Investigation of Sequence-Tagged Site (STS) in Chitinase Gene of Gamma Radiated Trichoderma viride Isolates
Journal Title: International Journal of Farming and Allied Sciences - Year 2014, Vol 3, Issue 11
Abstract
Trichoderma spp. have been identified as effective biological control agents of a number of soil borne pathogens through different strategies like cell wall degrading enzymes production (Chitinase and Glucanases). The most crucial boundaries to use Trichoderma as bio-fungicides are their low effective level in field condition. Thus, genetic modification using mutagenesis offers the potential for producing improved bio- protection. Thereby, effects of induced mutation via gamma irradiated with 250 Gry dose (in Agricultural and Medical Research School- Nuclear Science and Technology Institute, Karaj) on Trichoderma viridea conidia on chitinase activity by STS molecular marker have been evaluated in this study. Among 21 irradiated mutants which selected via improved antagonistic capability against Macrophomina phaseolina, PCR amplified chitinase mutated genes with their nested primers produced different sizes of amplified regions between 500- 700 bp, which showed polymorphism in compare with control (non-irradiated) isolate. In T. viridea irradiation induced considerable positive mutations and in 90.9% of mutants chitinase activity increased. Dual culture test showed that, 10 mutants of T. viridea have statistically more antagonistic capability against M. phaseolina than its parent isolate. Protein profile analysis of these mutants supported our hypothesis that: enhancement in fungal cell wall degrading enzymes production via irradiation resulted in most of antagonism capacity increasing. Other mutants which have better antagonism against M. phaseolina without enzyme activity increasing, maybe carry mutations in other bio-control mechanisms. STS analysis data proved that in considerable mutants, induced mutation placed in chitinase gene. According to these results, bio-control capability of T. viridea could be improved through gamma radiation.
Authors and Affiliations
Samira Shahbazi , Mahsa Karimi and Mohammad Ali Ebrahimi
Keywords
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