Journal of Chemical Health Risks

Journal of Chemical Health Risks ( ) ( ), –

ORIGINAL ARTICLE

Changes Germination, Growth and Anatomy ervilia in Response to Light Crude Oil Stress

Bahareh Lorestani, Nasrin Kolahchi, Mahshid Ghasemi*, Mehrdad Cheraghi

Department of Environmental, Hamedan Branch, Islamic Azad University, Hamedan, Iran

(Received: August Accepted: November )

ABSTRACT: Thepetroleum concentrations are contaminant important sources in the environment. Hydrocarbon’s contaminants depend on their type and concentration can cause variable toxicity in KEYWORDS soils, on the other hand, different kinds of also response concentration of contaminant Oil pollution differently, because of the diversity of physiological and morphological characteristics. In this study Germination the effect of different concentrations of light crude oil was investigated on stem length, germination Stem length and anatomical of Vicia ervilia. A factorial experiment was used with a randomized complete block Anatomy design (RCBD) with three replications. The results showed that the germination and stem lengths

Vicia ervilia decreased significantly (p≤ ) with increasing concentrations of petroleum. Anatomical studies the changes of parenchyma, vascular bundles, epidermal and increase crack showed.

INTRODUCTION Oil pollutions make disturbances in vegetations [ ]. The signs of pollution in vegetation are numerous [ ]. Increasing industrial productions are along with Petroleum cause chemically disturbances in metabolism increasing pollutant in the environment [ ]. Petroleum and growth of vegetation [ ], reduction of vegetation pollutant is an unavoidable consequence of population growth [ ], autumn of areal organs (leaves), variations increasing and industrialization process. Soil pollution of pile, chlorophyll destruction, delay in flowering [ ] by petroleum concentrations is an important and cell death [ ]. In studies which were conducted by environmental problem [ ]. Petroleum hydrocarbons are researchers, the performance of vegetation was studied harmful for the alive and no alive components due to in polluted soil by petroleum. The results showed that toxicity effects [ ]. Pollutant in soil can insert into food petroleum in some vegetations cause to decrease chain [ ] and this is dangerous for the health of human germination and growth ( , and ), delay or lack of and other animals [ ]. Petroleum concentrations can germination [ , ], increase root length and improve affects on vegetation growth through stimulate the soil the vegetation growth in polluted soil [ , and ]. bacteria, consume soil nutrients and reduce soil oxygen.

∗Corresponding author: gha [email protected](M. Ghasemi).

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In previous studies, the performance of vegetation was Where in this equation N is the number of planted seeds, studied in polluted soil by petroleum. The obtained n is the number of germinated seeds and G is the results showed that petroleum in some vegetations cause germination percentage [ ]. The length of stem from the following effects: decrease of germination and the soil surface to top of the stem was measured using growth ( , and ), delay or lack of germination [ , ruler in cm [ ]. The lower part of stem was sampled for ], increase of root length and improve the vegetation the anatomy experiments [ ]. Vegetation texture was growth in polluted soil [ , and ]. Results of placed in solution of glycerin and ethanol , because several researches showed that the petroleum vegetation texture can be protected in these concentrations produce different degree of toxicity consolidators in long term [ ]. The common blade was according to the type and their concentration. Mentioned used to prepare sections from vegetation; the sample items show that the petroleum pollution caused coloring was done using duplicated Carmen Stain. disturbances that are morphological, physiological and Prepared samples were assessed under the light anatomy variations in vegetations. The anatomic Moticmicroscope and IXUS HS canon camera was properties of vegetations are very important, but a little used to image samples. Excel was used to design attention has been paid to the effects of petroleum on diagrams and SPSS software was used to compare vegetation anatomy which have grown in polluted soil. means by Duncan test at probability level of . Therefore, in this study the effects of different RESULTS concentrations of light crude oilon germination, stem length and anatomy of Vicia ervilia was investigated in The results of this study showed that the existence of polluted soil. light crude oil at the different concentrations had effects

on germination of plants and the germination percentage MATERIALS AND METHODS decreased with increasing petroleum in soil. At Soil was prepared from the adjacent agricultural lands. concentration of it was observed a delay in Soil was polluted by different concentration ( , , and germination. germination was detected in control W/W %) of petroleum and a soil without pollution treatment. Germination at the concentrations of , , was considered as control. Manual method was used to and were , , and , respectively mix soil and petroleum [ ]. Seeds with germination (Figure ). potential were planted in vessels contained by polluted and unpolluted soil at the depth of cm. three Control %1 %2 %3 %4 replications were prepared from each concentration. All 20

18 vessels were placed in free location with even 16 14 temperature and environmental conditions. Control 12 treatment was for the vegetation grown in unpolluted 10 8 soil. 6 4 In this study the germination and stem height were (Number) Germination 2 measured. The germination rate in each vessel was 0 calculated according to the number of planted seeds and Day following equation: Figure . Germination of Vicia ervilia (Based on the counting of the number of germination per day)

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Statistical analysis of data showed the significant , but there was no significant difference among the difference among treatments. In comparison to control control and treatments of and (Figure ). and contaminant treatments, it was not observed a 16 a significant difference between the control and ba 14 ca treatments of and (Figure ). 12 dc 10 8 e 120 a 6 100 4 bac ea

80 2 Average stem length (Cm) length stem Average 60 0 cd Percent 40 Different concentrations of oil 20 bd Figure .mean height of Vicia erviliastem in cm(Columns with same 0 subscript shows that there isno significant difference at probability level of )

Different concentrations of oil The stem of the vegetation grown in contaminant soil Figure . Germination rate of Vicia ervilia (At least a common subscript shows that there isno significant difference at probability was different with the stem of vegetation grown in non- level of ) contaminant soil. These variations are shown in Figures

and . The height of stem was measured every days to determine the difference in growth variations. The in initial days had more height growth at concentration of as compared to other contaminant concentrations.

But at the end of growth period it showed significant reduction compared to treatment of and (Figure ).

Control %1 %2 18 %3 %4

16

14 12 10 8 6 Stemlength (Cm) Stemlength 4 2 0

Day

Figure . measuring stem height of Vicia ervilia (Every day in growth period)

The results of the statistical analysis of the mean of stem height and investigation of the effects of concentration on stem height showed that there was significant difference among the treatments at probability level of

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Figure . Increasing of pile in contaminant sample compared to control (Left sample is control and right sample is contaminant sample) e: Epidermal, ph: Phloem, X: Xylem, co: Collenchyma, p: Parenchyma, Pi: pile

Figure . comparison of the cross section of Vicia ervilia and Variation in contaminant sample (right) and control (left) e: Epidermal, ph: Phloem, X: Xylem, co: Collenchyma, p: Parenchyma

Anatomical studies showed the changes of parenchyma, samples was irregular and compacted as compared to vascular bundles, epidermal and increase crack. As it control. was shown in Figure , control sample of Vicia ervilia DISCUSSION has determined section and shape, in contaminant samples, it can be observed and an increase in swell and Germination is the most important vegetative stage of variations in cross section shape. In Figure (down plants [ ]. Germination rate was observed in all the figure) control sample had large parenchyma cells and concentrations of petroleum but the germination rate regular epidermal but in contaminant samples the size decreased with increasing petroleum concentration in of Parenchyma cells decreased but its frequency soil which this was in agreement with the findings of increased. Moreover the irregularity in epidermal size Njoku and colleagues, and Kirk and colleagues [ , can be observed. Phloem vascular of contaminant

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]. Decreasing of germination was happened due to grown plants. Vicia erviliais dicotyledonous species reducing oxygen and moisture [ ], poor accessibility which have epidermal layers, parenchyma and nutrient for plant [ ] and increasing toxicity materials vascular bundles in the structure of stem. In contaminant of petroleum compositions in soil [ ]. Delay in treatment the pile increased. Increasing pile can be a germination at concentration was in agreement with tool of sourcing more petroleum compounds. Moreover the findings of Kisic and colleagues, which reported that an irregularity was observed in vascular bundles. Xylem petroleum compositions provided a soft layer around the and phloem vascular were compacted [ ] and seed and prevented from access to oxygen and water decreased by increasing the concentration of petroleum [ ]. This caused delaying or lacking germination in which this was not in agreement with the finding of polluted soil by oil. The mean of stem length of Vicia Omosun and colleagues, [ ]. The response of erviliain three replications from control to oil vegetation to decrease nutrients and nitrogen of soil concentrations of were respectively equal to , reduced xylem and phloem vascular. Thus, reduction of , , and . The stem length was vascular bundles led to decrease transporting petroleum significantly decreased with increasing the into plant. Decreasing of the vegetation growth showed concentrations of light crude oil (P< ). Reduction of the disturbances in performance of xylem and phloem vegetation growth by contaminant soil was proved by vascular in providing water and nutrient under the stress several researchers [ , and ]. Daryabeigizand and of soil contaminant by petroleum. This result was in colleagues found that the light crude oil compounds agreement with the finding of Agbogidi and Eshegbeyi, hadn’t significant negative effects on germination but it and Agbogidi and Ofuoku [ , ]. Increasing of had significant effects during the vegetative stages [ ]. parenchyma was in agreement with the findings of Gill This result showed that the oil by its toxicity effects and colleagues, [ ]. This change has been occurred to prevented the natural growth of plant. Mendelssohn and produce stability in response to stress and/or preventing Hester reported that the decreasing of stem length was from the entering petroleum compounds in to cells. The due to physical, chemical and biological variations by cross section Vicia erviliahas changed. Nogueira and petroleum compounds in soil [ ]. Moreover reduction colleagues, in their research showed the changing of of stem length can be due to decreasing accessible cross section [ ]. Irregularity in epidermal was due to nutrients for plants [ ] or poor ventilation of soil which the disturbances in plant morphology. Epidermal caused that the growth prevented and its length thickness increased slowly which this was in agreement decreased [ ]. According to the reports and results of with the finding of Sharma and colleagues, and Gill and statistical investigations of different concentrations of colleagues [ , ]. They found the epidermal variations oil as compared to control showed that the toxicity in vegetation grown in contaminant soil. Epidermal compounds of light crude oil had negative effect on variations was occurred due to the preventing the germination and growth of Vicia ervilia and the severity reduction of inter tissue water [ ]. In Table of effects increased with increasing the concentration of observations of the present study are given compared to petroleum in soil. Species had degrees of sensitivity other studies. based on their morphological and physiological properties; therefore it is important to investigate the anatomical properties of plants. The observed variations were due to the effects of petroleum compounds in soil

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Table . Observations of the present study comparison with other studies The parameters Studied Observations in this study Correspondence Nonconcurrence

Germination rate of was observed in all the concentrations of petroleum but the germination rate Njokuet al., ; Kirk et al., Germination decreased with increasing petroleum concentration in —

soil.

Delay in germination was observed at a concentration of Delaygermination percent. Kisicet al., —

The stem length was significantly decreased with Okonokhuaat el., ; Averagestem length increasing the concentrations of light crude oil (P< ). Shahriariet al., ; — Ogboghodoet al., The pile was increased in contaminant treatments. — —

Irregularity was observed in vascular bundles. — —

Xylem and phloem vascular were compacted by increasing the concentration of petroleum. Kofidiset al., —

Xylem and phloem vascular were decreased by increasing the concentration of petroleum. — Omosunet al., Anatomical Decreasing of the vegetation growth shows the disturbances in performance of xylem and phloem vascular in providing water and nutrient under the stress Gill et al., — of soil contaminant by petroleum.

Increasedparenchymaltissue. Nogueiraet al., —

The cross section Vicia ervilia has changed. Sharma et al., ; Gill et al., —

CONCLUSIONS REFERENCES

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