Effective Biological Control of Carnation Fusarium Wilt Using a New Combination of Trichoderma Mutant Isolates

Document Type : Original Research

Authors
1 Department of Plant Production, Shirvan Agriculture Faculty, University of Bojnord, Bojnord, Islamic Republic of Iran.
2 2 Nuclear Agriculture Research School, Nuclear Science and Technology Research Institute, Alborz, Islamic Republic of Iran.
3 Department of Plant Protection, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Islamic Republic of Iran.
Abstract
Fusarium infection in carnation is the most important limiting factor for carnation production. We isolated 38 fungal isolates from infected carnation plants collected from Mahallat, Iran, and 15 Fusarium species were identified morphologically. Isolates of J14, k5, and k72 were the most pathogenic isolates in pathogenicity test on White Liberty cultivar. Molecular identification of J14, k5, and k72 isolates was done based on Elongation Factor 1-alpha (EF1-α) gene. To investigate the effect of Trichoderma and some of its mutants on reducing the Fusarium infection, six wild type isolates were examined. The species of T. harzianum, T. virense, and T. ghanens showed more inhabitation potential and were chosen for induced mutation via gamma irradiation at 250 Gy. The number of 270 mutants were screened morphologically and 60 mutants were screened using dual culture against J14, k5, and k72. Morphological and molecular identification of J14, k5, and k72 isolates recognized them as F. oxysporum. Three mutants, i.e. ThM7(67.17%), TgM1 (59.45%), and TvM17(57.55%) showed the highest efficacy and were selected. Evaluation test of efficacy in greenhouse by mixture of T. harzianum, T. virense and T. ghanens (TW) and mixture of mutant isolates ThM7, TgM1, and TvM17 (TM) showed that biological method had higher ability to control Fusarium infection on carnation plants in greenhouse condition, and mutation had no adverse effects on plants. The results of this experiment proved that the use of mutation in the Trichoderma genome with the use of gamma radiation could be an effective way to achieve isolates with better performance in this bio-control agent.

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Reference
1. Abbasi, S., Safaie, N. and Shams-bakhsh, M., 2014. Evaluation of gamma-induced mutants of Trichoderma harzianum for biological control of charcoal rot of melon (Macrophomina phaseolina) in laboratory and greenhouse conditions. Journal of Crop Protection, 3(4): 509-521.
2. Abbasi, S., Safaie, N., Shams-Bakhsh, M. and Shahbazi, S., 2016. Bio control activities of gamma induced mutants of Trichoderma harzianum against some soil borne fungal pathogens and their DNA fingerprinting. Iranian Journal of Biotechnology, 14(4): 260-269.
3. Akrami, M., Sabzi, M., Baghbani, F. and Khodadadi, E. 2012. Effect of seed Treatment with Trichoderma harzianum and Trichoderma asperellum species for controlling Fusarium rot of common bean. Annals of Biological Research, 3 (5): 2187-2189.
4. Altschul, S. F., Madden, T. L., Schäffer, A. A., Zhang, J., Zhang, Z., Miller, W. and Lipman, D. J. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic acids research, 25(17): 3389-3402.
5. Bani J., Seyed M. and Bayat, H. 2007. The effect of soil sunshine on the control of carnivorous Fusarium wilt and some chemical, biological and soil fertility properties, 10th Iranian Soil Science Congress, Karaj, Tehran University of Agriculture and Natural Resources Campus, https: // www .civilica.com / Paper-SSCI10-SSCI10_372.html.
6. Carver, C. E., Pitt, D. and Rhodes, D. J. 1996. Etiology and biological control of Fusarium wilt of pinks (Dianthus caryophyllus) using Trichoderma aureoviride. Plant pathology, 45: 618-630.
7. Castresana, J. 2000. Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis. Molecular Biology and Evolution, 17: 540-552.
8. Crous, P. W. et al. 2021. Fusarium: more than a node or a foot-shaped basal cell. Stud. Mycol, 98:100116. doi: 10.1016/j.simyco.2021.100116. PMID: 34466168; PMCID: PMC8379525
9. Dennis, C. and Webster, J., 1971. Antagonistic properties of species-groups of Trichoderma: III. Hyphal interaction. Transactions of the British Mycological Society, 57(3): 363-369.
10. Fattahi, B., Rahanandeh, H. Chavoshi, S. Zamanizadeh, H. R. Bayat, H. and Moshayedi, M. 2013. Report primary of Fusarium Proliferatum and Fusarium solani agents of Dianthus caryophyllus wilting in Markazi province in Iran, International Journal of Biosciences, 12(3): 1-7.
11. Geiser, D. M., Jime´nez-Gasco, M. M., Kang, S., Makalowska, I., Veeraraghavan, N., Ward, N., Zhang, T. J., Kuldau, G. A. and O’Donnell, K. K. 2004. FUSARIUM-ID v. 1.0: A DNA sequence database for identifying Fusarium. European Journal of Plant Pathology, 110: 473-479. https://doi.org/10.1023/B:EJPP.0000032386.
12. Ghasemi, S., Safaie, N., Shahbazi, S., Shams-Bakhsh, M. and Askari, H., 2020. The Role of Cell Wall Degrading Enzymes in Antagonistic Traits of Trichoderma virens Against Rhizoctonia solani. Iranian Journal of Biotechnology, 18(4):18-28.
13. Ghasemi, S., Safaie, N., Shahbazi, S., Shams-Bakhsh, M. and Askari, H., 2019. Enhancement of Lytic Enzymes Activity and Antagonistic Traits of Trichoderma harzianum Using γ-Radiation Induced Mutation. Journal of Agricultural Science and Technology, 21(4):1035-1048.
14. Hall, T.A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95-98.
15. Harman, G.E., Howel, C.R., Viterbo, A., Chet, I. and Lorito, M. 2004. Trichoderma species-Opportunistic, avirulent plant symbionts. Nature Reviews Microbiology, 2: 43-56.
16. Hirooka, Y., Rossman, A. Y., & Chaverri, P. 2011. A morphological and phylogenetic revision of the Nectria cinnabarina species complex. Studies in Mycology, 68, 35-56.
17. Katoh, K. and Standley, D. M. 2013. MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. Molecular Biology and Evolution, 30: 772-780.
18. Kermajany, Z. Jamali Zavareh, A. and Fadaei Tehrani, A. 2017. In vitro inhibitory effects of five strains of Trichoderma on the growth of Fusarium oxysporum f.sp. dianthi. Biological control of plant pests and diseases, 1: 121-125
19. Lava, A. and Babaeizad, V., 2021. Effects of Trichoderma and endophytic fungus Piriformospora indica on cucumber physiology and Fusarium wilt disease of Cucumber. Annals of the Romanian Society for Cell Biology, 25(4): 20742-20755.
20. Larget, B. and Simon, D. L. 1999. Markov chain Monte Carlo algorithms for the Bayesian analysis of phylogenetic trees. Molecular Biology and Evolution, 16: 750-759.
21. Leslie, J. F. and Summerell, B.A. 2006. The Fusarium Laboratory Manual. Blackwell Publishing. 1a. Ed. USA, 388 p. https://doi:10.1002/9780470278376.
22. Lombard L, Sandoval-Denis, M, Lamprecht SC and Crous PW. 2019, Epitypification of Fusarium oxysporum - clearing the taxonomic chaos. Persoonia. 43:1-47.
23. 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): 181-191.
24. Moutassem, D., Lakhdar, B. and Yuva, B. 2020. Efficiency of secondary metabolites produced by Trichoderma spp. in the biological control of Fusarium wilt in chickpea. Journal of Crop Protection, 9(2): 217-231.
25. Nylander, J. A. A. 2004. MrModeltest v2. Program distributed by the author. Evolutionary Biology Centre: Uppsala University.
26. O’Donnell, K., McCormick, S. P., Busman, M., Proctor, R. H., Ward, T. J., Doehring, G. and Rheeder, J. P. 2018. Marasas et al. 1984 “Toxigenic Fusarium species: identity and mycotoxicology” revisited. Mycologia, 110(6), 1058-1080
27. Ommati, F. and Masoud Z. 2012. Evaluation of some Trichoderma isolates for biological control of potato wilt disease (Fusarium oxysporum) under laboratory and greenhouse conditions." Journal of Crop Protection, 1(4): 279-286.
28. Orojnia, S., Habibi, D., Shahbazi, S., Paknejad, F. and Ilkaee, M.N. 2021. Investigation the Effect of Different Formulations of Mutated Trichoderma with Gamma Ray on Soybean Morphological Indices. Crop Physiology, 12(4): 97-114.
29. Pandey, A., Soccol, C.R., Nigam, P. and Soccol, V.T., 2000. Biotechnological potential of agro-industrial residues. I: sugarcane bagasse. Bioresource technology, 74(1): 69-80.
30. Prados-Ligero, A. M., Maria Jose, B.U., Carlos, J.H. and José María, M.V. 2007. Evaluation of susceptibility of carnation cultivars to Fusarium wilt and determination of Fusarium oxysporum fsp. dianthi races in southwest Spain. HortScience, 42 (3): 596-599.
31. Ronquist, F. and Huelsenbeck, J. P. 2003. MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics, 19:1572-1574.
32. Sahampoor, L., Zaker Tavallaie, F., Seyed Reza, F. and Shahbazi, S. 2020. In vitro efficiency of Trichoderma harzianum mutants in biocontrol of Fusarium oxysporum f. sp. radicis-cucumerinum. Journal of Crop Protection, 9 (2): 285-300.
33. Shahbazi, S., Zaker Tavallaie, F. and Daroodi, Z. 2021. Morphological and molecular identification of Fusarium spp. associated with carnation Dianthus caryophyllus in Mahallat, Iran. Journal of Crop Protection, 10(3): 461-471.
34. Shanmugam, V. and Sharma, V., 2008. Genetic relatedness of Trichoderma isolates antagonistic against Fusarium oxysporum f. sp. dianthi inflicting carnation wilt. Folia microbiologica, 53(2):130-138.
35. Sharma, S., and Nirupma, S. 2008. Carnation diseases and their management –A review. Agricultural Reviews, 29(1): 11-20.
36. Soufi, E., Safaie, N., Shahbazi, S. and Mojerlou, S., 2021. Gamma irradiation induces genetic variation and boosting antagonism in Trichoderma aureoviride. Archives of Phytopathology and Plant Protection, 54: 1649 - 1674
37. Van Hove, F., Waalwijk, C., Logrieco, A., Munaut, F., & Moretti, A. 2011. Gibberella musae (Fusarium musae) sp. nov., a recently discovered species from banana is sister to F. verticillioides. Mycologia, 103(3): 570-585.
38. Vaughan, T. G., IcyTree: Rapid browser-based visualization for phlogenetic trees and networks. Bioinformatics 2017. DOI: 10.1093/bioinformatics/btx155
39. Verma, M., Brar, S.K., Tyagi, R.D., Surampalli, R.Y. and Valero, J.R. 2007. Antagonistic fungi, Trichoderma spp.: panoply of biological control, Biochemical Engineering journal, 37: 1-20.