Allelopathic effect of Silene villosa on germination and seedling growth of Triticum aestivum L.

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1 Research Article ISSN: Available online through Allelopathic effect of Silene villosa on germination and seedling growth of Triticum aestivum L. Waheed Murad 1, Rehman Ullah 1, 3 *, Aziz Ullah 1, Muhammad Gul Riaz Khan 1, Hameed ur Rehman, Zulqarnain 3, Zain Ullah Khan 3 1 Department of Botany, Kohat University of Science and Technology (KUST) Kohat, Khyber Pakhtunkhwa, Pakistan Department of Chemistry, Kohat University of Science and Technology (KUST) Kohat, Khyber Pakhtunkhwa, Pakistan 3 Department of Botany, Government Post Graduate College Karak, Khyber Pakhtunkhwa, Pakistan Received on:7-1-16; Revised on: 1--16; Accepted on: ABSTRACT Background: The aim of the present study was to determine the allelopathy of Silene villosa Forssk collected from District Karak Khyber Pakhtunkhwa Pakistan. Materials and hods: For allelopathy stock solution of (1mg/ml) was prepared in 1% DMSO for each of n-ane, oroform and Ethyl acetate soluble fractions. For each stock solution 4 dilutions of,, and were prepared. The stock solution and dilutions were tested against Triticum aestivum L. variety Shahkar 13. The allelopathy was obtained by measuring the germination percentage, root shoot length and fresh and dry weight of root and shoot. Results: The result showed that, Silene villosa have significant inhibitory role against wheat. The germination percentage was reduced in all the fractions as the concentration was increased. Increasing concentration has definite inhibitory effect on root and shoots length, fresh and dry weight of root and shoot. Conclusions: The investigation shows that the plant has remarkable allelopathic activity against wheat plant. KEY WORDS: Allelopathy, Silene villosa, Triticum aestivum, n-ane, oroform, Ethyl acetate. *Corresponding author. Rehman Ullah Department of Botany, Kohat University of Science and Technology (KUST), Kohat, Khyber Pakhtunkhwa, Pakistan. INTRODUCTION Plants naturally prepare certain substances which have effect on germination and growth of other plants. Such reciprocal relation of plants is called allelopathy 1. These substances include phenol, tannins, flavenoides, alkaloides and terpenoides. There are various factors which are responsible for transfer of these substances to the environment, e.g. during exudation these substances are removed from the wound parts of plant. Some substances are transferred when certain parts of plants become decomposed, while other are transferred as volatile compounds 3. These compounds generally have indirect effect on the physiology of the plants. They are active against seed germination, causing injury to root growth and other meristems, or inhibiting seedling growth. For example the two quinines compounds, known as juglone and sorgoleone, inhibit chloroplast oxygen evolution and strongly affect mitochondrial functions. The chloroplast block by sorgoleone is in the photosystem II complex. Similarly Cinnamic and benzoic acid derivatives alter membrane potential resulting membrane impermeability 4. Flavonoides are regarded as plant defensive mechanism because it protects the plants from the attack of pathogens and herbivores 5. The phenolic compounds found in some plants where they regulate various enzymatic activities are having allelopathic affect on other plants 6. The serious problem faced by farmers is weeds control. There are some herbicides which control weeds but these substances are harmful for the environment. They remain in the environment for large time because the microorganisms cannot degrade them easily. Silene villosa Forssk is an annual herb. It is a member of family caryophyllaceae. The stem is erect, cylindrical, herbaceous and branched. The leaves are dense, glandular, inversely lanceolate, sessile, leaf apex acute to subacute. Inflorescence biparous cyme, flowers lie at junction of stem and petiole throughout the stem. Calyx 5, slightly petaloide, small densely glandular, hairy and constricted below with capsule of fruit. Petals 5 in number, polypetalous, white, adnate, limbs bilobed. Androcium, polyandrous, obdiplostaminous and Gynecium are tricarpellary, syncarpous, ovary superior, placentation is free central, and fruit is capsule 7. MATERIALS AND METHODS Plant collection Silene villosa was collected from Distric Karak Khyber Pakhtunkhwa, Journal of Pharmacy Research Vol. Issue 4 April 16

2 Pakistan. The plant was identified with the help of available literatures 8. Fresh sample of plant was washed with tap water to remove sand and dust particles. The plant was then shade dried at room temperature (5 C), grounded into fine powder by grinder and stored in plastic bags for further analysis. Preparation of crude extracts About 3 kg of plant powder was soaked in 8 L of analytical grade of methanol. The plant material was regularly stirred after 4 hrs till 15 days. The solvent containing the extracts was filtered with Muslin cloth to obtain the filtrate. The filtrate was passed through rotary evaporator at temperature of 55 C under reduce pressure to remove the extra solvent. The extract was then completely dried to obtain crude extracts. Preparation of stock solution (1mg/ml) was prepared in 1% Dimethyle Sulfoxide (DMSO) for each of n-ane, oroform and Ethyle acetate soluble fractions. Four dilutions with,, and were prepared for each of the fraction. Experimental procedure Healthy seeds of wheat variety shahkar 13 were dipped in ethanol for one minute and washed with distilled water three times to sterilize it. Double layered filter paper was placed in equal sized sterile Petri dishes. Ten seeds in each Petri dish were placed with equal distance and each Petri dish was labeled. 5 ml from each solution were poured into each respective Petri dish. The Petri dishes were placed in incubator at 5C for 6 days. The inhibitory effect was determined by measuring the germination percentage, root and shoots length, fresh and dry weight of both root and shoot. The experiment was performed three times to ensure the removal of possible errors. Data analysis Mean value of three replica was selected and One Way ANOVA was used for the analysis of data. RESULTS AND DISCUSSIONS Germination percentage The special effects of different concentrations of Silene villosa methanolic extract and its consequent fractions on the germination percentage of wheat seeds are discussed in the following headings. Journal of Pharmacy Research Vol. Issue 4 April 16 n- ane fraction of Silene villosa. No germination percentage was observed after 4 hrs and after 48 hrs maximum germination percentage was noted as the control which was 3.3%. At the same time germination percentage was % at, 16.6% at and and 13.3% at and. It was observed that germination percentage was increased with decreasing n-ane fraction. The result calibrated with the work of Mubeen et al 9, who s observed that the weedy plants inhibit the germination percentage of Oryza sativa. Statistical analyses suggest that there was no significant difference between the germination percentage at and the control while a significant difference of germination percentage was noted in comparison of with the control. Germination (%age) Control 4h 7h h Fig: 1 Germination percentage of wheat at various concentrations of the n-ane fraction of Silene villosa oroform fraction of Silene villosa. The germination percentage of wheat was recorded for hrs against and was lowest at higher concentration of the chloroform fraction. The same result was found by Saberi et al while studying Eucalyptus camadulensis extract on Trifolium rigidom and Vicia villosa. The increasing concentration of extract and its fraction with decreasing germination is the factor that plants contains phenolic compounds as proposed by Mousavi et al 11. Statistical analysis show no differences between Control and but show differences between Control and. Germination (%age) Control 4h 7h h Fig: Germination percentage of wheat at various concentrations of the oroform fraction of Silene villosa

3 Ethyl acetate fraction of Silene villosa The germination percentage of wheat was observed against various concentration of Ethyl acetate fraction till hours. The maximum germination percentage was noted in (86.7%) after control and the lowest was in which was 56.6%. The result obtained was same as noted by Khan et al 1 by applying aqueous extracts of Eucalyptus camadulensis against Zea mays. By applying statistical analysis it was observed that there are significant differences between the control and. Germination % Control 4h 7h h Fig: 3 Germination percentage of wheat at various concentrations of the Ethyl acetate fraction of Silene villosa hanol fraction of Silene villosa The crude (hanolic) extract of Silene villosa was tested on germination percentage of wheat. The maximum percentage was noted in which was 93.3 % and lowest 46.6 % was found in. The result obtained corresponds with the work of Ferguson and Rathinasabapathi 13. They proposed that seed germination of most plants is effected by allelochemical substances. Statistical analysis show significant difference between Control and. Germination % Control Fig: 4 Germination percentage of wheat at various concentrations of the hanol fraction of Silene villosa 4h 7h h Journal of Pharmacy Research Vol. Issue 4 April 16 Average shoot length The average shoot length of wheat is affected by hanolic extract and fraction of Silene villosa. The result showed that shoot length reduced with increasing concentration in all fractions. The maximum shoot length at was observed in Ethyl acetate fraction which was 7.3 cm. The lowest value at was observed at hanolic fraction. Seghatoleslami et al 14 observed that there is significant decrease of the shoot length with increasing Cardaria draba extract concentration Control Fig: 5: The effect of the crude extract and its fractions of Silene villosa on the average shoot length of Wheat seedlings Average root length The average root show remarkable decrease as the concentration of hanolic extracts and fractions increases. The shortest length (4.7cm) was observed in of hanolic extract followed by ane fraction which was 4.73cm. Femina et al 14 observed that Tridax procumbens extract inhibit the root elongation of Vigna radiata Control Fig: 6: The effect of the crude extract and its fractions of Silene villosa on the average root length of Wheat seedlings

4 Average fresh weight of shoot The average fresh weight of shoot was significantly decreased as the concentration of the fractions was increased. It was.7 gm in control while reduced to.19 at in ethyl acetate fraction. Statistical analysis shows that there is significantly a difference between control and. Kamal and Bano 16 have reported that the extracts and fractions of various plants have inhibitory effects on germination percentage, shoot and root length, fresh and dry weight of seedlings Control Fig: 7: The effect of the crude extract and its fractions of Silene villosa on the fresh weight of shoot of wheat seedlings Average dry weight of shoot The dry weight of shoot was decreased as the concentration of the fraction was increased. The lowest dry weight was observed in and highest in control. The value obtained was similar to the work of Khan et al 17 who described that testing the extracts of Woodfordia fruiticosa against radish that there was decreased reduction of dry weight of shoot with increasing concentration. Analysis of variance suggests that there is significance difference between control and and no significance difference between control and Control Average fresh weight of Root The average fresh weight of root decreased as the concentration increased. The lowest fresh weight was noted at in all the fractions. The highest fresh weight was in control. As the concentration increased, the fresh weight of root decreased as supported by the work of Babar et al 18 who describes that the root fresh weight of Cicer arietenum was decreased as the concentration of Asphodelus tunifolius extracts increased. Statistical analysis show no significance difference between control and and but show significance difference between control and. Fig: 9: The effect of the crude extract and its fractions of Silene villosa on the fresh weight of root of wheat seedlings Average dry weight of root The lowest dry weight was observed at in hanolic fraction followed by n-ane fraction, while the highest value was observed in control. When the concentration goes towards p from control side, the dry weight was decreased. The same result was obtained by Dongre and Yadav 19 who noted reduction of dry weight of root of Pea and wheat by using weeds extract. ANOVA show significance difference between control and. Fig: 8: The effect of the crude extract and its fractions of Silene Fig: : The effect of the crude extract and its fractions of Silene villosa on the dry weight of shoot of Wheat seedlings villosa on the dry weight of root of wheat seedlings Journal of Pharmacy Research Vol. Issue 4 April 16 Length (cm) Length (cm) Control Control

5 CONCLUSION The present study was performed to investigate the allelopathic affect of Silene villosa. The investigation shows that the plant has remarkable allelopathic activity against wheat plant. The Silene villosa have inhibitory effect on germination of seedlings, root and shoot length and fresh and dry weight of root and shoot. Further studies will suggest the isolation of these substances and their inhibitory effect on wheat. ACKNOWLEDGEMENT The author is thankful to Chairman, Department of Botany, Kohat University of Science and Technology (KUST), Kohat, for his persistent support and encouragement during research work. Source of support: Nil, Conflict of interest: None Declared REFERENCES 1. S Ahmad, M Arfan, A.L Khan, R Ullah, J Hussain, Z Muhammad, R Khan, N Khan, N Watanabe, (11) Allelopathy of Teucrium royleanum Wall. Ex. Benth. from Pakistan Journal of Medicinal Plants Research 5, S Abu-Romman, M Shatnawi, R.S Shibli, () Allelopathic effects of Euphorbia hierosolymitana on Triticum durum American-Eurasian Journal of Enviromental Sciences Oueslati, M Ben-Hammouda, M.H Ghorbal, M El Gazzeh, R.J Kremer, (9) Role of phenolic acids in expression of barley (Hordeum vulgare) autotoxicity Allelopathy Journal A.E Frank, (9) Allelopathy American Chemical Society D.C Oliveira, G.L.G Soares, R.M.S Isaias, (8) Allelochemicality of the extracts of Lonchocarpus muehlbergianus Hassl (Fabaceae) leaflets and galls on seed germination and early development of lettuce Acta bot. bras, R Capasso, (1997) The chemistry, biotechnology and ecotoxicology of the polyphenols naturally occurring in vegetable wastes Current Topics in Phytochemistry 1, A Shahina, Ghazanfar, Y.J Nasir, (197) Flora of Pakistan A Shahina, Ghazanfar, Y.J Nasir, (197) Flora of Pakistan K Mubeen, M.A Nadeem, A Tanveer, Z.A Zahir, (11) Allelopathic effect of aqueous extracts of weeds on the germination and seedling growth of Rice (Oryza sativa L.) Pakistan journal of life Sciences 9, M Saberi, A Davari, F Tarnian, M Shahreki, E Shahreki, (13) Allelopathic effects of Eucalyptus camaldulensis on seed germination and initial growth of four range species Annals of Biological Research 4, S.A Mousavi, F.R Tavakkol, A Asadi, M.H Gharineh, (11) Allelopathic effects of aqueous extract of leaf, stem and root of sorghum bicolor on seed germination and seedling growth of Vigna radiate Natural and Biological Science 3, M.A Khan, I Hussain, Khan, (7) Effect of aqueous extract of Eucalyptus camaldulensis on the germination and growth of Zea mays Pakistan Journal of Weed Sciences Research 13, J.J Ferguson, B Rathinasabapathi, (3) Allelopathy: How Plants Suppress Other Plants. (Horticultural Sciences Department, Florida Cooperative Extension Service, Institute of Food and Agricultural Sciences, University of Florida). 14. M.J Seghatoleslami, G.R Mousavi, H Akbari, (1) Studying the allelopathic effects of shoot and root water extracts of hoary cress (Cardaria draba) on germination and seedling growth of wheat Pakistan Journal of Weed Sciences Research 18, D Femina, P Lakshmiprya, S Subha, R Manonmani, (1) Allelopathic effect of the weed (Tridax procumbens) on seed germination and seedling growth of some leguminous plants International Research Journal of Pharmacy 3, J Kamal, A Bano, (8) Allelopathic potential of sunflower (Helianthus annus L.) on soil metals and its leaves extracts on physiology of wheat (Triticum aestivum L.) seedlings African Journal of Biotechnology 7, A.M Khan, R.A Qureshi, F Ullah, S.A Gilani, (11) Allelopathic effects of selected medicinal plants collected from Margalla Hills, Islamabad Pakistan Journal of Medicinal Plants Research 5, B.H Babar, A Tanveer, M Tahir, A Aziz, A.U.H Ahmad, M.A Nadeem, M.M Javaid, (9) Allelopathic potential of Asphodelus tenuifoliuson the germination and seedling growth of Cicer arietinum Weed Biology and Management 9, P.N Dongre, B Yadav, (5) Inhibitory allelopathic effect of weed leaf leachates on seed germination of pea (Pisum sativum L.) Crop Research Hisar 9, Journal of Pharmacy Research Vol. Issue 4 April 16