Author(s):

Gadambe Shilpad, Kale Sonam S, Kadu Tanvi P, Toshy Agrawal and As Kotasthane

Abstract: <em>Trichoderma</em> has gained immense importance since last few decades due to its biological control ability against several plant pathogens. Twenty isolates of <em>T. aureoviride</em> were collected from different geographical locations of Chhattisgarh. Molecular characterization of isolates of <em>T.</em> <em>aureoviride</em> revealed homogeneity with ITS primer (internal transcribed spacer). In the present study the genetic relatedness among twenty isolates of <em>Trichoderma aureoviride </em>was analyzed by using sixteen gene specific markers. PCR amplification with ITS primers showed homogeneity of all <em>T. aureoviride </em>isolates indicating that they belong to the same species. Six different chitinase gene specific primers produced amplification of chitinase genes in almost all <em>T. aureoviride </em>isolates except one of the exochitinase encoding gene (<em>N</em>-acetyl- -glucosaminidase), <em>nag</em>70 based primer Nag70F-R and two endochitinase encoding gene specific primers Ech42-2F-R and Ech42-3F-R that could not amplify any of the isolates of <em>T. aureoviride</em>. PCR amplification of fragments of cellulase gene using CBH1 primer amplified a nucleotide fragment of the expected size from some isolates while PR808-PRegl4 primer pair amplified 340, 140 &amp; 100 bp bands in all the isolates. Primer pair fRPB2-5F2- fRPB2-7cR2 generated major band above 1000 bp confirming the amplification of a 1.2-kb fragment of subunit 2 of the RNA polymerase B gene (RPB2) containing an ITS-like region. Almost all the isolates showed the presence of endochitinase and cellulose gene fragments which proves their ability to antagonize phytopathogens.

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