Matricaria Chamomilla L.) Populations

Full Length Research Paper

Effect of different levels of salinity on some morpho- physiological characteristics in German Chamomile (Matricaria chamomilla L.) populations

Yasamin, M.1*, Adel, D.M.N.1, Hossein, Z.2 and Rohallah, A.1 . 1Department of Plant Ecophysyology, Faculty of Agriculture, University of Tabriz, Tabriz, Iran 2Natural Science department, Isfahan Agriculture and Natural Resources, Research and Education Centre, AREO, Isfahan, Iran.

Accepted 16 December, 2017

The experiment was conducted to determine the effect of different levels of salinity on some of morpho- physiological characteristics in German chamomile (Matricaria chamomilla L.) cultivated for two years in Isfahan, Iran. The experimental treatments consisted of four salinity levels of the irrigation water (6, 9, and 12 dS m-1) compared to the control (2 dS m-1) and 3 chamomile (Isfahan, Tehran and Ardebil) populations. A field examination was conducted using randomized Split-Plot statistical designs with 3 replications. The seedlings were plannted in nursery then transplanted in early March in 2011 and 2013 to the main land. Salinity treatments were applied by weekly irrigation after transplanting. In this study plant height, number of flowers per plant, shoot, root and flower weight (fresh and dried), flower essential oil weights, Leaf relative water content, leaf relative water lose and 1000 seed weight were investigated. The results showed that the Tehran population in most characteristics was best. There were not significant difference in economically important traits such as fresh and dry weight of flower and essential oil weight among populations. Except of root fresh and dry weight, plant height and 100 seed weight, other traits were higher significantly at first year. All traits were highest in control Ec. Additionally by increasing in salinity levels, morphological traits showed different trend. Salinity could reduce RWL and increase RWC adversely. In conclusion we could demonstrate that using of different levels of salinity had significant effects on some morpho- physiological characteristics in Matricaria chamomilla L. Despite the non significant difference, Isfahan population had highest essential oil. According to the effect of salinity on economically important characteristics, the chamomile cultivation can be recommended by 9 dS m-1.

Key words: Matricaria Chamomilla, Salinity, irrigation, Morpho-physiological characteristics, Isfahan. Population.

INTRODUCTION

Iran is located in mostly arid and semi-arid areas and (Al-Khateeb, 2006). Salt stress and soil salinity creates water availability is a major problem in crop production. ionic and osmotic stress on many plants. Also, ionic Fifteen percent of total agricultural lands of Iran have salt toxicity generated from salt contaminated soil has in water or soil. Soil salinity is one of the worldwide negative effects on plant growth and development problems adversely affecting the agricultural production (Munns et al., 2006).There are many defense (Levitt, 1980). Salinity is a major environmental stress mechanisms in plants which are tolerant to water-deficit and is more conspicuous in arid and semi-arid areas and salt stresses, such as osmoregulation, ion where 25 % of the irrigated lands are aﬀected by salts homeostasis, antioxidant and hormonal systems (Mahajan and Tuteja, 2005), helping plants to survive and grow under severe environmental conditions prior to their reproductive stages. The defense mechanisms in *Corresponding author e-mail: [email protected]. sensitive plant species are weaker, leading to growth Scholarly J. Agric. Sci. 96

Table 1. Soil chemical characteristics of experimental field.

EC (dS/m) 5 pH 7.5 Cl (Meq/l) 23 S.Anions 28.6 Mg + Ca (Meq/l) 48 Na(Meq/l) 21 S.Cations 69

retardation and yield reduction. Biochemical and -nt of Natural Science at Agriculture and Natural physiological parameters in plants cultivated in salt Resources, Research and Education Centre of Isfahan conditions have been developed as effective indices for Province, Isfahan, Iran during 2013-2014. The tolerant screening in plant breeding programs (Ashraf geographical location of the station was “323616”N and and Foolad, 2007). Salinity adversely affects the process “513448”E with 1671 m altitude. The soil properties of the of germination, seedling establishment, plant growth and location are presented in Table 1. yield in almost all the cultivated crops by lowering the Seeds were planted in the 2011 autumn cases and the osmotic potential of water in the growing medium or by seedlings were transplanted in early 2013 March. causing specific ionic toxicity or both. Salinity has been Selected populations for planting were pointed from the reported to affect adversely the composition of result of genetic diversity plan during the previous years. photosynthetic pigments and the changes in pigment Several indices to measure each samples were selected composition depend upon the specific nature of ion, plant randomly from the area and finally they were averaged. species and the age of plant (Levitt, 1980). The Each experimental plot size was 3 m × 4 m and in each anatomical and morphological features typical of plot, the plants were grown in three equidistant rows with halophytes are usually considered to be adaptations to adjacent rows being 30 cm apart. After transplanting the salinity. There is very little experimental evidence to show seedling from the nursery to the main land, salinity whether the same features occur when the halophytes treatments were applied with weekly irrigation. The are not exposed to the effects of salinity. German experimental treatments consisted of four salinity levels chamomile (Matricaria chamomilla Linn) is an important of the irrigation water (6, 9, and 12 dS m-1) compared to medicinal herb belonging to the family Asteraceae the control (2 dS m-1) and 3 chamomile populations (Maheshwari, 1963). It naturally grows in west, northwest (Isfahan, Tehran and Ardebil) with 3 replication. The and southern part of Iran. The oil extracted from flower is salinity levels were obtained by addition of appropriate aphrodisiac and analgesic used for relieving all kinds of amount of NaCl to water and were adjusted by a portable pains. About 120 chemical constituents have been EC meter instrument. The different irrigation treatments identified in chamomile as secondary metabolites, started in the beginning of transplanting after the started including 28 terpenoids, 36 flavonoids and 52 additional their growth and development until harvest. compounds with potential pharmacological activity. It may be considered as an economic substitute for field crops Investigation some morpho-physiological traits irrigated with fresh water since it has adaptability to wide range of climate and soil (Abari et al., 2011; Bewley and At full flowering stage, five plants were selected from Black, 1982). The areas becoming unsuitable for middle row by eliminating border effects. Then, the traits; conventional crops may be used for growing such plants No. of flowering stems, plant height (cm), capitol diameter depending upon their suitability to the prevailing of flowers, flowers and root weight (wet or dried), plant environmental condition. The performance of chamomile height, number of branches per plant, number of flowers on saline water has not been well studied or documented per bushes, plant dry weight, number of flowering and there are only a few reports on the effect of water branches and essential oil weight were measured and salinity on morphological characters and oil composition investigated. The flowers were gradually harvested and of chamomile (Afzali et al., 2006). The aim of this study weighted for determination of fresh flower yield. Dry root was to determine the effect of different levels of salinity and flower weight of each plot was calculated after drying on some morpho-physiological characteristics in the flowers at room temperature (20–25 at 8 ºC). Plant Matricaria Chamomilla L cultivated in Isfahan, Iran. height was measured by using a ruler with an accuracy of 1mm of 5 plants and their averages were evaluated. The individual plant samples of each population were MATERIAL AND METHODS conditioned in plastic bags and transported to the laboratory under refrigeration. Samples (~200 g) were air The present experiment was carried out in the Departme- dried at room temperature (20 to 25ºC) and maintained in Yasamin et al., 97

Table 2- salinity effect on some morpho-physiological traits in Matricaria chamomilla populations.

Treatments Means Fresh hoot Dry shoot Fresh root Dry root Fresh flower Dry flower Number of weight (g) weight (g) eight (g) weight (g) weight (g/m2) weight (g/m2) flowers per plant Genotypes Isfahan 27.83b 9.53 64.21b 12.08b 2014 1117 40.47b* Tehran 32.46a 10.38 77.88a 16.73a 1980 1089 48.22a Ardebil 32.13a 9.78 76.55b 15.80b 2020 1125 47.40b P-value ** n.s ** ** n.s n.s ** Year 1 32.54a 17.11a 62.18b 13.77b 1980a 1528a 47.43a 2 29.07b 12.68b 83.56a 15.98a 1130b 692b 43.50b P-value ** ** ** ** ** ** ** EC 2 37.98a 10.99a 85.02a 18.40a 1440a 1190a 57.56a 6 25.47c 8.92bc 62.22c 13.61c 1307ab 1141ab 45.31b 9 30.86b 9.90b 78.00b 14.90b 1295ab 1130ab 40.52bc 12 38.92b 8.83c 66.27c 12.58c 1397b 1097b 38.34c P-value ** ** ** ** ** ** **

*Means within row with no common on letter are significantly different (p≤ 0.05).

a refrigerated chamber (10ºC) until extraction. Each Isfahan, Tehran and Ardebil selected genotypes in first sample used in this survey was deposited at the and second years of cultivation period. Herbarium, Islamic Azad University, Isfahan (Khorasgan) The statically model was: branch, Isfahan, Iran. The air-dried samples (100 g) were subjected to water distillation for 2 h using a Clevenger- yijk = µ + αi + βj + (αβ)ij + cik + eijk type apparatus. The leaf relative water content (LRWC) was determined in the fully expanded topmost leaf of the Whereas : µ is a population mean, αi is the main effect of main shoot. The fresh weight of the sample leaves was soil type (A) i, i = 1, . . . , 3, P3 i=1 αi = 0, βj is a main recorded and the leaves were immersed in distilled water effect of barley variety (B) j, j = 1, . . . , 4, P4 j=1 βj = 0, in a Petri dish. After 2 h, the leaves were removed, the (αβ)ij is the interaction effect of A and B, Pi(αβ)ij = 0 for surface water was blotted-off and the turgid weight each j and Pj(αβ)ij = 0 for each I cik ∼ iid N(0, σa2) is the recorded. Samples were then dried in an oven at 70°C to plot (pan) error distribution, k = 1, 2, eijk ∼ iid N (0, σe2) constant weight. Leaf relative water content was is the subplot (individual) error distribution, k = 1, 2. calculated using the following formula (Turner, 1981) : Data were analyzed by analysis of variance using the LRWC (%) = [(F.W – D.W) / (T.W – D.W)] × 100 SAS (2003) software program. The Whereas: F.W., Fresh weight; D.W., Dried weight; T.W., mean values were compared using the least significant Turgid weight. difference test (LSD) and P value less than 0.05 was To determine the relative water loss (RWL), A sample considered as significant. of 5 fresh flag leaves were taken from each plot and FW was measured. The leaves were then wilted at 35°C for 5 hrs and reweighed (W5H). Then the samples were oven- RESULT AND DISCUSSION dried at 70°C and weighed again (DW). RWL was calculated by the following formula (Farshadfar et al., The mean comparisons of some morphophysiogical traits 2001) : RWL (%) = [(F.W – W5H) / (F.W – D.W)] × 100 of Matricaria chamomilla due to salt stress are shown in Whereas: F.W., Fresh weight; W5H., The leaves were tables 1. There were significant differences in trial then wilted at 35°C for 5 hrs and reweighed: D.W., Dried groups. Fresh Shoot weight, root fresh and dry weights, weight; Number of flowering branches and flowers per meter were at the highest amount in Tehran population Statistical analysis compare to other ones. Except of root fresh and dry weight, plant height and 100 seed weight, other traits A field examination was conducted using randomized were higher significantly at first year. All traits were Split-Plot statistical designs with 3 replications at the highest in control Ec. The amount of Fresh and dry shoot experimental station of Isfahan training center of weight, Fresh and dry root weight, Essential oil weight agriculture. The main factors were different levels of and Number of flowering branches in plots that obtained salinity (2, 6, 9 and 12 dS m-1) and sub-factors were water Ec = 9 dS m-1, were near to them in control level. Scholarly J. Agric. Sci. 98

Table 3- salinity effect on some morpho-physiological traits in Matricaria chamomilla populations.

Means Treatments Essential oil Number of Flowers capitol 1000 seed plant RWC RWL weight (g) flowering branches diameter (cm) weight (kg) height (cm) Genotypes Isfahan 0.380 22.15b 2.14 0.028a 31.82 63.83b 18.06a Tehran 0.316 26.25a 2.23 0.027b 30.83 60.72c 17.58b Ardebil 0.341 23.79b 2.15 0.026c 31.50 65.30a 17.20b P-value n.s ** n.s ** n.s ** ** Year 1 0.345 36.22a 2.35a 0.027b 30.57b 63.29 17.61a 2 0.346 11.90b 1.98b 0.034a 32.77a 63.98 15.97b P-value n.s ** ** ** ** n.s ** EC 2 0.386a 30.02a 2.15 0.028a 33.91a 59.28c 18.50a 6 0.305b 19.16d 2.19 0.028a 30.49c 61.73b 18.27a 9 0.377a 25.22b 2.14 0.027b 29.10c 63.56b 16.51b 12 0.314b 21.84c 2.19 0.026b 32.08b 64.87a 16.14b P-value ** ** n.s ** ** ** n.s

*Means within row with no common on letter are significantly different (p≤ 0.05).

Fresh and dry flower weight, Number of flowers per plant, reduction in oil content was highest under sulphate 1000 seed weight and RWL were decreased significantly dominated salinity as compared to chloride dominated by increasing in salinity levels. Leaf relative water content salinity German chamomile appears to be highly salt (RWC) was in highest and leaf relative lose (RWL) in tolerant medicinal herb. Zeinali et al (2008) investigated least amount in Ardebil population. the regression model for dry flower yield as dependent Results revealed that flower essence weight was higher variable and the other traits as independent in German in Isfahan population and the year of cultivation had no chamomile. Results showed the traits days to 50% effect on weight of flower essence. Plant height was at flowering, No. of flower plant-1 and plant height entered the lowest when EC increased and it was at the highest to model and accounted for 73, 8 and 3% of dry flower in Isfahan population. yield variation, respectively. In the other research, has The result of 1000 seed weight (kg), RWC and RWL been reported the significant and positive relationship of are shown on table 2. As result revealed from this the traits, No. of flower plant-1, fresh flower yield, 100 data1000 seed weight (kg) was increased in second year. flower weight, days to flowering and plant height with Additionally different levels of salinity and genotypes essence percent (Pirkhezri et al., 2008). Salt stress is a could change it significantly. There were significant limiting factor of plant growth and yield, and becoming a differences between experimental groups toward RWC serious problem in the world (Chaum and Kirdmanee, and RWL.RWC was increased by salinity levels and RWL 2009). Salinity which is produced by root cells plays a was decreased instead. prominent role in various physiological processes such as plant's growth, ions uptake, photosynthesis and germination (Hayat and Ahmad, 2007, Raskin, 1992). In DISCUSSION the currents study there different types of salinity was changed flowers capitol diameter and number of flowers In this study data showed that there were significant per each plant significantly (P≤0.05). The results of differences in experimental groups about Shoot weight (Mahdikhani et al. 2007) revealed that the high (P≤0.05). Additionally there were significant differences phenotypic variation for the traits biological yield, dry between experimental groups by EC types (P≤0.05). flower yield, Number of flower plant-1 and essence Number of flowering branches, flowers capitol diameter percent. Omid (1999) showed the lower essence percent and number of flowers per each plant were at the highest and kamazulene of the local varieties in comparison with in Tehran population (P≤0.05). Results of (Deepika et al., the advanced cultivars. 2015) study indicated that increasing salinity from 0 to Pirkhezri et al (2008) observed significant differences 150 mM, decreased fresh weight of shoot (76.3%) and between chamomile genotypes for all the traits except increased of root fresh weight (53.8%). Also they shiwed stomata length. Zeinali et al (2008) investigated the effect that oil content of air dried flowers remained almost of planting date in chamomile. They reported that on 5th unaffected upto 8 dSm-1 EC levels but thereafter it March increase in the traits No. of flower plant-1, days to declined with increasing EC levels. They noted that this 50% flowering, dry flower yield and essence percent in Yasamin et al., 99

comparison with 20 March and 5 April. Indeed, delay Education Centre. planting and impact of flowering stage with temperatures above 23ºC causes florets infertility and finally flower yield deficiency. In (Ghanavati and Sengul, 2010) study there REFERENCES was a significant difference between genotypes studied Abari, A.K., Nasr, M. H., Hojjati, M. and Bayat, D. (2011). Salt effects on for all traits except for root relative water content. In seed germination and seedling emergence of two Acacia species. general, high NaCl concentrations produce water deficit, African Journal of Plant Science, 5(1), 52-56. Afzali, S.F., H. Shariatmadari, M.A. Hajabbasi and Moatar, F (2007). ion toxicity, nutrient imbalance and oxidative stress. Salinity and drought stress effects on flower yield and flavonol-O- These adverse effects cause modifications of root glycosides in chamomile (Matricaria chamomilla L.). Iranian Journal morphology and inhibition of plant growth and can result of Medicinal and Aromatic Plants, 23(3): 382-390. in plant death. Tolerance to abiotic stresses is associated Alexandra S (2005). German chamomile (Matricaria chamomilla L.) population morphological and chemical diversity. Budapest Doktorin with modifications of morphological and physiological thesis. Budapest university Department of Horticulture. traits (Edmeades et al., 2001). These results are highly Al-Khateeb, S.A. (2006). Effect of salinity and temperature on inconsistent with findings of Cirecelav et al (2008), Zeinali germination, growth and ion relations of Panicum turgidum Forssk. et al (2008), Alexandra (2005) and Marinho (2006) on Bioresource Technol, 97, 292-298. Ashraf M and Foolad M.R. (2007). Role of glycine betaine and proline in German chamomile genotypes and accessions. improving plant Ghanavati et al (2011) showed from salt application trials abiotic stress resistance. Environ. Exp. Bot.206-216. indicated that dry matter yield was decreased with Bewley, J.D. and Black, M. (1982). Physiology and Biochemistry of increasing NaCl doses. Pirzad et al (2011) showed that seeds in Relation to Germination, Springer-Verlag, Berlin. Blokhina O., Virolainen, E. and Fagerstedt, K.V. (2003). Antioxidants, leaf relative water content (LRWC), proline and total oxidative damage and oxygen deprivation stress. A review. Ann. Bot. soluble sugar were not affected by irrigation levels. They 91: 179–194. showed that in their experimental condition, LRWC had Chaum S. and Kirdmanee, C. (2009). Effect of salt stress on proline no changes in range of our treatments. Despite being accumulation, photosynthetic ability and growth characters in two Maize cultivars.Pak. J. Bot. 41(1): 87-98. same, LRWC in all treatments were in high amounts, Cirecelav G, De MG, D'Andrea, L. and Nano, G.M. (2002).Comparison showing a high level maintenance of water in water deficit of chamomile biotypes (Chamomilla recutita L. Rauschert). ISHS and excess water conditions. The rate of RWC in plants Acta Horticulturae 330: WOCMAP I – Med. Arom. Plants Conf., p. 44. with high resistance against drought is higher than Darzi M .2002. Evaluation of agro-ecological problems in chamomile and fennel. Zeitun, 152: 43-49. others. In other words, plant having higher yields under Deepika, U.K., Varshney, P.S. and Manogya, S.S,P.(2012). Influence of drought stress should have high RWC. Under water salinity on morpho-physiological characters of German chamomile. deficit, the cell membrane is subjected to changes such World journal of pharmacy and pharmaceutical sciences.4, 10, 762- as increase in penetrability and decrease in sustainability 775. Edmeades, G.O., Cooper, M., Lafitte, R., Zinselmeier, C., Ribaut, J.M., (Blokhina et al., 2003). Habben, J.E., Löffler, C. and Bänziger, M.2001.Abiotic stresses and staple crops. In Crop Science: Progress and Prospects. Proceeding of the Third International Crops Science Congress, 17–21. CONCLUSION Farhoudi, R.; Lee, D.-J.; Hussain, M. Mild Drought Improves Growth and Flower Oil Productivity of German Chamomile (Matricaria recutita L.).2014.J. Essent. Oil Bear. Plants.17, 26–31. In conclusion we could demonstrate that using of different Farshadfar, E., Farshadfar, M. and Sutka, J. (2000). Combining ability levels of salinity had significant effects on some morpho- analysis of drought tolerance in wheat over different water regimes. physiological characteristics in Matricaria chamomilla L. Acta Agronomica Hungarica.48(4): 353-361. Ghanavati, M. and Sengul, S. (2010). Salinity effect on the germination populations. Although the Tehran populations in most and some chemical components of Chamomilla recutita L. Asian J. characteristics were best but despite the non significant Chem., 22(2): 859-866. difference, Isfahan population had highest essential oil. Hayat, S., Fariduddin, Q., Ali, B. and Ahmad, A. (2005). Effect of According to the effect of salinity on economically salicylic acid on growth and enzyme activities of wheat seedlings. Acta Agron. Hung. 53: 433-437. important characteristics such as fresh and dry weight of Levitt J (1980). Salt stress. In: Responses to Environmental Stresses. flower and essential oil weight, the chamomile cultivation Vol. II. Academic Press, New York can be recommend till 9 dS m-1 water Ec. Regarding to Mahajan, S. and Tuteja, N. (2005). .Salinity and drought stresses: an these results and non significant effect of salinity on overview. Arch. Biochem. Biophys.444: 139–158. Mahdikhani, H., Solouki, M., Zeinali, H. and Imam, J.A. (.2007). Genetic RWC, it can be concluded that chamomile is a salt diversity assessment in German chamomile populations base on tolerant plan. Our primary recommendation is that this morphological traits. 3rd Medicinal Plants Congress. Shahed experiment should do in other field with other types of University, Tehran, Iran, p. 17. genotype and EC .Also further tests are needed to Maheshwari, J.K. (1963). The flora of Delhi. CSIR publications, New Delhi. explore and more detail explanation. Marinho, M., Larissa, P., De BV., Klezia, M.S.B., Cid AMS. and Almeriane, M.W.S. (2006. Comparison of chemical constituents of Chamomilla recutita L. Rauschert essential oil and its anti- ACKNOWLEDGMENTS chemotactic activity.Braz. Arch. Boil. Technol., 49: 234-240. Munns, R., James, R.A. and La Uchli, A. (2006). Approaches to increasing the salt tolerance of wheat and other cereals. J. Exp. This research project has been supported by Isfahan Bot.57: 1025–1043. Agriculture and Natural Resources, Research and Omid, B.R. (1999). Evaluation of Iranian chamomile chemotypes and Scholarly J. Agric. Sci. 100

comparison with improved cultivars. J. Agric. Sci. Modarres, 1: 45-53. Raskin, I. (1992). Role of salicylic acid in plants. Annu. Rev. Plant Pirkhezri, M., Hasani, M.A. and Tabatabaie M.F. (2008). Evaluation of Physiol. Plant Mol. Biol. 43 439–463. genetic diversity of some German chamomile populations (Matricaria SAS, 2003. Statistical analysis system version 9.1 for windows. SAS chamomilla L.) using some morphological and agronomical Institute, Inc., Cary, NC, USA. characteristics. J. Hortic. Sci., 22: 87-99. Zeinali H, Bagheri M, Golparvar AR, Jafarpour M, and Shiranirad AH. Pirzad A.R., Shakiba, M.R., Zehtab-Salmasi, S., Mohammadi, S.A., (2008). Effect of different planting time and nitrogen fertilizer rates on Darvishzadeh, R. and Samadi A (2011. Effect of water stress on leaf flower yield and its components in German chamomile (Matricaria relative water content, chlorophyll, proline and soluble carbohydrates recutita L. Rausch). Iran. J. Crop Sci., 10: 220-230. in Matricaria chamomilla L. Journal of Medicinal Plants Research 5(12): 2483-2488.