid green peach (individuals) 5 cm-2 (disk leaf) from the leaf for different crops. Myzus persicae. The reason for the disappearance of the Analysis of chemical compounds in the host plant cotton aphid may be due to feeding competition between the leaves: The fourth and fifth leaves were collected under the apical meristem for a number of plants and it selected 3 y = 1.401x - 0.060 randomly for the three crops. The leaves were transferred to 2.5 R² = 0.996 the laboratory and washed with water to remove the soil, 2 t 540 nm insects, and honeydew. Estimating total phenols (Hus et al 1.5 2003), total carotenoids based on (AOAC 1975) and total 1 carbohydrates (Dubois et al 1956) were done as per standard 0.5

method. Absorbance a 0 Assessing the activity of α-amylase: To study the effect of 0 0.5 1 1.5 2 2.5 maltose concentration (mg ml-1) the host plants on the activity of α-amylase in a cotton aphid, cucumber, zucchini, and pepper were cultivated in pots (the Fig. 1. Standard curve to the determination of maltose height of 23 cm and their diameter of 26 cm), with an average released in the α-amylases assay 198 Hind Ibrahim AL-Khazraji

two aphid species. As for the cucumber crop, the highest cucumber (Engliah, Kaam, Katrina Verities) amounted to population density for the insect amounted to (21.3 (2.45, 2.13, 2.41 mg g-1 plant tissue). Gateh (2016) reported individuals disk leaf-1) was in mid-March and gradually that the content of carotenes in pepper leaves (Fruetescenc decreased to (4 individuals disk leaf-1) at the end of April with Varity) amounted to (0.62 mg 100 g-1). As for the role of the end of the growing season for cucumber crop. As for the phytochemical compounds in the insect resistance, Wojcicka zucchini crop, the population density was very low at the and Leszczynski (2005) indicated that there is a positive beginning of the crop growth and the infestation continued, correlation between plant resistance to aphids and high but low for most of the growing season. This may be due to phenol content. In a study conducted by Wojcicka (2010) on the availability of suitable and preferred hosts of cotton aphid the transgenic wheat plants, it was less preference to cereal to feeding and eggs lying, which is a cucumber crop and then aphid (Sitobion avenae and Rhopalosiphum padi) than non- began to increase gradually and the highest increase transgenic plants, because the total phenol in flag leaves of amounted at the beginning of May after yellowing and death the transgenic wheat plants amounted to (9.34 mg g-1 dry of the cucumbers crop. Saleh et al (2017) indicated in their weight) compared to (4.62 mg g-1 dry weight) in flag leaves of study for the seasonal abundance of sucking insects on the non-transgenic wheat. Czerniewicz et al (2016) indicated cucumber crop and for the two seasons 2014-2015 and that phenolic compounds isolated from Juglans regia L. 2015-2016 that a cotton aphid started to appear with the plants caused deterrent effect, where a deterrence index beginning of crop cultivating and then gradually increased amounted to (41.3%) for Myzus persicae and 34.8% for with the developing plant growth, the infestation was during Rhopalosiphum padi, as well as negative effects of phenols the Autumn season is higher than during the summer season. on insect life representing by decreasing fertility. Murata et al Effect of chemical compounds found in host plant leaves (2019) showed that the biological activity of some on cotton aphid population density: The cucumber crop carotenoids derivatives and their role in resistance to insect was more preference by cotton aphid compared to zucchini infestation, which caused a decrease in the survival time and pepper crops (Table 1). The highest average of period of both Tetranychus urticae and Spodoptera litura. population density for cotton aphid amounted to (9.18 (Hegab et al 2014) reported that the results of the chemical individuals disk leaf-1) compared to (5.21 and 4.43 individuals analysis indicated that there was a direct correlation between disk leaf-1) for the zucchini and pepper crops, respectively. the host plant content of carbohydrates and the percentage The food preference may be due to the cucumber leaves of infestation of cotton aphid, whitefly and leafhoppers on all content of secondary compounds, the results indicate to varieties cultivated for eggplant and pepper. Bayoumy et al reducing the cucumber leaves content of total phenols, which (2017) explained that the population density of Aphis spp. is amounted to (1.4 mg 100 g-1) compared to (2.77 and 3.73 mg increased by increasing the percentage of total 100 g-1) for the zucchini and pepper crops, respectively. In carbohydrates in the host plant where the average of aphids addition to the reduction of the cucumber leaves content of spp. (35.2, 39.3 and 29.6) individuals for eggplant, squash total carotenoids which amounted to (0.93 mg 100 g-1), while and common bean crops and the percentage of the leaves content of total carotenoids amounted to (0.98 and carbohydrates amounted to 45.6, 47.1 and 41.2% in the three 1.11 mg 100 g-1) on zucchini and pepper crops, respectively. crop leaves, respectively. The table also indicated to increase the percentage of total Activity of α-amylase in cotton aphid: Figure 3 shows that carbohydrates in cucumber leaves which amounted to the highest activity of α-amylase enzyme amounted to (1.01 (8.29%) compared to zucchini and pepper crops. Alawami and Abosnena (2013) indicated that the phenol content in 25 C. sativus C. pepo C. annum 20

Table 1. Relationship between chemical compounds of host 15 plant leaves and cotton aphid population density

tion density 10 Host plants Average of Total Total Per cent total population phenols carotenoids carbohydrates 5 -1 popula

density of (mg 100 (mg 100 g ) individuals/5cm² leaf cotton aphid g-1) 0

C. sativus 9.18 1.43 0.93 8.29 2017 2017 2017 2017 / / / / 4 9 6 / / / 16 5 4 / C. pepo 5.21 2.77 0.98 7.57 3 3/2/2017 3 4/20/2017 4/27/2017 5/11/2017 5/18/2017 3/23/2017 3/30/2017 4/13/2017 C. annum 4.43 3.73 1.11 5.98 Fig. 2. Seasonal abundance of cotton aphid (nymphs and CD (p=0.05) 2.931 1.287 0.267 1.77 adults) on host plants Amylase Activity in Cotton Aphid 199

1.2 the use of plant crops as an attractive insect traps depending 1 on the contents of plant compounds and their inclusion in integrated pest management programs. 0.8

0.6 REFERENCES adult Alawami AMY and Abosnena TAS 2013. Effect of treatment with 50 0.4 plant regulators on susceptibility of cucumber cultivars to ylase activity (U/ml) infection by powdery mildew. Libyan Journal of Plant Protection

am 0.2 - 13:1-15. α 0 Ali HI and Ahmed RF 2009. Effects of some insect growth regulators C. sativus C. pepo C. annum and bio-insecticide (Abamectin) in reproductive potential of the cotton leaf worm. The Iraqi Journal of Agricultural Sciences Host plants 40(3): 98-107. Fig. 3. Relationship between activity of α-amylase on cotton Al-Khazraji HI, Ahmed RF and Al-Jorany RS 2016. Effect of feeding treatment with some extracts of black pepper on some aphid and host plants biological aspects of cotton leaf worm. The Iraqi Journal of Agricultural Sciences 47(3): 856-864. U ml-1) on the aphid adults which feed on cucumber crop and AL-Khazraji HI, Abed NS, AL-Mharib MZ and AL-DarrajiS 2018. Effect of of potassium fertilization and organic nutrient (Reef reduced on the aphid adults which feed on zucchini crop to Amirich) in the population density of of Bemisia tabaci (Genn.) reach (0.90 U ml-1), the lowest activity was on the aphid adults and Thrips tabaci (L.) on cucumber. Journal of Biodiversity and Environmental Sciences 12(2):11-18. which feed on the pepper crop which amounted to (0.73 U ml- AOAC 1975. Official methods of analysis. Association of official 1). The reduction in the activity of the enzyme may be due to analytical chemists.13 ͭ ͪ ed. Washint. D. C. the reduction in the percentage of carbohydrates in the Applebaum SW, Jankovic M and Birk Y 1961. Studies on the midgut amylase activity of Tenebrio molitor L. larvae. Journal of Insect leaves of pepper crop. The results agree with (Darvishzadeh Physiology 7: 100–108. et al 2014) who mentioned that the activity of α-amylase for Bandani AR, Amiri B, Butt TM and Gordon-Weeks R 2001. Effects of cotton aphid feed on cucumber crop amounted to (1.225 U efrapeptin and destruxin, metabolites of entomogenous fungi, -1 on the hydrolytic activity of a vacuolar type ATPase identified on ml ). The benefiting from carbohydrates in insects depends the brush border memebrane vesicles of Galleria mellonella on hydrolysis, absorption, and metabolism, which is a midgut and on plant membrane bound hydrolytic enzymes. complex relationship, the increase in the content of both Biochemica ET Biophysica Acta 1510: 367-377. Bayoumy MH, Awadalla SS, El-Gendy MA and El-Lawatay NE 2017. starch and glycogen in insect food indicates the presence of Comparative morphology and chemical composition of plant digestive enzymes in the insect's gut. Bandani et al (2001) leaf and their relation with population density of certain piercing-sucking insect pests. Journal of Plant Protection and and Applebaum et al (1961) explained with increasing the Pathology 8(2):31-37. need for digestion and high carbohydrate metabolism, we Beinger CW, Abou-Zaid MM, Kistner AL, Hallett RH, Iqubal MJ, notice an increase in the number of enzymes responsible for Grodzinski B and Hall JC 2004. A flavanone and two phenolic acids from Chrysanthemum morifolium with phytotoxic and carbohydrate digestion, thus the importance of amylase insect growth regulating activity. Journal of Chemical Ecology appear in the survival, growth, and development of insects. 30: 589-606. Bernfeld P 1955. Amylase, α and β. Methods in Enzymology 1: 149- 158. CONCLUSIONS Blackman RL and Eastop VF 2000. Aphids on the World's Crops: An Insects differ in their food preference depending on their Identification and Information Guide, 2nd ed, John Wiley and basic needs of different nutrients for their growth and Sons, Chichester, UK. 414 p. development, they prefer nutrition over another, therefore Cáceres LA, Lakshminarayan S, Yeung KKC, McGarvey BD, Hannoufa A, Sumarah MW, Benitez X and Scott IM 2016. insects be have different behavior to choose the best Repellent and attractive effects of a-, b-, and dihydro-b-ionone nutrition, depending on several factors, including the traits of to generalist and specialist herbivores. Journal of Chemical Ecology 42: 107–117. the host plant, where many plants have the traits and Cartwright B 1992. Assessment of damage to watermelon by Aphis morphological characteristics make them a plant host gossypii (Homoptera: Aphididae). Proceedings of the 19th resistant to insects infestation including the presence of international congress of entomology, June 28-July 4,1992, Beijing, China. trichomes, glandular bristles, leaf area, the thickness of Czerniewicz P, Chrzanowski G, Sytykiewicz H, Sprawka I and cuticle layer and plant age or depending on the secondary Leszczynski B 2016. Aphidicidal and deterrent activity of plant compounds and the variation of the contents of phenolic acid extracts from some herbal plants towards Myzus persicae Sulz. and Rhopalosiphum padi L. Fresenius nutrients, including soluble proteins and carbohydrates, as Environmental Bulletin 25(12a): 5714-5721. well as the different nature of plant sap and pH, which play a Darvishzadeh A, Bandani AR and Mousavi SQ 2014. Biochemical characterisation of α-amylase in two aphid species, Aphis major role in the behavior of nutritional preference for insects. fabae Scopoli (Hemiptera: Aphididae) and A. gossypii Glover The results obtained from the current study can be utilized in (Hemiptera: Aphididae). Plant Protection Science 50: 84-89. 200 Hind Ibrahim AL-Khazraji

Dhaliwal GS, Jindal V and Dhawan AK 2010. Insect pest problems 179: 1822–1833. and crop losses: changing trends. Indian Journal of Ecology 37: Nisar N, Li L, Lu S, Khin NC and Pogson BJ 2015. Carotenoid 1-7. metabolism in plants. Molecular Plant 8:68-82. Douglas AE 2006. Phloem sap feeding by animals: Problems and Pinto ZV, Rezende JAM, Yuki VA and Piedade SMS 2008. Ability of solutions. Journal of Experimental Botany 57: 747-754. Aphis gossypii and Myzus persicae to transmit cucumber Dubois M, Gilles KA, Hamilton JK, Rebers PN and Smith F 1956. mosaic virus in single and mixed infection with two Poty viruses Colorimetric method for determination of sugars and related to zucchini squash. Summa Phytopathologica Botucatu substances. Analytical Chemistry 288(3): 350-356. 34(2):183-185. Egea I, Barsan C, Bian W, Purgatto E, Latché A, Chervin C, Pyati P, Bandani AR and Fitches E 2011. Protein digestion in cereal Bouzayen M and Pech JC 2010. Chromoplast differentiation: aphids (Sitobion avenae) as a target for plant defence by Current status and perspectives. Plant and Cell Physiology endogenous proteinase in hibitors. Journal ofInsect Physiology 51:1601-1611. 57: 881-891. Gateh HJ 2016. A study on the effect of salinity in germination, Saleh AAA, El-Sharkawy HM, El-Santel FS and Abd El-Salam RA chemical and ionic contents of pepper plant (Capiscum 2016. Seasonal abundance of certain piercing sucking pests on annuum L.). Thi-Qar University Journal for Agricultural cucumber plants in Egypt. Egyptian Academic Journal of Researches 5(1): 361-374. Biological Sciences 10(7): 65-79. HegabMA, Ibrahim AE, Shahein AA and Abdel-Magid JE 2014. Satar S, Kersting U and Yokomi R 2012. Presence of two host races Susceptibility of certain solanaceous plant varieties to some of Aphis gossypii Glover (Hemiptera: Aphididae) collected in homopterous insects infestation. Journal of Entomology 11(4): Turkey. Annals of Applied Biology 1-10. 198-209. Wojcicka A2010. Cereal phenolic compounds as biopesticides of Hus CL, Chen W, Weng YM and Weng CY 2003. Chemical cereal aphids. Polish Journal of Environmental Studies 19(6): composition, physical properties and antioxidant activities of 1337-1343. yam flours as affected by different drying methods. Food Wojcicka A and Leszczynski B 2005. Secondary plant compounds Chemistry 83: 85-92. may serve as biopesticides for the grain aphid Sitobion avenae MurataM, Nakai Y, Kawazu K, Ishizaka M, Kajiwara H, Abe H, (F.) Pesticides 4: 163-170. Takeuchi K, Ichinose Y, Mitsuhara I, Mochizuki Aand Seo S Wool D, Hales D and Sunnucks P 1995. Host plant relationships of 2019. Loliolide, a carotenoid metabolite, is a potential Aphis gossypii Glover (Hemiptera: Aphididae) in Australia. endogenous inducer of herbivore resistance. Plant Physiology Australian Journal of Entomology 34: 265-271.

Received 28 October, 2019; Accepted 28 November, 2019 Manuscript Number: S-281 Indian Journal of Ecology (2019) 46 Special Issue (8): 201-208 NAAS Rating: 4.96

Effect of Foliar Feeding on Copper and Molybdenum in Some OF the Physiological, Productivity and Quality Traits for Two Genotypes Mung Bean (Vigna radiata L.)

Riyadh L. Attieh and Basheer H. Abdallah Department of Field Crops, College of Agriculture, University of Anbar, Iraq E-mail: [email protected]

Abstract: A field experiment was carried out in the autumn season of the years 2017 and 2018 in Anbar province, the alsharqia Husaybah region- Located on the (a longitude of 43.46o and a latitude of 33.41o width circuit), to determine the effect of leave feeding on three concentrations of copper (0, 8 and 16 mg Cu l-1) and molybdenum (0, 30 and 60 mg l-1). In some of the physiological, productive and qualitative characteristics of two genotypes of Mung bean (Indian and local). Concentration of copper (8, 16 mg Cu L -1) showed the highest efficacy of the nitrate reduction enzyme (NR) in the second season of the experiment. The concentration also gave 8 mg Cu l-1 the highest average of leave area and percentage of fertile in bods and seeds of plant (23.97 and 27.17 gplant-1). While concentration gave 16 mg Cu l-1 highest percentage of molybdenum in leaves and number of bods plant and ratio of carbohydrates in seeds (21.44 and 22.05%) seasons respectively. Concentration of molybdenum given 30mg l-1, highest average leave area, number of pods plant, bods fertilization ratio, plant seed yield (28.15 and 31.70 g), carbohydrate ratio in seeds (22.10 and 22.69%) and seasons respectively. While the concentration of 60 mg l-1 gave the highest efficacy of the NR in leaves the second season (137.11 μm NO2 gm-1 hour-1) and the highest molybdenum ratio in both seasons (13.9 and 21.57 mg kg-1) dry matter, respectively. Indian genotype superiority (NR) in season and in all other traits and both seasons except for the leave area.

Keywords: Copper, Molybdenum, Physiological trait, Quality trait, Mung bean

Moong (Vigna radiata L.) is an important fabaceae crop, Based on the importance of the above, this study aims to which is used in most of the countries produced mainly as a know the best concentration of the copper and molybdenum food for humans or as a protein source, which ranged in seeds component achieves a significant increase in growth, seed from (19-29%), carbohydrates (62-65%) and fat (1- 5%), yield and some specific traits. Determine the best genotype These ratios depend on the genotypes used and the that responds to the foliar feeding of these two elements and prevailing environmental factors (AL-Fahdawi 2016, Ali 1990). gives the highest seed yield with improved quality of the yield. The plant is also grown for green animal feed and to improve soil properties, especially in reclaimed land. The problem of MATERIAL AND METHODS this crop is the increase in the percentage of flowering falling A field experiment was carried out in the autumn season that reaches 60-80% of the total flowering in the plant (Ali of 2017 and 2018 in Anbar province in the alsharqia 1990). This high percentage leads to a decrease in yield Husaybah region located near of the Habbaniyahon the right productivity. One of the treatments of this problem is the use of bank of the Euphrates, on a longitude of 43.46o and a latitude nutrients related to increasing fertility rate flowers, seeds and of 33.41o, in soil with a mixture of green clay (Table 1). The pods, especially balanced foliar feeding with molybdenum target of study is the effect of three concentrates of copper (0, and copper. Copper is a small nutrient that performs this role, 8 and 16 mg Cu l-1) and molybdenum (0, 30 and 60 mg Mo l-1) The lack of it causes an imbalance in plant growth through its in some physiological, productive and qualitative participation in photosynthesis in photosynthesis and characteristics of two genotypes of Mung bean (Local and oxidation and reduction processes in the electronic transport Indian VCV 89A10). The experiment was carried out in the chain (Electron transport chain) in aerobic respiration (Abu order of split-split plots according to the design of randomized Dahi and al-Yunis 2017, Ara and Hasan 2009, Black 1965). complete block design and four replicates, The genotypes The molybdenum component also plays an important role in included main plots, while the molybdenum component photosynthesis processes within the plant, It is the key to the included sub plots and copper the sub-sub plots, use biological metabolism of the nitrogen element, and the ammonium molybdate 6MO 7O24 4H2O (NH4) (12.88% MO) process of biologically stabilizing atmospheric nitrogen by as a source of molybdenum, copper sulfate CuSO4 5H2O micro-soil regeneration stimulates and increases efficiency in (39.28% Cu) as a copper source. The molybdenum and the availability of this element (Ali 1990). copper concentrations were sprayed on the first two steps at 202 Riyadh L. Attieh and Basheer H. Abdallah

Table 1. Some chemical and physical properties of pre- the Indian variety and on 30/10/2017 for the local variety. In planting experiment soil for both 2017 and 2018 the second season, on 13/10 for the Indian variety and 27/10 Elements \ unit 2017 2018 for the local variety for 2018. The number of bods was pH 8.5 7.8 calculated as the average number of plants in 10 plants Electrical conductivity EC MG m-3 Soil 4.25 3.85 randomly taken from the mid lines of each experimental unit. Water 2.4 2.7 Plant seeds were calculated from the ten plants' bods that N 42% 40.40% were dried in an electric furnace at an appropriate P 10.70% 10.30% temperature, then weighed by a sensitive balance and then K 194% 185.20% take the averaged. The ratio of carbohydrates in seeds was estimated according to the official method of chemical Zn 0.32% 0.42% analysts (AOAC 1990). The statistically data was analyzed Cu 0.26% 0.29% using Genstat and the least significant difference (L.S.D) was Sand 50.2 ppm 55.7 ppm used to compare the averages at the probability level of 0.05 Clay 460.4 ppm 458.8 ppm (Al-Rawi and Khalafallah1980). Silt 480.4 ppm 485.5 ppm texture Silt clay Silt clay RESULTS AND DISCUSSIONS Total efficacy of enzyme nitrate-reduced NR (micromol NO g-1 hr-1): Figure 1a shows that the effectiveness of this the beginning of the vegetative growth and the second at the 2 beginning of the flowering. The comparison treatment was enzyme increased with the increased concentration of sprayed with distilled water only. Plant the sprayed in the copper-based leave feeding, increasing from 120.85 when -1 evening using a 16 liter and add 0.15 ml L-1 of the zahi as a compared to 131.96 at the concentration of 8 mg Cu l . And -1 diffuser to reduce the surface tension of the water and ensure then to 133.35 μm NO2 gm hour-1 hourly weight at the high -1 complete wetness of the leaves and increase the efficiency of concentration of copper (16 mg Cu ) but the increase was not -1 the spray solution to penetrate the outside surface of the leaf significant after copper concentration 8 mg Cu l . This (Abu Dahi et al 2001). The experimental land was divided increase in the effectiveness of the enzyme by adding the into experimental units area 7.5 m2 (3m x 2.5m) and element of copper is due to the that the mineral elements, contained of six lines along the length of the line 3m between especially the copper element, is a stimulant for the the line and another 40cm and between hole and another effectiveness of the enzymes, As well as it is enter in the 25cm to obtain plant density 100,000 plants h-1 (Ashwini composition of the metal part of the enzyme (non-protein) 2005). Seedlings were planted for both varieties which was and this is reflected positively on the performance of the obtained from the Department of Agricultural Research - enzyme and its effectiveness. Ministry of Agriculture on 26/7 of 2017 and 2018 at a depth of Figure 1b, there was a significant increase in the 2-3 cm and after the complete of germination plants, keep effectiveness of the reduced enzyme of nitrate with increased one plant one plant in hole. Land of experiment with concentration of foliar feeding with molybdenum until it phosphate and nitrogen fertilizers according to reached the highest concentration at 60 mg l-1, which reached -1 -1 recommendations for the amount of fertilizer and the date of 137.11 Micromol NO2 gm Light weight hour , with an addition (Ali 2012). The efficacy of nitrate reduction enzyme increase of 13.71 and 6.73% compared to its effectiveness in (NR) according to the method proposed by Ashwini (2005). the treatment of the control (Mo 0) and the concentration The concentration of molybdenum in the leaves after 10 days plants 30 mg l-1 respectively. The role of molybdenum in the of the second spray of both elements (copper and efficacy of NR is due to the fact that it is a key component of molybdenum) was determined by a Flame Photometer (PGI this enzyme and, it worked as a catalyst or stimulation and Automatic 2000) of English origin in the manner cited by thus increases the efficiency of the enzyme in nitrate (Black 1965). Leave area measurement after 14 days of the reduction process. measurement of the enzyme effectiveness, according to the Figure 1c, the efficacy of the enzyme was high and way disk, where take 30 leave disk from 10 plant leave. Dry significantly in the plants of the Indian genetics, which -1 -1 plant and disk leaves, then estimated leave area in the amounted to 130.99 μm NO2 gm hr . As compared to its following equation. Leave area plant (Dry weight plant leaves effectiveness in local genotypes, which reached 126.45 μm -1 -1 /dry weight for 30 disk)* area of 30 disk (Johnson and NO2 gm hr . Arris1967). Leave area plant (cm2 plant-1): Table 2 shows that feeding The harvest was for the first season on 20/10/2017 for plants with a concentration of 8 mg Cu l-1 gave the highest Mung Bean Effectiveness by Foliar Feeding on Copper and Molybdenum 203

plant-1 respectively. This results agree with (AL-Khafaji 2015, Al-Mahmoudi 2012). Interaction between copper and molybdenum the plants with concentration of 8mg L-1 with a concentration of 30 mg l-1 significantly superiority all other interference coefficients in this characteristic, in which the comparison plants gave the two plants the lowest mean of the two seasons. The results of the double and triple interactions between the studied factors were significant in this and both seasons (Table 2). Molybdenum ratio in the leaves (mg kg-1 dry matter): The results of Table 3 showed a significant increase in the ratio of molybdenum in leaves with an increase in concentrations of copper and both seasons. The high concentrations of copper (16 mg Cu l-1) showed the highest ratio of the elements in both seasons of 10.81 and 15.70 mg kg-1 dry matter, respectively, compared to the control treatment that gave the lowest ratio of the two seasons amounted 8.71 and 12.07 mg kg-1 dry matter, respectively. The percentage of molybdenum in leaves increased with the addition of its level. The plants achieved concentration (60 mg Mo l-1). The highest ratio in both seasons was 13.90 and 21.57 mg kg-1 dry matter respectively, while the control plants recorded the lowest Fig. 1. Effect of Cu and Mo paper feeding on the total ratio of 5.59 and 5.79 mg kg-1 dry matter and for the two effectiveness of nitrates reduction enzyme NR (μm -1 -1 seasons respectively, this results agree with (Kandil and NO2 gm weight hr ) for two genotypes of mung bean for the season (2018) Abdelhamid 2013) on the bean crop. The results of Table 3 showed that the plants of the Indian genotype gave the highest ratio of molybdenum in their leaves in both seasons -1 average leave area in both seasons of 437.46 and 445.37 of 9.81 and 15.01 mg kg dry matter respectively, and cm2 plant-1 respectively, significantly superiority the high significantly superior the local genotype plants which concentration of copper (16 mg Cu-1). Comparative plants recorded the lowest ratio in the seasons of 9.70 and 12.77 mg -1 that gave the lowest mean for both seasons were 369.67 and kg dry matter, respectively. The results of the triple 378.27 cm2, respectively. The reason for the increase of interference between the studied factors showed that they leave area at the concentration of 8 mg Cu l-1 may be due to its were significant in this trait and for both seasons. superiority in plant height and the number of plant branches Number of bods plant: Table 4 shows that plants with high -1 and thus increase the number of leaves in the plant and then concentration of copper (16 mg Cu l ) gave the highest mean -1 increase the leave area (AL-Fahdawi 2016, Kalaikandhan et of 54.25 and 55.36 bod plant and differed significantly from -1 al 2014, Mohammadi 2011). the concentration plants of 8 mg Cu l and the control plants -1 It is clear from the same table that plants with that gave the lowest average 49.24 and 49.38 bod plant and Molybdenum concentration (30 mg l-1) gave the highest for the two seasons, respectively. This result agree with average leave area in both seasons of 442.38 and 451.67 others researches (AL-Fahdawi 2016, Mohammadi 2011, cm2 plant-1 respectively and significantly superiority the Mohsen et a l2013). The same table shows that the plants -1 concentration plants (60 mg l-1). The comparison plants that were sprayed with a concentration of 30 mg Mo L gave recorded the lowest average of 365.12 and 375.00 cm2 plant-1 the highest average in the seasons of 59.41 and 59.87 bod -1 and for the two seasons, respectively. This result agree with plant respectively and significantly superior the control Awomi et al (2012), Malik and Singh (2015). plants that gave the lowest average of the seasons in 44.53 -1 The results of Table 2 showed that the local genotype and 45.90 bod plant respectively. The results of this study significantly superiority the mean leave area in both seasons were agreed with (Awomi 2012, Hazili and Fajr 2016, Tahir of 455.03 and 463.57 cm2 plant-1 respectively, and increased and Muhammad 2014). by 29.76 and 28.68% for the Indian genotype, which gave the The results of Table 4 indicate that the Indian genotype lowest mean in the seasons was 350.67 and 360.26 cm2 was significantly superiority higher average in the two 204 Riyadh L. Attieh and Basheer H. Abdallah

Table 2. Effect of foliar feeding on Cu and Mo in the average leave area of the plant (cm2 plant-1) for two genotypes of mung bean for the seasons (2017 and 2018) 2017 2018

Genotypes Concentrations Concentrations Cu (mg l-1) VxMo Concentrations Cu (mg l-1) VxMo of MO (mg l-1) 0 8 16 0 8 16

Indian (V1) 0 287.19 344.70 309.39 313.76 299.42 355.77 322.38 325.86 30 352.79 415.10 374.63 380.84 364.16 424.94 383.40 390.83 60 337.26 379.25 355.76 357.42 345.33 383.12 363.77 364.08 Local (V2) 0 389.07 453.43 406.93 416.48 392.30 460.82 419.29 424.14 30 444.42 557.29 510.06 503.92 448.92 565.80 522.81 512.51 60 407.29 475.01 451.74 444.68 419.50 481.74 460.94 454.06 LSD 5% 5.048 2.867 8.466 5.228 Average (V) Average (V) VxCu V1 325.75 379.69 346.59 350.67 356.52 387.95 336.30 V2 413.59 495.24 456.24 455.03 467.68 502.79 420.24 LSD 5% 2.713 1.694 4.956 4.241 Average (Mo) Average (Mo) MoxCu 0 338.13 399.06 358.16 365.12 345.86 408.30 370.84 375.00 30 398.61 486.20 442.34 442.38 406.54 495.37 453.10 451.67 60 372.27 427.13 403.75 401.05 382.42 432.43 412.36 409.07 LSD 5% 3.711 2.328 6.051 3.856 Average (Cu) 369.67 437.46 401.42 378.27 445.37 412.10 LSD 5% 2.155 3.484

Table 3. Effect of foliar feeding on Cu and Mo in the average Mo ratio in leaves (mg kg-1 dry matter) for two genotypes of mung bean for seasons (2017 and 2018) 2017 2018

Genotypes Concentrations Concentrations Cu (mg l-1) VxMo Concentrations Cu (mg l-1) VxMo of MO (mg l-1) 0 8 16 0 8 16

Indian (V1) 0 5.91 6.51 6.87 6.43 6.11 6.67 7.10 6.62 30 8.57 9.22 11.57 9.79 12.83 15.40 17.84 15.36 60 12.61 12.81 14.26 13.22 20.74 22.94 25.49 23.06 Local (V2) 0 4.20 4.86 5.18 4.75 4.42 5.05 5.40 4.96 30 7.44 10.67 11.20 9.77 10.63 13.82 15.38 13.28 60 13.56 14.37 15.80 14.57 17.69 19.55 23.02 20.08 LSD 5% 0.1661 0.0973 0.2023 0.1103 Average (V) Average (V) VxCu V1 9.03 9.51 10.90 9.81 13.23 15.00 16.81 15.01 V2 8.40 9.97 10.72 9.70 10.91 12.80 14.60 12.77 LSD 5% 0.0924 0.0681 NS 0.0499 Average (Mo) Average (Mo) MoxCu 0 5.05 5.68 6.02 5.59 5.26 5.86 6.25 5.79 30 8.00 9.95 11.38 9.78 11.73 14.61 16.61 14.32 60 13.08 13.59 15.03 13.90 19.21 21.24 24.25 21.57 LSD 5% 0.1208 0.0761 0.1499 0.0925 Average (Cu) 8.71 9.74 10.81 12.07 13.90 15.70 LSD 5% 0.0700 0.0878 Mung Bean Effectiveness by Foliar Feeding on Copper and Molybdenum 205 seasons amounted 55.57 and 56.58 bod plant-1 respectively, interference in the same table showed significant differences with an increase of 16.52% and 17.14% for the local between the study factors for two seasons. genotype which gave the lowest average of 47.69 and 48.30 Bods fertility ratio (%): It is clear from the results of Table 5 bod plant-1 and for the two seasons respectively, That the that the copper spray at the concentration of 8 mg Cu L-1 superiority of the Indian genotype in the ratio of molybdenum achieved the highest average for this trait in both seasons of in the leaves as well as the effectiveness of the enzyme NR 83.03 and 84.05% respectively, compared with the average was high in this genotype was reflected positively in control treatment of 77.49 and 78.58% both seasons increasing the ratio of fertility in flowers and then increase the respectively. The increase in the ratio Fertility pods may be number of bods. This results agree with (Ara and Hasan due to the role of copper in the improvement of the plant 2009, Mohammadi 2011, Yusuf et al 2014). The overlap biological processes, which are reflected in the improvement between the copper and genotypes Table 4 shows that foliar of this trait. This result was agreed with Al-Fahdawi (2016). feeding at the concentration of 16 mg Cu l-1 for Indian Table 5 shows that plants with a concentration of 30 mg MO l-1 genotypes achieved the highest average number of plant had the highest average for this trait at 82.16 and 83.27% bods in the two seasons of 57.52 and 58.95 pod plant-1, both seasons, respectively. The Indian genotype was respectively, compared to local genotype control plants Less significantly superiority in both seasons (82.88 and 83.98%) average of both seasons was 43.80 and 43.75 bod plant-1 while in the local genotype less ratio amounted (77.58 and respectively. Table 4 shows that the Indian genotype under 78.65%) in both seasons. This result was agreed with (AL- the spraying effect of 30 mg L-1 gave the highest average for Dabbagh 2017 and AL-Fahdawi 2004). The results of Table both seasons of 65.44 and 66.24 bod plant-1, respectively, 5 show that the plants of the Indian genotype with while the lowest average was obtained in local genotype non- concentration feeding (8 mg Cu l-1) achieved the highest ratio sprayed molybdenum In both seasons it reached 39.89 and fertility pods in the first season of 85.27%, while the 40.79 bod plant-1 respectively. The results of triple comparative treatment of the local genotype gave the lowest

Table 4. Effect of foliar feeding on Cu and Mo in the average number of pods plant-1 for two genotypes of mung bean for the seasons (2017 and 2018) 2017 2018

Genotypes Concentrations Concentrations Cu (mg l-1) VxMo Concentrations Cu (mg l-1) VxMo of MO (mg l-1) 0 8 16 0 8 16

Indian (V1) 0 42.84 48.75 55.92 49.17 44.37 51.37 57.29 51.01 30 62.82 63.91 69.59 65.44 63.99 64.10 70.64 66.24 60 58.37 50.88 47.06 52.11 56.64 51.92 48.91 52.49 Local (V2) 0 35.50 39.98 44.19 39.89 35.29 41.37 45.71 40.79 30 45.28 54.21 60.64 53.38 44.51 55.55 60.42 53.49 60 50.61 50.68 48.12 49.80 51.44 51.19 49.20 50.61 LSD 5% 3.617 2.796 3.035 2.149 Average (V) Average (V) VxCu V1 54.68 54.51 57.52 55.57 55.00 55.80 58.95 56.58 V2 43.80 48.29 50.98 47.69 43.75 49.37 51.78 48.30 LSD 5% 2.230 2.297 1.656 1.464 Average (Mo) Average (Mo) MoxCu 0 39.17 44.36 50.05 44.53 39.83 46.37 51.50 45.90 30 54.05 59.06 65.12 59.41 54.25 59.82 65.53 59.87 60 54.49 50.78 47.59 50.95 54.04 51.55 49.06 51.55 LSD 5% 2.597 2.048 2.249 1.693 Average (Cu) 49.24 51.40 54.25 49.38 52.58 55.36 LSD 5% 1.239 1.137 206 Riyadh L. Attieh and Basheer H. Abdallah ratio fertility pods reached 74.25%. respectively. Compared to the local genotypes that gave the Plant seeds yield (g): Table 6 shows that plants with copper lowest average of 11.94 and 14.00 g and for the two seasons, concentration of 8 mg Cu l-1 gave the highest average for this respectively. Agree this results with the (Mohammadi 2011 trait in the seasons of 23.97 and 27.17 g, respectively and and Sadeghipour 2008). The results of the Table 6 indicate a significantly superiority the concentration plants 16 mg Cu l-1. significant effect of the triple interference between the study The control plants that gave the lowest average in the factors. seasons were 19.42 and 22.94 g, respectively. Carbohydrate ratio in seeds (%): Table 7 shows that plants Concentration superiority of 8 mg l-1 in bods fertilization ratio with a concentration of copper 8 and 6 mg Cu l-1 achieved the (Table 5) may have been reflected in increasing the number highest percentage of carbohydrates in both seeds and both of seeds bods and thus increasing the number of seeds in the seasons, and significantly superior the control plants in the plant and thus increasing the seed yield of the plant. The two seasons, which gave the lowest percentage of these same table shows that the 30 mL MO l-1 concentration feeding traits at 18.67 and 19.18%, respectively. The superiority of plants gave the highest average yield of seeds of the two these two concentrations in the leave area (Table 2) was seasons at 28.15 and 31.70 g, respectively. And significantly positively reflected in increased interference and absorption superior the high concentration plants of the element, The of light and thus increased photosynthesis products, which control plants that gave the lowest average in both seasons are the main source of carbohydrates. The results of Table 7 were 18.05 and 21.16 g, respectively. The concentration of showed that the plants with a concentration of 30 mg l-1 had 30 mg l-1 in the number of bods in the plant was positively the highest average in both seasons of 22.10 and 22.69%, reflected in the increase in plant yield of seeds. The results respectively, and significantly superiority in both seasons on agree with (Bhuiyan et al 2008 and Kandil et al 2013). The the high concentration and the control plants which gave the results of Table 6 indicate that the superiority of the plants of lowest average of 19.09 and 19. 63%, respectively. The the Indian genotype significantly with the highest average results of the same table show that the plants of the Indian plant yield in both seasons was 35.26 and 40.02 g, genotype significantly superior the percentage of

Table 5. Effect of Cu and Mo foliar feeding in pods fertility ratio (%) for two genotypes of mung bean for the seasons (2017 and 2018) 2017 2018

Genotypes Concentrations Concentrations Cu (mg l-1) VxMo Concentrations Cu (mg l-1) VxMo of MO (mg l-1) 0 8 16 0 8 16

Indian (V1) 0 78.84 82.77 80.64 80.72 79.88 83.84 81.87 81.86 30 82.46 87.38 84.63 84.82 83.74 88.20 85.78 85.91 60 80.99 85.67 82.68 83.11 82.04 86.58 83.88 84.17 Local (V2) 0 72.67 78.73 75.38 75.59 73.58 79.67 76.23 75.49 30 75.72 83.37 79.41 79.50 76.92 84.51 80.45 80.62 60 74.36 80.27 78.32 77.65 75.34 81.52 79.62 77.83 LSD 5% NS NS NS NS Average (V) Average (V) VxCu V1 80.73 85.27 82.65 82.88 81.89 86.21 84.84 83.98 V2 74.25 80.79 77.70 77.58 75.28 81.90 78.77 78.65 LSD 5% 0.7991 0.8571 NS 0.9511 Average (Mo) Average (Mo) MoxCu 0 75.70 80.75 78.01 78.15 76.73 81.75 79.05 79.18 30 79.09 85.37 82.02 82.16 79.33 85.35 83.11 83.27 60 77.67 82.97 80.50 80.38 78.69 84.05 81.75 81.50 LSD 5% NS 0.4665 NS 0.5415 Average (Cu) 77.49 83.03 80.18 78.58 84.05 81.30 LSD 5% 0.3792 0.4551 Mung Bean Effectiveness by Foliar Feeding on Copper and Molybdenum 207

Table 6. Effect of Copper and Molybdenum foliar feeding on the average plant seeds yield for two genotypes of mung bean for the seasons (2017 and 2018) 2017 2018

Genotypes Concentrations Concentrations Cu (mg l-1) VxMo Concentrations Cu (mg l-1) VxMo of MO (mg l-1) 0 8 16 0 8 16

Indian (V1) 0 22.13 29.72 33.40 28.22 28.22 34.90 37.76 32.81 30 39.93 48.59 47.24 45.28 45.28 52.21 52.31 50.41 60 34.31 34.75 30.12 33.18 33.18 37.74 36.01 37.44 Local (V2) 0 08.04 10.55 11.06 9.85 9.85 12.29 12.85 11.60 30 11.75 16.24 16.65 14.83 14.83 19.00 18.40 11.90 60 09.79 12.57 11.58 11.29 11.29 14.58 13.07 13.60 LSD 5% 0.1436 0.0889 0.1436 0.0281 Average (V) Average (V) VxCu V1 31.74 37.32 36.76 35.26 36.74 41.31 42.02 40.02 V2 09.90 13.05 13.03 11.94 12.09 15.22 14.71 14.00 LSD 5% 0.0829 0.198 0.0262 0.0335 Average (Mo) Average (Mo) MoxCu 0 14.42 19.16 20.82 18.05 17.24 22.43 23.82 21.16 30 23.74 30.46 30.43 28.15 27.41 34.05 33.64 31.70 60 20.41 22.84 20.28 21.21 24.56 25.58 23.58 24.57 LSD 5% 0.102 0.0629 0.0321 0.0198 Average (Cu) 19.42 23.97 23.70 22.94 27.17 26.47 LSD 5% 0.0017 0.0185

Table 7. Effect of foliar feeding on Cu and Mo in the ratio of carbohydrates in seeds (%) for two genotypes of mung bean for the seasons (2017 and 2018) 2017 2018

Genotypes Concentrations Concentrations Cu (mg l-1) VxMo Concentrations Cu (mg l-1) VxMo of MO (mg l-1) 0 8 16 0 8 16

Indian (V1) 0 18.24 20.21 21.76 20.07 18.51 20.91 22.20 20.54 30 19.25 24.30 26.29 23.28 19.65 25.17 27.12 23.98 60 21.18 22.84 19.56 21.19 21.95 23.29 20.12 21.78 Local (V2) 0 16.58 17.78 20.01 18.12 17.04 18.52 20.59 18.71 30 17.60 21.89 23.30 20.93 18.03 22.23 23.95 21.40 60 19.18 21.16 17.71 19.35 19.93 21.94 18.34 20.07 LSD 5% 0.1841 0.1237 0.1535 0.0946 Average (V) Average (V) VxCu V1 19.55 22.45 22.54 21.51 20.03 23.12 23.14 22.10 V2 17.79 20.28 20.34 19.47 18.33 20.89 20.96 20.06 LSD 5% 0.1218 0.1210 0.0262 0.0937 Average (Mo) Average (Mo) MoxCu 0 17.41 18.99 20.88 19.09 17.77 19.71 21.39 19.63 30 18.42 23.09 24.79 22.10 18.84 23.70 25.53 22.69 60 20.18 22.00 18.63 20.27 20.94 22.61 19.23 20.93 LSD 5% 0.1249 0.0772 0.1045 0.0578 Average (Cu) 18.67 21.36 21.44 19.18 22.01 22.05 LSD 5% 0.0731 0.0641 208 Riyadh L. Attieh and Basheer H. Abdallah

carbohydrates in their seeds and both seasons reached Regulators on Physiology and Yield in French bean (Phaseolus Vulgaris L.). Master Thesis, Department of Crop Physiology, 21.51 and 22.10% respectively, and the ratio of superiority to University of Agriculture Science, Dharwad, India. pp: 16-22. 10.48 and 10.17% compared to the local genotype, which Awomi TA, Singh AK, Kumar M and Bordoloi LJ 2012. Effect of gave the lowest ratio of the trait amounted to 19.47 and phosphorus, molybdenum and cobalt nutrition on yield and quality of mungbean (Vigna radiata L.) in acidic soil of 20.06% for the two seasons, respectively. Table 7 reveals Northeast India. Indian Journal of Hill Farming 25(2): 22-26. the double and triple overlap between the study factors was Bhuiyan MMH, Rahman MM, Afroze F, Sutradhar GNC and Bhuiyan significant in this and both seasons. MSI 2008. Effect of phosphorus, molybdenum and rhizobium inoculation on growth and nodulation of mungbean. Journal of Soil Nature 2(2): 25-30. REFERENCES Black CA 1965. Methods of Soil analysis. Amer. Soc. of Agron. Inc. AOAC 1990. Official Methods of Analysis of the Association of official USA. Analysis Chemists, Washington, D. C: pp 1015. Boardman NK 1975. Trace elements in Photosynthesis. P: 199-212. Abu Dahi YM, Lahoud AM and Al-Kawaz GM 2001. Effect of foliar In: Trace elements in Soil – Plant – Animal System. Nicholas, E. feeding on maize yield and its components. Iraqi Journal of Soil D. , Egan , D. J. D. and Egan , A. R. Academic press . Science 1(1): 122-138. Hazili KH and Fajr JF 2016. Effect of molybdenum and boron on Abu Dahi YM and Al-Yunis MA 1988. Index of Plant Nutrition. Ministry some growth traits in the faba bean (Vicia faba L.). Basrah of Higher Education and Scientific Research - University of Journal of Agricultural Sciences 29(1): 201-213. Baghdad. Johnson BJ and Arris HB 1967. In fluence of plant poulation on yield AL-Dabbagh IJJ 2017. Effect of foliar feeding on boron and salicylic and other characteristics of Soy bean. Agronomy Journal 59: acid in the physiological, productive and quality characteristics 447-449. of two genotypes of the (Vignaradiata L.). M.Sc. Department of Kalaikandhan P, Jayareng PV, Sivasankar R and Mathivanan S Field Crops – collage of Agriculture - Anbar University. 2014. The effect of copper and on the morphological parmeters AL-Fahdawi AIAF 2016. Effect of spraying copper and potassium of Sesuvium pprtulacastrum L.Interntion Journal of Current levels in the growth and yield of the faba bean (Vicia faba L.). Research and Academic Review 2347-3215: 105-120. Master of Field Crops Department – Collage of Agriculture - Kandil Hala, Nadia Gad Magdi A and Abdelhamid T 2013. Effects of University of Anbar. different rates of phosphorus and molybdenum application on AL-Fahdawi AIH 2004. Effect of Potassium spraying and Phosphate two varieties common bean of (Phaseolus vulgaris L.). Journal Fertilizer in Some Growth, yield and Quality Characteristics of of Agriculture and Food Technology 3(3): 8-16. Several Mung bean genotypes. M.Sc. Collage of Agriculture _ Malik K, Kumar S, Arya A and Singh KP 2015. Effect of zinc, Anbar University. molybdenum and urea on chlorophyll and protein content of Ali HJ, Issa TA and Jadan HM 1990. Legumes crops. Ministry of mungbean (Vigna radiata L. Wilczek). International Journal of Higher Education and Scientific Research Baghdad University Plant Sciences 10(2): 152-157. Box 259. Mohammadi S and Nema I 2011. Response of the growth and yield of Ali NAS 2012. Fertilizers and their applications. Ministry of Higher some varieties of bread wheat (Triticum astivum L.) for copper Education and Scientific Research. University of Baghdad. foliar feeding. Anbar Journal of Agricultural Sciences 8(4) Special Issue: 417-431. AL-Khafaji AHAG 2015. Effect of addition of potassium and iron and zinc spray in some growth and yield characteristics of Mung Mohsen A, Dowidar S, Abo-Hamad S and Khalaf B 2013. Role of bean (Vigna radiata L.). Ph.D. Thesis. Field Crops Department, cyanobacteria in amelioration of toxic effects of copper in Collage of Agriculture, University of Baghdad. Trigonella Fenum gracum. Australian Journal of Crop Science 7(10): 1488-1493. Al-Mahmoudi MSH 2012. Effect of Levels of Nitrogen and Potassium Sadeghipour O 2008. Response of mung bean Varieties to different Fertilizer on the Growth and yield of Two genotype Mung bean sowing dates. Pakistan Journal of Biological Sciences 11(16): of (Vigna radiata L.). M.Sc., Field Crops Department, Collage of 2048-2050. Agriculture, Anbar University. Singh AK, Khan MA and Srivastava A 2014. Effect of boron and AL-Mashhadani AIA 2015. Effect of the date of agriculture and plant molybdenum application on seed yield of mungbean. Asian density in the growth and yield of Mung bean (Vigna radiata L.). Journal of Biological Sciences 9(2): 169-172. M.Sc. Field Crops Department – collage of Agriculture - University of Anbar. Tahir M, Alam S and Muhammad AM 2014. Effect of Molybdenum on Yield and quality of black gram (Vigna mungo L.). Pakistan Al-Rawi KM and Khalafallah AA 1980. Design and analysis of Journal of Life and Social Sciences 12(2): 101-105. agricultural experiments. Directorate of Dar Al Kutub for Printing and Publishing. University of Al Mosul.P.487. Togay Y, Togay N and Dogan Y 2008. Research on the effect of phosphorus and molybdenum applications on yield and yield Ara N, Rokonuzzaman M and Hasan M 2009. Effect of brady parameters in lentil. African Journal of Biotechnology 7: 1256- rhizobium and Azotobacter on growth and yield of mung bean 1260. varieties. Bangladesh Journal of Agricultural Research 7(1): 7- 13. Yusuf M, Fariduddin Q, Ahmad I and Ahmad A 2014. Brassinosteroid- mediated evaluation of antioxidant system and nitrogen Arnon DI 1950. Functional aspects of copper in plants. P: 89-114. In metabolism in two contrasting cultivars of (Vigna radiata L.) copper metabolism. The Jone Hopkins press, Baltimore. under different levels of nickel. Physiology and Molecular Ashwini GM 2005. Effect of Organic Nutrients and Plant Growth Biology of Plants DOI 10.1007/s12298-014-0259.

Received 08 October, 2019; Accepted 20 November, 2019 Manuscript Number: S-282 Indian Journal of Ecology (2019) 46 Special Issue (8): 209-213 NAAS Rating: 4.96

Evaluation of the Efficiency of Ultraviolet Radiation and Hot Peppermint Extract in the Protection of Some Biological Systems of Male Albino Mice from the Toxic Effects of Aflatoxin B1 and Fumonisin B1 Toxins

Ali F. Jubair and Sabah L. Alwan1 Department of Plant Protection, Agriculture College, Al-Muthanna University, Iraq 1Department of Plant Protection, Agriculture College, Kufa University, Iraq E-mail: [email protected]

Abstract: The study was carried out to determine the efficacy of ultraviolet radiation and hot peppermint extract in minimizing the toxic effects of aflatoxin B1 and fumonisin B1 in some biochemical and histological parameters in male mice. The animals were divided into 8 groups of 5 animals each and dosed with aflatoxin B1 and fumonisinB1 (500 mg kg-1 body weight each) for three weeks. The study showed that blood hemoglobin concentration decreased in the treatment of aflatoxin B1 toxin only. It was 7.10 g 100 ml-1 and significantly different from the control which was 10.70 g 100 ml-1. The treatment of FB1 + peppermint had a positive role in raising the blood hemoglobin level, reaching 13.17 g 100 ml- 1 compared with treatment of fumonisin B1 only, which was 8.27 g 100 ml-1. The study also found that there was a variation in the number of leukocytes in the blood of treated animals, where an increase in the number of leukocytes was observed for aflatoxin B1 and fumonisinB1 toxins, which reached 14.00 and 12.20 cells mm-3compared with the control of 4.83 cells mm-3. Preventive treatments (FB1 + peppermint, FB1 + uv-c, AFB1 + peppermint and AFB1 + uv-c) showed that the number of leukocytes decreased to a normal level in the blood of animals where it gave 4.93, 6.60, 7.30 and 6.27 cells mm-3, respectively. The results showed that the effect of aflatoxin B1 and fumonisin B1 was negative on the volume of PCV cells in the blood of animals (31.37 and 22.43%, respectively), compared to the control which gave 37.17%.While the treatment of just peppermint and other protective treatments (FB1 + peppermint, FB1 + uv-c, AFB1 + peppermint and AFB1 + uv-c) showed significant differences (41.00, 34.00, 31.30, 39.00 and 33.13%, respectively),with toxin coefficients AFB1 and FB1.The results of the histological study showed clear changes in liver tissue, namely acute cell decomposition and disappearance of the nucleus, i.e., the loss of body structure and decomposition in the cells of Kupfer. While the protective treatments, which used ultraviolet radiation and hot peppermint extract with aflatoxin B1 and fumonisinB1 toxins showed a significant improvement with some slight cell decomposition and haemorrhage.

Keywords: Aflatoxin B1, Fumonisin B1, Hemoglobin, Leukocytes, PCV, Albinomale mice

Mycotoxins are products of secondary metabolism, for the formation of mycotoxins (Sultuna, Hanif 2009, Reddy produced by a certain species of fungi (Kabak and Dobson et al 2010, Khatib and Khalesi 2011) as well as drought 2009, Chulze 2010, Zain 2011). This damage poses a health conditions that can strain plants and make them susceptible risk to humans and animals due to the formation of mycotoxins to contamination with the Aspergillus species (Robertson (Bryden 2007). The most common mycotoxins are aflatoxins 2005). Preventing contamination with mycotoxins and (a strong human carcinogen and classified as a carcinogen of preventing their entry into the food chain makes it difficult to the group A), which have received great attention from avoid in most developing countries despite the necessary specialists in many regions of the world as a result of their precautions (Markovic et al 2010). impact on human and animal health, as recorded in many Humans and animals are always exposed to mixtures health disasters due to exposure to these toxins. As per the rather than to individual compounds. Therefore, future risk American Drug and Drug Administration FDA (Food and Drug assessments should consider mixture toxicity data Administration), the allowed level of fumonisins (FB1, FB2 and (Heussner et al 2006). Three different effects were observed FB3) in foods should be less than 4 mg kg-1 in food and feeds when there were toxins together in food or feed, synergistic or consumed by humans and animals based on information of additive or antagonistic effects. The effect of synergy is that researches conducted in many animals (FDA, 2001). the effect of two toxins together is greater than the effect of Levels of fungi contamination and their toxins in their alone, the additional effects are the effect of the agricultural crops vary by geographic and environmental combined toxins equal to the sum of their effects both and variability, as developing countries are more susceptible to individually, and in the antagonistic effect, both toxins fungal poisoning than developed countries. Due to the high interfere with the impact of the each other and make them humidity and high temperatures that provide ideal conditions less toxic (Khatoon 2004). 210 Ali F. Jubair and Sabah L. Alwan

Different methods of decontamination of fungal toxins in shaker) for 6 hours for the purpose of mixing it and give more cereals and food have been proposed and maintained within space to extract the active substance in the sample. The the permissible limits, but one method cannot be successfully solution filtered by filter paper several times, and the applied to all fungal toxins (Kabak et al 2006). Many methods solutions prepared were sterilized using micro-filters have been used to prevent infection and contamination of (Millipore filters) with a diameter of0.22 μm and then stored in various foods with fungi producing fungal toxins, including the refrigerator at 4°C until use (Al Obeidi 2000). physical methods of treating fruits with hot water or exposing Animal division: 40 male albino mice were divided in eight them to various rays as well as the use of some groups of five animals each. preservatives, and plant extracts (Huwig et al 2001). Treatment of animals: The following treatments were applied to male albino mice (Table 1). MATERIAL AND METHODS The Dosing process lasted three weeks, during which Laboratory animals preparation: In this study, male time the clinical symptoms that could appear in the treated laboratory animals (Mus musculus) of the Balb/c strain were animals were observed. After the end of the experiment, the used, ranging in age from 12 to 14 weeks. Procured from the animals were sacrificed after being anesthetized with Iraqi Center for Research on Cancer and Medical Genetics / chloroform and explained by opening the abdominal cavity Baghdad, placed in plastic cages andmice were kept for a and withdrawing blood by heart stapling and placing the week for the purpose of adaptation to food and environmental blood drawn in test tubes containing anticoagulant (EDTA) conditions (23-28ºC and appropriate lighting). The animals for physiological blood tests. Parts of the liver of male rats were fed a concentrated pellet (Pellets) placed in the food were then taken and fixed in 10% formalin to study area in the breeding cages and given free food and water to histological changes. all mice. Study standards: The physiological parameters ofblood Preparation of aflatoxin B1 toxins and Fumonisin B1: were measured using a Japanese Hematology analyzer Aflatoxin B1 toxin was prepared from the isolation of A. (Sysmex Kx-21) in the blood bank in Muthanna province and parasiticus producing aflatoxin B1, while FumonisinB1 was the results were confirmed using another device (mindray secreted from F. proliferatum isolation by using thin layer CBC) of Chinese origin. chromatography (TLC) technique. Preparation of hot Histological study: Attended the tissue sections in the peppermint extract 15%. histopathology lab of Al-Hussein Teaching Hospital in Weighed 200g of peppermint leaves to prepare the Muthanna province following the method of Stevens and extract and washed under running tap water then distilled Bancroft (1982) which was as follows: water and left to dry, leaves cut into small pieces and air dried Dehydration: The samples were passed in upward away from sunlight, the ingredients were crushed and milled concentrations of ethyl alcohol (70, 80, 90, 95 and 100%) for with an electric grinder, to prepare 15% concentration adding 1.5-2 hours at each concentration to remove water from 15 g to 100 ml boiling distilled water in a 250 g sterile glass them. beaker and mixing well put on an electric vibrator(Flask Clearing: The samples were stirred with xylene twice for 1-

Table 1. Treatment of male albino mice Treatments Description of treatments Control Sound animals Treatment of aflatoxin B1 toxin Dosing animals of AflatoxinB1 (500 microgram kg-1 animal weight) Treatment of fumonisinB1 toxin Dosing animals of fumonisinB1 (500 microgram kg-1animal weight) Hot peppermint extract with a concentration 15% only Dosage of animals with hot peppermint extract at a rate of 1 ml kg-1 animal weight Treatment offumonisinB1 + hot peppermint extract at a Dosage of animals with hot peppermint extract at a concentration of 15% concentration 15% at a rate of (1 ml kg-1of animal weight) followed by dosing of animals with the fumonisin B1(500 microgram kg-1of animal weight) after 24 hours Treatment offumonisinB1 toxin exposed to ultraviolet radiation Dosage of animals with fumonisinB1 toxinexposed to UV-c wavelength 254 nm for 15 minutes at a rate of (500 microgram kg-1 animal weight) Treatment of aflatoxin B1 + hot peppermint extract with 15% Dosage of animals with hot peppermint extract at a concentration of 15% concentration at a rate of (1 ml kg-1animal weight) followed by dosing of animals with aflatoxin B1at a rate of (500 microgram kg-1 animal weight) after 24 hours Treatment of aflatoxin B1 exposed to ultraviolet radiation Dosage of animals with aflatoxin B1 exposed to UV-cwavelength 254 nm for 15 minutes at a rate of (500 microgram kg-1 animal weight) Protection of Albino Male Mice from Toxins by Ultraviolet Radiation and Hot Peppermint Extract 211

1.5 hours at a time to remove the solution from the tissue. including aflatoxin, stop the action of the enzyme Infiltration: The samples were covered with molten paraffin Hemoxygenase and that the action of this enzyme works to wax at 56-58°C by placing the samples twice and for 1-1.5 increase the breakdown of erythrocytes and convert them to hours each time. bilirubin, which is reflected on the concentration of Embedding: Samples were buried in special molds hemoglobin (Sakhara et al 2007). containing molten paraffin wax and left to harden. Rate of white blood cell counts (WBC): The results of table Sectioning: Tissue sections with thickness 5 micrometer 2 showed a variation in the number of leukocytes in the blood were cut using rotary microtome, the models were fixed to of treated animals, An increase in erythrocyte preparation glass slides using the Meyer's albumin adhesive. The slides was observed for aflatoxin B1 and fumonisin B1 treatments, were placed in the oven at a temperature of 56-58ºC reaching 14.00 and 12.20 cells mm-3 compared to the control vertically for 20 minutes to remove excess wax. treatment of 4.83 cells mm-3. On the other hand, Just Staining: The sections were dyed with the hematoxylin dye Peppermint and other protective treatments (FB1 + as the slides were passed in (xylene-100% absolute ethyl Peppermint, FB1 + uv-c, AFB1 + Peppermint and AFB1 + uv- alcohol - 95% alcohol - 90% -70% - hematoxylin dye) for two c) were reduced of white blood cells (5.50, 4.93, 6.60, 7.30 minutes each, the sections were washed with tap water, and 6.27 cells mm-3, respectively), compared with the toxin immersed in the eosin dye for 2 minutes and then washed treatment AFB1 and FB1. Reason for this is because some with tap water and passed in upward concentrations in ethyl mycotoxins stimulate the immune response in animals alcohol (70% -90% -95% -100%) and xylene for two minutes treated with these toxins, and that metabolic products of a per concentration. number of fungi cause an increase in the number of Mounting: Cover slip was put on the sections using leukocytes and anemia in laboratory animals This is because candabalsm. The tissue sections were then examined to these products stimulate the body's immune system by record histological changes. increasing the number of lymphocytes to try to neutralize foreign substances in the body (Peraica et al 1999). RESULTS AND DISCUSSION The increase in the number of leukocytes may be The amount of hemoglobin Hb (g 100 ml-1): Results in attributed to the fact that the fungi have the ability to secrete Table2 showed decrease in hemoglobin concentration of metabolic products that induce excessive secretion of AFB1 only was reached 7.10 g 100 ml-1 significantly different immune response factors, including T.N.F( Tumor necrosis from the control treatment which was 10.70 g 100 ml-1, factor), Interleukin-6 (IL-6)and Cytokinase especially the treatment of FB1 + Peppermint had a positive role in cytokines responsible for the occurrence of primary increasing of hemoglobin was reached 13.17 g 100 ml-1 inflammation and the cytokines responsible for regulating compared with treatment of FB1 only which was 8.27 g 100 immunity (Kemacki et al 1998). Excessive production of ml-1. The treatment of Peppermint only was not significantly these ingredients is likely to be an important reason for different with control treatment that noticed hemoglobin increasing the number of sour white blood cells that concentration in animals blood was normal. On the other neutralize or remove toxic substances from the body. hand, there is no significant differences were observed Packed cell volume rate: Effect of aflatoxin and fumonisin between the treatment of AFB1 which reduced the negatively on the volume of compacted cells (PCV) in the hemoglobin concentration and treatments (AFB1 + blood of treated animals AFB1 and FB1 showed a significant Peppermint and AFB1 + uv-c), which were (9.63 and 8.83 g decrease in the size of the compacted cells (31.37 and 100 ml-1 respectively). 22.43%), respectively. AFB1 and FB1 showed a significant The decrease in hemoglobin concentration in the decrease in the size of the compacted cells (31.37 and treatments of AFB1 and FB1 may be due to oxidative damage 22.43%), respectively, T1, which gave 37.17%, while Just of the activity of mycotoxins, which increased the rate of Peppermint and other protective treatments (FB1+ oxygen oxidation in blood hemoglobin as well as hemolysis Peppermint, FB1+uv-c, AFB1+Peppermint and AFB1+uv-c) (Abdel-wahab et al 2014). significant differences were observed it reached (41.00, The reason for the low hemoglobin belong to presence 34.00, 31.30, 39.00 and 33.13%), respectively, compared to of aflatoxin in blood leads to pathological changes in various the toxin treatments AFB1 and FB1 (Table 2). The reason for parts of the body, including the bone marrow which leads to the significant decrease in blood accumulation rates can be fatty degeneration and this process reduces bone marrow attributed to the metabolic products produced by these fungi, tissue and these tissues is responsible for the formation of including mycotoxins, Which negatively affects the sources erythrocytes thus, hemoglobin, or sometimes some toxins, of erythrocyte production in the bone marrow, since there is a 212 Ali F. Jubair and Sabah L. Alwan

and haemorrhage (Fig. 1b). These pathological changes may be due to the role of aflatoxinin generating free radicals, which oxidize the unsaturated fats that make up liver walls, leading to cell death and liver tissue damage (Ross et al 1992). The mechanism of the effect of aflatoxins on organs is not yet understood and its toxicity is reported to be generated by the formation of free radicals. These compounds attack cells and attack membranes, eventually leading to a decrease in cell function with cellular degeneration (Berg et al 2004). The role of peppermint extract in reducing the effect of mycotoxins is due to the fact that it contains phenolic compounds as it has a role in the activation and detoxification processes (Chattopadhyay and Kumar 2007). The liver is affected by mycotoxins due to the effect of toxins on the cell membrane and shrinkage, thereby increasing permeability and extracellular enzymes (Coles and Cox 1981). Also, the cause of bleeding in the liver tissue is due to hemorrhage of the blood vessels, and the departure of blood-forming cells into the tissues maturity process and this causes inflammation in the tissues (Macsween and Whaley 1992), While the other protective treatments, which used ultraviolet radiation and hot peppermint extract with toxins of aflatoxin

Fig. 1a. Effect of ultraviolet rays and hot peppermint extract in the liver of male mice treated with aflatoxin B1 and fumonisin B1 direct correlation between the total enumeration of erythrocytes and the values of each blood stack through reduction of the number of red blood cells produced in the bone marrow, which in turn is reflected in the values of blood accumulation in the blood They are below normal levels (Groopman et al 2003). In a study by Sakhare et al (2007), aflatoxins had an effect on the physiological and biochemical parameters of treated animals, where the numbers of white blood cells were higher and the packed cell volume decreased compared to the control treatment. Histological effects: The results of microscopic examination of the liver tissue sections of male mice that were dosed with aflatoxin B1 and fumonisin B1 toxins showed significant changes in liver tissue represented by acute cell decomposition and disappearance of the nucleus. Infiltration was observed in liver cells and the occurrence of the phenomenon of vacuolation and apparent hemorrhage and clear blood full of sinusoidal blood (Fig. 1a) When compared with the control treatment, the tissue appears normal with the presence of nuclei, the central vein, the cover cells and the presence of blood sinusoid .While the male mice dosed with spicy peppermint extract with aflatoxin B1 and also spicy peppermint extract with fumonisin B1 showed Fig. 1b. Effect of ultraviolet rays and hot peppermint extract in the liver of male significant improvement with some slight cell decomposition mice treated with aflatoxin B1 and fumonisin B1 Protection of Albino Male Mice from Toxins by Ultraviolet Radiation and Hot Peppermint Extract 213

Table 2. Effect of ultraviolet rays and hot peppermint extract Coles RJ and Cox RH 1981. Toxic fungal metabolites. Academic on physiological blood standards of male mice press, New York, pp: 233-256. treated with aflatoxin B1 toxins and fumonisin B1 Effiuvwevwere BJO 2000. Microbial Spoilage Agents of Tropical and Treatment Physiological blood standards Assorted fruits and Vegetables. Port Harcourt: Paragraphics Publishing Company: p. 1‒39. The amount of Rate of white Packed FDA 2001. U.S. food and drag administration center for applied hemoglobin Hb blood cell counts cell volume nutrition center for veterinary medicine. ground in support of (g 100 ml-1) (WBC) (cells rate (%) fumonisin level in corn and corn products. Intended for human mm-3) ×1 03 consumption. Control 10.7 4.83 37.17 Groopman AM, Stevan MA and Cole MN 2003. A study about effect of Aspergillus flavus and Aspergillus fumigates AFB1 7.1 14 31.37 inchiken. Medicine 45(9): 5-2. FB1 8.27 10.2 22.43 Heussner AH, Dietrich DR and Brien EO 2006. In vitro investigation of individual and combined cytotoxic effects of ochratoxin A and Just Peppermint 12.3 5.5 41 other selected mycotoxins on renal cells. Journal of Toxicology FB1+ Peppermint 13.17 4.93 34 20(3): 322- 341. Huwig A, Freimund S, Käppeli O and DutlerH 2001. FB1+uv-c 8.7 6.6 31.3 Mycotoxindetoxication of animal feed by different adsorbents. AFB1+Peppermint 9.63 7.3 39 Toxicology Letters 122: 179-188. Kabak B, Dobsonand AD and VarI 2006. Strategies to prevent AFB1+uv-c 8.83 6.27 33.13 mycotoxin contamination of food and animal feed: A review. LSD (p=0.05) 3.002 1.843 1.761 Critical Reviews in Food Science and Nutrition 46: 593-619. Kabak B and Dobson AD 2009. Biological strategies to counteract Values are means of three replicates. Differences among treatments were the effects of mycotoxins. Journal of Food Production 72: 2006- calculated based on least significant difference LSD (P≤0.05) 2016. Kemacki A, David M and Broti M 1998. Histopathology, O. Chuchil B1 and fumonisin B1 showed a significant improvement (Fig. Living stone. New York, pp: 12-14. 1b). This result was comparable to Oujda Khalidi (2010) and Khalesi M and Khatib N 2011. The effects of different ecophysiological factors on ochratoxin a production. Saadi (2014). Environmental Toxicology and Pharmacology 39: 113-121. Khatoon S 2004. Interactions between mycotoxins. Romer Labs. CONCLUSIONS Pakistan, Rawalpindi . The study concludes that the use of ultraviolet radiation Kumar A and Chattopadhyay S 2007. DNA damage protecting activity and antioxidant potential of pudina extract. Food and hot peppermint extract reduced the toxic effects of Chemistry 100(4): 1377-1384. aflatoxin B1 and fumonisinB1 in some biochemical and MacSween RWM and Whaley K 1992. Murices development and histological parameters in male albino mice. Preventive survey. Foodaddit. Cutam 16:253-265. Markovic R, Sefer D, Radulovic S and Speranda M 2010. Prisustvo I treatments, using UV-c and hot peppermint extract van be znacajmikotoksina u hranizasninje, veterinarskiglasnik, used to reduce the toxicity of AFB1 and FB1 toxins, in male Fakultetveterinarskemedicine. Beogard 64: 83-92. albino mice. Peraica M, Radic B, Lucie A and Pavlovic M 1999. Toxic effect of mycotoxins in human. Bulletin of The World Health Organization. REFERENCES Reddy KRN, Salleh B, Abbas HK, Abel CA and Shier WT 2010. An Abdel-Wahhab KG, Mannaa FA and Abdel-Wahhab MA 2014. Panax overview of mycotoxin contamination of food and its implication ginseng C.A. Meyer Extract protects rat erythrocyte from the for human health. Toxin Reviews 29: 3-26. oxidative damage induced by the synergistic effect of Robertson A 2005. Risk of aflatoxin contamination increases with hot subchronic treatment with aflatoxin B1 and fumonisin. British and dry growing conditions.Integrated Crop Management, IC Journal of Medicine and Medical Research 4(9):1883-1901. 494: 185-186. Al-Khaldi, Bahija Obais Hammoud 2010. And to detect some of its Ross R, Yuan JM and Yu M 1992. Urinary aflatoxin biomarkers and functional and histopathological effects in male white rat. Ph.D. risk of hepatocellular carcinoma. Lancet. 339: 943-946. Thesis, Faculty of Science - University of Kufa. Saadi Zulfiqar Abdul Sattar Jabbar 2014. Integrated resistance to Al-Obeidi, Lamia Abdul-Razzaq 2000. Effect of pinch extract on A.niger and A. parasiticus producing mycotoxins contaminated growth of some pathogenic microorganisms. Master Thesis. with apples and pears. Master Thesis, Plant Protection College of Science. Baghdad University, Iraq Department, Faculty of Agriculture. Karbala University, p. 117. Bancroft JD and Stevens A 1982. Theory and Practice of histological Sakhara PS, Haren SDDR, Kalorey D, Warke SR, Bhandarkar AG technique. Churchill Living stone, New York., pp:1 17. and Kurkure NV 2007. Effect of toxiroakpolyherbal feed Berg D, Youdim MB and Riederer P 2004."Redox imbalanc". Cell and supplement during induced aflatoxicosis, Ochratoxicosis and Tissue Research 318: 201-213 combined mycotoxicosis in broilers. Veterinary Archive 77: 129- 146. Bryden WL 2007. Mycotoxins in the food chain: human health implications. Asia Pacific Journal of Clinical Nutrition 16: 95-101. Sultana N and Hanif NQ 2009. Mycotoxin contamination in cattle feed and feed ingredients. Pakistan Veterinary Journal 29: 211- Chulze SN 2010. Strategies to reduce mycotoxin levels in maize 213. during storage: A review food additives and contaminations part a, chemistry analysis. Control, Exposure and Risk Assessment Zain ME 2011. Impact of mycotoxins on humans and animals. 27: 651-657. Journal of Saudi Chemical Society 15: 129-144.

Received 22 September, 2019; Accepted 04 November, 2019 Manuscript Number: S-283 Indian Journal of Ecology (2019) 46 Special Issue (8): 214-218 NAAS Rating: 4.96

Effect of Ozone Gas in Milk Treatment on Chemical and Microbial Properties of Soft Cheese

Azhar J. ALmosowy and Zenib H. ALmosowy1 Department of Food Science, College of Agricultural Engineering Sciences, University of Karbala, Iraq 1Department of Animal Production, College of Agriculture, University of Karbala, Iraq E-mail: [email protected]

Abstract: Ozone gas was used in the treatment of raw milk for the purpose of use in the manufacture of soft cheese, where three treatments of raw milk were presented to the weight gas at a concentration of 0.5 g hr-1 and for 5 minutes for the first treatment of milk and 15 minutes for the second treatment and 30 minutes for the third treatment with the preparation of treatment Milk was processed at a temperature of 63°C / 30 minutes for adoption as a control treatment. Soft cheese was manufactured from these treatments and the four treatments were kept in a refrigerator at a temperature of 7±2°C. The % of acidity and pH were estimated. Total bacterial numbers and coliform bacteria were estimated. And cluster Golden psychrotrophic and analyst fat and analyst of proteins as well as the preparation of yeast and molds, and the results showed that exposed to ozone treatments were good and close to the control treatment for preservation, since the rise in the rate of acidity in the cheese transactions was less with the increase in the period of exposure to ozone. The results of microbiological tests indicated that ozone had a clear effect in reducing the growth and inhibition of microorganisms, and this effect increased with increasing exposure period up to 30 minutes, and continued its effect up to 5 days of conservation at a temperature of 7 ± 2°C.As for the best treatment of ozone gas recommended to be adopted for the preservation of milk with ozone was the treatment in which milk was presented to the concentration of 0.5 g hr-1 and for 30 minutes is better than the treatment in which milk was presented to the gas for 15 minutes. Much better treatment is offered for ozone for 5 minutes.

Keywords: Ozone, Milk, Microbiology, Cheese

Ozone (O3) is the second strongest common oxidizing less expensive than other methods. Researchers in the field agent after fluorine (Guzel Seydim et al 2004, Hu and Xia and of ozone uses that ozone can be generated on demand on 2017) and leaves no harmful chemical residue behind. It was site, and unlike traditional chemical disinfectants, it requires discovered by Sch € onbein in 1839 and was first used neither transport nor storage. The use of ozone requires less commercially for drinking water treatment in France more energy to operate than other methods. Moreover, the costs of than a century ago (Hill and Rice 1982, Rubin 2001). Ozone, operating ozone-depleting systems are low because they a bluish gas with a pungent odor, is a highly variable and consume a limited amount of electricity. unstable oxygen (O2) gas that has a high oxidation potential In a study conducted in April 2016, scientists studied the of -2.07 V, which carries highly antimicrobial properties. It is impact of ozone use on sanitation practices in dairy farms, produced by the interaction of free oxygen radicals with O2 and concluded that ozone treatment is a cost-effective and molecules. There are many ways of generating ozone, environmentally friendly food treatment technology including electron discharge, ultraviolet radiation, thermal, (Vargaand Sigeti 2016), although it is an antiseptic. Highly chemical, electrical and chemical methods (Khadre et al effective, however, ozone leaves no chemical residue on any 2001, Patil and Bourke 2012). surfaces in contact with food or food because it is rapidly Ozone treatment is a cost-effective and environmentally transferred to non-toxic products, thereby reducing friendly food processing technology. It has been successfully environmental impacts and company costs (Cullen and used to remove residues of milk and biofilm-forming bacteria Norton 2012, O 'donnell et al 2012). from stainless steel surfaces. Ozone can also be used to Heating processes are commonly used to treat milk to sterilize a wide variety of dairy products. Ozone is liquid milk, make it fit for consumption, but this may negatively affect its powdered milk and cheese. Ozone has been shown to inhibit nutritional value and sensory properties. On the other hand, the growth of mold on cheese and disrupt its growth in air in scientists found that ozone did not adversely affect the cheese ripening and storage chambers. Ozone treatment nutritional value or taste of liquid milk studied, and other uses has been found to be a promising way to reduce of ozone in farms Dairy use at very low concentrations, in the contaminants in dairy waste water (Ozone Systems 2014, barn atmosphere to destroy airborne pathogens and Varga and Sigeti 2016). eliminate the smell of manure in the barn, Rojek et al (1995) The use of ozone in the treatment of dairy products is used compressed ozone (5-35 mg l-1 for 5-25 minutes) to Ozone Gas Effect on Milk 215 preserve skim milk by Reduce their microbial concentrations APHA (1978) was used to estimate the total count of bacteria, Sander (1985) obtained a patent for prescribing a method for total coliform bacteria, count of yeasts and molds, and the treating liquids using low concentrations of ozone, such as count of Staphylococcus aureus. Staphylococcus Ssp. milk and liquid dairy products, thereby reducing their Blatimore Biological Laboratory (1973) used salt agar potential degradation, and it was found that the use of ozone Mannitol and incubated dishes at 32°C for 48 hours, reduces the number of psychrotrophs by more than 99%. depending on the count of golden colonies surrounded by a Sheelamary and Muthukumar (2011) completely removed yellow halo representing colonies of Staphylococcus sp Listeria monocytogenes from both raw milk samples and Proteolytic bacteria and Lipolytic bacteria were used in branded samples using ozone. Therefore, the main objective Harrigan and McCence (1976), while the count of bacteria of this study is to know the impact or role of weight gas in growing in the psychrophilic bacteria was calculated using prolonging the period of raw milk in the conditions of Iraq, the method described in APHA (1978), using nutrient agar. especially in hot climates. The lap temperature is 7°C for 10 days. Chemical tests: The pH and titration acidity according to the MATERIAL AND METHODS method described in AOAC (2000), was estimated as the The raw milk supplied by the dairy was used by the total nitrogen count according to the method described by College of Agricultural Engineering Sciences of the Joslyn (1970), the soluble nitrogen and non-protein nitrogen University of Baghdad, based in Abu Ghraib. The ozone (NPN) according to the method described in Ling (1956), the system was used for the purpose of providing the ozone gas Acid Degree Value (ADV) was estimated in the Bureau of required for the milk treatment. The milk was divided into four Dairy Industry (BDI) cited by Deeth and Fitz-Gerald 1976). sections, each representing a treatment. The first was Statistical Analysis: The Statistical Analysis System (SAS) treated with ozone for 5 minutes and the second treatment (2012). was 15 minutes while the third treatment was 30 minutes, and the fourth treatment was left untreated for adoption as a RESULTS AND DISCUSSION control treatment. Raw milk samples were treated with ozone at a Manufacturing soft cheese: Four soft cheese treatments concentration of 0.5 g hr-1 for different times of 5, 15 and 30 were made from milk treated with ozone. The first treatment minutes to determine the effect of these treatments on the included the use of 5 kg of raw milk treated with ozone 0.5 g h- effectiveness of microorganisms that are naturally present in 1 for 5 minutes (M1). The second treatment used the same these models. The concentration of 0.5 g ozone hr-1 for 5, 15 amount of milk and exposed to the same dose of ozone but and 30 minutes was selected. for 15 minutes (M2). In the third treatment, the duration of After fixing the concentration of ozone used, the test was exposure to ozone was 30 minutes (M3). The milk was then conducted by exposing the raw milk treatments to this heated to a temperature of 34°C. And the restraint and move concentration for different times according to the and then summon the whey and salting 2% salt, sodium manufacturing conditions in the country and also taking into chloride and then fill the clot in the templates and save them account that the time of exposure to gas is not enough for the in the refrigerator any second day, as extracted cheese emergence of oxidative action, the results showed that the molds and divided into several parts preserved at pH ranged between 0.18 - 0.19% in the four treatments. temperature 7 ± 2°Cbags of polyethylene for the purpose of Immediately after processing, when the treatments were 7 days) there was a gradual evolution in/ conducting chemical and sensory tests and micro biological refrigerated (7±°C٢ them. The treatment (M4) milk pasteurization to a the pH of all treatments with the duration of preservation and temperature of 63°C/30 minutes and then cooled to a that the least evolution in acidity was in the treatment of temperature of 34°C and completed the necessary steps for cheese that introduced milk from it to ozone for 30 minutes, processing as in the three treatments ozone treatment. The highest development in the proportion of acidity Vka In Microbiological tests: All agricultural media were prepared the first treatment, which was introduced to ozone gas for in the laboratory and according to the instructions of the only 5 minutes (Table 1). These results can be explained that company equipped, sterilized in autoclave at a temperature ozone gas has an effect on the work of bacteria that ferment of 121ºC and pressure 121 pounds in-2 for 15 minutes, used lactose, which leads to reduce the production of lactic acid, Peptone water for the decimation of the decimals of bacterial which in turn is reflected in low values pH. farms and attended to dissolve 1 g of Peptone and 1000 ml Proteolysis: Table 2 shows increase in the ratio of dissolved distilled water. In a 9 ml test tube per tube and sterilize at an nitrogen to total nitrogen in the three treatments of cheese. It autoclave at 121°C for 15 minutes, the method described in is noted that the ratio of dissolved nitrogen to total nitrogen 216 Azhar J. ALmosowy and Zenib H. ALmosowy was higher especially in the final stages in the treatment M1 source as well as the decomposition of fat by Lipases compared to the treatments M1 and M2. The same table also enzymes analyst fat produced by bacteria and other initiator shows increase in NPN ratios in three treatments. This is due and lactose fermentation mainly in charge of the basic to the properties of the ozone against microorganisms composition of acetic acid and propionic. The results also especially protein-neutral bacteria. show that the greater the time of exposure to ozone, the lower Lipolysis: ​​Table 3 shows that the values ​​of the ADV of the the amount of development in ADV values. M2, also note soft cheese treated with the ozone ranged from 0.71 to 0.80 from the table that there are significant differences in the mm 100 g-1 lipid, indicating that there were no significant values of ADV between treatments treated with ozone and differences between them. This was confirmed by the results control treatment at the end of the conservation period, ie at of the statistical analysis. After the storage of the treatments the age of 5 days, but no such differences were observed at 7±2°C for 5 days, the ADV. of all the treatments increased between the four factors during the treasury ages 0 and 3 with the age of storage. This increase is due to the lipid days. degradation factors caused by the lipid-producing enzymes Microbiological tests: The results also show that the and others. With what Abdle Baky et al (1982) reported ADV greater the time of exposure to ozone, the lower the amount of fat increases the progress of a period of storage where the of development in ADV values. M2, also note from the table that there are significant differences in the values ​​of ADV between treatments treated with ozone and control treatment Table 1. Results of pH and acidity tests (%) for soft cheese at the end of the conservation period, ie at the age of 5 days, treatments treated at 0.5 g ozone h-1 for different but no such differences were observed between the four times during the preservation period at 7±2°C and 5 days factors during the treasury ages 0 and 3 days. Test Storage Treatments While the results of the microbiological tests conducted period (day) for milk treatments treated with weight as shown in Tables 2 M1 M2 M3 M4 and 3 show that ozone has a clear effect in the inhibition of pH 0 6.6 6.5 6.6 6.6 different microorganisms addressed in this study, as it is 1 6.5 6.4 6.6 6.6 clearly noted the development of the total numbers of 3 6.2 6.2 6.5 6.4 bacteria as well as the numbers of bacteria, yeasts, molds 5 5.7 6 6.3 6.1 and Staphylococcus aureus. As well as the number of cold- loving bacteria and lipophilic and protein-decomposing Acidity (%) 0 0.18 0.19 0.18 0.18 bacteria in the treatment of cheese M1, which was the 1 0.2 0.2 0.19 0.18 treatment of milk to ozone for the least period, which was 5 3 0.24 0.24 0.21 0.19 minutes, and the evolution of these numbers increased with 5 0.28 0.28 0.23 0.21 the preservation period despite the cooling conditions These treatments were then saved, and with an increase of 15 and Table 2. Percentage of soluble nitrogen and non - protein 30 minutes of exposure to ozone gas, colon bacteria, as well nitrogen for soft cheese treatments treated at 0.5 g as molds and yeasts, were completely eliminated in M2 and ozone h-1 for different times during the preservation period at 7±2°C and 5 days M3 treatments and no new growth was observed within 7 Treatment Cheese NPN/ TN NPN (%) SN/ TN SN (%) days of refrigerated preservation, indicating the effective age (day) (%) (%) effect of ozone. In reducing the growth and inhibition of the M1 0 14.21 0.56 6.67 0.263 3 15.44 0.63 18.67 0.762 Table 3. Acid degree value for soft cheese treatments treated at 0.5 g ozone h-1 for different times during the 5 20 0.84 20.4 0.857 preservation period at 7±2°C and 5 days M2 0 15.15 0.57 7.47 0.281 Treatment ADV 3 16.49 0.63 20.44 0.781 0 3 5 LSD 5 20.05 0.77 25.67 0.986 M1 0.71 1.87 3.28 * 0.703 M3 0 19.83 0.72 7.46 0.271 M2 0.8 1.59 3.01 0.793 * 3 23.77 0.87 23.41 0.857 M3 0.71 1.49 2.79 0.694 * 5 25 0.94 26.96 1.014 M4 0.7 1.19 1.5 0.844 * LSD 6.31 * 3.783 * 0.338 * 6.31 * LSD 0.398 NS 0.623 * 0.856 * - P< 0.05 NS, (P<0.05)* Ozone Gas Effect on Milk 217 activity of many types of microorganisms in the treatments Table 5. Results of tests for protolytic and lipolytic bacteria for -1 exposed to it, which makes it close to the treatment of control soft cheese treatments treated at 0.5g ozone h and different exposure times during the preservation in which milk was treated for pasteurization. period at 7± 2°C for 5 days Previous studies have shown that the minimum ozone Treatments Count of bacteria (cfu gm-1) concentration to limit microbial growth is 1 ppm, while other Psychrophilic Protolytic Lipolytic studies indicate that values ​​between 20 ppm and 40 ppm are M1 0 6× 102 1.2× 102 2× 102 effective (Saleh et al 2009). Dyas et al (1983) found that 1 1 77× 102 1.3× 103 4.1× 102 ppm of ozone was sufficient to reduce the numbers of most 2 3 2 species tested by more than 95%. 3 92× 10 5× 10 8.6× 10 3 3 4 To identify the role of lipolytic and proteolytic bacteria 5 98× 10 8× 10 2× 10 and follow the development in the growth of cold-loving M2 0 5× 101 9× 102 5× 101 bacteria, which also have a role in the production of 1 7× 102 7× 102 2× 102 lipoproteins and proteins in milk and milk products (Kubaisi, 3 73× 102 32× 102 5× 102 2001). Refrigerated, the results of these tests showed that 5 5× 103 1.2× 103 25× 102 soft cheese that was not treated with ozone gas, which is 2 2 M3 0 Nil 3.0× 10 5 × 10 made from pasteurized milk, the numbers of these bacteria 1 Nil 9.0× 102 1.3× 103 ranged in numbers 5 × 210, 3 × 210 and 7 × 210 m ml-1, 3 Nil 12× 102 32× 103 respectively after manufacturing, but As the preservation 5 Nil 8.9× 102 50× 103 period progressed, a clear development was observed The 1 1 development of these three bacteria continued to the end of M4 0 Nil 9.1× 10 5.1× 10 2 1 the preservation period of 7 days, while we find that the 1 Nil 3.2× 10 8× 10 development in the preparation of these three types of 3 Nil 9.3× 101 21× 101 bacteria was more in cheese treatments that were treated 5 Nil 9.0×102 7×102 with ozone during the period of preservation, as the results of these tests show that the greater the duration of milk exposure The number of lipophilic and proteolytic bacteria Table 4. The results of microbiological tests for soft cheese treatments treated at 0.5 g ozone h-1 and different decreased further, with the total elimination of cryophilic exposure times during the preservation period at bacteria in treatments that were shown for 15 and 30 minutes 7±2 °C for 5 days of ozone gas. Treatments Microbiological tests (cfu/gm) These findings are consistent with what many Staphylococcus Mould & Coliform Total researchers have pointed out about the ozone inhibiting Sp. yeast count action against many microorganisms, especially pathogens. bacteria This gas reacts with the double binding of fatty acids in the M1 0 4 × 101 Nil Nil 41× 105 wall of Cram-negative cells (GuzelSeydim et al 2004), as well 1 6 1 12× 10 Nil Nil 72× 10 as the effect of gas in carbohydrates and proteins in the wall 2 1 8 3 19× 10 4 × 10 Nil 21× 10 of Cram-positive and negative bacteria in different 5 32× 103 4 × 101 Nil 37×108 proportions. On breaking a Nuclear interaction with them M2 0 Nil Nil Nil 14× 103 when acids, and when the study of the impact of ozone in the 1 Nil Nil Nil 23× 103 nuclear acids in bacteria E. coli, it was noted that the Pthoia 3 Nil Nil Nil 78× 105 DNA plasmid from the circular shape to open the closed form,

8 leading to the loss of his job. In a study by Cavalcante et al 5 Nil Nil Nil 92× 10 (2013) the effectiveness of inactivation of microbial activity in M3 0 Nil Nil Nil 2× 103 raw milk was assessed by ozone treatment at 1.5 mg l-1 for 15 1 Nil Nil Nil 76× 103 min to reduce the numbers of bacteria and fungi by up to a 4 3 Nil Nil Nil 24×10 logarithmic cycle. 5 Nil Nil Nil 95× 104 M4 0 Nil Nil Nil 4× 103 REFERENCES 1 Nil Nil Nil 8.9× 103 Abdel Baky AA, El Fak AM,Rabia AM and El-Neshewy AA 1982.

3 Cheese slurry in the acceleration of Cephalotyre “Ras” 3 Nil Nil Nil 23× 10 cheese. Ripening Food Port 45: 894-897. 5 Nil Nil Nil 7.8×104 American Public Health Association (APHA) 1978. Standard 218 Azhar J. ALmosowy and Zenib H. ALmosowy

Methods For The Examination of Dairy. 14th Ed. E.H. Marth, Chapman and Hall Ltd., London. Washington, D.C Khadre MA, Yousef AE and Kim JG 2001. Microbiological aspectsof Association of Official Analytical Chemists International 2000. ozone applications in food: a review. Journal of Food Science Official Methods of Analysis. 17th ed. A.O.A.C. Arlington, VA. 66:1242-1252. Blatimore Biological Laboratory (BBL) 1973. Manual of products and Kubaisi NA 2001.The use of lipase-backed mortar in the ripening of laboratory procedures. 5th ed. Division of Becton, Dickinson oshary-like cheese. Master Thesis, College of Agriculture, and company. University of Baghdad Cavalcante MA, Leite Junior BRC, Tribst AAL and Cristianini M 2013. O'Donnell C, Tiwari BK, Cullen PJ and Rice RG 2012. Status and Improvement of the raw milk microbiological quality by ozone trends of ozone in food processing. In Ozone in Food treatment. International Food Research Journal 20: 2017- Processing, p. 1-6. O'Donnell C, Tiwari B K, Cullen P J and Rice 2021. R G, eds. Oxford: Wiley-Blackwell. Cullen PJ and Norton T 2012. Ozone sanitisation in the food industry. Ozone Systems 2014. Ozone benefits for dairy farmers. [Internet In Ozone in Food Processing, pp 163–176. O'Donnell C, Tiwari document URL http://www.puricare.co.za/UserFiles/Ozone% BK, Cullen PJ and Rice RG, eds. Oxford: Wiley-Blackwell. 20Benefits% Deeth HC and Fitz-Gerald CH 1976. Lipolysis in dairy products: A Patil S and Bourke P 2012. Ozone processing of fluid foods. In Nove review. Australian Journal of Dairy Technology 31: 53-64. Thermal and Non-Thermal Technologies for Fluid Foods, pp Dyas A, Bj. Boughton and Das BC 1983. Ozone killing action against 225-261. bacterial and fungal species; microbiological testing of a Rojek U, Hill AR and Griffiths M 1995. Preservation of milk by domestic ozone generator. Journal of Clinical Pathology 1102- hyperbaric ozone processing. Journal of Dairy Science 1104. 78(Suppl. 1): 125. Guzel-Seydim ZB, Greene AK and Seydim AC 2004. Use of ozone in Rubin MB 2001. The history of ozone: the Sch€onbein period, 1839- the food industry. LWT - Food Science and Technology 37: 453- 1868. Bulletin for the History of Chemistry 26: 40-56. 460. Saleh DS, Habib KA and Razouqi NS 2009. Study of the effect of Harrigan WF and McCance ME 1976. Laboratory method in food and different concentrations of ozone dissolved in water in two dairy microbiology. Acadmic press INC. New York. U.S.A. types of pathogenic bacteria and two types of pathogenic Hill AG and Rice RG 1982. Historical background, properties and fungi of the skin in vivo. The Iraqi Journal of Science 50(3): applications. In Handbook of Ozone Technology and 303-310. Applications Sander M 1985. Process for the gentle ozone treatment of liquids, Hu L and Xia Z 2017. Application of ozone micro-nano-bubbles to such as fruit juices, milk, liquid milk products, wine, oils, liquid groundwater remediation, J. of Hazardous Materials, 304. medicaments, blood and/or similar products (In German). John S, Novak JT C and Yuan 2007. Chapter 10. The Ozonation Patent No. DE 3325568A1. Concept: Advantages of Ozone Treatment and Commercial Sheelamary M and Muthukumar M 2011. Effectiveness of ozone in Developments. Advances in Thermal and Non-Thermal Food inactivating Listeria monocytogenes from milk samples. World Preservation. (eds G. Tewari and V. K. Juneja), Blackwell Journal of Young Researchers 1: 40-44. Publishing, Ames, Iowa, USA. rd Varga L and Szigeti J 2016. Use of one in the dairy industry: A review. Ling ER 1956. A Textbook of Dairy Chemistry. Vol. 2.practical. 3 Ed. International Journal of Dairy Technology 69: 157-168.

Received 12 September, 2019; Accepted 04 November, 2019 Manuscript Number: S-284 Indian Journal of Ecology (2019) 46 Special Issue (8): 219-223 NAAS Rating: 4.96

Negative Effects on Some Internal Organs of Rats Infected by Salmonella typhimurium

Fatima Aziz Mahdi Al-badry Biology Department, College of Education for Pure Sciences, University of Thi-Qar, Iraq E-mail: [email protected] .iq

Abstract: The present study was conducted to detect the harmful effects of experimentally infection of Salmonella typhimurium in some organs of laboratory female rats. Thirty two of rats were used in the current study which divided into four groups, the first group administrated normal saline considered as control group. The second, third and fourth groups were infected experimentally by Salmonella typhimurium at dose (1*108) for periods (one, seven and fourteen days, respectively). The current results found many histopathological changes in liver and kidney of all treated groups with bacteria. These histological changes in liver were included congestion, inflammation clearly, enlargement of central vein and sinusoids, hypertrophy of hepatocytes. Also, some histological damages were observed in kidney of infected groups like hemorrhage, shrinkage, absence and death of glomeruli, enlargement of Bowman's space, infiltration of inflammatory cells and congestion.

Keywords: Salmonella typhimurium, Experimental infection, Histopathological changes

Salmonellosis is one of a serious zoonotic diseases First group (Control group): These animals were caused by Salmonella spp. Salmonella is a gram negative administrated normal saline. bacteria cause gastroenteritis access to typhoid fever, this Second group (Infected group): These animals were disease occur by virulence factors injection of it into cells of infected orally by bacteria (Salmonella typhimurium) at dose host by a type III protein secretion system (Woc-Colbum and (1* 108) for one day according to method of Takeuchi (1966). Bobak 2009), 2500 serotypes were isolated from different Third group (Infected group): These animals were infected hosts which related to human infections (Callaway et al 2008). orally by bacteria (Salmonella typhimurium) at dose (1* 108) Also, Salmonella is responsible of infecting other animals as for one week according to method of Takeuchi (1966). birds, cattle and pigs, Non-typhoidal cases infected annually Fourth group (Infected group): These animals were by Salmonella were estimated about 1.4 million in over 1000 infected orally by bacteria (Salmonella typhimurium) at dose deaths ( Scallan et al 2011). Symptoms as gastroenteritis, (1* 108) for two week according to method of Takeuchi diarrhea and some systemic disorders are associated with (1966). infection which appear in animals and human (Jawetz et al Histo-pathological study: Liver and kidney of control and 2007). In experimental infections as mice, the symptoms are treated groups were isolated then put in 10% formalin similar to that in of typhoid fever in human (Tuin et al 2005). solution for fixation. Sections were made according to Serotype of Salmonella and host species are two factors method of Bancroft and Gamble (2008). determine the immune responses for Salmonella (Moreau 2006). The current study was carried out to assessment the RESULTS AND DISCUSSION histological changes of internal organs as liver and kidney of The results of present study were demonstrated rats which infected by Salmonella typhimurium experimentally. histopathological changes in liver after experimentally infection by Salmonella typhimurium, these changes MATERIAL AND METHODS included congestion , inflammation clearly , enlargement of Isolates of bacteria: The isolates of Salmonella central vein and sinusoids , hypertrophy of hepatocytes typhimurium were brought from laboratory of Al-Hussein (picture 2-8) . These damages may be related to presence of hospital in Thi-Qar province. These were previously collected bacteria in blood which will carry into internal organs of body from stool of persons with diarrhea. The method of Quinn et al as liver and kidney, this agree with Ma¨kela¨ and Hormaeche (1998) was used to determination of biochemical properties. (1997) who reported to Salmonella arrive to circulatory Thirty two of albino rats (Rattus norvegicus) were used in system by efferent lymph causing bacteremia. Everest this study, which supplied from animal house of biology (1999) suggested gastroenteritis due to Salmonella department, university of Thi-Qar . Female rats were divided typhimurium infection. Also, the histopathological changes into four groups (n=8) as: were observed after (one and two weeks) from bacterial 220 Fatima Aziz Mahdi Al-badry

Fig. 1. Section in the liver of control group showing central Fig. 2. Section in the liver of second group showing vein (A) hepatocytes (B) sinusoids (C) (H&E) (100X) infiltration of inflammatory cells (A) congestion (B) (H&E) (100X)

Fig. 3. Section in the liver of second group showing Fig. 4. Section in the liver of third group showing infiltration congestion (H&E) (100X) of inflammatory cells (A) enlargement of central vein (B) enlargement of sinusoids (C) (H&E) (100X)

Fig. 5. Section in the liver of third group showing congestion Fig. 6. Section in the liver of fourth group showing (A) enlargement of sinusoids (B) (H&E) (100X) congestion (A) infiltration of inflammatory cells (B) enlargement of sinusoids (C) (H&E) (100X) Negative Effects on Some Internal Organs of Rats Infected by Salmonella typhimurium 221

Fig. 7. Section in the liver of fourth group showing Fig. 8. Section in the liver of fourth group showing congestion (A) hypertrophy of hepatocytes (B) enlargement of sinusoids (H&E) (100X) enlargement of sinusoids (C) (H&E) (100X)

Fig. 9. Section in the kidney of control group showing Fig. 10. Section in the kidney of second group showing glomerulus (A) renal tubules (B) (H&E) (100 X) hemorrhage among renal tubules (H&E) (100 X)

Fig. 11. Section in the kidney of second group showing Fig. 12. Section in the kidney of third group showing shrinkage (A) absence of glomerulus (B) (H&E) hemorrhage (A) death of glomerulus (B) (H&E) (100 X) (100 X) 222 Fatima Aziz Mahdi Al-badry infection belong to numerous numbers of Salmonellae which inflammatory cells, infiltration of kupffer cells and other form colonies inside intestinal epithelium compared with damages of internal organs after mice inoculation with (one day) infection, this is similar to Cirillo et al (1998) who bacteria (Salmonella). Zedan (2011) mentioned to revealed to colonize of Salmonella in small intestine after degeneration, necrosis and infiltration of inflammatory cells ingestion orally. were histological damages observed in experimentally Enlargement of central vein and sinusoids, hypertrophy infection with Salmonella typhimurium. A previous another of hepatocytes and other histological changes may be back study by Malik et al (2015) was confirm the histological to cytotoxic effect of bacterial infection on hepatocytes lesions in liver of Salmonella infection such as necrosis, causing harmful effects on liver structure, this finding is degeneration of hepatocytes and inflammation. Another accordance with Zghair (2012) who noticed histological reason of hepatic damage may be associated to hazard damages of liver mice which infected by Salmonella bacterial infection which caused disturbances in liver typhimurium. Both infiltration of inflammatory cells and enzymes as ALT and AST in blood, Olaleye et al (2014) who congestion were found , they are agree with Zghair (2012) exhibited that liver enzymes act signs of hepatic injury. This is who demonstrated congestion of blood vessels, infiltration of accordance with Tala et al (2015) who noted elevated of ALT

Fig. 13. Section in the kidney of third group showing Fig. 14. Section in the kidney of fourth group showing enlargement of Bowman's space (A) absence (B) infiltration of inflammatory cells (A) congestion (B) shrinkage of glomerulus (C) (H&E) (100 X) shrinkage of glomerulus (C) (H&E) (100 X)

Fig. 15. Section in the kidney of fourth group showing Fig. 16. Section in the kidney of fourth group showing enlargement of Bowman's space (A) death of absence of glomerulus (A) hemorrhage (B) glomerulus (B) congestion (C) (H&E) (100 X) shrinkage of glomerulus (C) (H&E) (100 X) Negative Effects on Some Internal Organs of Rats Infected by Salmonella typhimurium 223 and AST in serum of infected rats resulting damage in Rats. International Journal of Contemporary Research and Review 9(4): 20239-20250. histological architecture of liver. Jawetz J, Melnick E and Adelbergs E 2007. Enterobacteriaceae In: Kidney was affected internal organ with bacterial Review of medical microbiology. 24th ed. by Geo F, Janet S infection, microscopic examination of histological sections and Stephen Mc Graw–Hill companies, U.S.A., Pp: 256-261. showed hemorrhage, shrinkage, absence and death of Mahdi NR and Mohammad AJ 2018. Antibacterial effect of grape seeds polyphenols against Salmonella typhimurium infection glomeruli, enlargement of Bowman's space, infiltration of in mice. Al-Anbar Journal of Veterinary Sciences 11(1): 46-59. inflammatory cells and congestion (pictures 10-16). These Ma¨kela¨ PH and Hormaeche CE 1997. Immunity to Salmonella. In results were come agreement with Mahdi and Mohammad Host Response to Intracellular Pathogens (SHE Kaufmann, ed.), Austin, TX: R.G. Landes (2018) who their study explained the potent inflammatory 143-166. response (comprised in infiltration of inflammatory cells) and Malik MA, Kotwal SK, Rahman S, Singh M and Sharma N 2015. the histological signs in organs as liver and intestine, this is Pathomorphological changes in various organs of experimentally induced Salmonellosis in mice. Journal of similar to Parry et al (2013) who observed severe Animal Research 5(3): 605-609. gastroenteritis in human infected with Salmonella Moreau MC 2006. Immune Modulation by the Intestinal Microbiota in Gastrointestinal Microbiology. Taylor and Francis Group, LLC. typhimurium. Al-Joboury (2007) revealed to congestion, Muchtaromah B, Maslikah SI, Mufarrichah L and Fitriasari PD 2019. interstitial nephritis and mild micro thrombi in kidney of mice Histological description of small intestine and kidney of white which exposed for Salmonella infection. rats (Rattus norvegicus) infected with Salmonella typhi by giving earthworm flour. International Conference on Biology Hemorrhage was observed in kidney of Salmonella and Applied Science (ICOBAS) 700031- 700037. infected groups is identical to two studies by Malik et al (2015) Olaleye MT, Amobonye AE, Komolafe K and Akinmoladun AC 2014. and Yehya and Salman (2008) which their studies had Protective effects of Parinari curatellifolia flavonoids against acetaminophen-induced hepatic necrosis in rats. Saudi showed bleeding and histological signs as degeneration, Journal of Biological Sciences 21(5): 486-492. necrosis, infiltration of inflammatory cells of renal tubules of Parry CM, Thomas S, Aspinall EJ, Cooke RP, Rogerson SJ, Harries kidney. There are study by Muchtaromah et al (2019) AD and Beeching NJ 2013. A retrospective study of secondary bacteraemia in hospitalised adults with community acquired included histological description of kidney that infected by non-typhoidal Salmonella gastroenteritis. BMC Infectious Salmonella occurred karyolysis, dilation of Bowman's space Diseases 13: 107. and pyknosis of nucleus. Also, experimentally infection by Quinn PJ, Carter ME, Markey B and Carter CR 1998. Clinical Veterinary Microbiology. Pp: 261-267. M. Wolfe, London. Salmonella typhimurium caused disturbance of kidney Scallan E, Hoekstra RM, Angulo FJ, Tauxe RV, Widdowson MA, Roy structure comprised necrosis, inflammation and edema of SL, Jones JL and Griffin PM 2011. Foodborne illness acquired in the United States-major pathogens. Emerging Infectious renal tubules which act a pathogenesis of Salmonella Diseases 17(1): 7-15. infection (Isirima and Siminialayi 2018). Takeuchi A 1966. Electron microscope studies of experimental Salmonella infection. Journal of American Society for Microbiology 5(1): 109-136. REFERENCES Tala DS, Gatsing D, Fodouop CP, Fokunang C, Kengni F and Djimeli Al-Joboury KH 2007. A study on the bacterial dissemination and MN 2015. In vivo anti-salmonella activity of aqueous extract of experimental pathology of Salmonella paratyphi. An infection in Euphorbia prostrata Aiton (Euphorbiaceae) and its white mice. Iraqi Journal of Medical Sciences 5(3): 23-30. toxicological evaluation. Asian Pacific Journal of Tropical Al-Joboury KH 2007. A study on the bacterial dissemination and Biomedicine 5(4): 310-318. experimental pathology of Salmonella paratyphi. An infection in Tuin A, Huizinga-Van der Viag A, Van Loenen-Weemaes AM, Meijer white mice. Iraqi Journal of Medical Sciences 5(3): 23-30. DK and Poeistra K 2005. On the role and fate of LPS- Bancroft JD and Gamble M 2008. Theory and practices of dephosphorylating activity in the rat liver. American Journal of histological technique. 2nd ed. Churchill Elsevier. London, pp Physiology-Gastrointestinal and Liver Physiology 290(2): 377- 56. 385. Callaway TR, Edrington TS, Anderson RC, Byrd JA and Nisbet DJ Woc-Colbum L and Bobak DA 2009. The expanding spectrum of 2008. Gastrointestinal microbial ecology and the safety of our disease due to Salmonella an international perspective. food supply as related to Salmonella. Journal of American Current Infectious Disease Reports 11: 120-124. Society of Animal Science 86: 163-172. Yehya MM and Salman AM 2008. Immunological and pathological Cirillo DM, Valdivia RH, Monack DM and Falkow S 1998. studies on the efficacy of porin proteins of Salmonella Macrophage dependent induction of the Salmonella enteritidis isolated from diarrheic calves. Egyptian Journal of pathogenicity is l and 2 type III secretion system and its role in Comparative Pathology and Clinical Pathology 21(2): 300-325. intracellular survival. Molecular Microbiology 30: 175–188. Zedan ZK 2011. Hepatoprotective effect of Lactobacillus Plantarum Everest P 1999. Evaluation of Salmonella typhimurium mutants in a against Salmonella typhimurium in mice. Journal of Al-Nahrain model of experimental gastroenteritis. Infection and Immunity University 14(4): 126-131. 67(6): 2815-2821. Zghair ZR 2012. Histopathological study of Salmonella typhimurium Isirima JS and Siminialayi IM 2018. Effects of the methanol leaf infection in laboratory mice by using the light and electron extract of Chromolaena Odorata on blood lipids and the microscope. Kufa Journal For Veterinary Medical Sciences histology of the kidney in Salmonella Typhi Infected Wistar 3(1): 124-131.

Received 12 September, 2019; Accepted 04 November, 2019 Manuscript Number: S-285 Indian Journal of Ecology (2019) 46 Special Issue (8): 224-228 NAAS Rating: 4.96

Effect of the Isolated Alkaloid from Teucrium polium L. on Dna Isolated from Pathogenic Bacteria (Escherichia coli and Staph aureus) of Urinary Tracts

Rawaa A. Kushaish, Bushra A.M. AL-Salem* and Mouayed A.A. Hussein Chemistry Department, University of Basra, Iraq E-mail: [email protected]

Abstract: Four compounds 1, 2, 3 and 4 have been isolated from Teucrium polium L. using the thin layer chromatography and column chromatography techniques. Phytochemical analysis showed that all the compounds are of alkaloids category. The suggested chemical structures of isolated compounds were determined by 1HNMR, 13CNMR, and GC-Mass spectroscopy. The nuclease activity of the compounds was investigated against DNA cleavage isolated from Escherichia coli and Staph aureus, respectively. Gel electrophoresis revealed that all the isolated compounds have a noticed ability to cleave DNA. Compounds 1, 2, and 3 started to cleave the DNA showing the linear form at concentrations 15 µM, 13.5 µM, and 18 µM, respectively. Whereas Compound 4 showed different cleavage efficiency, it can be nicked one strand produced the slow migration form, related to the open circular relaxed form.

Keywords: Teucrium polium L., Alkaloids, DNA cleavage, Gel electrophoresis, Linear DNA form, Column chromatography

Medicinal plants are universally considered as important investigate the medicinal effect of alkaloid compounds sources of new chemical substances with potential isolated from Teucrium polium L., on DNA isolated from therapeutic effects. Teucrium polium L. (syn Teucrium pathogenic bacteria of urinary tracts. capitatum L. or felty germander) is one of the 300 species of the genus Teucrium from Lamiaceae family (Bahramikia and MATERIAL AND METHODS Yazdanparast 2012). It is believed that T. polium contains Teucrium polium L. was collected from Basrah, Iraq. The several subspecies and varieties including aragonense, aerial parts were separated, dried for one week in the shade aurasiacum, capitatum cylindricum, expansum, at room temperature and then ground to a fine powder, and gnaphalodes, pilosum, polium, vincentinum, yalentinum and stored at 7°C in glass containers until used. The powdered many others Teucrium polium L. (Lamiaceae) is a perennial plant material (100 gm) was extracted three times with shrub, 20–50 cm high, distributed widely in the dry and stony ethanol 95% (3×2L) at room temperature. The extracted places of the hills and deserts of almost all Mediterranean plant was concentrated under reduced pressure and countries, South Western Asia, Europe, and North Africa. dissolved with 2% sulfuric acid. After filtration, the acidic Sessile, oblong, or linear leaves have a length of about 3 cm aqueous solution was extracted three times with chloroform (Haghshenas et al 2018). T. polium has been used for to remove the lipophilic impurities. The pH of is adjusted to different pathological conditions such as gastrointestinal 10.0 using aqueous ammonia, and then extracted three disorders, inflammations, diabetes, and rheumatism.During times with CHCl3 (Harborne1984). Finally, chloroform was the past 40 years, different classes of compounds have been totally evaporated from the organic phase to form the alkaloid isolated from various parts of T. polium of which the main powder. groups are terpenoids and flavonoids (Bahramikia et al2 Thin layer chromatography (TLC): TLC technique was 012). used for separation of alkaloids compound and It has been found that these compounds possess a determination of their purity. One hundred microliters of broad spectrum of pharmacological effects including alkaloids extract were toulerenced on silica gel plate (2×10 antioxidant, anticancer, anti-inflammatory, hypoglycemic, cm) and were put in glass Jar then (n-hexane: di chloro hepatoprotective, hypolipidemic, antibacterial and methane: ethyl acetate) solvent was used as eluent system antifungal. with a ratio equal to (9:9:1) for 10 min. The glass plate was There is increasing evidence for T. polium effect on driedandthe compounds were developed by UV-light at 366 cardiovascular such as positive inotropic and chronotropic, nm, Dragendroff reagent and steam iodine (Harborne1984). reducing blood pressure, and lowering blood lipid Rates of flow (Rf) values were measured for each alkaloids (Mahmoudabady et al 2018). In this study aimedto compound. Urinary Tracts effectiveness by Teucrium polium L. alkaloid 225

Column Chromatography of alkaloids: Column Compound 4: Anal. Calcd for C7H9N: 1H NMR (CDCl3, Chromatography (CC) technique was carried out to separate ppm):1.29 (CH3), 7.52 (=CH), 7.72 (CH=). 1C NMR (CDCl3, each alkaloidic compound by using a column has length and ppm): 14.13 (CH3), 29.81(CH=N) 29.70 (CH=), 37.11 (=CH). diameter equal to 30 cm, 2 cm, respectively. The glass wood GC-Mass (CDCl3, m/z): 108 (C7H9N), 42 (C2H3N), 68 put in the down and of the column and was filled with silica gel (C4H6N).

(230-400 mesh; RDH), then (n-hexane, CH2Cl2 and ethyl Bacterial strains: Two bacterial species used in this study. acetate with increases the polarity to 100% CH2Cl2 and then The gram–positive species was Staph aureus (S. aureus) to 15% ethyl acetate in CH2Cl2) was used as an eluent and Gram negative species bacteria was Escherichia coli (E. (Elmasri 2015). coli) which were clinically isolated from the urine of patients Preliminary qualitative tests: Isolated alkaloid was and identified by using device Vitek 2 biomerieux Automated undergone to several tests: Microbiology System (Biomerieux-Francee). Alkaloids test was carried out by using Dragendroff DNA cleavage study: The DNA cleavage of the isolated reagent and Mayer's reagent (Harborne 1984). compounds was investigated against one type of Gram- Carbohydrates test was done by using Molish reagent positive bacteria and one type of Gram-negative bacteria by (Harborne 1984). Glycosides test was made by using using the procedure of genomic DNA mini kit (Geneaid, Benedict reagent (Al-Khazraji 1972). Korea). The stock solution of DNA was prepared by Amino acids test was made by using ninhydrine (1%) dissolution of a suitable amount of DNA in 6.3 (Harborne1984). mMTris–HCl/50 mMNaCl buffer (pH 7) at room temperature. Saponins test was made by using Mercuric chloride A solution of DNA in the buffer gave a UV absorbance ratio at (5%) (Harborne1984).Phenols test was made by using Ferric 260 and 280 nm of ca. 1.9: 1, indicating that the DNA was chloride (1%) (Ribéreau-Gayon1972). sufficiently of free protein. The DNA concentration was Diagnosis of isolated alkaloids by NMR and GC-Mass estimated by UV absorbance at 260 nm with known molar Nuclear Magnetic Resonance (NMR): The 1H and 13C absorption coefficient of 6600 M-1 cm-1(Reichmann et al spectra for the alkaloidic compounds were obtained in 1954). The UV-Vis spectra were scanned from 230 to 600 nm deuterated solvent CDCl3 by using Bruker 500 MHz. with a Tris/HCl buffer solution as reference. Chemical shifts were reported in part per million (ppm) by The ability of the isolated compounds to cleavage DNA using tetramethyl silane (TMS) as standard. was studied agarose gel electrophoresis in Tris/EDTA buffer Gas Chromatography-Mass Spectroscopy: Gas solution (Hussein et al 2014). The Samples were incubated Chromatography-Mass Spectrometry (GC-MS) analysis was at 37 °C for 2 h, treated with loading dye, and performed with an auto Mass Hunter\GCMS instrument electrophoresed for 1 h at 50 V on 1% agarose gel consisting using a SPB 5 (30 m/ 0.25 mm/ 0.5 µm) capillary column. 1µL of 9 lanes: lane 1, DNA (10 µL); lane 2, DNA (10 µL) + H2O2 (5 of each isolated alkaloid dissolved in chloroform was injected µM ); lane 3, DNA (10 µL) + H2O2 (5 µM) + sample (3µM); lane in the following conditions: injector temperature, 300 °C; 4, DNA (10 µL ) + H2O2 (5 µM) + sample (6.2 µM); lane 5, DNA carrier gas, helium; pressure, 11.962 psi. Compounds were (10µL) + H2O2 (5 µM) + sample (9.3 µM); lane 6, DNA (10 µL) identified on the basis of their mass spectral data. + H2O2 (5 µM ) + sample (10.5 µM ); lane 7, DNA (10 µL) + 1 Compound 1: Anal. Calcd for C8H17N: HNMR (CDCl3, ppm): H2O2 (5 µM ) + sample (13.5 µM); lane 8, DNA (10 µL) + H2O2

0.84 (CH3), 1.26 (CH2), 1.28 (CH-N), 1.33 (-CH2), 1.43 (NH). (5 µM) + sample (15 µM); lane 9, DNA (10 µL) + sample (18 13 CNMR (CDCl3, ppm): 14.13 (CH3), 22.70 (CH2), 30.19 (C- µM). The gel was then stained with ethidium bromide before

N), 31.44 (-CH2). GC-Mass (CDCl3, m/z): 128 (C8H18N), 69 being photographed under UV light.

(C4H7N), 55 (C3H5N), 43 (C3H7). Compound 2: Anal. Calcd for C7H15N: 1H NMR (CDCl3, RESULTS AND DISCUSSION ppm): 0.91 (CH3), 1.26 (CH2), 1.28 (CH3-N), 1.33 (CH-N). 1C In the current study, alkaloids extract was isolated and NMR (CDCl3, ppm): 14.13 (CH3), 22.70 (CH3-N), 29.71 purified. Table 1 indicates the qualitative chemical analysis of (CH2), 31.94 (C-N). GC-Mass (CDCl3, m/z): 114 (C7H16N), alkaloids isolated from Teucrium polium. The results show 56 (C3H7N), 70 (C4H9N). the presence of alkaloids only but carbohydrate, glycosides, Compound 3: Anal. Calcd for C6H13NO: 1H NMR (CDCl3, amino acids, saponins, and phenol were not found. ppm): 0.87 (CH3-O), 1.25 (CH-O), 1.33 (CH2-N) 1.28 (-CH2), Figure 1 and Table 2 represent thin layer 1.43 (NH). 1C NMR (CDCl3, ppm): 29.71 (OCH3), 22.70 (- chromatography (TLC) results of alkaloids extracted from T. CH2), 31.44 (-CH2). GC-Mass (CDCl3, m/z): 116 polium L. There are four spots were separated have rates of

(C6H14NO), 43 (C2H3O), 29 (C2H5). flow (Rf) value equal to 0.9, 0.85, 0.75 and 0.7. This ensures 226 Rawaa A. Kushaish, Bushra A.M. AL-Salem and Mouayed A.A. Hussein presence of four alkaloidic compounds in the extract and determined by 1HNMR, 13CNMR, and Mass spectroscopy these components were tested and developed by which are well revealed their chemical structures. Dragendroff reagent as a qualitative and characteristic The Mass spectra of compounds are shown in Figure 1. developer for alkaloids, also by using iodine vapour. The Compounds 1 and 2 revealed three common mass spectral alkaloids separated show brown colour, and by using litmus signals at 56 attributed to CH3CH2CH=NH and CH3C=NCH3 paper; the colour has changed from red to blue. fragments; 70 attributed to CH3CH2 (CH2)C=NH and CH3

Therefore, this ensures that these compounds are (CH2) C=N- CH3 fragments and, 84 attributed to CH3CH2 (CH2 nitrogenous having basic basic features. Column CH2)C=NH and CH3 (CH2 CH2)- C=NCH3 fragments,

Chromatography (CC) technique was used to separate each respectively. The signal at 43 is attributed to CH3CH2CH2 alkaloid compounds alone with time equal to 0.8 ml min-1. fragment. Whereas the signals at 128 and 114 are attributed Then TLC results of alkaloids compounds separated from to m+1 for 1 and 2, respectively. Compounds 3 and 4 showed column, showed a rate of flow (Rf) equal to 0.9, 0.85, 0.75 three mass spectral signal at 29, 43 116 and 42, 68, 108 and 0.7. attributed to the fragments of CH3CH2, CH2CH=O, m+1 and

Analysis: Four compounds, 1, 2, 3, and 4 were isolated from CH3CH=NH, CH=CHC (CH3)=NH, respectively. 1 Teucrium polium. The identity of isolated compounds was HNMR spectra were scanned using CDCl3 as a solvent that shows a singlet signal at 7.26 ppm (Fig. 2). The compound 1 shows a triplet signal at 0.84 ppm attributed to Table 1. Preliminary qualitative test for isolated alkaloid from the protons (1) of the terminal CH groups. The multiplet T. polium 3 signal that emerged in the range 1.26 ppm to 1.33 ppm Test Test Notes Conclusion result attributed to the protons 2, 4 and 5, whereas, the broad signal Dragendroff + Formation of orange Presence of that emerged at 1.43 ppm attributed to the proton 3 of NH precipitate alkaloid group. 1HNMR spectrum of compound 2 shows a duplet Mayer + Formation of white Presence of signal at 0.91 ppm attributed to the protons 1 of the CH precipitate alkaloid 3 groups. The singlet signal emerged at 1.5 attributed to Molish - No violet ring No carbohydrate protons 2. Whereas, the multiplet signal and duplet signal, Benedict - No red precipitate No glycosides emerged at 1.7 ppm and 1.28 ppm, attributed to the protons 3 Ninhydrine - No violet colour No amino acid and 4, respectively. In compound 3, the protons 1 of CH3 HgCl (5%) - No white precipitate No saponin 2 group appears a single signal at 0.87 ppm, the multiplet

FeCl3 (1%) - No blusich- green colour No phenol signal that appeared at the range 1.28 ppm to 1.35 ppm is attributed to 2, 3 and 4 protons, whereas the broad signal at 1.43 ppm back to the proton 5 of NH group. The aromatic compound 4 revealed singlet signal at 1.29 ppm is attributed

to the protons 1 of CH3 groups. The aromatic protons 2 and 3 are emerged duplet signal at 7.72 ppm and triplet signal at 7.35 ppm, respectively. The structures of the isolated compounds were also confirmed by 13CNMR spectroscopy. The spectra that shown in Figure 3, reveal the compound 1 emerged four signals at 14.13 ppm, 22.76 ppm, 31.44 ppm, and 30.19 ppm attributed to the carbons 1, 2, 4 and 3, respectively. Compound 2show its carbons 1, 2, 4 and 3 at 15.83 ppm, 22.70, 31.94 ppm and 29.71, respectively. The carbons 1, 2, 3 and 4 of compound 3 appeared at 14.98 ppm, 31.44 ppm, 22.70 ppm and 29.71 ppm, respectively. Whereas, the carbons that emerged at 14.73 ppm, 29.70 ppm, 29.81 ppm and 37.11 ppm, are attributed to 1, 3, 4 and 2 of compound 4, respectively. DNA cleavage activity: The DNA cleavage of the isolated compounds was investigated against Esherichia coli and Staph aureus, respectively. Agarose gel electrophoresis experiments using (10 µL) of DNA were investigated for a Fig. 1. TLC of alkaloid fractions developed by UV light Urinary Tracts effectiveness by Teucrium polium L. alkaloid 227

Fig. 2. Mass spectra of compounds 1, 2, 3 and 4 micro-concentrations 3, 6.2, 9.3, 10.5, 13.5,15, and 18 µM (in respective, lanes: 3-9) of isolated compounds in aqueous buffer solution 6.3 mMTris-HCl/50 mMNaCl buffer (pH= 7) in

presence of fixed amount (90 µM) of 30% H2O2 as an oxidant, and the solution was incubated at 37 0C for 2h. The agarose gel electrophoresis patterns (Fig. 4) display that all the isolated compounds have high efficiency to cleave DNA. Compound 1 cleaves DNA at concentrations of 15 and 18 µM (lanes; 8 and 9) showing the linear form (form III) as well as the natural fast migration form related to the closed circular supercoiled form (SC, form I), lanes; 4 to 7. Compound 2 shows the linear form at concentrations of 13.5, 15 and 18 µM (lanes; 7, 8 and 9). The compound 3 exhibits the linear form atconcentration of 18 µM (lane; 9). Although the compounds 1, 2 and 3are produced the linear DNA form, the cleavage efficiency is totally different. The migration of linear form produced by the compound 3 is faster from that for the compound 2 which is faster than that for the compound 1; this explains the small molecular size of linear DNA form produces by 3 compared to 1 and 2. Compound 4 and unlike the previous compounds display a different DNA cleavage efficiency. It can be nicked one strand of DNA producing the slow migration form related to the open circular relaxed form Fig. 3. 1HNMR spectra of the compounds 1, 2, 3 and 4 (OC, form II) (Gao et al 2010). 228 Rawaa A. Kushaish, Bushra A.M. AL-Salem and Mouayed A.A. Hussein

Table 2. TLC results of isolated alkaloids from T. polium The DNA cleavage activity was investigatedin presence Eluent system Reagent Spots No. Rf Conclusion of H2O2 as an oxidant. The agarose gel electrophoresis

n-hexane: di I2-vapour 4 0.90 Presence of patterns show that the DNA cleavage is not taking place in chloro methane: 0.85 nitrogenous presence of H O only (lane; 1) or the presence of compound ethyl acetate 0.75 organic 2 2 (9:9:1) 0.70 compound only (lane; 2) which displays the natural SC form, this Dragendroff 4 0.90 Presence of explains that the DNA cleavage depends on the presence 0.85 alkaloid both of the H O and the compound. These results give 0.75 2 2 0.70 evidence that the compounds produce hydroxyl radical from

H2O2, and then the free radicals share in the oxidation of deoxyribose moiety, followed by hydrolytic cleavage of the sugar-phosphate backbone (Babu and Krishna 2007, Pogozelski and Tullius 1998).

CONCLUSION The DNA cleavage study exhibits that the isolated compounds of T. polium can cleave the DNA in presence of

H2O2. The agarose gel electrophoresis patterns for compound 1, 2, and 3 shows the linear form with the closed circular super coiled, whereas the compound 4 show the linear form with the open circular relaxed.

REFERENCES Babu MS, Reddy KH and Krishna PG 2007. Synthesis, characterization, DNA interaction and cleavage activity of new mixed ligand copper (II) complexes with heterocyclic bases. Polyhedron 26(3): 572-580. Bahramikia S and Yazdanparast R 2012. Phytochemistry and medicinal properties of Teucrium polium L.(Lamiaceae). Phytotherapy Research 26(11): 1581-1593. Elmasri W 2015. Teucrium Polium Phytochemical Analysis as a Source of Antibacterial and Cancer Cytotoxic Agents. Gao EJ 2010. New pH-dependent complexes, from mononuclear Pd (II) monomer to heteronuclear [Pd (II), K (I)] Polymer: DNA cleavage and cytotoxicity in vitro. European Journal of Medicinal Chemistry 45(3): 1034-1041. Harborne JB 1984. Phytochemicaal method 2nd. ed Chapman and Fig. 4. 13CNMR spectra of compounds 1, 2, 3 and 4 Hall, New York. USA. Hussein MA 2014. Structures, DNA binding, DNA cleavage, and antitumor investigations of a series of molybdenum(VI) complexes with some N(4) methyl and ethyl thiosemicarbazone ligands. Journal of Coordination Chemistry 67(4): 714-727. Mahmoudabady M, Haghshenas M and Niazmand S 2018. Extract from Teucrium polium L. Protects rat heart against oxidative stress induced by ischemic–reperfusion injury.Advanced Biomedical Research 7: 34-38. Pogozelski WK and Tullius TD 1998. Oxidative strand scission of nucleic acids: Routes initiated by hydrogen abstraction from the sugar moiety. Chemical Reviews 98(3): 1089-1108. Reichmann M 1954. A further examination of the molecular weight and size of desoxypentose nucleic acid. Journal of the American Chemical Society 76(11): 3047-3053.

Received 08 October, 2019; Accepted 20 November, 2019

Fig. 5. Agarose gel electrophoresis patterns of DNA for the compound 1, 2, 3 and 4 in 6.3 mMTris-HCl/50 mMNaCl buffer (pH= 7) at 37oC Manuscript Number: S-286 Indian Journal of Ecology (2019) 46 Special Issue (8): 229-232 NAAS Rating: 4.96

Evaluation of Amino Acid Profile for Freshwater Fishes Yellow Barbell (Carasobarbus luteus) and Common Carp (Cyprinus Carpio) of Euphrates River, Iraq

Kadhim O.M. AL. humairi, Hamza G. Al-Agidi and Sadeq M. Al-Haider1 Al-Furat AL- Awsat Technical University, AL- Musaib Technical College, Babylon-51009, Iraq 1College of Veterinary Medicine, Al-Qasim Green University, Iraq E-mail: Com.kdm @atu.edu.iq

Abstract: The present investigation was carried out to evaluate the amino acids composition of two important commercial freshwater fish yellow barbell (Carasobarbus luteus) and common carp (Cyprinus carpio) of Euphrates River. The chemical analysis was conducted and the results showed that total protein content (% dry weight) for C. luteus and for C. carpiowas 26.12 and 22.59%, respectively. The total fat content was 2.17 % and 4.04 % for C. luteus and C. carpio, respectively. The ash content for C.luteus was 1.5% and C. carpio was 2.96% while moisture content 71.15% for C. luteus and 71.0% for C. carpio, Amino acids ranged from 0.56 - 14.03% for C.luteus and 0.7- 13.11% for C. carpio fish. The most present amino acids were glycin, aspartic, tryptophan, and glutamic, which ranged from 8.22 to 14.03%. The essential amino acids in the two species were arginine, histidine, isoleucine, leucine, phenylalanine, threonine, tryptophan, and valine were detected. The percentage ranged from 0.56 - 11.69% for essential amino acids in two species to improve good health, protection, and reduce of diseases in humans and therefore an important source of nutrients.

Keywords: Amino acids, Carasobarbus luteus, Cyprinus carpio, HPLC, Protein

Fisheries and aquaculture play an important role in the Cyprinus carpio belong to the same family Cyprinidiae. economy of many developing countries around the world, Omnivorous feedings and dominant groups present in the particularly in Asia and Africa. Fishes and fish products are Euphrates during the seasons of the year (Amari et al 2012). considered to be the most widely traded commodities in the They are considered to be economically important species world (Martini and Lindberg 2013). Freshwater fish are an and are widespread in many freshwater bodies, especially in important source of protein and unsaturated fatty acids central and southern Iraq (Al-Tamimi 2004). C. luteus is an (USFA) (Ljubojevic et al 2013) and are protein rich in Iraqi fish found in inland waters (Coad 2017). While C. carpio essential amino acids like (lysine, methionine, cysteine, was introduced to Iraq during 1955-1956 by the first of threonine, and tryptophan) (Sikorski 1996). Indonesia (Pantene carp) and the second from the Humans have used fish as a food throughout the ages as Netherlands (Japanese multi -color carp) for breeding at the an easily digestible food; as it is rich in protein and fat and also Zaafarania fish farm in Baghdad and then launched into the for treat numerous diseases, like prevention of pulmonary natural water bodies in the Alhammar and Sunni marshes and tuberculosis and weak liver function (Craig and Helfrich the lakes of Habbaniyah and Tharathar n 1965 (AL Hamed 2002). Fish muscles are the main and important part of 1971, Al-Rubaie 1986). In 2009, the C. carpio was imported human consumption, so their composition of proteins, from Henkaria by private sector companies considered as especially their essential amino acids, should be known in the one of the most important economic fish cultivated in Iraq, it is human diet (Ackman 1990). Limin et al (2006) noted that the the first in the field of cultivated (Hassouni and Janabi 2005). total of amino acid content in fish is influenced by internal Al-Amari et al (2012) has reported that, algae are the main factors such as (species, size, sexual maturity) and other food for C. luteus, followed by aquatic plants, organic crumbs, external factors for example (food type, fishing season, and diatoms with a tendency to plant species. In the study of salinity, temperature). Also, it is affected by season and Mohammed et al (2015) pointed out the C. luteus in the East location, there is a strong relationship between the pattern of Alhammar marsh adoption in feeding on twelve different the content of essential amino acids found in fish body tissue types of natural food, the diet included diatoms, and the pattern of dietary requirements for each species phytoplankton, algae, and aquatic plants. C. carpio feed on (Wesselinova 2000). There are several studies on the natural foods in their external environment and commercial quantitative and qualitative properties of amino acid diet in different farming systems (Kocour et al 2007). The composition in many species (Ovissipour et al 2009, Kenari et common commercial diet used to feed C. carpio made from al 2009, Dezhabad et al 2012). Carasobarbus luteus and cereals, soybeans, fishmeal and supplemented with 230 Kadhim O.M. AL. humairi, Hamza G. Al-Agidi and Sadeq M. Al-Haider vitamins, minerals and possibly some feed additives, which calculated on the dry matter of the two freshwater species C. are in the form of Peale at, such as sinking and floating. These carpio and C. luteus shown in Table 1. feeds are largely used in intensive culture systems (cages) Content of protein: Content of protein for C. carpio ranged and semi-intensive culture, clay ponds (Deutsch et al 2007). from 22.18 to 22.67% While we noted that C. luteus spread Natural foods are a valuable source of proteins from 25.87% to 26.45%. The mean Content of protein spread (essential amino acids), fatty acids and vitamins, which from 22.59% forC. carpio to 26.12% for C. luteus. Note determine the nature of the nutritional value of each type of significant differences (p< 0.05) between the species. fish meat used in human nutrition (Kibria et al Record the highest protein content in C. lute. 1997).Therefore, it is necessary to study and measure the Content of lipid: The lipid ratio of C. carpio ranged from content of amino acids in the tissues and muscles of each 3.76-4.2% and for C. luteus 2.03-2.4%. The average total type of fish, to determine the optimal type of fish in human content of lipid ranged from 4.04- 2.17% with significant nutrition. The current study aimed to know the composition of differences (p< 0.05) between species. The highest lipid amino acids for two types of freshwater fish including content was recorded in C. carpio. Carasobarbus luteus and Cyprinus carpio located in the Content of ash: The content of ash for C. carpio ranged from inland waters of Iraq. 2.87- 3.09 % and 1.22-1.66% in C. luteus. The average total content of ash for 2.69-1.5%. The highest ash content was MATERIAL AND METHODS noted in C. carpio. Collection of samples: Carasobarbus luteus was obtained Content of moisture: The content of moisture ranged from from the fishermen of the Euphrates River, and Cyprinus 70.58 to 71.18% with average of 71.0% for C. carpio and carpio was obtained from the floating cages farms on hours. 70.85 to 71.53% and average of 71.15% for C. luteus. No The total protein in the muscles of the fish was determined by significant differences were observed (p< 0.05) between the Kjeldahl method. Measure the total nitrogen content in species. the sample after digesting a known weight with the sulfuric Relative volume: Relative volume ranged from 0.37 to 0.43 acid (H2SO4), the distillation with per chloricacid (HCLO4) and with average of 0.403 in C. carpio and 0.33 to 0.375 and the hydrochloric acid (HCL) correction, then multiply the average 0.359 for C. luteus. No significant differences were nitrogen value by a factor (6.25N) to convert total nitrogen to observed (p< 0.05) between species. crude protein (CP)in all fish samples. Ash was determined in Amino acid content: Eighteen different amino acids were the dry sample by taking 5g of the sample and then placed in found from fish under study. The total amino acids for C. furnace at 550°C for 8 hours. The crude fat was extracted carpio was 225.39 mg g-1 and in C. luteus was 261.21 mg g-1 from the fish muscles by the organic solvent petroleum ether (Table 2). Eight of the essential amino acids that are very by using the Soxlet. important to human health are present in the fish understudy. Analysis of amino acids: The amino acid profiles of raw fish These essential amino acids are arginine, histidine, muscles are determined by (HPLC) High-Performance isoleucine, leucine, phenyl al., tryptophan, threonine Liquid Chromatography analysis according to Vidotti et al andvaline. Total essential amino acids 36.87% for C. carpio (2003) by using the cationic exchange resin column and ninhydrin post-column derivation in auto analyzer (Shimadzu SPD-6AV UV-visual detector). Samples were analyzed using Table 1. Estimation of ingredients (dry matter) of C. carpio and C. luteus HCl 6N, for 22 h at 110°C to determine amino acids. Parameters C. carpio C. luteus Sinceacidic hydrolysis destroys tryptophan, the samples Weight (g) 250.66 ± 26.59 76.366 ± 3.43 were also analyzed using lithium hydroxide 4N to determine Total length (cm) 25.433 ± 1.23 18.066 ± 0.52 tryptophan. When you combine the replicas together, one Standard Length (cm) 20.233 ± 2.13 15.233 ± 0.54 value for each material was obtained and used. The tests Depth of body (cm) 8.10 ± 0.58 5.466 ± 0.12 were conducted at the laboratories of the Ministry of Science and Technology, Baghdad. Relative Volume 0.403 ± 0.01 0.359 ± 0.01 Statistical analysis: Use the statistical program Statistical Crude protein (% w/w) 22.593 ± 0.2b 26.121 ± 0.17a Analysis System (SAS 2012). The mean differences Crude fat (% w/w) 4.044 ± 0.13a 2.173 ± 0.11b between the averages were calculated by Duncan (1955). Crude Ash (% w/w) 2.961 ± 0.06a 1.503 ± 0.13b Dray matter 28.993 ± 0.21a 28.846 ± 0.19a RESULTS AND DISCUSSION Moisture (% w/w) 71.006 ± 0.21a 71.153 ± 0.19a Proximate composition: The proximate composition a, b The different letters in column means that significant differences (P <0.05) Amino Acid Profile for Freshwater Fishes Yellow Barbell and Common Carp 231

Table 2. Amount of amino acids of total protein Table 3. Proportion of amino acids in total amino acids -1 No. Amino acids mg g C. carpio C. luteus No. Amino acids (%) C. carpio C. luteus 1 Arginine 5.94a 10.95a 1 Arginine 2.626 ±0.41a 4.192±0.87a 2 Histadine 8.12a 10.41a 2 Histadine 3.597±0.21a 3.986±0.78a 3 Isoleucine 10.37b 12.85a 3 Isoleucine 4.591±0.17a 4.922±0.05a 4 Leucine 15.8a 16.085a 4 Leucine 6.996±0.19a 6.157±0.63a 5 Phenyl al. 5.00a 7.78a 5 Phenyl al. 2.214±0.38a 2.980±0.96a 6 Threonine 14.03a 13.4a 6 Threonine 6.209±0.28a 5.131±0.93a 7 Tryptophan 22.39a 30.54a 7 Tryptophan 9.910±0.82a 11.691±1.70a 8 Valine 1.64a 1.47a 8 Valine 0.727±0.04a 0.563±0.11a Sum ΣEssential amino acids (EAA) 83.31a 103.51a Σ EAA 36.875±0.99a 39.626±3.21a 9 Aspartic 16.72b 36.65a 9 Aspartic 7.403±0.50b 14.031±2.24a 10 Serine 13.11a 15.31a 10 Serine 5.802±0.88a 5.864±0.36a 11 Glutamine 18.57a 16.99a 11 Glutamine 8.222±0.60a 6.504±0.81a 12 Glycine 29.63a 20.78b 12 Glycine 13.115±0.82b 7.955±0.16a 13 Alanine 7:00 AM 8.9a 13 Alanine 3.099±0.15a 3.409±0.26a 14 Tyrosine 8.62a 6.59a 14 Tyrosine 3.817±0.31a 2.526±0.54a 15 Glutamic, A. 13.15a 9.95b 15 Glutamic,A. 5.821±0.24a 3.811±0.37b 16 Ornithine 8.84a 7.55a 16 Ornithine 3.916±1.26a 2.893±0.62a 17 Asparagin 12.82a 15.54a 17 Asparagin 5.674±0.37a 5.952±0.51a 18 Taurine 14.12a 19.39a 18 Taurine 6.250±0.32a 7.424±0.79a Sum Σ Non Essential amino acids 142.62a 157.7a Σ Non EAA 63.124±0.99a 60.373±3.21a Sum Total amino acids 225.39 261.21 Sum EAA/NonEAA 0.584±0.02a 0.666±0.09a EAA/NonEAA 0.584a 0.666a a, b The different letters in column means that significant differences (P <0.05) a, b The different letters in column means that significant differences (P <0.05) and 39.62% for C. luteus showed significant difference at 5% (Deutz et al 1992). The results showed that the amount of level of significance between species percentage of each amino acids in C. luteus is higher than in C. carpio and this is amino acid of total amino acids are shown in Table 3. The due to the nature of nutrition of each species. Where C. luteus highest percentage of amino acids were aspartic acid, relies heavily on natural food while C. carpio feeds on glycine, tryptophan and glutamine spread from 8.22-14.03% commercial foods (Fadaee 2012). of total protein. While the ratios of other amino acids 0.72 to 13.11% in C. carpio and 0.56 to 14.03% in C. luteus. CONCLUSION Essential amino acids cannot be manufactured within To sum-up, fishes are considered an important source of the human body so they get them in a large and balanced human food in Iraq, because it contains a high percentage of intake of fish meat, they are important for healing wounds protein and a good source of amino acids, especially (Mat Jais et al 1994). The present study indicated that both essential amino acids, which is in good quantity for good types contain most of the essential amino acids. health and prevention of diseases in humans. Obstruction in the healing process occurs when there is a lack of essential amino acids (Mat Jais et al 1994). To REFERENCES ensure the formation of hemoglobin and regulate the stability Ackman RG 1990. Seafood lipids and fatty acids. Food Reviews International 6(4): 617-646. of sugar and energy in the blood, the body needs Isoleucine. Al-Daham NK 1977. Fishes of Iraq and the Arabian Gulf. Volume 1. While leucien plays a major role in the healing of skin, Squaliformes to Atheriniformes. Publication 9, Centre for Arab boneand muscle tissue, and for the formation of human Gulf Studies, University of Basrah. 546 pp. (In Arabic). glycogen, Glycine is considered to be a key ingredient Al-Temimy LM 2004. Ecology, biology and assessments of fish community in Euphrates River Near Al-Mussaib Power Station. (Heimann 1982, Witte et al 2002). Ph.D. Thesis, College of Agriculture, University of Basrah. 147 The importance of Glutamine is concentrated in its work pp. (In Arabic) as an important carrier of ammonia (nitrogen) in the immune Al-Abbadi and Khalil Ibrahim 2010. Practical applications on the artificial propagation of fish and the management of hatcheries system of the muscles in the case of dangerous diseases. book,182 p. 232 Kadhim O.M. AL. humairi, Hamza G. Al-Agidi and Sadeq M. Al-Haider

Al-Amari, Moayad Jasim Yas, Al-Tai, Meason Mahdi Saleh and Al- Kibria G, Nugegoda D, Fairclough R, Lam P and Bradly A 1997. Sultani and Dargham Ali Abbas 2012. Studying the composition Zooplankton it's biochemistry and significance in aquaculture. of the fish community and some environmental evidence in the Naga International Center for Living Aquatic Resources Euphrates River in the city of Hindi / Iraq. Journal of Babylon Management 20: 8-14. University Pure and Applied Sciences 20(5): 35-39. Kocour M, Mauger S, Rodina M, Gela D, Linhart O and Vandeputte M Al-Amari, Moayad Jassim; Al-Tai, Meason Mahdi Saleh, Rashid and 2007. Heritability estimates for processing and quality traits in Karim Hamid 2012. Some life and environmental aspects of common carp (Cyprinus carpio L.) using a molecular pedigree. Barbusluteus (Heckel) fish in the Hilla River. Journal of Aquaculture 270(4-1): 43-50. Babylon University of Pure and Applied Sciences 22(1): Limin L, Feng X and Jing H 2006. Amino acids composition Al-Hamed MI 1971. Salinity tolerance of common carp (Cyprinus difference and nutritive evaluation of the muscle of five species carpio L.) Bulletin of the Iraq Natural History Museum 64:1-30. of marine fish, Pseudosciaena crocea (large yellow croaker), Al-Rubaie.Khalaf Hanoun 1986. The first scientific experiment to Lateolabrax japonicas (common sea perch), Pagrosomus raise carp fish in the area of Basra. University of Basrah Journal major (red seabream), Seriola dumerili (Dumeril's amberjack) 2(3-4): 237-244. and Hapalogenysnitens (black grunt) from Xiamen Bay of China. Aquaculture Nutrition 12(1): 53–59. Coad WB 2017. Freshwater fishes of Iraq. Retrieved from www.briancoad.com . Ljubojevic D, Cirkovic M, ĐorĊevic V, Puvaca N, Trbovic D, Vukadinov J and Plavsa N 2013. Fat quality of marketable Craig S and Helfrich A 2002. Understanding fish nutrition, feeds, and fresh water fish species in the Republic of Serbia. Czech feeding. *Extension Specialists, Virginia-Maryland College of Journal of Food Sciences 31(1): 445-450. Veterinary Medicine, and Department of Fisheries and Wildlife Sciences, Virginia Tech, respectively. doi=10.1.1.559.6121 Martini R and Lindberg C 2013. Fishing for Tomorrow: Managing &rep=rep1&type=pdf fisheries for sustainable development. Organization for Economic Co-operation and Development 2: 12. Deutsch L, Graslund S, Folke C, Troell M, Huitric M, Kautsky N and http://dx.doi.org/10.1787/888932860218. Lebel L 2007. Feeding aquaculture growth through globalization: exploitation of marine ecosystems for fishmeal. Mat Jais AM, McCulloh R and Croft K 1994. Fatty acid and amino Global Environmental Change 17: 238-249. acid composition in haruan as a potential role in wound healing. General Pharmacology 25: 947-950. Deutz NEP, Reijven PLM, Athanasas G and Soeters PB 1992. Post- operative changes in hepatic, intestinal, splenic and muscle Jais AM, McCulloch R and Croft K 1994. Fatty acid and aminoacid fluxes of amino acids and ammonia in pigs. Clinical Science 83: composition in haruan as a potential role in wound healing. 607-614. General Pharmacology 25(2): 947-950. Dezhabad A, Dalirie MS and Toudar S 2012. Amino acid profile of Mohammed AM, Hussein SA and Mutlaq MF 2015. Some biological kutum (Rutilus frisii), silver carp (Hypophthalmicthys molitrix) traits of Barbus luteus (Heckel 1843) in Al Hammarmarshe, and rainbow trout (Oncorhynchus mykiss). African Journal of Iraq. Jordanian Journal of Agricultural Sciences 11(2): 551- Agricultural Research 7(34): 4845-4847. 563. Duncan DB 1955. Multiple range and multiple F-test. Biometrics 11: Ovissipour M, Abedian AM, Motamedzadegan A, Rasco B, Safari R 4-42. and Shahiri H 2009. The effect of enzymatic hydrolysis time and temperature on the properties of protein hydrolysates from Fadaee R 2012. A review on earthworm Esieniafetida and its the Persian sturgeon (Acipenser persicus) viscera. Food applications. Annals of Biological Research 3(5): 2500-2506. Chemistry 115(1): 238-242. FAO United Nations Food & Agriculture Organization 2005. SAS 2012. Statistical Analysis System, User's Guide. Statistical. Nutritional elements of fish. FAO, Rome. Version 9.1th ed. SAS. Inst. Inc. Cary. N.C. USA Hasan MR, Hecht T, De Silva SS and Tacon AGJ 2007. Study and Sikorski ZE 1996. Charakterystyka białek – glownych surowcow analysis of feeds and fertilizers for sustainable aquaculture żywnosciowych. W: Chemiczne i Funkcjonalne Własciwosci development. FAO Fisheries Technical Paper No. 497. Rome, Składnikow Żywnooci. Red. Sikorski Z. E., WNT, Warszawa, FAO. 510 pp. (available at: www. extension.org 286-318 (in polish). /mediawiki/files/5/54/Studyandanalysis.pdf). Stankovic BM, Dulic PZ and Markovic ZZ 2011. Protein sources and Hassouni Khalid Mohammed and Janabi Ali Hadi 2005. The reality of their significance in carp (Cyprinus carpio). Nutrition Journal of fish culture in Basrah Governorate. Iraq, Iraqi Journal of Agricultural Science 56(1): 75–86. Agriculture 2: 155-164. Vidotti RM, Viegas EMM and Carneiro DJ 2003. Amino acid Heimann W 1982. Fundamentals of Food Chemistry, pp. 27-233. composition of processed fish silage using different raw Avi Pub. Co., West Port, Connecticut, U.S.A. materials. Animal Feed Science and Technology 105(1-4): Kenari AA, Regenstein JM, Hosseini SV, Rezaei M, Tahergorabi R, 199–204. Nazari RM, Mogaddasi M and Kaboli SA 2009. Amino acid and Witte MB, Thornton FJ, Tantry U and Barbul A 2002. L-Arginine fatty acid composition of cultured beluga (Husohuso) of supplementation enhances diabetic wound healing: different ages. Journal of Aquatic Food Product Technology Involvement of the nitric oxide synthase and arginase 18(3): 245-26. pathways. Metabolism 51(10): 1269-1273. Received 30 September, 2019; Accepted 04 November, 2019 Manuscript Number: S-287 Indian Journal of Ecology (2019) 46 Special Issue (8): 233-236 NAAS Rating: 4.96

Using Mixed Model to Detect the Effect of Genetic and Fixed Factors on Milk Production in Awassi Sheep

M.A. AL-Shaikh, W.J. AL-Khazraji, WafaaI. AL-Samaraai and L.J. AL-Jaryan Department of Animal Production, College of Agricultural Engineering Science, University of Baghdad, Iraq E-mail: [email protected]

Abstract: The study was conducted on 100 Turkish and local Awassi ewes reared in the ruminants' research station, which belong to state of Agricultural Fundamental, Ministry of Agriculture, Iraq. Data were available during the season 2017-2018to determine the relationship of some genetic and non-genetic factors on milk quantity and lactation period length in Turkish and Local Awassi ewes. Results showed a significant variance of dam age and dam weight on lactation period (LP), the mean squares were 2853.36 and 2658.48, respectively while a high significant effect of dam age and weight on total milk yield 8944.27 and 4462.92, respectively. Significant effect of breed on total milk yield (TMY) and daily milk yield (DMY), the high values were noticed in Turkish Awassi breed (94.64 and 0.904 kg) compared with Local Awassi breed (78.41 and 0.784 kg). Damage at lambing effected significantly on lactation period, total milk yield and daily milk yield, the highest values were noticed in dams with fife year of age namely, 114.84 day, 103.58 kg and 0.952 kg, respectively. Total milk and daily milk yield were differed significantly according to lambing type, the yield increased in dams with twin birth lambs (102.16 and 0.933 kg, respectively) compared with dams with single birth lambs. Heritability coefficients estimated for DMY, TMY and LP were low and ranged from 0.22 to 0.08 while a significant and positive genetic correlation among these traits and ranged from 0.96 to 0.67. Significant and positive phenotypic correlations among milk traits and lactation period ranged from 0.98 to 0.81.

Keywords: Sheep, Milk, Genetic/non-genetic parameters, Heritability, Correlation

Awassi sheep breed is one of most famous and Dam age, parity, lactation stage, sex of birth and type of important sheep breeds around the word, its widespread in birth were effected significantly on the major milk the most of countries in Asia and middle east (FAOSTAT components such as protein and fat content in Awassi sheep 2007). Awassi is contributing about 58.2% from Iraqi sheep breed reared in Iraq (AL-Khuzai and AL-Anbari 2018). In and the productive performance is faced with many addition to environmental factors, genetic variation in sheep difficulties under the classical breeding systems (Ishaq and breeds has been widely documented and the evidences Ajeel 2013). Sheep milk is characterized by lower cholesterol show substantial differences among breeds and in a number content, more beneficial composition of fatty acids and better of cases, exceptional variations within breeds (Ghaffari et al hygiene (lower somatic cell content) compared with cow of 2009, AL-Khuzai and AL-Anbari 2018.) buffalo or cow milk (Molik 2008). The major aim of this study is to investigate the effect of Many studies referred that the milk yield of this sheep is environmental and genetic factors, which effect on milk yield affected by many factors and these factors can be classified and components in Awaasi breeds reared in Iraq and uses in to two parts: first part is environmental factors such as food the results as a guidelines to improve the Animals and which effect significantly in sheep performance and the increase the outcome for benefits. weather factors such as temperature and humidity which play a crucial role in sheep survival and activity. Abeia et al (2017) MATERIAL AND METHODS reported that the milk yield is associated metrological The study was conducted on 100 Turkish and local conditions. Second part is genetic factors represented in Awassi ewes reared in the ruminants research station which many genes which make a large genetic variance among belong to state of Agricultural fundamental - Ministry of flock individuals and this variance can be exploited in Agriculture, Iraq. Data were available during the season selection programs (Hamann et al 2004, Selvaggi et al 2017.) 2017-2018. Semi open sheds were used to housing animals Sheep will begin for milking from lambing to fall when the and management program was used to feeding and shortening day length decreases milk production and many preparing to reproduction. Flock can be fed on the studies indicated that the sheep milk yield and components concentrated ration consuming about 500-600 gm head-1 are different considerably among sheep breeds (Belo 2009). day-1, for the period from mating season to the last six weeks Management factors are also effect in milk yield such as of pregnancy. Ration is normally containing 37% yellow corn, nutrition (Gilbert 1992.) 40% wheat bran, 10% hulled barley, 5-10% soybean meal, 234 M.A. AL-Shaikh, W.J. AL-Khazraji, WafaaI. AL-Samaraai and L.J. AL-Jaryan

1% NaCl and 1% CaCO3 and green roughages such as Table 1. Analyses of variance of factors that effect on milk Alfalfa and clover can be added throughout the season. traits S.O.V. d.f Mean squares Annual routinely operations on sheep are dipping and washing with chemicals in order to kill extra parasites so LP TMY DMY sheep will be ready to mating after hand wool shaving. Sires Breed 1 136.49 NS 6261.84 * 0.4473 * and dams will be recorded in breed records. Lambs are Dam age 1 2853.36 * 8944.27 ** 0.8038 ** weighed directly after parturition and tagged with plastic tags. Lambing type 1 72.49 NS 4477.91 * 0.3485 * Lambs stays with their dams up to 90 days (weaning Lamb sex 1 19.82 NS 1306.50 NS 0.1652 NS age).The health status of the flock must be under regular Lambing status 1 158.63 NS 179.72 NS 0.0239 NS observations. Dam weight 2 2658.48 * 4462.92 ** 0.4178 ** Milk measures: Milk yield was measured twice-daily every Lambing month 1 52.73 NS 2592.03 * 0.4048 * week. On the milk recording day, lambs were isolated from Exp. Error 91 571.487 369.694 0.02572 their dams, and dam's udders were evacuated handily from ** (P<0.01), * (P<0.05) the surplus milk post lambs suckling, and milk collected was weighed and recorded. 0.904 kg) compared with Local Awassi breed (78.41 and Statistical analysis: General liner model (GLM) was used 0.784 kg). The results are similar with those of Hamdon by statistical analysis system- SAS (2001) computer program (2005), Iñiguez and Hilali (2009) and Abdallah etal (2013) to detect the effect of environmental and genetic factors on who reported that the milk yield and components are affected milk yield, lactation period length and milk components under significantly by breed differences. In addition, the Turkish the mixed model below: Awassi individuals characterized as bigger in size than Local Yijklmnopq= µ+ Bi + Aj + Tk + Sl + Hm + Wn + No + Rp + Awassi breed which reflex positively on milk production. eijklmnopq Dame age at lambing effected significantly on lactation Where: period, total milk yield and daily milk yield, the highest values µ: over all mean were noticed in dams with fife year of age namely, 114.84 Bi: effect of breed (Turkish Awassi and Local Awassi) day,103.58 kg and 0.952 kg, respectively. The results are in Aj: effect of dam age at lambing (3 and 4 year) accordance with the results of Alkudsi and Ibraheem (2014). Tk: effect of lambing type (single and twin) Many possible causes could be suggested to explain the Sl: effect of lamb sex (male and female) increase of milk yield as the damage progressed. Firstly, the Hm: effect of lambing status (normal or difficult) increase body weight of the ewes with a greater number of Wn: effect of dam weight at lambing (less than 50 kg, 50- lactations, which leads to a greater availability of body 60 kg or more than 60 kg) reserves for the synthesis of milk. Secondly, the greater No: effect of lambing month (January or March) development of the udder glandular tissue as the number of Rp: effect of sire. lactations rises could also result in an increased synthesis of eijklmnopq: is a random error. milk constituent, the results came to support the past results of van der Linden et al (2009). Total milk and daily milk yield RESULTS AND DISCUSSION were differed significantly according to lambing type, the Results represented in Table 1 indicated that a yield increased in dams with twin birth lambs (102.16 and significant variance (P<0.05) of dam age and dam weight on 0.933 kg, respectively) compared with dams with single birth lactation period (LP), the mean squares were 2853.36 and lambs. The current results came harmonic with many results 2658.48 respectively while a high significant effect (P<0.01) that demonstrate the effect of birth type on many economical of dam age and weight on total milk yield (TMY) namely, traits in sheep such as milk production, lambs weight or 8944.27 and 4462.92, respectively. In the same direction, livability (Molik et al 2008). Although there was no significant dame age and weight effected significantly on daily milk yield difference between dams with male or female lambs in milk (DMY) namely, 0.8038 and 0.4178, respectively. Results production but there is a theoretical difference towards dams showed a significant effect (P<0.05) of breed, lambing month that produced male lambs. Lambing status did not affect and lambing type on lactation period, total milk yield and daily significantly on milk yield in both Awassi sheep breeds. Dm milk yield. weight effected significantly on LP, TMY and DMY, dams with Results represented in Table 2 showed a significant 50-60 kg recorded the highest values namely, 111.30 day, effect of breed on total milk yield and daily milk yield, the high 104.95 kg and 0.943 kg, respectively. The significant values were noticed in Turkish Awassi breed (94.64 and relationship of dam weight with milk production was Milk Production in Awassi Sheep 235

Table 2. Effect of factors that studded on lactation period and milk yield Factors d.f Means ±S.E LP (day) TMY (kg) DMY (kg) Overall mean 100 103.18 ± 8.75 89.97 ± 19.51 0.872 ± 0.26 Breed Local Awassi 38 100.01 ± 8.66 a 78.41 ± 17.75 b 0.784 ± 0.26 b Turkish Awassi 62 104.69 ± 10.61 a 94.64 ± 25.16 a 0.904 ± 0.28 a Dam age (year) 4 65 94.52 ± 11.78 b 73.06 ± 14.49 b 0.773 ± 0.35 b 5 35 114.84 ± 7.66 a 103.58 ± 23.45 a 0.952 ± 0.26 a Lambing type Single 71 97.46 ± 8.19 a 74.45 ± 17.84 b 0.764 ± 0.19 b Twin 29 109.50 ± 9.46 a 102.16 ± 18.43 a 0.933 ± 0.24 a Lamb sex Male 56 106.58 ± 8.38 a 88.57 ± 16.52 a 0.831 ± 0.16 a Female 44 99.61 ± 8.50 a 85.27 ± 21.25 a 0.856 ± 0.22 a Lambing status Normal 91 104.92 ± 7.39 a 90.33 ± 17.54 a 0.861 ± 0.19 a Difficult 9 98.43 ± 9.02 a 82.48 ± 15.27 a 0.838 ± 0.17 a Dam weight Less than 50 kg 24 89.25 ± 7.35 b 69.26 ± 11.70 b 0.776 ± 0.14 b 50-60 kg 52 111.30 ± 10.59 a 104.95 ± 26.92 a 0.943 ± 0.16 a More than 60 kg 24 105.48 ± 8.80 a 98.83 ± 18.38 a 0.937 ± 0.16 a Lambing month January 41 98.37 ± 7.62 a 82.92 ± 20.17 b 0.843 ± 0.18 b March 59 105.92 ± 7.55 a 102.32 ± 17.69 a 0.966 ± 0.18 a Values within each subclass with different superscripts differ significantly. demonstrated in many past studies (Hernández et al 2018), Table 3. Genetic parameters of milk yield and lactation period who reported that that the milk production increased in rate in Awassi sheep between 2.99 to 3.91 liter as body weight increased one Traits DMY TMY LP kilogram. Results reveal inequality in total milk and daily milk DMY 0.22 0.96** 0.67** according month of birth, dams that lambed in March TMY 0.98** 0.22 0.80** produced milk more than dams that lambed in January and LP 0.81** 0.86** 0.08 this result is in accordance with the results of Abdul-Rahman Heritability-h2 (diagonal), genetic correlation (above diagonal) and phenotypic and Abbo (2013) and AL-Samari et al (2014). The results are correlation (below diagonal) in accordance with the finding of Raoof et al (2017) who as Mavrogenis(1996) and AL-Samarai and AL-Anbari (2009), reported that the non-genetic factors such as dam age, parity, who reported that heritability estimates were 0.47 and 0.56, dam weight and birth type are correlated with milk yield in respectively. The yield traits were a highly genetically and sheep. phenotypic correlated one another and the results were in Results represent in Table 3 showed a heritability accordance with Carta et al (2009) and Murphy et al (2017). coefficients estimated for DMY,TMY and LP were low and In conclusion, local Awassi sheep breed is characterized with ranged from 0.22 to 0.08, while a significant (P<0.01) and low milk production in Iraq therefore, detecting the actual milk positive genetic correlation among these traits and range yield with high accurate records and diagnosis the main from 0.96 to 0.67.Significant (P<0.05) and positive environmental and genetic factors that effect on milk yield will phenotypic correlations among milk traits and lactation contribute for developing the breed performance and period ranged from 0.98 to 0.81. Similar results were found. A continue to become a main breed to compete strongly with higher heritability estimates of TMY in the same breed such the standard sheep milk breeds. 236 M.A. AL-Shaikh, W.J. AL-Khazraji, WafaaI. AL-Samaraai and L.J. AL-Jaryan

REFERENCES Hamann H, Horstick A, Wessels A and Dist O 2004. Estimation of Abd Allah M, Abass SF and Allam FM 2013. Factors affecting the milk genetic parameters for test day milk production, somatic cell yield and composition of Rahmani and Chios sheep. Global score and litter size at birth in East Friesian ewes. Livestock Journal of Dairy Science 1: 53-59. Production Science 87: 153-160. Abdul-Rahman FY and Abbo NY 2013. Non-genetic factors affecting Hernández G, Schilling SR, Vera Arias MA, Lonso RAA, Ramírez milk yield, some constitute and lactation period of Awassi ewes. Pérez AH and González Ronquillo M 2018. Effect of live weight Tikrit Journal for Agricultural Sciences 13(2): 61-69. pre- and post-lambing on milk production of East Friesian sheep. Italian Journal of Animal Science 17(1): 184-194. Abeia JA, Garcia A and Riocerezo CP 2017. The effect of weather on milk production in dairy sheep vary by month of lambing and Ingiguz L and Hilali M 2009. Evolution of Awassi genotypes for milk lactation phase. Journal of Animal Behaviour and selection in Syria. Livestock Science 120(3): 232-239. Biometeorology 5: 56-65. Ishaq MA and Ajeel HM 2013. Reproductive traits of local and Turkish AL-Khuzai HM and AL-Anbari NN 2018. Relationship of POU1F1 Awassi sheep in semi intensive breeding circumstances. The gene polymorphism with some of economical traits in Iraqi Iraqi Journal of Agricultural Sciences 5: 615-623 Awassi ewes. Journal of Entomology and Zoology Studies 6(2): Mavrogenis AP 1996. Estimates of environmental and genetic 2082-2085. parameters influencing milk and growth traits of Awassi sheep Alkudsi NH and Ibrahim AH 2014. Effect of age on economic traits of in Cyprus. Small Ruminant Research 20: 141-146. improved Awassi sheep. The Iraqi Journal of Agricultural Molik E, Murawski M, Bonczar G and Wierzchoś E 2008. Effect of Science 45(3): 29-294. genotype on yield and chemical composition of sheep's milk. Al-Samarai FR, Abdul-rahman YK, Mohammed FA, Al-Zaidi FH and Animal Science Papers and Reports 26: 211-218. Al-Anbari NN 2014. Influence of some genetic and non-genetic Murphy TW, Berger YM, Holman PW, Baldin M, Burgett RL and factors on total milk yield and lactation period in Iraqi Awassi Thomas DL 2017. Estimates of genetic parameters, genetic sheep. Advances in Animal and Veterinary Sciences 2(12): trends, and inbreeding in a crossbred dairy sheep research 662-667. flock in the United States. Journal of Animal Science 95: 4300- Al-Samarai FR and Al-Anbari NN 2009. Genetic evaluation of rams 4309. for total milk yield in Iraqi Awassi sheep. ARPN Journal of Pettigrew EJ, Hickson RE, Morris ST, Lopez-Villalobos N, Pain SJ, Agricultural and Biological Science 4(3): 54-57. Kenyon PR and Blair HT 2019. The effects of birth rank (single Belo AT, Belo C, Fontes AL, Martins AP, Pereira EA and Vasconcelos or twin) and dam age on the lifetime productive performance of MM 2009. Effect of sheep breed on milk composition and female dual purpose sheep (Ovisaries) offspring in New coagulation properties. Options Méditerranéennes: Série A. Zealand. PLoS ONE 14(3): e0214021. Séminaires Méditerranéens 91: 221- 225. Raoof SO, MahmudKI, Abdul Kareem AA and Mouhammad KG Carta A, Casu S and Salaris S 2009. Invited review: Current state of 2017. Estimation of the best linear unbiased prediction (BLUP) genetic improvement in dairy sheep. Journal of Dairy Science of Kurdi rams for average daily milk yield depending on the 92: 5814-5833 progeny. The Iraqi Journal of Agricultural Science 6: 1399- FAOSTAT: Online: http:// faostat.fao.org/. FAOSTAT© FAO Statistics 1404. Division 2007, Accessed: 23 August 2007. SAS 2001. SAS / STAT `Users` Guide for Personal Computers. Ghaffari M, Nejati JA and Rahimi G 2009. Detection of polymorphism Release 6.12.SAS Institute Inc., Cary, NC., USA. in BMPR IB gene associated with twining in Shal sheep using Selvaggi M, D'Alessandro AG and Dario C 2017. Environmental and PCR-RFLP method. International Journal of Agricultural genetic factors affecting milk yield and quality in three Italian Biology 11: 97-99. sheep breeds.​ Journal of Dairy Research 1: 27-31. Gilbert G 1992. Principles of machine milking of dairy sheep. In Van der Linden DS, Kenyon PR, Blair HT, Lopez-Villalobos N, sheep dairying. The manual. Editors S.T. Dawe and Dignand Jenkinson CM, Peterson SW and Mackenzie DD 2009. Effects NSW Agriculture. of ewe size and nutrition on fetal mammary gland development Hamdon HAM 2005. Chios and Farafra sheep conditions. Ph.D. and lactational performance of offspring at their first lactation. Thesis. Journal of Animal Science 87(12): 3944-3954. Received 24 September, 2019; Accepted 15 October, 2019 Manuscript Number: S-288 Indian Journal of Ecology (2019) 46 Special Issue (8): 237-241 NAAS Rating: 4.96

Seasonal and Physiological Changes on Renal Function Parameters in Iraqi Cats

Ahmed Noaman Awad AL-Ani, Taha Yassin Ghani Al-Badrawi, Basil Razook Faraj Razook1 and Naseir Mohammed Badawi Department of Internal and Preventive Veterinary Medicine, 1Zoonoses Unit, College of Veterinary Medicine, University of Baghdad-3, Iraq E-mail: [email protected]

Abstract: This study was conducted on 90 healthy local Iraqi cats and recorded the normal range of serum chemical values (blood urea nitrogen and serum creatinine) and urine parameters in the pets and stray cats, and studied the effects of sex, age and seasons on these parameters. The results showed there were no effect of the housing mode or seasons on the specific gravity, PH, total protein, WBC, RBC, ketones, urobilinogen and glucose in urine of cats. While significant increase (p ≤ 0.05) in RBC of urine occurred in the female compare to male. Seasonally the blood values were significantly higher (p ≤ 0.05) in creatinine and total protein values in summer compare to winter. On other hand the serum chemical results (creatinine, urea, total protein and albumin) of local cats are classified according sex and age which was not recorded any significant changes. In conclude we found renal function changes are affected by seasons, and the blood urine showed high frequency occurrence in female cats. This study was determining normal range of kidney function test computable for local breed and environmental conditions in Iraq.

Keywords: Renal function, Seasons, Age, Sex, Pets and stray cats, Iraqi cats

Renal problems diagnosed as a part of more common diagnose CKD in early stage when serum creatinine is feline diseases (Wakeling 2009). In spite of desert root of cats normal (Hall et al 2019). Zenad et al (2015) confirmed on the increased its urinary system resistance due to highly urine role seasonal variation induced some diseases in the Iraqi specific gravity (Brown 2010). Domestic cats had incorporeal cats and the seasons considered essential factor effect on value to owners so it reach 600 million that's lead them to the healthy dogs and cats in Iraq. Quantification of glomerular taken care of pets even with untreatable health issue. filtration rate (GFR) is considered the most appropriate Chronic kidney disease (CKD) is a degenerative and method for evaluating kidney function and it is reflected progressive disease that is common in cats, no known amount of normal kidney mass. Nevertheless, due to effective treatments that treated this disease progression difficulty of sample colleting and requires specific equipment and repair kidneys diseases have been identified (Thomson that make GFR measurement isn't practical techniques (Kerl et al 2019). Diagnosed renal disease is traditionally done with and Cook 2005). Its need to collect urine for a day with blood urea nitrogen, serum creatinine concentration (sCr), several blood samples to compare renal with plasma and urinalysis findings. Becker et al (2008) showed the clearance rate that make GFR expensive and unwieldy (Von importance of blood and biochemical studies as basic sings Hendy-Willson and Pressler 2011). Cheap and Easily to reflect health and physiological state of animal. Cats with measuring the concentration of serum creatinine (sCr) in kidney disease normally appear non-specific signs, routine serum make it standard method to assess function of kidney, laboratory tests as blood creatinine and blood urea may be sCr is a derivative of muscle endogenous metabolism make but not sensitive and enough for CKD detection, removed by the kidneys, due to the tubules didn't reabsorbed ultrasonography for kidney can showed information on it. So used as marker of kidney dysfunction (Hall et al 2014). kidney parenchyma that is very important for detecting the Urea is as a waste product and excrete rate interfere with primary causes of chronic kidney disease (Thanaboonnipat dehydration, protein loss, catabolism and food intake et al 2019). Recently, Many cats kidney healthy evaluated (Cannon 2016). Azotemia (elevation of sCr and BUN levels with Symmetric Dimethylarginine (SDMA) concentrations together) with decrease urine concentrating capability occurs that indicated on prevalence of CKD with 28 percent, high in state of chronic kidney disease (CKD) (Hall et al 2014). prevalence of percent with age (67 in cats >18 years), cats Persistence of increase sCr with decrease urine specific can be measured with serum SDMA concentrations in CKD gravity (USG) under 1.03 for several months it remarks CKD rather than creatinine concentrations alone, serum SDMA (Cannon 2016). The International Renal Interest Society 238 Ahmed Noaman Awad AL-Ani et al

(IRIS) with the American and European Societies of statistical program (SPSS program version 20), One-Way Veterinary Nephrology and Urology developed outline stage ANOVA for compare the data at level significance of of Feline CKD (Hall et al 2019). Acute uremia occurs in (P<0.05). sudden damage of kidney parenchyma that called Acute kidney injury and may followed by acute renal failure RESULTS AND DICUSSION ((Langston 2010), and cause increase morbidity and The results of chemical and physical urine examination mortality rates due to late disease diagnosis (Kovarikova of cat showed the mean value of specific gravity, pH, total 2015). This study was aimed to determine the normal range protein, WBC, RBC, ketones, urobolliongen and glucose in of kidney function in local breed and effect the environmental urine has no significant difference between stray and pet cats conditions on mentioned parameters in Iraq by study serum as well as no influence of the season on these parameters. chemical values (blood urea nitrogen and serum creatinine) The glucose in urine cat revealed non-significant increase in and urine parameters in Iraqi cats. the summer compare to the winter (Table 1). The chemical and physical examination of urine in (Table MATERIAL AND METHODS 2) according sex and age groups showed significant increase Specimen collection and analysis: The study was in RBC μl-1 in males urine as compared to females. The conducted on 90 local cats in Baghdad province. The cats did ketones in female and in catsup to 5 years, were also non- not showed any clinical signs of acute or chronic kidney significant .The non-significant increase in glucose value was diseases, and subdivided to groups according to housing observed in urine of male cats compare to female and in mode (pet and stray), season (summer, April to October and young age group (1-5 years) as compared to other age group winter, November to March), sex (male and female), and age (6-15 years). groups (1-5 year and more than 6 year). The blood values were significantly higher in creatinine Animals and samples: Simple local trap techniques were and total protein values in summer compared to winter. made to cat catching for cats in Baghdad city, over 12 months Results of serum chemical values according to housing from. Cats were anesthetized by inhalation (10 ml of mode and season were recorded the range data in total cats Halothan 100% BP stabilized by 0.01% thymol) in closed included blood creatinine, blood urea, blood urea nitrogen, special box from glass (Al-Ani 2012). After cat anesthetized, blood total protein, blood albumin. All these tests showed the neck area was sterilized then 6 ml blood withdrawal non-significant difference between stray and pet cats. directly from jugular vein and puts blood in plain tube and Although, the serum creatinine values in summer and total centrifuged to obtain serum. The urine collected directly by protein values were recorded significant differences (p ≤ disposable aspiration syringe from urinary bladder after 0.05) in summer compare to winter. shaving and sterilization the area using 22 gauge needle. On the other hand, the effect of age and sex on Cats were classified to subgroups according housing mode creatinine, urea, total protein and albumin in serum did not (stray cats 47) pets cat (43), according seasons (in summer have any effect on Iraqi cats. The results of cats were 44) and (winter 46), according sex (male 43) and (female 47), classified according sex and age (Table 4), which not showed and aged groups 50 cats (1-5 years) and 40 cats (6-15 any significant differences between males and females cats years), the age estimated by teeth formula according as well as between age group of 6-15 years and 1-5 years (American Association of Feline Practitioners). (Table 4). Urine commercial strips were used in chemical and The study not recorded any significant difference physical analysis of urine. Protein specific gravity, glucose, between age groups, there are indicator not influence of the pH, urobilinogen, blood, ketones, and bilirubin were done age on values of the serum creatinine and the blood urea using ordinary dipstick strips (Combur urine test/Roche, (Table 4). These results disagree with earlier workers German).Total protein was measured by refractometer (Kovarikova 2015. (Wang et al 2017. Elliott and Barber system(Coles 1986), while other parameters were measured (1998) recorded significant disorder of renal function in cats according to commercial kits.A lbumin measured using with the progress of age especially after 13 years. This may (Biolabo Reagent Company- France), Creatinine by be due to the non-significant differences in results with a (Biomaghreb Company- Tunisia) andblood urea and blood progressive of the age in cats because variation on urea nitrogen in serum by Linear Chemicals Company- environment and nutrition habits of Iraqi cats The low Spain. These were measured by cholorimetric methods in phosphate and low protein foods in the cats decrease the spectrophotometer. renal disease (Geddes et al 2016). The increase protein and Statistical analysis: The results were analyzed with salt on the dietary intake increase the chance of the renal Seasonal and Physiological Changes on Renal Function in Iraqi Cats 239

Table 1. The chemical and physical appearances of urine in the total cat and classified according housing type and seasons (Mean +SE) Parameters Total cat Stray cat Pet cat Summer Winter (n=90) (n=47) (n=43) (n=44) (n=46)

Specific gravity 1.03+0.00 1.03+0.00 1.032+0.00 1.032+0.00 1.030+0.00 pH 6.56+0.07 6.68+0.10 6.43+0.09 6.44+0.08 6.67+0.11 Total protein (g l-1) 447.4+38.6 423.8+51.4 473.3+58.3 420.9+52.9 472.8+56.3 WBCμl-1 446.41+9.4 425.5+24.5 469.3+30.4 441.0+28.6 451.7+26.5 RBCμl-1 2.67+1.10 2.55+1.50 2.79+1.64 2.73+1.60 2.61+1.54 Ketones (mg dl-1) 0-5 0.17+0.1 0.32+0.18 0 0 0.33+0.18 Urobillinogen (mg dl-1) 1.39+0.06 1.32+0.08 1.47+0.08 1.33+0.09 1.46+0.07 Glucose (mg dl-1) 10.0+5.87 17.0+11.10 2.3+1.62 14.8+11.56 5.4+3.20

Table 2. The chemical and physical appearances of cat urine classified according to sex and age groups (Mean+SE) Parameters Sex Age /year Male (n=43) Female (n=47) <1-5.(n=50) <6-15 (n=40) Specific gravity 1.030+ 0.001 1.032+0.001 1.032+0.001 1.030+0.001 pH 6.59+0.096 6.53+0.106 6.54+0.104 6.58+0.096 Total protein (g l-1) 527.0+57.4 374.7+50.2 420.4+50.9 481.3+59.3 WBCμl-1 474.4+26.9 420.9+27.5 432.0+26.3 464.5+28.8 RBCμl-1 0 5.11+2.06 A 2.60+1.42 2.75+1.75 Ketones (mg dl-1) 0.35+0.20 0 0 0.38+0.21 Urobillinogen (mg dl-1) 1.47+0.09 1.32+0.07 1.38+0.07 1.41+0.09 Glucose (mg dl-1) 14.0+11.69 6.4+3.60 8.0+3.60 12.5+12.50 Differences in capital letters horizontally refer to presence of significant deference at (P<0.05).

Table 3. The serum chemical values in the total cat and classified according housing mode and seasons (Mean+SE) Parameter Total cat Stray cat Pet cat Summer Winter (n=90) (n=47) (n=43) (n=44) (n=46)

Creatinine (μ mol l-1) 119 +3.2 118.2+3.4 119.0+5.64 127.9+4.91A 110.5+3.81B Urea (m mol l-1) 8.80+0.23 9.16+0.35 8.41+0.30 9.25+0.34 8.38+0.32 Urea nitrogen mg dl-1 24.70+0.66 25.70+0.99 23.60+0.85 25.94+0.95 23.50+0.90 Total protein (g l-1) 74.14+0.69 74.12+1.02 74.16+0.92 77.22+0.001 71.19+0.84 Albumin (g l-1) 32.94+0.45 33.82+0.60 31.99+0.64 33.30+0.75 32.61+0.51

Table 4. Serum chemical values of cat classified according sex and age groups (Mean+SE) Parameters Sex Age (year)

Male (n=43) Female (n=47) <1-5 (n=50) <6-15(n=40)

Creatinine (μ mol l-1) 122+4.8 116+4.3 117+4.34 120+4.8 Urea (m mol l-1) 8.70+0.35 8.89+0.32 8.87+0.29 8.71+0.39 Urea nitrogen mg dl-1 24.41+0.99 24.96+0.91 24.90+0.82 24.44+1.10 Total protein (g l-1) 74.14+0.69 74.14+0.69 74.14+0.69 74.14+0.69 Albumin (g l-1) 32.94+0.45 32.94+0.45 32.94+0.45 32.94+0.45 240 Ahmed Noaman Awad AL-Ani et al

disease in cats (Reynolds et al 2013), although chronic Journal of veterinary Internal Medicine 30(5): 1690-1702. disease lead to poor appetite especially in cats with CKD Hall JA, Yerramilli M, Obare E, Yerramilli M and Jewell DE 2014. Comparison of serum concentrations of symmetric (Quimby et al 2019). The influence of the sex is agree with dimethylarginine and creatinine as kidney function biomarkers King et al (2007) and Dickerson et al (2017) observed light in cats with chronic kidney disease. Journal of veterinary Internal Medicine 28(6): 1676-1683. enough effect of castration on the renal function and the Hall JA, Yerramilli M, Obare E, Li J, Yerramilli M and Jewell DE 2017. urinary tract normal function. The normal range of total Serum concentrations of symmetric dimethylarginine and protein in this is study (63-91 g l-1), this range very close with creatinine in cats with kidney stones. PLoS One 12(4): 1-11. standard references range (Elliott and Barber 1998), Segev Hall JA, Fritsch DA, Jewell DE, Burris PA and Gross KL 2019. Cats with IRIS stage 1 and 2 chronic kidney disease maintain body -1 et al (2013) recorded 35-89 g l of total protein in acute weight and lean muscle mass when fed food having increased uremia. On the other hand, the albumin present study was caloric density, and enhanced concentrations of carnitine and essential amino acids. Veterinary Recored 184(6): 190. 24.7-43.6 g l-1), while the Bijsmans et al (2015) recorded 30- Jepson RE, Syme HM and Elliott J 2014. Plasma renin activity and 34 g l-1 in chronic kidney diseases and considered normal aldosterone concentrations in hypertensive cats with and level. without azotemia and in response to treatment with amlodipine besylate. Journal of veterinary Internal Medicine 28(1): 144- 153. CONCLUSION Kerl M E and Cook CR 2005. Glomerular filtration rate and renal scintigraphy.Clinical Techniques in Small Animal Practice In conclusion this study was no significant difference 20(1): 31-38. between stray and pet cats as well as no influence of the King JN, Tasker S, Gunn-Moore DA and Nther SG 2007. Prognostic season on the parameters whereas significant increase RBC Factors in Cats with Chronic Kidney Disease and the BENRIC (benazepril in renal insufficiency in cats) Study Group: 906- urine of females. Also, the results concluded increase serum 916pp. creatinine and total protein of cats in summer. Serum Kovarikova S 2015. Urinary biomarkers of renal function in dogs and chemical results of cats were classified according sex and cats: a review. Veterinarni Medicina (Praha) 60(11): 589-602. age not showed any significant differences. In Iraqi future Langston CE 2010. Acute uremia In: Ettinger SJ, editor., Feldman EC, editor., eds. Textbook of Veterinary Internal Medicine. researches must compared their results with this study Philadelphia: Saunders WB: 1955-2115p. instead of foreign universal ranges. Lefebvre HP 2010. Normal kidney function: from small to extra-large. In Proceedings of the 35th World Small Animal Veterinary Congress. June (Vol. 2) https://www.vin.com/doc/ REFERENCES ?id=4516273. Al-Ani AN 2012. Some hematological and biochemical values as O'neill DG, Elliott J, Church DB, McGreevy PD, Thomson PC and relate to anemia in local cats. M.Sc. Thesis, College of Brodbelt DC 2013. Chronic kidney disease in dogs in UK Veterinary Medicine University of Baghdad: 45-46. veterinary practices: prevalence, risk factors, and survival. Al-Kalidi JAO, Hasso SA and Al-Ani ANA 2017. Levels of five Journal of veterinary Internal Medicine 27(4): 814-821. elements in the serum of cats in Baghdad city. Al-Anbar Journal Quimby JM, Benson KK, Summers SC, Saffire A, Herndon AK, Bai S of Veterinary Science 10(1): 34-37. and Gustafson DL 2019. Assessment of compounded Brown SA 2010. Linking treatment to staging in chronic kidney transdermal mirtazapine as an appetite stimulant in cats with disease. In August JR: Consultations in Feline Internal chronic kidney disease. Journal of Feline Medicine and Surgery Medicine. St. Louis: Elsevier Saunders: 475-482 p. 1098612X19851303: 1-8. Becker M, Moritz A and Giger U 2008. Comparative clinical study of Reynolds BS, Chetboul V, Nguyen P, Testault I, Concordet DV, canine and feline total blood cell count results with seven in- Samoedrano CC and Lefebvre HP 2013. Effects of Dietary Salt clinic and two commercial laboratory hematology analyzers. Intake on Body Fluid Volume and Renal Function in Healthy Veterinary Clinical Pathology 37(4): 373-384. Cats. Journal of veterinary Internal Medicine 27: 507-515. Bijsmans ES, Jepson RE, Chang YM, Syme HM and Elliott J 2015. Schrock JW, Glasenapp M and Drogell K 2012. Elevated blood urea Changes in Systolic Blood Pressure over Time in Healthy Cats nitrogen/creatinine ratio is associated with poor outcome in and Cats with Chronic Kidney Disease. Journal of Veterinary patients with ischemic stroke. Clinical Neurology and Internal Medecine 29(3): 855-861. Neurosurgery 114(7): 881-884. Cannon M 2016. Diagnosis and investigation of chronic kidney Segev G, Nivy R, Kass PH and Cowgill LD 2013. A retrospective disease in cats. In Practice 38(3): 2-9. study of acute kidney injury in cats and development of a novel clinical scoring system for predicting outcome for cats managed Coles EH 1986. A Text Book of Veterinary Clinical Pathology. 4-Ed by hemodialysis. Journal of Veterinary Internal Medicine 27(4): WB Sounders Company Philadelphia. London, Toronto: 331- 830-839. 333 p. Sent U, Gössl R, Elliott J, Syme HM and Zimmering T 2015. Dickerson VM, Rissi DR, Brown CA, Brown SA and Schmiedt CW Comparison of efficacy of long-term oral treatment with 2017. Assessment of acute kidney injury and renal fibrosis after telmisartan and benazepril in cats with chronic kidney disease. renal ischemia protocols in cats. Comparative Medicine 67(1): Journal of veterinary Internal Medicine 29(6): 1479-1487. 56-66. Thanaboonnipat C, Sutayatram S, Buranakarl C and Elliott J and Barber PJ 1998. Feline chronic renal failure: Clinical Choisunirachon N 2019. Renal shear wave elastography and findings in 80 cases diagnosed between 1992 and 1995. urinary procollagen type III amino-terminal propeptide Journal of Small Animal Practice 39(2): 78-85. (uPIIINP) in feline chronic kidney disease. BMC Veterinary Geddes RF, Biourge V, Chang Y, Syme HM and Elliott J 2016. The Research 15(54): 1-10. effect of moderate dietary protein and phosphate restriction on Thomson AL, Berent AC, Weisse C and Langston CE 2019. Intra- calcium-phosphate housingostasis in healthy older cats. arterial renal infusion of autologous mesenchymal stem cells Seasonal and Physiological Changes on Renal Function in Iraqi Cats 241

for treatment of chronic kidney disease in cats: Phase I clinical Wang IC, Hsu WL, Wu PH, Yin HY, Tsai HJ and Lee YJ 2017. trial. Journal of veterinary Internal Medicine 33(3): 1353-1361. Neutrophil gelatinase-associated lipocalin in cats with naturally Von Hendy-Willson VE and Pressler BM 2011. An overview of occurring chronic kidney disease. Journal of Veterenary glomerular filtration rate testing in dogs and cats The Veterinary Internal Medicine 31(1): 102-108. Journal 188(2): 156-165. Zenad MM, Badawi NM and Wali AR AJ 2015. Cross sectional study Wakeling J 2009. Feline kidney disease: its symptoms and on cutaneous mycotic infections of dogs and cats in Baghdad. management. The New Zealand Veterinary Nurse 49(1): 6-9. World's Veterinary Journal 5(4): 66-73.

Received 01 December, 2019; Accepted 18 December, 2019 Manuscript Number: S-289 Indian Journal of Ecology (2019) 46 Special Issue (8): 242-245 NAAS Rating: 4.96

Development Perspectives for Biogas Production from Animal Manure in Iraq

Ali Alhelal and Inas Abdulsattar A. Jabbar 1 Ministry of Oil, Iraq; 1Al-Musaib Tech. College, Al-Furat Al-Awsat Tech. University, Iraq E-mail: [email protected]

Abstract: Both the developing and developed worlds have witnessed the rapidly increasing in energy demand since the energy crisis in the beginning of 1970s. Since then many countries, which do not have large reserve of crude oil or natural gas have put much effort into utilizing renewable energy as an alternative. Agricultural operations and livestock production are among the important economic activities in the world. But the great majority of energy demand of the agricultural enterprises is still supplied by conventional energy sources. One of the main renewable energy resources is biomass which has been utilizing by people for over 200 years. The main uses of biogas are for cooking, lighting homes, manure and as fuel for cars. This study aims to investigate the biogas potentials which can be produced by the waste of livestock and poultry in Iraq. Also, giving reliable information to both decision makers and industry developers is another goal for this research. In this study, the amount of animal manures, have been analyzed. Then, the amount of energy that can be generated from this resource has been calculated based on their availability in certain geographical areas.

Keywords: Biogas production, Animal manure, Green environment

Energy demand has been increasing in the last five be used as sources of biogas in the farms (Ayhan 2015). decades which was the catalyst to push other countries to With the case of Iraq, biomass resources are take giant steps in utilizing renewable energy. Climate available in most regions. These resources are change, the main cause of green gas emissions, is another agricultural residues, urban wastes, animal manures, reason of pushing up sustainable development. In addition to and forests residues and by products. Discussed that that, the increasing in oil price, the possibility of oil depletion, there are different sources of renewable energy in Iraq and the exponentially growth of the world pollution have which includes on-shore and off-shore wind power, solar further accelerated the utilization of renewable energy power, and biomass. With regards to biomass resources, (Alhelal 2015). There is increasing consensus in both the also addressed the possibility of producing bio-ethanol scientific and political communities that significant reductions from dates, sugar cane, and corn (Chaichan 2012). As an in greenhouse gas (GHG) emissions are necessary to limit initial investigation, it is worth noting that a portion of the data the magnitude and extent of climate change. Renewable used in this paper has been collected from the energy systems already reduce GHG emissions from the literatures published in the areas of agriculture energy sector, although on a modest scale. Most long-term resources, crops production, animal manure, forests energy projections show that renewable energy will play a residues, and domestic and industrial waste (Jaradat major role in the global energy supply in the second half of the 2002, FAO 2014, Magid 2003, Spencer and Schnittker 2009, century, with capacity increasing gradually in the first three Annepu 2015). Methane (CH4), Carbon dioxide (CO2), some decades (Bilen et al 2008) other minor constituents like nitrogen, ammonia (NH3), sulfur

Biomass, which is defined as organic materials derived dioxide (SO2), hydrogen sulfide (H2S) and hydrogen are the from plants and animals that can be renewed over time main components of biogas. The more methane percentage (Energy 2005), is amongst the available renewable energy in biogas means more useful energy can be produced. resources. This type of energy is the product of Various parameters have a major influence on methane photosynthesis process in which plants utilizes solar energy formation such as moisture content, percentage of organic to breakdown carbon dioxide, release oxygen, and store matter, pH and temperature. This means a good quality gas carbon in the mass. Animal manure can also be considered for energy generation can be obtained by controlling these as a biomass as well. The environment friendly biogas characteristics (Wright and Overend 2007). production which reduces the production costs in agricultural enterprises, is becoming more and more important. Animal MATERIAL AND METHODS manure, energy plants and agricultural organic wastes can Animal manure resources: With the case of Iraq, Biogas Production from Animal Manure in Iraq 243 biomass resources are available in most regions To convert animal manure to biogas, the coefficients of (Chaichan 2012). These resources are agricultural 33 m3 year-1 t-1 for cattle/horse/camel, 58 m3 year-1 t-1 for residues, urban wastes, animal manures, and forests sheep/goat, 78 m3 year-1 t-1 for poultry, and 26 m3 year-1 t-1 for residues and by products. As an initial investigation, it is donkey were used as conversion coefficients (adopted from worth noting that a portion of the data used in this paper Tatlidil et al 2009). has been collected from the literatures published in the The following coefficients were used for calculation of areas of animal manure (Jaradat 2002, FAO 2014, Magid biogas equivalents (adopted from Tatlidil et al 2009) gas oil: 2003, Spencer and Schnittker 2009, Annepu 2015). 0.62 L, charcoal: 1.46 kg, wood: 3.47 kg, butane: 0.43 kg, dry Livestock is one of the major features and has always cow manure: 12.3 kg, electricity: 4.7 kWh, diesel fuel:0.66l, been an important source of income for the traditional fuel: 0.75l, and propane: 0.25m3 farmers in Iraq, as a result of that many animal farms have Table 2 shows the numbers of animals in Iraq and been set up all over Iraq since the ancient time. However, the equivalent amount of electricity that may be the number of animals stock have showed a downward produced from the manure. It is worthy of mentioning that, trend since the second golf war in 1990. In further details, Table 2 has been calculated based on several reliable the number of camels, chickens and buffalos has fallen assumptions, which has been utilized by many researchers significantly by almost 87%, 72% and 57%, respectively. (Tatlidil 2009). While the number of other type of castles such as horses, asses, mules, goats and sheep have also gradually RESULTS AND DISCUSSION decreased (Magid 2003). There are several factors which There are various advantages of utilizing animal affect the amount of waste and biogas obtainable during manures such as producing biogas, organic fertilizer which livestock operations. These factors include the type and age can then be used to enrich soil composition, socio-economic of animal, body weight, type of breeding, total solids ratio, impact, and environmental impact. Based on the tables volatile solids ratio and the availability of waste and biogas above, about 892,263,400 m3 of biogas can be produced in yields (Antonia et al 2013). compliance with the number of livestock and poultry waste. Calculation method: In order to obtain the amount of biogas Cattle and sheep manures have the biggest proportion in the and the other products that can be produced from animal potential of biogas production. Then, poultry and goats follow manure, the related data and conversion coefficients have this, recording 65,208,000 and 64,960,000 m3 of biogas, been used. These data are the number of cattle, sheep, respectively. goats, camels, horses, chicken, asses, buffaloes, and mules. As shown in Table 2, biogas coefficients such as gas oil, To calculate the potential biogas that can be produced from butane, diesel fuel, fuel, and propane have been calculated animal manures, various conversion coefficients, which are based on the conversion coefficients which adopted from listed below, were employed. (Tatlidil et al 2009). Thus, 553203308l of gas oil, 383673262 -1 -1 Cattle/horse/camel: 3.6 t unit year ; sheep/goat: 0.7 t kg of butane, 588893844l of diesel fuel, 669197550l of fuel, -1 -1 -1 -1 -1 - unit year ; poultry: 0.022 t unit year ; donkey 2.9 t unit year and 223065850 kg of propane can be obtained from biogas 1 (adopted from Tatlidil et al 2009). using conversion coefficients. For electricity sector,

Table 1. Biogas production potentials Animal type Population Manure ton Animal manure Coefficient of Biogas m3 Electricity KWh (head-1) year-1 (head-1) (ton year-1) biogas (ton m-3) Cattle 2780000 3.6 10008000 33 330,264,000 1552240800 Sheep 8250000 0.7 5775000 58 334,950,000 1574265000 Goats 1600000 0.7 1120000 58 64,960,000 305312000 Camels 65000 3.6 234000 33 7,722,000 36293400 Horses 52000 3.6 187200 33 6,177,600 29034720 Asses & Mules 391500 3.6 1409400 33 46,510,200 218597940 Poultry 38000000 0.022 836000 78 65,208,000 306477600 Buffaloes 307000 3.6 1105200 33 36,471,600 171416520 Total 892,263,400 4193637980 244 Ali Alhelal and Inas Abdulsattar A. Jabbar

Table 2. Biogas equivalents (Ozturk 2005) addressed that each 1 m3 of biogas can be lighten lamb for 6 hours, to cook three meal for a family, to start an engine for 6 h, and to generate 1.25 KWh electricity Animal type Biogas m3 Gas oil (l) Butane (kg) Diesel fuel (l) Fuel (l) Propane (kg) Cattle 330,264,000 204763680 142013520 217974240 247698000 82566000 Sheep 334,950,000 207669000 144028500 221067000 251212500 83737500 Goats 64,960,000 40275200 27932800 42873600 48720000 16240000 Camels 7,722,000 4787640 3320460 5096520 5791500 1930500 Horses 6,177,600 3830112 2656368 4077216 4633200 1544400 Asses 46,510,200 28836324 19999386 30696732 34882650 11627550 Poultry 65,208,000 40428960 28039440 43037280 48906000 16302000 Buffaloes 36,471,600 22612392 15682788 24071256 27353700 9117900 Total 892,263,400 553203308 383673262 588893844 669197550 223065850

According to the International Energy Agency (2012), the biomass resources such as offset greenhouse gases electricity consumption in Iraq in 2010 was approximately emissions, utilize the waste, and control the erosion.

57 terawatt-hours and the demand is predicted to (Overend 2011) stated that roughly 1.6 ton of CO2, which increase roughly three-fold in 2035, which noticeably causes global warming and increases its concentration in the shows a sharp upward trend during the first quarter of this atmosphere, can be displaced for every MWh of biomass century. The equivalent electricity which can be generated power. Therefore, if the use of the biomass resources was from the total amount of biogas production is 4.193 TWh. This under a proper management, that would have a great role in means that utilizing the total amount of animal manures can reducing CO2 emissions. It is worthy of mentioning that, cover about 6.9% of the total electricity consumption in Iraq. Iraq has witnessed an increasing in the amount of CO2

Socio-economic impact: The utilisation of biomass emissions recently. Figure 11 shows how the amount of CO2 resources will certainly attract both domestic and foreign emissions has been increasing since the beginning of the investments to the country. As a result, this will help Iraq twenty-first century. The graph shows an upward trend from increasing its job opportunities and reducing the national 1980 to 2012. Also, the amount of CO2 emissions is sharply unemployment rate, which is currently 15.01% (Iraq increased during the period from 2000 and 2012. Unemployment Rate n.d). Furthermore, Iraq's dependency On the other hand, using improper managed on foreign energy sources can be reduced once a local biomass resources could also lead to the negative biomass to energy production takes place. Moreover, environmental influences such as deforestation, utilizing biomass resources in rural areas could have positive biodiversity limitation, and soil erosion. Reverse impacts on rural regions development as well as boosting the greenhouse gas emissions can be effected by clearing the agricultural development which has not received much land from the crops (deforestation), which is usually done by investment since 1990. This development may include burning and logging. The burning process releases the CO2 utilising new technologies in agriculture as well as which is the principal cause of global warming. Also, after improving the farmer's life quality. In addition to that, the the land being removed, crops and trees will no longer general public may also access biofuels, which has been contribute in carbon storage. Furthermore, deforestation proven to produce less emission compared to fossil contributes in both soil erosion and reduces the biodiversity. fuels. On the other hand, despite the fact that there are a various advantages of having a diversity of biomass CONCLUSION resources, the increasing of this development could Although both bovine and hen rearing is developed affect adversely on countryside societies. According to province, the existing potential cannot be effectively used in (Milbrandt 2009) quoting from World watch institute terms of benefitting from biogas production capacity. study, plantations, which have poor wages and working Considering animal waste potential and agricultural land conditions, have had most jobs. Also, local people who live in holding of Iraq, making biogas facilities widespread, extensive monoculture plantations could also be affected especially in rural areas, will bring economic benefits for the due to rarity of the wood and food resources. farmers. The results show a promising opportunity to utilize Environmental impact: It is undeniable that many this type of clean energy. Unfortunately, Iraq still gives a environmental benefits can be achieved from the use of back-seat for renewable energy due to its dependency on Biogas Production from Animal Manure in Iraq 245 fossil fuel plants. The chief job for this study was to shed the International Energy Agency 2012. Iraq Energy Outlook. Paris: International Energy Agency. light on the possibility of utilizing animal manure in producing Iraq Unemployment Rate (n.d). http://www.tradingeconomics.com/. biogas and its equivalents. Also, various advantages can be Retrieved from Trading Economics: http://www.tradinge obtained in several sectors such as agricultural, socio- conomics.com/iraq/unemployment-rate economic and environmental sectors. Iraq MO 2013. Crops and vegetable production in 2012. Baghdad: Ministry of planning. Iraq PM 2009. National development plan 2010 - 2014. Baghdad: REFERENCES Planning ministry of Iraq. Magid AM and Authores 2003. Country Report. Baghdad. Iraqi ministry of planning 2009. National development plan 2010- Alhelal A 2015. Solar Energy as an Alternative Energy than the 2014. Baghdad: Iraqi ministry of planning. Conventional Means of Electricity Generation in Iraq. Jaradat AA 2002. Agriculture in Iraq. Washington. International Journal of Inventive Engineering and Sciences 11- Jenkins W 2008. An Assessment of Biomass Resources in 14. California, 2007. California. Alnajjar AY 2014. http://www.ecomena.org/. Retrieved from Echoing Kida E (n.d.). lehigh. Retrieved from http://ei.lehigh.edu/: Sustainability: http://www.ecomena.org/swm-iraq/ http://ei.lehigh.edu/eli/energy/resources/readings/biomass.pdf Annepu R 2014. http://www.bioenergyconsult.com/. Retrieved from Milbrandt A 2009. Assessment of Biomass Resources in Liberia. Bioenergy Consult: http://www.bioenergyconsult. Colorado: National Renewable Energy Laboratory. com/tag/waste-management-strategy. Molland J 2012. http://www.care2.com/. Retrieved from Care 2: Annepu R 2015. http://www.bioenergyconsult.com/. Retrieved from http://www.care2.com/causes/5-top-countries-leading-the- Bioenergy Consult: http://www.biengeryconsult.com/tag/ world-in-renewable-energy.html waste-management-strategy. Naji I 2014. www.newsabah.com. Retrieved from Alsabah Aljadeed Antonia D, Cividino SRS, Carlino A, Gubiani R and Pergher G 2013. newspaper: http://www.newsabah.com/wp/newspaper/31758 Development perspectives for biogas production from agricultural waste in Friuli Venezia Giulia (Nord-East of Italy). Overend M 2011. Assessment of Biomass Resources in Regione Friuli Venezia Giulia, Via Sabbadini, 31 – 33100 Udine, Afghanistan. Colorado: NREL is a national laboratory of the U.S. ITALY. Department of Energy, Office of Energy. Ayhan A 2015. Biogas production potential from animal manure of Sasaki S 2003. Rice Husk Power Project in Thailand Invested by bursa province. Journal of Agricultural Faculty of Uludag Japanese Company. DHP P 259 Sustainable Development. University 29(2): 47-53. Spencer and Schnittker 2009. Persistent Animal Feed Shortage Bilen K, Ozyurt O, Bakırcı K, Karslı S, Erdogan S, Yılmaz M and Taxes Livestock Producers. USDA. Comaklı O 2008. Energy production, consumption, and Tatlidil BA 2009. Animal manure as one of the main biogas production environmental pollution for sustainable development: A case resources in Turkey. Journal of Animal and Veterinary Advances study in Turkey. Renewable and Sustainable Energy Reviews 32: 2473-2476. 12(6): 1529-1561. U.S Energy Information Administration. (n.d). http://www.eia.gov/. Chaichan 2012. Status and future prospects of renewable energy in Retrieved from Energy Kids: http://www.eia.gov/kids/energy. Iraq. Elsevier, 6007-6012. cfm?page=biomass_home-basics-k.cfm Duku 2011. A comprehensive review of biomass resources and U.S Energy information adminstration 2013. http://www.eia.gov/. biofuels potential in Ghana. Elsevier 404-415. Retrieved from Independent statistic and analysis/U.S Energy Energy UD 2005. What is Biomass? Washington, DC.: U.S. information adminstration: http://www.eia.gov/countries/ Department of Energy (DOE) and U.S. Department of country-data.cfm?fips=IZ#tpe Agriculture (USDA). U.S. Energy Information Administration 2015. Monthly Energy FAO 2014. Global Information and Early Warning System on Food Review. Retrieved from U.S. Energy Information and Agriculture. Food and agriculture organization of the united Administration: http://www.eia.gov/totalenergy/data/ nations. monthly/pdf/sec10_3.pdf Fuaad Mansur 2010. Investment in Date Palm & Wood Waste U.S. Office 2007. Integrated Strategic Plan Needed to Help Restore Recycling Projects. Dubai: Khalifa international date palm Iraq's Oil and Electricity Sectors. Washington, DC: United award. States Government Accountability Office. Grossman Z 2003. http://academic.evergreen.edu/. Retrieved from Wilkie M 2010. GLOBAL FOREST RESOURCES ASSESSMENT Biomass Fuels: http://academic.evergreen.edu/g/grossmaz 2010: country report: Iraq. Rome: Forestry Department / Food /thomdd.html and Agriculture Organization of the United Nations. Halder 2014. Assessment of biomass energy resources and related Wright and Overend 2007. Biomass Energy. Taylor & Francis Group, technologies. Elsevier, 444–460. LLC.

Received 24 September, 2019; Accepted 28 October, 2019 Manuscript Number: S-290 Indian Journal of Ecology (2019) 46 Special Issue (8): 246-250 NAAS Rating: 4.96

Impact of Speed and Extraction Depths on Performance of Potato Digger

Ghaleb Abdul Kadhim Muhaibis Department of Mechanization, Shatra Technical Institute, Southern Technical University, Basrah, Iraq E-mail: [email protected]

Abstract: An experiment was conducted to study the effect of tractor-practical speed, conveying chain -speed and lifting depths on some performance indicators of a potato digger. Indicators involved, two levels of tractor- practical speed of 2.5 and 5 km hr-1,two levels of conveying chain-speed of 29, 55.8 m min.-1 and two levels of lifting depths of 20, 25 cm. The results showed that the 2.5 km hr-1 tractor - practical speed recorded less damaged tubers (3.11 % ),less scratched tubers(1.07%), less yield loss (0.95%) and less slippage percentage (5.15 %).

Keywords: Conveying - chain speed, Damage tubers, Scratched tubers, Slippage percentage, Extraction depth

Some of the crops that grow under the surface of the soil, importance of this study is to evaluate the newly fabricated are in need of equipment, that differs in their design, from the and developed potato digger implement. equipment that is used for harvesting the crops that grow on the soil and all of which require means of dealing with the soil MATERIAL AND METHODS to disassemble it, without damaging the crop, optimizes crop An experiment was conducted on the field of the institute care operations The incorporation of mechanization in the of Shatre, Iraq in spring season in sandy soil (2019). Nested cultivation and harvesting of the crop, will help to reduce the design under randomized block design (RBD) with three labor and production costs, nevertheless, shortening time replications were used in this experiment. The treatment and effort to carry out all operations that results in production were randomly distributed within the experimental units the increase along with a better pace in conducting all main plot comprised of two secondary plot comprised of two operations, which are required in carrying out any agricultural levels of chain- conveyor speed of 29 and 55.8 m min-1 and process. The damage caused by harvesting operations that two levels of extraction depths of 20 and 25 cm using a affects the quality of potato tubers varies according to the potato digger with a chain conveyor (Fig. 1) and a conveyor equipment used in the harvest besides the nature of the soil. side and rear (Fig. 2). Damaged tubers levels of tractor- To ensure the highest quality of potato crop extraction, practical speed of 2.5 and 5 km hr-1 and, scratched tubers, improvements must be made to the extraction/ lifting loss of yield, slippage percentage, and implement implements under all prevailing conditions of the production productivity were measured in this experiment using the MF areas. There are several factors limiting the damage caused 650 tractor type. to the tubers, which include conditions of tuber, its Implement installation: The implement is comprised of a temperature, and the process of harvesting itself (Johnson triangular rail located in the front, thus the shear/weapon rail and Bohl 2010). In the specialty of ​​potato extraction/ digging/ is located in the front of the digger. That's enables the soil with lifting, mechanically, the primary objective is to collect the the tubers to drift and be surged upward, where it falls on a crop in an optimal manner, with the aim of avoiding the closed that runs sideways/ laterally along the width of the scratching the tubers or even leaving it in the ground without digger. The chain conveyor is equipped with interstices extraction (Al-obade 2012). The damage can be minimized allowing the potato tubers, which have been extracted ,to by calibrating the harvesting shears/weapons that are used transfer into the rear, while the soil blocks and residues fall in the harvest to below the level of the tubers in the soil, to out through the distances between the plates of the chain avoid cutting it and moreover to ensure that they are conveyer, when rotates backwardly. The conveyer is transferred to the implement on a soil cushion, thus reducing installed on a set of toothed wheels distributed on both sides the rate of scratching, taking into account that forward speed of the conveyor, as a result of this installation a vibrating of the harvesting implement along with the speed of the movement is generated that helps to expel the blocks of the conveying chain. It is necessary to reduce slippage, because soil that are attached to the tubers. The conveyer receives the increase in slippage leads to a significant loss of tractor the motion from the PTO shaft, by a set of gears. The potato power along with the reduction in field productivity. The digger is hitched to the tractor from its front, into the rear of the Speed and Extraction Depths of Potato Digger 247 tractor by the three hitching points. When the potato tubers 1recordedless damaged tubers (3.11%), and the highest ratio reach the back of the conveyor, the tubers fall directly of damaged tubers was of 3.38% with the tractor practical sideways laterally, on a sideway conveyer, which takes its speed of 5 km hr-1 conveying chain speed of 29 m min-1 had movement from the PTO shaft, through a set of pulleys, and less damage tubers (3.07%) and chain-conveyor speed of belts. Thus the large tubers of potatoes is transferred into a 55.8 m min-1 had the highest ratio of damaged tubers (3.42 bag. In addition, closed conveyor belt was also developed %.) The lifting depth of 20 cm recorded the least ratio of that which rotates backwardly, in a similar rotation damaged tubers (2.28 %) and depth of 25 cm recorded the movement to the chain conveyer and the conveying belt highest damaged tubers (4.21%). The practical speed of 2.5 transfer the potato tubers of small sizes and soft ones of an km hr-1 with chain - conveyor speed of 29 m min-1 and lifting 8 cm radius and less into a tank of a capacity of 30 kg that depth of 20 cm, caused the least damaged tubers and the tank is attached to the rear of the implement and beneath the interaction among the tractor-practical speed of 5 km hr-1 chain conveyer, thus the process of the extraction of the with the chain- conveyor speed of 55.8m\ min and lifting tubers was done completely, mechanically, without the need depth of 25 cm recorded the highest damaged. The reason to use manpower to collect the potatoes from the ground. of the increased damage tubers, maybe due to the increasing Slippage percentage:% The theoretical and practical speed in tractor-practical speed. These results are inconsistent with was calculated according to the following equations AL- Obaidi (2012). provided by zoz,1972. Scratched tubers (%): ​​Tractor- practical speed of 2.5 km VT= (ST/Tt)*3.6…------………1 hr-1 with chain-conveyor speed of 29 m min-1 and lifting depth Where, VT= theoretical speed (km hr-1), ST= Theoretical of 20 cm recorded the less scratched tubers of 1.07, 0.51 distance(m), Tt= theoretical time(S) and 1.21 % and maybe due to the decreasing the tractor- VTp=(SP/TP)*3.6………------…2 forward speed besides the presence of the chain conveyer Where, VP= practical speed (km hr-1), Sp= practical distance that direct connection with the potato tubers (Sidiyq 2012) (m), TP= practical time (s) = (VT-Vp/VT)*100………------Lifting depth of 25 cm recorded scratched tubers of 1.52%. -----…3 The impact of interaction among tractor practical speed of 2.5 S= slippage percentage (%), VT= theoretical speed (km hr-1), km hr-1 and chain-conveyor speed of 29 m min-1 with lifting VP= practical speed (km hr-1) depth of 20 cm resulted in less scratched tubers of (0.32%), in comparison with the interaction among tractor-practical RESULTS AND DISCUSSION speed of 5 km hr-1 and chain- conveyor speed of 55.8 m min-1 Damaged tubers (%): Tractor-practical speed of 2.5 km hr- with lifting depth of 25.

Fig. 1. Potato digger with a chain conveyor Fig. 2. Side and the rear 248 Ghaleb Abdul Kadhim Muhaibis

Loss of yield (%): The tractor practical speed of 2.5 km hr-1 speed of 55.8 m min-1 recorded more yield loss of1.63 resulted less yield loss of 0.95, practical speed of 5 km hr-1 %.Lifting depth of 20 cm recorded yield loss of 3.0%, recorded yield loss of 2.14% Chain-conveyor speed of 29 m whereas, lifting depth of 25 cm recorded less yield loss of min-1 recorded less yield loss of 1.10%, chain- conveyor 1.24%. interaction among practical speed of 2.5 km hr-1 with

Table 1. Impact of practical speed, chain-conveyor speed and lifting depths on damaged tubers (%) Indicators Damaged tubers (%) Interaction between practical speed and Practical speed Conveyor chains speed m Interaction among practical speed and conveyor conveyor chain's speed min-1 chain's speed and lifting depths 20 cm 25 cm 2.5 km hr-1 29 1.90 3.95 2.92 5 km hr-1 55.8 2.50 4.10 3.30 29 2.30 4.15 3.22 55.8 2.45 4.65 3.55 LSD=0.05 0.66 0.03 Mean of lifting depths 2.28 4.21 LSD=0.05 0.86 Conveyor chains speed m Interaction among practical speed and conveyor chain's speed and lifting depths Mean of conveyor speed min-1 29 2.10 4.05 3.07 55.8 2.47 4.37 3.42 LSD=0.05 0.42 0.04 Practical speed Interaction among practical speed and conveyor chain's speed and lifting depths Mean of practical speed 2.5 km hr-1 2.2 4.02 3.11 5 km hr-1 2.37 4.4 3.38 LSD=0.05 NS

Table 2. Impact of tractor practical speed, chain- conveyor speed and lifting depths on scratched tubers (%) Indicators Scratched tubers (%) Interaction between practical speed and Practical speed Conveyor chains speed m Interaction among practical speed and conveyor conveyor chain's speed min-1 chain's speed and lifting depths 20 cm 25 cm 2.5 km hr-1 29 0.32 0.56 0.44 5 km hr-1 55.8 1.66 1.74 1.70 29 0.42 0.76 0.59 55.8 2.45 3.65 3.05 LSD=0.05 0.58 1.05 Mean of lifting depths 1.21 1.52 LSD=0.05 0.027 Conveyor chains speed m Interaction among practical speed and conveyor chain's speed and lifting depths Mean of conveyor speed min-1 29 0.37 0.66 0.51 55.8 2.05 2.69 2.37 LSD=0.05 0.03 0.06 Practical speed Interaction among practical speed and conveyor chain's speed and lifting depths Mean of practical speed 2.5 km hr-1 0.99 1.15 1.07 5 km hr-1 1.43 2.20 1.81 LSD=0.05 0.02 0.04 Speed and Extraction Depths of Potato Digger 249

chain - conveyor speed of 29 m min-1 and lifting depth of 25 yield loss of 2.65%. The reason of increased yield loss maybe cm recorded yield less loss of 0.44% in comparison with the due to the increasing in the layer depth 20 cm. These results interaction among practical speed of 5 km hr-1 with chain - are in consistent with Sidiyq (2006). conveyor speed of 55.8 and lifting depth of 20 cm recorded Slippage percentage (%): Table 4 showed the impact of the

Table 3. Impact of practical speed, chain-conveyor speed and lifting depth on yield loss (%) Indicators Loss of yield (%) Interaction between practical speed and Practical speed Conveyor chains speed m Interaction among practical speed and conveyor conveyor chain's speed min-1 chain's speed and lifting depths 20 cm 25 cm 2.5 km hr-1 29 0.62 0.44 0.53 5 km hr-1 55.8 0.82 0.50 0.66 29 1.92 1.45 1.68 55.8 2.65 3.65 2.61 LSD=0.05 0.03 0.05 Mean of lifting depths 3.00 1.24 1.39 LSD=0.05 0.017 Conveyor chains speed m Interaction among practical speed and conveyor chain's speed and lifting depths Mean of conveyor speed min-1 29 1.27 0.94 1.10 55.8 1.73 1.54 1.63 LSD=0.05 NS 0.06 Practical speed Interaction among practical speed and conveyor chain's speed and lifting depths Mean of practical speed 2.5 km hr-1 1.44 0.47 0.95 5 km hr-1 2.28 2.01 2.14 LSD=0.05 0.034 0.062

Table 4. Impact of practical speed, chain-conveyor speed and lifting depth on slippage percentage (%) Indicators Slippage percentage Interaction between practical speed and Practical speed Conveyor chains speed m Interaction among practical speed and conveyor conveyor chain's speed min-1 chain's speed and lifting depths 20 cm 25 cm 2.5 km hr-1 29 4.57 5.22 4.89 5 km hr-1 55.8 4.66 6.18 5.42 29 7.42 9.02 8.22 55.8 10.45 11.66 11.05 LSD=0.05 0.024 1.06 Mean of lifting depths 6.77 8.02 7.39 LSD=0.05 0.72 Conveyor chains speed m Interaction among practical speed and conveyor chain's speed and lifting depths Mean of conveyor speed min-1 29 5.99 7.12 6.55 55.8 7.55 8.92 8.23 LSD=0.05 0.63 0.092 Practical speed Interaction among practical speed and conveyor chain's speed and lifting depths Mean of practical speed 2.5 km hr-1 4.61 5.70 5.15 5 km hr-1 8.93 10.34 9.63 LSD=0.05 0.73 1.04 250 Ghaleb Abdul Kadhim Muhaibis

Table 5. Impact of practical speed and chain-conveyor speed and lifting depth on productivity (hr) Indicators Productivity (hr) Interaction between practical speed and Practical speed Conveyor chains speed m Interaction among practical speed and conveyor conveyor chain's speed min-1 chain's speed and lifting depths 20 cm 25 cm 2.5 km hr-1 29 1.22 1.27 1.24 5 km hr-1 55.8 1.18 1.25 1.21 29 1.05 1.02 1.03 55.8 1.01 1.00 1.05 LSD=0.05 0.06 0.024 Mean of lifting depths 6.77 1.18 1.21 LSD=0.05 0.062 Conveyor chains speed m Interaction among practical speed and conveyor chain's speed and lifting depths Mean of conveyor speed 29 1.13 1.14 1.135 55.8 1.09 1.12 1.10 LSD=0.05 0.03 0.92 Practical speed Interaction among practical speed and conveyor chain's speed and lifting depths Mean of practical speed 2.5 km hr-1 1.20 1.26 1.23 5 km hr-1 1.03 1.01 1.02 LSD=0.05 1.43 1.07

tractor practical speed, chain-conveyor speed and lifting Al-Qazzaz KM 1989. Agriculture and Agricultural Equipment, College of Agriculture, University of Baghdad, Ministry of depths in the slippage percentage. Tractor- practical speed of Higher Education and Scientific Research. Bohl WH and 2.5 km hr-1 recorded less slippage percentage (5.15 %). as Johnson SB 2010 Commercial Potato production in compared 5 km chain-conveyor speed of 29 m min-1 resulted NorthAmerica.pp.75-80. AL-Obaidi OMT 2012. Effect of harvesting machine type on loss in less slippage (6.55 %). Lifting depth of 20 cm resulted in quality of potato crop in different lifting dates and speeds. M.Sc. less slippage (6.77). Interaction among the tractor-practical Thesis Agricultural Machines and Equipment. speed of 2.5 km hr-1 with chain- conveyor speed of 29 m min-1 Abedallah IAI 2001. Quantitative and qualitative loss potato sown mechanically. M.Sc. Thesis, Department of Agricultural and lifting depth of 20 cm recorded less slippage percentage Mechanization, Faculty of Agriculture and Forestry, University (Table 4). Reason of the increased slippage percentage of Mosul. maybe due to the increasing, in practical speed. Al-sadoon AA 1991. Potato crop in Saudi Arabia. Ministry of Agriculture, Saudi Program for Agricultural Development, Productivity (less time to lift the potato): Tractor - practical Riyadh, Saudi Arabia. speed of 2.5 km hr-1 recorded time productivity of 1.23 hr-1 Erdogan D 1987. A Study on Field Performance of Small Tractors, whereas, tractor - practical speed of 5 km hr-1 recorded less Agri. Machine Asia, Higher Education and Scientific Research, Baghdad University, Faculty of Agriculture, pp 267– 268. time productivity of 1.02 hr. Chain-conveyor speed of 55.8 m Kasmire RF 1983. Influence of mechanical harvesting on quality of min-1 required less time productivity of 1.10 %, in comparison non-vegetables. Hortscience 18: 421-423. with chain- conveyor speed of 29 m min-1 recorded time Shebi JG, Oni KS and Braide FG 1986. Tractate performance of three tractors. Agricultural Mechanization in Asia, Africa and productivity of 1.135 hr. The lifting depth of 20 cm recorded Latin America 17(2).122-127. productivity time of 1.25 hr, whereas, lifting depth of 25 cm McGechan MB 1981. A comparison of some studies of damage recorded less time productivity of 1.18 hr. Interaction among susceptibility of different potato varieties. Journal of Agricultural Engineering Research 26: 161-170. tractor-practical speed of 2.5 km hr-1 with chain-conveyor Salih M and Karim SE 2012. Potato - Production - Storage – and speed of 29 m min-1 and lifting depth of 20 cm required less Manufacturing, Part II, University of Mosul Press. time productivity of 1.22 hr, in comparison with interaction Sidiyq AMA and Natiq SH 1998. Effect of mechanical extraction on -1 quantitative and qualitative loss of potato yield in Ninewi among practical speed of 5 km hr with chain-conveyor Governorate. M.Sc. Thesis, Department of Agriculture speed of 55.8 m min-1 and lifting depth of 25. Mechanization, Faculty of Agriculture and Forestry, University of Mosul. Sidiyq AMA 2006. Development of the separation system in the REFERENCES potato and evaluation of performance. Ph.D. Thesis. ASAE, Standard 2003. Agricultural Machinery data, D 497.4 Feb 03, Department of Agricultural Machinery and Machinery, Faculty of pp 372-380. Agriculture and Forestry University of Mosul.

Received 15 September, 2019; Accepted 04 November, 2019 Manuscript Number: S-291 Indian Journal of Ecology (2019) 46 Special Issue (8): 251-253 NAAS Rating: 4.96

Effect of Foliar Nano-Fertilizers of Marine Algae Extract and Boron on Growth and Yield of Faba Bean (Vicia faba L.)

Ali R. Alhasany, Dargham S.K. Altai1 and Ali H. Noaema College of Agticulture, Al-Muthanna University, Iraq 1College of Agticulture, University of Missan, Iraq E-mail: [email protected]

Abstract: The study used a split plot design in RCBD with three replicates to study the effect of three foliar concentrations of nano- marine algae extract (0, 50 and 100 ml l-1) and nano-boron (0, 1 and 2 ml l-1) on chemical, physiological, growth and yield characters of broad bean (Vicia faba L.). Foliar applications of nano- algae extract 100 ml l-1 gave the highest values of (N and chlorophyll content in plant leaves), plant highest, no pod plant-1 and seed yield were 48.02 mg kg -1, 40.07, 102.70 cm, 16.33 pod. plant-1 and 5277 kg h-1, respectively compared to control treatment. The results showed that increasing the concentration of nano- boron to 2 ml l-1 in the spray solution resulted in an increase respectively compared to ,٤٣٫١١ mg kg -1 and ٧١٫٨١ ,in concentration of N, Bo and chlorophyll content in plant leaves were 48.70 mg kg -1 control treatment. The increasing rate no pod plant-1 was 18.82 %, weight of 100 seed 17.01% and seed yield 46.64%, respectively compared to control treatment. As for the interaction between (nano-algae extract and boron), it was significant in (N, Bo content in plant leaves), no of pod plant-1 and weight of 100 seed.

Keywords: Nano- algae extract, Nano-boron, weight of 100 seed, Foliar application, Faba bean

Faba bean (Vicia faba L.) is one of the most important 2004). Mineral nutrition of plants is important for controlling legume crops and is a good source of many nutrients, such physiological and biochemical processes of plants. Its as potassium, calcium, magnesium, iron and zinc (Khalil et al deficiency may lead to changes in these processes and 2015, Neme et al 2015). It has an important role in improving disturbed plant growth and yield. Boron is an essential chemical and physical properties of soil the by stabilizing micronutrient for plant growth and development Shireen et al atmospheric nitrogen because of their symbiosis with (2018). Khattab et al (2016) indicated that foliar application of rhizobia root nodule bacteria (Abbas 2012). Nanotechnology boron (150 ppm), potassium sulfate (2%) and potassium is one of the new techniques that contribute to improve and phosphate (1.5%) significantly increased growth, yield increase agricultural production widely and are promising components and chemical composition of faba bean. applications in production quantity and because it increases Therefore, in present study on the effect of foliar Nano- the absorption of water and nutrients which help to improve fertilizers of marine algae extract and boron on growth and plant growth and productivity (Al-Ramadi et al 2016). Nano yield of faba bean (Vicia faba L.) was determined. applications are play in agricultural in increasing the efficiency of fertilizers and reducing costs used in agriculture MATERIAL AND METHODS (Singh 2016). Nano compounds rapidly and completely Field trials were carried out during seasons 2017-2018 absorbed by the plant to meet nutritional needs and in Rumaytha district, 25 km north of Muthanna Governorate, deficiencies (www.Nano.ir). Presently, organic farming is Iraq. The experimental soil was classified as a silty clay loam priority to reduce the use of chemical fertilizers and relying on (180 Kg-1s and, 465 Kg-1silt and 355 Kg-1 clay) with pH 7.6 and natural alternatives of organic fertilizers that are non-toxic, E.C. 4.2 dS m-1. The rates of N, P, K and B available were 27, non-harmful and non-polluting (Oad et al 2004). 9, 157 and 1.1 mg. kg-1 soil, respectively. Treatments Foliar application of algae extracts, nutrients and growth consisted of three Nano- algae extract (0, 50 and 100 ml l-1) regulators are economical and provide clear response during and Nano-boron (0, 1 and 2 ml l-1). A split-plot in RCBD design vegetative and proliferative phase (Ali et al 2014). Spray with three replicates was used to study. The experimental organic compounds such as algae extracts, which are a good unit area was 6 m2 (3 m long and 2 m wide). Experiment unit source of growth regulators, macro and micronutrients, contained 4 lines; spacing between them was 35 cm and 20 which contribute to increasing the ability of roots to growth cm between hills. The seed was planted on October 14 by and absorption so thus increase the leaf area and this is hand drilling (Al-Tawki 2015). Irrigation and weeding were reflected positively on the increase in productivity (Jensen done uniform as per requirement. The nitrogen rates were 60 252 Ali R. Alhasany, Dargham S.K. Altai and Ali H. Noaema kg N ha-1 that applied through urea (46% N). Half of the N was increase the nitrogen absorbed from the soil and thus applied at sowing and the other half after 15 days after increase its concentration in plants (Bahkuni et al 2010). The sowing. The experimental area received 80 kg P ha-1 from accumulation of boron in the leaves due to increased triple superphosphate and 80 kg K2O ha-1 from potassium concentration of Nano-boron in the spray solution led to chloride as basal fertilizers at the time of sowing. (Abedi improved efficiency of the absorption mechanism of boron by 2011). Two-time spray was used from Nano- algae extract, the plant, which increased its concentration in the leaves. the first one-month after germination, and the second at The increase in leaf chlorophyll ratio by important role flowering. One spray was used from Nano-boron at 50% boron's in the activities various enzymes most well-known flowering. use involves the building of chlorophyll molecules also the role of this compounds biological processes such as RESULTS AND DISCUSSION photosynthesis reaction. The Nano-boron play an important Effect on N, B and chlorophyll content in leaf: The foliar role in the activities various enzymes involved in this application of 100 ml.l-1 Nano-algae extracts significantly biochemical reaction than increased leaf chlorophyll content increased of N and chlorophyll content in the leaves (Table 1). are in agreement with the findings (Amanullah et al 2010). The of spraying by Nano-algae extracts might have led to The foliar application of 100 ml l-1 Nano-algae extracts improved root growth, contributing to greater efficiency in combined to 1 ml l-1 Nano-boron significantly increased the absorbing some nutrients from the soil, including nitrogen, content of N in the leaves. The highest boron content was which reflected positive effect on nitrogen content in leaves. recorded with Nano (2 ml l-1 algae extract and 0 boron). Almosawy et al (2018) also reported similar findings. Effect on Growth and Yield However, the spray 2 ml l-1 Nano-boron, resulted in maximum The foliar application of 100 ml l-1 Nano-algae extracts N, B and chlorophyll content in the leaves, which did not significantly increased plant height, no. of pod plant-1 and significantly different from 1 ml l-1 in N content in leaves. This grain yield (Table 2). These are in agreement with findings of may be due to the spraying of boron which resulted in earlier workers (Al-Atabi 2018, Almosawy et al 2018). The

Table 1. Effect of Nano-fertilizers (Algae extracts / Boron) on nitorgen, boron and chlorophyll in leaves Nano-algae Nitrogen content (mg kg-1) Boron content (mg kg-1) Chlorophyll content extract ml l-1 Nano-boron ml l-1 Mean Nano-boron ml l-1 Mean Nano-boron ml l-1 Mean

0 1 2 0 1 2 0 1 2

0 41.13 44.87 48.27 44.76 56.80 59.63 68.54 61.67 35.90 35.33 38.13 36.52 50 44.07 49.47 49.17 47.57 63.13 77.57 75.30 71.67 34.88 40.32 44.27 39.82 100 46.30 49.10 48.67 48.02 64.67 62.97 72.57 66.73 33.70 39.58 46.93 40.07 Mean of boron 43.83 47.81 48.70 61.53 66.72 71.81 34.83 38.48 43.11 Algae Boron Algae x Algae Boron Algae x Algae Boron Algae x Boron Boron Boron CD (p=0.05) 2.23 1.34 2.56 NS 2.88 8.20 2.55 4.60 NS

Table 2. Effect of Nano-fertilizers (Algae extracts / Boron) on in plant height, number of pod plant-1 and seed pod-1 Nano-algae Nitrogen content (mg kg-1) Boron content (mg kg-1) Chlorophyll content extract ml l-1 Nano-boron ml l-1 Mean Nano-boron ml l-1 Mean Nano-boron ml l-1 Mean

0 1 2 0 1 2 0 1 2

0 80.50 86.80 91.00 86.10 11.50 12.36 15.16 13.01 4.65 4.95 5.30 4.96 50 95.40 86.10 100.50 94.00 13.10 15.90 16.23 15.07 4.65 4.55 4.53 4.57 100 98.70 105.60 103.70 102.70 16.36 15.36 17.26 16.33 4.88 4.63 5.13 4.88 Mean of boron 91.50 92.80 98.40 13.65 14.25 16.22 4.71 4.72 4.98 Algae Boron Algae x Algae Boron Algae x Algae Boron Algae x Boron Boron Boron CD (p=0.05) 11.08 NS NS 1.70 0.53 1.67 NS NS NS Growth and Yield of Faba Bean 253

Table 3. Effect of Nano-fertilizers (Algae extracts / Boron) and interaction between them in weight of thousand seed (g) and grain yield kg ha-1 Nano-algae extract Thousand seed weight (g) Grain yield (kg ha-1) ml l-1 Nano-boron ml l-1 Mean Nano-boron ml l-1 Mean 0 1 2 0 1 2 0 103.33 116.67 122.67 114.22 3552 4577 5407 4512 50 104.67 107.33 128.00 113.33 3690 4888 5541 4706 100 109.33 114.00 120.67 114.67 4490 5087 6253 5277 Mean of boron 105.78 112.67 123.78 3910 4851 5734 Algae Boron Algae x Boron Algae Boron Algae x Boron CD (p=0.05) NS 3.80 8.00 540.80 357.80 NS spray 2 ml l-1 Nano-boron produced the highest number of agricultural experiments. Dar Al Kutub Printing & Publishing -1 Est. University of Al Mosul. Ministry of Higher Education and pod plant , thousand seed weight and grain yield. Boron also Scientific Research. The Republic of Iraq. p 7-13. plays a role in determining the direction of pollen tube growth Al-Tawki WBA 2015. Response of genetic structures from the crop of ,thus flowering stages increased and pod number per plant the others (Vicia Faba L.) to the dates of cultivation in Muthanna Governorate. Master Thesis, Faculty of Agriculture, Muthanna was increased directly effected the yield. Similar University. p 41-46. observations were reported by(Ali and-Juthery (2017). The Amanullah M, Sekar S and Vincent S 2010. Plant growth substances enhanced effect of Nano-boron spraying on leaf chlorophyll in crop production: A review. Asian Journal Plant Science 9(4): 215-222. content reflected then better development of photosynthesis Bahkuni G, Khurana N and Chatterjee C 2010. Impact of boron processes (Table 1). The foliar application of 100 ml l-1 Nano- deficiency on changes in biochemical attributes, yield and seed algae extracts combined to 2 ml l-1 Nano-boron significantly reseves in Chickpea . Communications in Soil Science and Plant Analysis 42(2): 199-206. -1 increased no. of pod plant . The highest values of thousand Jbara AK 2018. Effect of adding of humic and fulvic acid on the seed weight were with combined Nano (50 ml l-1 algae extract availability of N.P.K, in the soil and number, weight of the root -1 nodules and yield of broad been (Vicia Faba L.). M. Sc. Thesis, and 2 ml l boron). Fac. of Agric. Univ of Muthanna., Iraq. Jensen E 2004. Seaweed; fact or fancy. Published by Moses the REFERENCES Midwest- Organic And Sustainable Education. From the broad Caster 12(3): 164-170. Abbas SH 2012. Analysis of the performance of genetic structure traits in the rest under the influence of different levels of Khalil NA, Metwally AM, Magdy MS and Morshedi OA 2015. Cereal fertilization NPK. Kufa Journal. of Agricultural Sciences crops and pulses. Faculty of Agriculture. Cairo University. P. 4(2):13-21. 186. Abedi JA 2011. Directory of Chemical and Organic Fertilizers in Iraq. Khattab EA , Elham AB and Afifi M H 2016. Response of some General Authority for Extension and Agricultural Cooperation - varieties of faba bean (Vicia faba L.) to boron and potassium. Ministry of Agriculture - Iraq Agronomy 38(1):-21. International Journal of Chemtech Research 9(8): 98-103. li NS and Al-Juthery HWA 2019. Nanoscience in soil and pant Al-Atabi AKJ 2018. Effect of Humic Acid, Fulvic Acid and Marine systems. ministry of higher education and scintific research. Algae Spraying on N. P. K Soil Growth and Growth and Bean Harvest Vicia faba L. faculty of Agriculture. Al-Muthanna Nair R, Varghese SH, Nair BG, Maekawa T, Yoshida Y and Kumar DS University. 2010. Nanoparticulate material delivery to plants. Plant Scienses 179: 154-163. Al-Hasany ARK 2018. Effect of foliar nutrition of proline and mixture of nutrients on growth and yield of different varieties of Broad Neme K, Bultosa G and Bussa N 2015. Nutrient and functional Bean (Vicia faba L.). Ph.D. Thesis, Fac. of Agric. Univ of properties of composite flours processed from pregelatinised Muthanna., Iraq. barley, sprouted faba bean and carrot flours. International Journal of Food Science Technology. 50: 2375-2382. Ali NS and Al-Juthery HWA 2017. The application of nanotechnology for micronutrient in agricultural production (review article). The Shireen F, Nawaz MA, Chen C, Zhang Q, Zheng Z, Sohail H, Sun J, Iraqi Journal of Agricultural Sciences: 48(4): 984-990. Cao H, Huang Y and Bie Z 2018. Boron: Functions and Approaches to Enhance Its Availability in Plants for Sustainable Ali NS, Rahi HS and Shaker AA 2014. Soil fertility. Scientific Book Agriculture. International Journal of Molecular Science 19(7): House. College of Agriculture - University of Baghdad. 1856-1870. Almosawy AN, Alamery AA, Al-Kinany FS, Hasan MMH, Alkinani LQ Singh A and Prasad SM 2016. Nanotechnology and its role in agro- and Jawad NN 2018. effect of optimus nanoparticles on growth ecosystem: astrategic perspective. International Journal and yield of some broad bean cultivars (Vicia faba l.). Environmental Science Technology DOI: 10.1007/s13762- International Journal Agriculure Statatics Science 14(2): 257- 016 -1062-8. 262. SultanT 2010. Nanotechnology in fertilizers. Nature Nanotechnology Al-Ramadi HR and Nehmeh AA, Tareq AS, Al-Saeedi AH, Al-Sikhan 5:91. doi: 10.1038/nnano.2. MS and Shalabi TA 2016. Botany and Environmental Zodape ST, Mukhopadhyay S, Eswaran K, Reddy M and Chikara J Nanotechnology. Faculty of Agriculture - Kafr El-Sheikh 2010. Enhanced yield and nutritional quality in green gram University. First Edition. P. 189. (Phaseolus radiate L.). Journal of Scientific and Industrial Al-Rawi KM and Khalaf Allah AM 2000. Design and analysis of Research 69: 468-471. Received 01 November, 2019; Accepted 23 November, 2019 CONTENTS

S-280 Effect of Host Plant on a Amylase Activity in Cotton Aphid Aphis Gossypii (Glov.) (Hemiptera: Aphididae) 196 Hind Ibrahim AL-Khazraji S-281 Effect of Foliar Feeding on Copper and Molybdenum in Some OF the Physiological, Productivity and 201 Quality Traits for Two Genotypes Mung Bean (Vigna radiata L.) Riyadh L. Attieh and Basheer H. Abdallah S-282 Evaluation of the Efficiency of Ultraviolet Radiation and Hot Peppermint Extract in the Protection of Some 209 Biological Systems of Male Albino Mice from the Toxic Effects of Aflatoxin B and Fumonisin B Toxins Ali F. Jubair and Sabah L. Alwan S-283 Effect of Ozone Gas in Milk Treatment on Chemical and Microbial Properties of Soft Cheese 214 Azhar J. ALmosowy and Zenib H. ALmosowy S-284 Negative Effects on Some Internal Organs of Rats 219 Infected by Salmonella typhimurium Fatima Aziz Mahdi Al-badry S-285 Effect of the Isolated Alkaloid from Teucrium polium L. on Dna Isolated from Pathogenic Bacteria 224 (Escherichia coli and Staph aureus) of Urinary Tracts Rawaa A. Kushaish, Bushra A.M. AL-Salem and Mouayed A.A. Hussein S-286 Evaluation of Amino Acid Profile for Freshwater Fishes Yellow Barbell (Carasobarbus luteus) and Common 229 Carp (Cyprinus Carpio) of Euphrates River, Iraq Kadhim O. M. AL. humairi, Hamza G. Al-Agidi and Sadeq M. Al-Haider S-287 Using Mixed Model to Detect the Effect of Genetic and Fixed Factors on Milk Production in Awassi Sheep 233 M.A. AL-Shaikh, W.J. AL-Khazraji, WafaaI. AL-Samaraai and L.J. AL-Jaryan S-288 Seasonal and Physiological Changes on Renal Function Parameters in Iraqi Cats 237 Ahmed Noaman Awad AL-Ani, Taha Yassin Ghani Al-Badrawi, Basil Razook Faraj Razook and Naseir Mohammed Badawi S-289 Development Perspectives for Biogas Production from Animal Manure in Iraq 242 Ali Alhelal and Inas Abdulsattar A. Jabbar S-290 Impact of Speed and Extraction Depths on Performance of Potato Digger 246 Ghaleb Abdul Kadhim Muhaibis S-291 Effect of Foliar Nano-Fertilizers of Marine Algae Extract and Boron on Growth and Yield of Faba Bean 251 (Vicia faba L.) Ali R. Alhasany, Dargham S.K. Altai and Ali H. Noaema CONTENTS

S-260 Effect of Biofertilizer on Growth and Yield of Wheat Triticum aestivium L 93 Iktifaa N. Jassim, Ali A.J. Al-Haidary and Sophia J. Jassim S-261 Effect of Nano Biological and Mineral Fertilizers on Growth and Yield of Wheat (Triticum aestivum L.) 97 Basim K. Hasan and Turki M. Saad S-262 Evaluation of Maize Hybrids, their Inbred Lines and Estimation of Genetic Divergence Based on Cluster 102 Analysis Wajeeha Abed Hassan, Banan Hassan Hadi and Majid S.H. Hamdalla S-263 Evaluation of Seed Viability and Vigour of Maize coated with Local GYPSUM (CASO / HO) and AG-NPS 108 for Resistance of Fusarium verticillioidis Saddam H. Cheyed, A. Ch. Al-Fahad, Eman A. Al-Qaisy and Aalaa K. Hassan S-264 Effect of Glutathione Foliar Spray on Some Growth Traits in Maize Varieties 113 Ruqaya H. Mahmood and Najat H. Zeboon S-265 Effectiveness of Oxidation Enzymes in the Ratio of Gluten to Wheat Bread Via Different treatments of 119 Weeds Control Alaa A.J. AL-Behadili and Ziyad A. Abed S-266 Effect of Ratooning of Sorghum Cultivars on Growth and Yield Characters 123 Adawiya S. Mustafa Al-rawi, Sadam H. Chiad and Arige M. Saleh S-267 Genetic Parameters and Hybrid Vigor of Single and Double Hybrids of Maize (Zea mays L.) in Two Plant 128 Densities Banan Hassan Hadi and Ahmad Nama Abdel-Amir S-268 Effect of Water Stress on Growth and Green Forage 139 Yield of Ratoon Sorghum Waleed A.T. El.Fahdawi, Adwea M. Al-Rawi, Saddam H. Cheyed, Areej M.S. AL-Mohammedi and Adwiya S.M. Al-Rawi S-269 Response of Growth and Yield Characters of Three Sorghum Cultivars to the Biological Catalyst (Amino 145 triplex) Ahmed H. Ahmed and Sinan A. Abbas S-270 Effect of Zero Tilled Dibbled and Phosphorus Fertilizer on Growth and Yield of Sunflower (Helianthus 151 annuus L.) Edrees S. Najm, Malath A. Hamed, Q. Snaa and S.H. Cheyed S-271 Management of Wheat Weeds using Sorghum bicolor (L.) Leaves Extract and Effect on Yield Components 157 Watheq Falhi Hammood and Suhad Mathkoor A. Safi S-272 Effect of Foliar Application of Urea and Boric Acid on Growth Parameters and Yield of Eggplant Solanum 161 Melongena L Aqeel Kareem Hasan Al-Tufaili, Jamal Ahmed Abbass, Mansoor Abed Aboohanah and Ali Razzak Jasim Mohammed Hraen S-273 EFficacy of Rigenal P and Liqhumis 8 in Rooting and Vegetative Growth of Olive, Olea Europaea Cuttings 165 G.B.A. Al-Abbasi, F.S.S. Alabbasi and M.A.A. Abdulhussein S-274 Effect of Genotype and Nano-Fertilizers on Some Traits and Yield of Potato 168 Abbas K. Mijwel and Hawraa H. Muhsin S-275 Effect of Niacin (Nicotinamide) and Humic Acid on Growth and Chemical Traits of Pelargonium hortorum L. 172 Thaer Y. Khudair, Fakhria A. Abdul Albbas and Khansaa A. Abdul Kreem S-276 Investigation of Intestinal Parasites in Fruits and Vegetables in Local Market of Tikrit, Iraq 173 Ashraf Jamal Mahmoud Zangana S-277 Effectiveness of Brassinolide and Dry Yeast Extract Spraying on Growth Parameters and the Chemical 179 Content of the Grape Seedlings Sabaa Jawad Abdulkadhim and Akram Abd-Alkadem Hadi S-278 Effect of Salicylic Acid on Chlorophyll A, B and Carotene Content of Petunia sp. 183 Zahraa H. Ali and Faiq H. Ali S-279 Effect of Integrated Pest Management on Mortality of Whitefly Parasitoid Eretmocerus eremicus 192 (Hymenoptera: Aphelinidae) on Egg Plant S.A. Alani and Z.A.A. Al-Gadban Indian Journal1 Indian Journal of Ecology (2019) 46 Special Issue (8) of Ecology CONTENTS

S-242 Analysis of the Climate Data (Earth Observation Stations and Space Stations) Through Geographic 1 Information System Balsam Shaker Shnichal S-243 Risks of Rainfall Intensity on Geomorphic Processes of Duhok Governorate in Northwest Iraq using RS 7 and GIS Ruqaya Ahmed M. Amin and Balsam Shaker Shnichal S-244 Digital Modelling of Geomorphic Hazards due to Lunar Gravity in Lake of Mosel Dam using Remote 15 Sensing Data and GIS Ruqaya Amin S-245 Effect of Supplementation with High Levels of Phytase Enzyme Compared with Commercial Dose in Corn- 21 Soybean based Diets on Productive and Physiological Performance in Broiler Husam H. Nafea, Firas M. Hussein and Bahaa I. Hamid S-246 Evaluation of Heavy Metals in Nassiryah City, Iraq 28 Shroog Hameed Hlail and Sanaa Talib Jaweed S-247 Effect of Copper Concentrations on Some Biochemical Properties of the Cladophora 31 Wameedh A.K. AL-Yasari, Rasool I. Al-Nafie and Bassam M. Al-Yaseen S-248 Ameliorative Effect of Ascorbic Acid on Hematological Parameters Due to Cadmium Toxicity in Cyprinus 36 carpio Hamza G. Al-Agidi, Kadhim O.M. AL-Humairi and Sadeq M. Al-Haider S-249 Effect of Crude Oil Pollution and Water Strom on Some of the Physical and Chemical Properties of Soil 40 Mohannad I. Khuder, Ehab M. Amen and Noor Fanar Abdulbaqi S-250 Detection of Organochlorine Pesticides in Typha domingensis in the Euphrates River - Middle of Iraq 44 Amirah Imran, H. AL-Janabi, Hala Jumaah Asree, Ali Abed Gatea, Nawar AL-Janabi and Ahmed Ali Khirallah S-251 Effect of Organic and Chemical Fertilizers on Growth and 50 Yield of Cucumber, Cucumis sativus L. Nasser Jubair Radhi Alhasnawi, Amer Neama Kareemand Zina Mohsin Abdullah S-252 Effect of Application of Pseudomonas fluorescens and Nitrogen Fertilizer on the Growth and Yields of 54 Cultivars of Bread Wheat Ali Salim Hussein, Hussein Razzaq Nayyef and Bassim Hassan Abdulrudha S-253 Evaluation of Hydrochemical Properties of Groundwater and Suitability to Irrigation Purpose and Fixed 60 Sand Dunes in Al-Barsa Area, Eastern Al-Dour City, Iraq Ehab Mohammad Amen, Mayada Ahmed Abdullah and K. MohannadIssa S-254 Effect of Heavy Metals on the Protein and Chlorophyll Content of Phragmitus australis and Typha 65 domingensis Qassim Ammar Ahmood AL-Janabi, Ziad Bassim Hameed and Saad Kadhim Ala Allah Al- Kalidy S-255 Effect of Soil Fertilization with Nutrition Solution (Enraizal) on Fruits Growth and Yield for the Two Hybrids 72 of Cucumber Cultivated in unheated Plastic Houses Dhafer H. Jawad, Fadhil Jawad Kadhim Altaee, Mohammed J.H. AL- Shareefi and Hussein Ali K. Al-Hasnawi S-256 Effect of Foliar Application with Novalon on Growth and 77 Grain Yield of Three Genotypes of Maize Nassr M. Jasim and Basheer H. Abdullah S-257 Effect of Iron Oxide Nanoparticle and Humic Acid Spray on Growth and Yield of the Fenugreek (Trigonella 82 Foenum Graecum L.) and Fixed Oil Content in Seeds Qasim A.S. Al-Zyadi and Emad Abud-Alkareem Mohammedreda Al-Thahab S-258 Effect of Spraying with Gibberellic Acid on Growth and Yield of Three Cultivars of Broad Bean (Vicia faba 85 L.) Ahmed Hasan Fadhil and Mohammed Mehdi Muhsen Almasoody S-259 Effect of Weed Management on Growth Parameters in Sunflower (Helianthus annuus L ) 90 Mahmoud T. Al-Jayashi and Hayder R. Laibi

Date of Publication 15.12.2019