Morphological and Molecular Characterization of an Antagonistic Fungus Trichoderma asperelloides TDOAE002
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Abstract
Trichoderma spp. are potential fungal biocontrol agents for plant disease control and plant growth promotion. An antagonistic fungus Trichoderma sp. TDOAE002 isolate has been promoted to control several plant diseases in Thailand, including rice diseases by the Department of Agricultural Extension (DOAE). Nevertheless, Trichoderma sp. TDOAE002 has not been classified into the current taxonomy of Trichoderma spp. This study contributed to the taxonomy of Trichoderma sp. TDOAE002 based on morphological characteristics and molecular analysis. The fungus grew rapidly on potato dextrose agar (PDA) at 25±2 °C, with white, fluffy mycelia, and pale green to dark green conidial masses forming within 48 hours. Conidiophores branched, central axis from which secondary branches arose, the branches terminating in a single ampulliform phialide or a whorl of 2-4 divergent phialides. Conidia were subglobose, 1.74-3.11 x 2.20-3.84 μm in size, with irregular warts on the conidial surface, and pale green in masses. Chlamydospores were globose to subglobose, surface smooth, and solitary. In addition, to confirm the species identification of Trichoderma sp. TDOAE002, the molecular analysis was conducted on Multilocus Identification System for Trichoderma (MIST) program based on the internal transcribed spacer (ITS) regions of the rDNA cluster (ITS1 and ITS2), partial sequences of the translation elongation factor 1 alpha (tef-1), and the RNA polymerase II subunit (rpb2) nucleotide sequences. The phylogenetic tree from the combined three loci revealed the Trichoderma sp. TDOAE002 was grouped into the T. asperelloides group. Consequently, we identified the antagonistic fungus TDOAE002 isolate as T. asperelloides.
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Abbey, J.A., D. Percival, L. Abbey, S.K. Asiedu, B. Prithiviraj and A. Schilder. 2019. Biofungicides as alternative to synthetic fungicide control of grey mould (Botrytis cinerea)-prospects and challenges. Biocontrol Science and Technology 29: 207-228.
Cai, F. and I.S. Druzhinina. 2021. In honor of John Bissett: authoritative guidelines on molecular identification of Trichoderma. Fungal Diversity 107: 1-69.
Carbone, I. and L.M. Kohn. 1999. A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia 91(3): 553-556.
Chamswarng, C. 2006. Biological Control of Plant Diseases. Department of Plant Pathology, Faculty of Agriculture at Kamphaeng Saen, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom province. 323 p. (in Thai)
Chamswarng, C. 2017. Trichoderma, the fugus play a roles in future crop production. pp. 14-17. In: Kasetapirom 4(9). Faculty of Agriculture at Kamphaeng Saen, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom province. (in Thai)
Chamswarng, C. 2020. Trichoderma: Antagonistic Fungus for Plant Disease Control. Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom province. 566 p. (in Thai)
Chamswarng, C., W. Intanoo, R. Dhitikiattipong and P. Charoenrak. 2012. The use of Trichoderma powder formulation isolate 01-52 for reducing dirty panicle disease and increasing yield of rice in paddy fields. pp. 460-464. In: The 2nd National Conference on Rice “a new dimension of Thai rice research, ready to take action on climate changes and open Asian free markets”. December 21-23, 2012. Swissotel Le Concorde, Bangkok. (in Thai)
Chaparro, A.P., L.H. Carvajal and S. Orduz. 2021. Fungicide tolerance of Trichoderma asperelloides and T. harzianum strains. Agricultural Sciences 2 (3): 301-307.
Charoenrak, P. and C. Chamswarng. 2015. Application of Trichoderma asperellum fresh culture bioproduct as potential biological control agent of fungal diseases to increase yield of rice (Oryza sativa L.). Journal of the International Society for Southeast Asian Agricultural Sciences 12(2): 67-85.
Department of Agricultural Extension. 2020. Application of Microorganisms (Biopesticides) for Plant Pest Control. 2nd ed. Bureau of Technology Transfer Development, Bangkok. 31 p. (in Thai)
Dou, K., Z. Lu, Q. Wu, M. Ni, C. Yu, M. Wang, Y. Li, X. Wang, H. Xie, J. Chen and C. Zhang. 2020. MIST: a Multilocus Identification System for Trichoderma. Applied and Environmental Microbiology 86(18): 1-13.
Druzhinina, I.S. and C.P. Kubicek. 2005. Species concepts and biodiversity in Trichoderma and Hypocrea: from aggregate species to species clusters. Journal of Zhejiang University SCIENCE 6B(2): 100-112.
Druzhinina, I.S., C.P. Kubicek, M. Komoń-Zelazowska, T.B. Mulaw and J. Bissett. 2010. The Trichoderma harzianum demon: complex speciation history resulting in coexistence of hypothetical biological species, recent agamospecies and numerous relict lineages. BMC Evolutionary Biology 10: 1-14.
ICTT. 2022. Taxonomy 2022. Available source: https://trichoderma.info/trichoderma-taxonomy-2020/. (February 21, 2003)
Jaklitsch, W.M., M. Komon, C.P. Kubicek and I.S. Druzhinina. 2005. Hypocrea voglmayrii sp. nov. from the Austrian Alps represents a new phylogenetic clade in Hypocrea/Trichoderma. Mycologia 97: 1365-1378.
Karnpakdee, S., O. Piasai, W. Serewan, P. Karnpakdee and N. Khewkhom. 2021. The application of Trichoderma asperellum powder to control sheath blight disease of rice caused by Rhizoctonia solani. Khon Kaen Agriculture Journal 49(1): 155-166. (in Thai)
Kumar, S. 2013. Trichoderma: a biological weapon for managing plant diseases and promoting sustainability. International Journal of Agricultural Sciences and Veterinary 1: 106-121.
Liu, Y.J., S. Whelen and B.D. Hall. 1999. Phylogenetic relationships among ascomycetes: evidence from an RNA polymerse II subunit. Molecular Biology and Evolution 16(12): 1799-1808.
Qiao, M., X. Du, Z. Zhang, J.P. Xu and Z.F. Yu. 2018. Three new species of soil-inhabiting Trichoderma from southwest China. MycoKeys 44: 63-80.
Ruangwong, O., P. Wonglom, N. Phoka, N. Suwannarach, S. Lumyong, S. Ito and A. Sunpapao. 2021. Biological control activity of Trichoderma asperelloides PSU-P1 against gummy stem blight in muskmelon (Cucumis melo). Physiological and Molecular Plant Pathology 115: 101663.
Safavi, S.A. 2010. Isolation, identification and pathogenicity assessment of a new isolate of entomopathogenic fungus, Beauveria bassiana in Iran. Journal of Plant Protection Research 50(2): 157-163.
Saitou, N. and M. Nei. 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution 4: 406-425.
Samuels, G.J., A. Ismaiel, M.C. Bonn, R. De Respinis and O. Petrini. 2010. Trichoderma asperellum sensu lato consists of two cryptic species. Mycologia 102(4): 944-966.
Samuels, G.J., E. Lieckfeldt and H.I. Nirenberg. 1999. Trichoderma asperellum, a new species with warted conidia, and redescription of T. viride. Sydowia 51: 71-88.
Tamura, K., G. Stecher and S. Kumar. 2021. MEGA11: molecular evolutionary genetics analysis version 11. Molecular Biology and Evolution 38(7): 3022-3027.
Unartngam, J., B. Srithongkum, W. Intanoo, P. Charoenrak and C. Chamswarng. 2020. Morphological and molecular based identification of Trichoderma CB-Pin-01 biological control agent of plant pathogenic fungi in Thailand. International Journal of Agricultural Technology 16(1): 175-188.
White, T.J., T. Bruns, S. Lee and J. Talor. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. pp. 315-322. In: Gelfand MAIDH, Sninsky, J.J. and White, T.J. (eds.) PCR Protocols: A Guide to Methods and Applications. Academic Press, New York.
Wongcharoen, A. 2014. Screening of endophytic fungi from rice (Oryza sativa L.) against rice pathogenic fungi. Khon Kaen Agriculture Journal 42(3): 385-396. (in Thai)
Yu, Z.F., M. Qiao, Y. Zhang and K.Q. Zhang. 2007. Two new species of Trichoderma from Yunnan, China. Antonie van Leeuwenhoek 92: 101-108.
Zheng, H., M. Qiao, Y. Lv, X. Du, K.Q. Zhang and Z. Yu. 2021. New Species of Trichoderma isolated as endophytes and saprobes from southwest China. Journal of Fungi 7: 467.