In vitro efficiency of Trichoderma harzianum mutants in biocontrol of Fusarium oxysporum f. sp. radicis-cucumerinum

Volume 9, Issue 2
June 2020
Pages 285-300

Document Type : Original Research

Authors

1 Plant Biotechnology, Higher Education Complex of Shirvan, Shirvan, Iran.

2 Plant Protection Research Department, Yazd Agricultural and Natural Resources Research and Education Center, AREEO, Yazd, Iran.

3 Plant Protection Research Department, Nuclear Science and Technology Research Institute (NSTRI), Atomic Energy Organization of IRAN (AEOI), Karaj, Iran.

Abstract
Trichoderma is very important as a bio control agent and probably a good alternative for chemical fungicides. Fusarium oxysporum is a plant pathogen that causes wilt in a wide range of plants. The use of gamma irradiation can be employed to increase Trichoderma efficiency against F. oxysporum. The induced mutation provides genetic changes in Trichoderma and in some of the mutated isolates the efficiency of bio control may be improved. In this study the efficiency of mutated T. harzianum CS5 against the F. oxysporum f.sp. radicis-cucumerinum was evaluated. Among 16 wild isolates of Trichoderma, CS5 isolate was chosen based on synchronous and nonsynchronous mutual cultivation with pathogen. The spore suspensions of T. harzianum CS5 were irradiated in a cobalt- 60 γ- irradiator at a dose rate of 0.23 Gy/Sec in Nuclear Agriculture Research School, (NSTRI, AEOI). Then antagonist screening of 76 mutants was investigated in the experiments of synchronous and nonsynchronous dual culture with pathogen. The results showed that only 17 mutants were able to control the pathogen better than wild type, and YFTM80 isolate had the most prevention. Using the ERIC-PCR marker, the distinction was investigated between the wild type and the 17 selected mutants. The results showed that the gamma ray is able to improve biocontrol efficiency of Trichoderma and the ERIC-PCR marker can differentiate among derived mutants, but it does not have the ability to distinguish mutants from wild type. In terms of antagonistic superiority, mutations might have occurred in antagonistic sites, which have led to improved antagonistic efficiency. Probably the ERIC-PCR marker has failed to replicate these areas.

Keywords

Subjects
1. 1. Abasi Iranog, S., Safaiei, N., Shams Bakhsh, M. and Shahbazi, S. 2014. Investigation of antagonistic properties of several mutated Trichoderma harzianum against a number of plant pathogenic fungi under in vitro conditions. Scientific Journal of Agriculture, 37 (4): 91-102.
2. Abasi Iranog, S., Safaiei, N., Shams Bakhsh, M. and Shahbazi, S. 2012. Investigation of genetic diversity of mutated isolates of Trichoderma using ERIC-PCR marker. Third National Iranian Biotechnology Conference, Mashhad, Ferdowsi University of Mashhad.
3. Abdel-lateif, K.S., 2017. Trichoderma as biological control weapon against soil borne plant pathogens. African Journal of Biotechnology, 16(50), pp.2299-2306.
4. Ashrafi zadeh, A., Etebarian, H. and Zamani zadeh, H. 2003. Evaluation of Trichoderma isolates for biocontrol of Fusarium wilt of melon. Plant disease, 41(1):39-58.
5. Arora, D.K. 2004. Fungal biotechnology in agricultural, food and environmental applications, New York:147-148.
6. Behbodi, K., Sharifi Tehrani, A., Hajaroud, gh. and Zad, G. 2003. Antagonistic effects of Trichoderma spp. On Phytophtora capsici, antagonist of pepper root and crown rot. Plant Pathology, 41(3):345-362.
7. Darvishnia, M. 2013. Morphological and phylogenetic studies of Fusarium species in Iran. Journal of Novel Applied Sciences, 2:1137-1142.
8. Dorneles, E.M.S, Santana, J.A., Ribeiro, D., Dorella, F.A., Guimara, A.S., Moawad, M.S., Selim, S.A., Garaldi, A.L.M, Miyoshi, A., Ribeiro, M.G., Gouveia, A.M.G , Azevedo, V., Heinemann, M.B. and Lage, A.P. 2014. Evaluation of ERIC-PCR as Genotyping Method for Corynebacterium pseudotuberculosis Isolates. PLOS ONE. 9(6): 1-10.
9. Elahi nia, S. A. 2008. Plant pathology and recognition of fungi and other pathogens. Third edition. Gilan university press, Gilan. PP: 647.
10. Fani, S. R., Moradi Ghahdarijani, M., Alipour Moghadam, M., Sherafati, A., Mohamadi Moghadam, M., Sedaghati, E. and Khodaygan, P. 2013. Efficacy of Native Strains of Trichoderma harzianum in Biocontrol of Pistachio Gummosis, Iranian Journal of Plantprotection Science, 44(2): 243-252.
11. Haggag, W. M. 2008. Induction of hyperproducing chitinase Trichoderma mutants for efficient biocontrol of Botrytis cinerea on tomato and cucumber plants growing in plastic houses. Arab Journal of Biotechnology, 5 (2): 151-164.
12. Haggag, W.M. and Mohamed, H.A.A., 2002. Enhanecment of antifungal metabolite production from gamma-ray induced mutants of some Trichoderma species for control onion white disease. Plant Pathology Bulletin, 11, pp.45-56.
13. Harman, G.E. 2006. Overview of Mechanism and uses of Trichoderma spp. Journal of the American Phytopatholog society.190-192.
14. Heydari faroughi, M., Etebarian, H. and Zamani zadeh, H. 1992. Evaluation of Trichoderma isolates for biological control of cucurbit wilt disease ( Phytophthora drechsleri) in greenhouse. Plant Pests and Disease, 72(2). 113-134.
15. Jamdar, Z., Mohammadi, A. H. and Mohammadi, S. 2013. Study of Antagonistic Effects of Trichoderma Species on Growth of Verticillium dahliae, the Causal Agent of VerticilliumWilt of Pistachio under Laboratory Condition. Journal of Nuts,4(4):53-56.
16. Karlsson, I., Edel-Hermann, V., Gautheron, N., Durling, M.B., Kolseth, A.K., Steinberg, C., Persson, P. and Friberg, H. 2016. Genus-Specific Primers for Study of Fusarium Communities in Field Samples. Applied and Environmental Microbiology. 82(2): 491-501
17. Khatib, A., Moradzadeh Eakandari, M., Derakhshan, A. and Mahmoudi, H. 2017. Evaluation of some commercial and native isolates of Trichoderma harzianum for controlling Melon Fusarium wilt using seed coating method, Biological Control of Pest and Plant Disease, 6(2): 165-172.
18. Khodayary, M., Safaiei, N. and Shams Bakhsh, M. 2008. Comparison of molecular markers PAPD, ISSR and ERIC in the study of genetic diversity of Rhizoctonia solani AG1-IA. New Genetic,3(2):37-46.
19. Molavy, A., Aminian, H., Etebarian, H. and Shahriary, D. 2009. Investigation the resistance of greenhouse-cucumber cultivars to Fusarium stem and root rot disease (F. oxysporum f. sp. radicis-cucumerinum). Journal of Agriculture,11(1):177-189.
20. Mohamadi, A.S., Shahbazi, S. and Askari, H. 2014. Investigation of γ-radiation on morphological characteristics and antagonist potential of Trichoderma viride against Rhizoctonia solani, International Research Journal of Applied and Basic Sciences, 8(3):329-336.
21. Mohamed, H.A.L.A. and Haggag and W.M., 2006. Biocontrol potential of salinity tolerant mutants of Trichoderma harzianum against Fusarium oxysporum. Brazilian Journal of Microbiology, 37(2), pp.181-191.
22. Moradi, M., Oerke, E.-C., Steiner, U., Tesfaye, D., Schellander, K. & Dehne, H.-W. 2010. Microbiological and Sybr®Green Real-Time PCR detection of major Fusarium Head Blight pathogens on wheat ears. Microbiology, 79 (5), 646–654.
23. Moradi, R., Shahbazi, S., Ahari Mostafavi. H., Ebrahimi, M. A., Askari, H. and Mir Majlesi, M. 2013. Investigation of Gamma Radiation Effects on Morphological and Antagonistic Characteristics of T. harzianum. Crop Biotechnology, 3(4):109-117.
24. Mostafavi H (2009). Application nuclear technology in management parasites and plant disease. The second congress national application nuclear technology in agriculture. Nuclear Science and Technology Research institute of Karaj, p. 331-335.
25. Mukherjee, P.K. and Raghu, K., 1997. Effect of temperature on antagonistic and biocontrol potential of shape Trichoderma sp. on Sclerotium rolfsii. Mycopathologia, 139(3), pp.151-155.
26. Norouzi, S., Rahnama, K., Rabbani Nasab, H. and Taqi Nasab, M. 2014. Evaluation of efficacy of Trichoderma and Bacillus isolates in biological control of melon fusarium wilt, Bicontrol in Plant Protection, 2(1): 43-56.
27. Okhovat, M., Zafari, D., Karimi rozbahani, A. and Rohani, H. 1996. Antagonistic effects of some Trichoderma pers. ex on Colletotrichum coccodes(Wallr)Hughes, isolated from potatoes. Plant pathology, 32:208-219.
28. Persoon, C. H. 1794. Dispositio methodica fungorum. Romer's neues Magazin Botanische, 181-128.
29. Poozeshi Miab, M., Razavy, M., Zamani Zadeh, H., Zare, R. and Rezaiei, S. 2014. Comparison of genetic diversity and pathogenicity among F. culmorum isolates, the causal agent of wheat root and crown rot disease in Varamin fields. Plant Pests and Disease. 82(1). 67-80.
30. Rahdar, M. 2012. Biological control of bean Rhizoctonia root and crown rot by some bacterial isolates and Trichoderma isolates alone and in combination. Msc thesis. Ferdowsi university of Mashhad.
31. Raja, H.A., Miller, A.N., Pearce, C.J., Oberlies and N.H. 2017. Fungal identification using molecular tools: a primer for the natural products research community. Journal of Natural Products. 756-770.
32. Samuels. G. j. and Seifert, K. A. 1987. Kinds of pleoanamorphy in the Hypocreales. In Pleomorphic Fungi: The Diversity and its Taxonomic Implications (ed. J. Sugiyama), Kodansha Ltd: 29-56.
33. Samouel, G.J. 1996. Trichoderma: a review of biology and systematics of the genus. Journal of Mycological Research, 100(8):923-935.
34. Seidl V and Seiboth B (2010). Trichoderma reesei: genetic approaches to improving strain efficiency. Biofuels 1(2):343-354.
35. Shahbazi, S., Askary, H., Ahary Mostafavy, H. and Mirmajlesi, M. 2012. Increases Trichoderma antagonistic potential by including mutation in the glucanase enzyme. Third Iranian Agriculture Biotechnology Conference, 4-6 of September, Mashhad.
36. Shahriari, D., Molavy, A., Aminian, H. and Etebarian, H. 2011. Histopathological Response of Resistant and Susceptible Cultivars of Cucumber to F. oxysporum f.sp. radicis-cucumerinum, the Causal Agent of Fusarium Stem and Root Rot, Seed and Plant Improvement Journal, 27(3). 375-391.
37. Singh, R., Maurya, S. and Upadhyay R.S. (2010). Improvement of antagonistic capability of Trichoderma harzianum by UV irradiation for management of Macrophomina phaseolina. Arch. Phytopathology and Plant. Protection. 43:1579-1588.
38. Singh, R., Maurya, S. and Upadhyay R.S. (2016). The improvement of competitive saprophytic capabilities of Trichoderma species through the use of chemical mutagens. Brazilian Journal of Microbiology. 47:10-17.
39. Szekeres, A., Kredics, L., Antal, Z., Kevei, F. and Manczinger, L. 2004. Isolation and characterization of protease overproducing mutants of Trichoderma harzianum. FEMS Microbiology Letters, 233: 215-222.
40. Waghunde, R.R., Shelake, R.M. and Sabalpara, A.N. 2016. Trichoderma: A significant fungus for agriculture and environment. African Journal of Agricultural Research. 11(22): 1952-1965.