Inhibitory Effect of Sambucus ebulus Extracts on Growth of Macrophomina phaseolina and Extraction of their Bioactive

پژوهش های حفاظت گیاهان ایران

Article 4, Volume 30, Issue 4 - Serial Number 34, December 2017, Pages 622-628   

Document Type: Research Article

DOI: 10.22067/jpp.v30i4.50722

Authors

Maede Shahiri Tabarestani^{* 1}; kamran Rahnama²; Alireza Amiri³

¹Payame Noor University, Tehran

²Gorgan University of Agricultural Sciences & Natural Resources

³Babol Noshirvani University of Technology

Abstract

Introduction: Macrophomina phaseolina is the causal agent of soybean charcoal rot. This disease is one of the most prevalent and destructive of soybean in the north of Iran. This pathogen has broad host range and invades more than 500 plant species of 75 families, such as Caprifoliaceae. Symptoms on stems appear as silver-gray lesions near the base, which eventually decay the stem. Plants show poor seed fill, premature ripening, and undersized heads. Seed yield and oil content are decreased. Numerous tiny black bodies called sclerotia are formed on the decayed tissues giving the stalks a charred appearance. This fungus exists in the soil as sclerotia, a compact mass of hardened mycelia structures, which can remain dormant for many years. As the causal agent, is a soil-borne pathogen, chemical fungicides are not effective for its control. This matter has led to use a large amount of fungicides that are harmful to human health and lead to environmental pollution. There is not any registered fungicide against the charcoal rot pathogen. Some medicinal plants have a potential for controlling various phytopathogenic fungi due to the variety of compounds. Scientists are trying to achieve some plant-derived compounds for diseases control. Natural plant products are biodegradable, show structural diversity and rarely consist halogenated atoms. These can act directly as pesticides or may supply structure lead to pesticidal discovery. The aim of this study was to investigate chemical composition and antifungal activity of alcoholic and aqueous extracts of Sambucus ebulus for control of M. phaseolina and replace the chemical fungicides.
Materials and Methods: The plant leaves were collected during July - August 2014 in Babol (Mazandaran province- Iran) and washed thoroughly with tap water and then rinsed with distilled water and shade dried at room temperature. The dried plant material was finely powdered using an electric grinder and used for aqueous and organic solvent extraction. The aqueous extract was prepared via maceration method. Extraction with ethyl alcohol lasted 8 hours in a soxhlet extractor. The organic solvent was evaporated by rotary evaporated shaker untile 5 ml of solvent was reached. Then GC-MS analysis was used. For investigation of antifungal effects of Sambucus ebulus extracts, different concentrations of aqueous and alcoholic extracts in PDA (autoclaved) have been prepared. Concentrations of %10, %20 and %30 have been used for aqueous and alcoholic extracts, individually.
Results and Discussion: The aqueous extract did not show any antifungal effect, but different concentrations (%10,%20,%30) of alcoholic extracts showed significant inhibitory effects. Alcoholic extract at %30 concentration by %100 was the most effective one and at %10 concentration by 39.25 was the least effective one, on inhibiting of mycelial growth and formation of sclerotia of the pathogen. Statistically, there was the significant difference in %1 and %5 levels, between different concentrations of alcoholic extract for control of mycelial growth and formation of sclerotia. Alcoholic leaf extract was more effective than water extract, hence it shows that presence of antifungal agent released in alcohol due to its solubility. The chemical compounds present in the alcoholic extract were identified by using the GC-MS device with non-polar column (HP-5). According to mass spectra library of this device, in extracted organic phase with ethyl alcohol solvent, 27 chemical compounds were identified, which the main components were included as phthalates (%54/3), fatty acids and derivatives (%26/61), terpenoids (%2/65), diterpenes alcohol (%2/09), phenolic derivatives (%1.58), Phytosterols (%3.38) and cycloalkanes (%0.38). The highest abundance of identified chemical compounds was, Mono (2-ethyl hexyl) phthalate (%54.3), palmitic acid (%8.24), α - Linolenic acid (%7.78), Isovaleric acid (%4.33), dihydro Stigmasterol (%3.38), Neophytadiene (%2.65), 1-Butanol (%2.49), Phytol (%2.09), Octadecadienoic acid (%1.97) respectively. Some of these compounds have not previously been reported. Because of the antifungal effects of these constituents, the high potential of antifungal activity for alcoholic extract is justified.
Conclusion: The selected medicinal plant extract used in this experiment are easily and locally available and effective as an antifungal agent, so can be recommended to the farmer after the field test. Leaf extract not only has an adverse effect on the environment but also improve the quality of seed and the soil fertility. Our results showed that S. ebulus has the potential for controlling soybean charcoal rot and can be suitable alternative for chemical toxins. This is the first time that this research studies in Iran.

Keywords

GC/MS; Macrophomina phaseolina; Plant extract; Sambucus ebulus

References

1- Ahmadi S.B., Jalili Sendi J., Khodaparast S.A., Ghadamyari M., Hasanzadeh N., and Padasht dehkaee F. 2007. Comparison of some plant extracts with Edifenphos and Tricyclazole fungisides on the control of rice blast disease agent in field condition. Journal of Agricultural Researches, 7(4): 133-142.

2- Akpuaka A., Ekwenchi M.M, Dashak D.A., and Dildar A. 2012. Gas Chromatography-Mass Spectrometry (GC/MS) Analysis of Phthalate Isolates in n-Hexane Extract of Azadirachta indica A. Juss (Neem) Leaves. Journal of American Science, 8(12): 146-155.

3- Al–Abed A.S., Qasem J.R., and Abu-Blan H.A. 1993. Antifungal effect of some common wild plant species on certain plant pathologenic fungi. Dirasat (Pure Applied Science), 20: 149-158.

4- Ashraf H., and Javaid A. 2007. Evaluation of antifungal activity of Meliaceae family against Macrophomina phaseolina. Mycopath, 5(2): 81-84.

5- Askari F., Sharifi Ashorabadi E., Mirza M., Teimouri M., and Ehsani E. 2014. Chemical composition and antimicrobial effects of the essential oil of Thymus pubescens Boiss.& Kotschy ex Celak from different localities. Iranian Journal of Medicinal and Aromatic Plants, 30(5): 756-770.

6- Behdad M., Etemadi N.A., Behdad E., and Zeinali H. 2013. Antifungal effect of three plant essential oils against Rhizopus stolonifer, the cause of soft rot on strawberry fruit. Iranian Journal of Medicinal and Aromatic Plants, 29(2): 399-411.

7- Channa A.R., Jiskani M.M., and Nizamani Z.A. 2008. Effect of plant extracts on yield and mortality of plants due to root rot caused by Macrophomina phaseolina. Pakistan Journal of Agriculture Science, 24(2): 40-45.

8- Carolina H.P., Kock J.L.F., and Thibane V.S. 2011. Antifungal free fatty acids: A Review: 61-71. In Mendez-Vilas A.,(Eds.). Science against microbial pathogens: communicating current research and technological advances, Microbiology Series, 2(3), Spain: Formatex, 691p.

9- Chirigiu L., Bubulica M.V., and Chirigiu R.G. 2010. GC-MS Analysis of Chemical Compounds from Stems of Sambucus Ebulus L. Acta Medica Marisiensis, 56(6): 522-525.

10- Chirigu L., Chirigu R.G., Tircomnicu V., and Bubulica M.V. 2011. GC-MS analysis of chemical composition of Sambucus ebulus leaves. Chemistry of Natural Compounds, 47(1):126-127.

11- Chirigu L., Bubulica M.V., and Averi L.M.E. 2012. Investigations of Three Phytopharmaceutical Products from Caprifoliaceae Family Using GC-MS and LC-MS. Revista De Chimie, 63(8): 764-768.

12- Dwivedi R.S., and Dubey R.C. 2009. Effects of volatile and none-volatile fractions of two medicinal plants on germination of Macrophomina phaseolina. Transactions of the British Mycological Society, 87(2): 326-328.

13- Ghesmati M. 2007. Investigation of antibacterial activity of sambucus ebulus on staphylococcus aureus and Pseudomonas aeruginosa. Journal of Biology Science, 1(3): 73-82.

14- Gujar J., and Talwankar D. 2012. Antifungal activity of leaf extract on growth of Macrophomina phaseolina on soyabeen seed. Indian Streams Research Journals, 2(6): 1116.

15- Hadian S.H., Shamloo P., Monazm K., and Khandooz E. 2011. Effect of some aqueous plant extracts against Fusarium oxysporum f.sp.Lycopersici causal agent of tomato. Journal of Plant Science Researches, 21(6): 68-77.

16- Javid A., and Amin M. 2009. Antifungal activity of methanol and n-hexan extracts of three Chenopodium species against Macrophomina phaseolina. Natural Products Research, 23(12): 1120-1127.

17- Javid A., and Rehman H. 2011. Antifungal activity of leaf extracts of some medicinal trees against Macrophomina phaseolina. Journal of Medicinal Plants Research, 13: 2868-287.

18- Lin D., and suzuki E. 2003. Effect of methanol extracts from Ophiopogon japonicuson rice blast fungus. Pest Science and Management, 28(2): 27-28.

19- Mashhadian N.V., and Rakhshandeh H. 2005. Antibacterial and antifungal effects of Nigella sativa extracts against S.aureus, P.aeroginosa and C.albicans. Pakistan Journal of Medicinal Science, 21(1): 47-52.

20- Mazandarani M., Jamshidi M., and Azad A. 2011. Investigation of secondary metabolites of Sambucus ebulus L. in two natural regions of Mazandaran province, North of Iran. Journal of Plant Science Researches, 21(6): 58-67.

21- Meric Z.I., Bitis L., Birteksoz-Tan S., Turan S., and Akbuga J. 2014. Antioxidant, antimicrobial and anticarcinogenic activities of Sambucus ebulus L. flowers, fruits and leaves. Marmara Pharmaceutical Journal, 18(1): 22-25.

22- Pandey D.K., Tripathi N.N., Tripathi R.D., and Dixit S.N. 1982. Fungitoxic and phytotoxic properties of essential oil of Hyptic sauceolens. Plfkrankh Pflschutz. 89: 344-349.

23- Rahnama K., Montazernia B., and Hemati KH. 2008. Antifungal effects of some medicinal plants on Macrophomina phaseolina in-vitro. Journal of Plant Protection and Food, 3(4): 46-52.

24- Raman V., La S., Saradhi P., Rao N., Krishna N.V., Sudhakar M., and Radhakrishnan T.M. 2012. Antibacterial, antioxidant activity and GC-MS analysis of Eupatorium odoratum. Asian Journal of Pharmaceutical and Clinical Research, 5(2): 99-106.

25- Rayatpanah S., and Alavi S.V. 2006. Study on soybeen charcoal rot disease in Mazandaran. Journal of Agricultural Sciences Natural Resources, 13(3): 107-114.

26- Sayad S., Hassanzadeh N., Ghasemi A., and Nazerian E. 2013. The management of soft rot disease of syngonium caused by Pectobacterium carotovorum using some essential oils and antibiotics under laboratory and greenhouse conditions. Iranian Journal of Medicinal and Aromatic Plants, 28(4): 730-740.

27- Sing R., Dar S.A., and Sharma P. 2012. Antibacterial activity and toxicology evaluation of semi purified hexan extract of Urtica dioica leaves. Research Journal of Medicinal Plant, 6(2): 123-135.

28- Usha R., Udayakumar R., and John D. 2009. Bio-efficacy of plant extracts and bio-control agents against Macrophomina phaseolina. Annual Plant Protection Science, 17(2): 389-393.

29- Wendlbo P., and Rechinger K.H. 1987. Flora Iranica. Graz, Akademische Druck-und Verlagsanstalt.