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Keywords: n Trichoderma harzianumn , Morphological, Antagonistic activity, ITS-rRNA, Phylogenetic analysis, n Fusarium oxysporumn
Trichoderma spp. isolates are widely recognized for their efficacy as antagonists against a range of soil-borne fungal phytopathogens. In this study, ten isolates of Trichoderma were obtained from rhizosphere soil samples and characterized on the basis of cultural, morphological, and molecular analysis. Based on the colony growth rate, isolate SVPRT-56 demonstrated the highest growth rate (30.3 mm/day), while isolate SVPRT-59 displayed the lowest growth rate (18.7 mm/day). The colonies developed dense fluffy mycelium and a floccose to arachnoid texture that looked like tufts on PDA medium. The coloration on the reverse side varied among the Trichoderma spp. isolates, ranging from transparent to a watery white. Amplification of genomic DNA using internal transcribed spacer (ITS)-PCR resulted in a ~ 600 bp amplicon for all Trichoderma isolates, and the amplicons were subsequently validated through sequencing. The ITS sequences from different Trichoderma isolates were analyzed to confirm their molecular identity and homology using the NCBI-BLASTn. Out of 10 Trichoderma isolates, 9 were classified as T. harzianum (Accession numbers; OL604503.1–OL604511.1), while one isolate was identified as T. brevicompactum (OL604512.1). The phylogenetic analysis of ITS sequences demonstrated that the isolates were clustered into distinct groups corresponding to their respective species. The confrontation assay demonstrated that T. harzianum isolate (SVPRT-56) effectively inhibited the radial growth of Fusarium oxysporum up to 86.7%, whereas the lowest inhibition (59.7%) was observed with T. brevicompactum (SVPRT-60). Overall, this study suggests that T. harzianum strain (SVPRT-56) has the potential to serve as an effective biological control agent (BCA) and could be developed into Trichoderma-based bio-pesticides after stringent field trials for the eco-friendly management of Fusarium oxysporum diseases.
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Department of Biotechnology, IILM University, Greater Noida, India