Vol. 8, Issue 2 (2019)

Author(s):
Gudia Punia, Neeraj Tondan, Ashwani Kumar and CS Prasad

Abstract:
Molecular characterization in present work involved slight modification in standard CTAB method and ommision of poly vinyl pyrrolidone (PVP). Genetic variability present among different strains of B. bassiana can be studied through different PCR based molecular markers such as RAPD (Random Amplified Polymorphic DNA). A Known isolate, MTCC 4492 from (Institute of Microbial Technology), Chandigarh was included as standard in the experiments. Microscopic studies revealed presence of conidiophores consisting of whorls and dense cluster of short and globose cells with apical zig-zag appearance and one celled spherical conidia. Fungal colonies on SDYA media were white to pale yellow; some were red pigmented in reverse. Eight RAPD primers were used to characterize the genetic diversity present among isolates; they showed a total of 96 reproducible bands. Each of the primer varied greatly in their ability to resolve variability among the genotypes. The individual primer produced bands in a range of nine (LC103) to 18 (LC-89) bands with an average of 9.6 bands per primer. Out of the 96 bands, 1 band was monomorphic i.e. they were present in both isolates. The amplified fragment ranged from 280bp (primer LC-72) to 4000bp (primer LC-103). The polymorphism percentage ranged between 91.6% (primer LC-73) to 100%. The average polymorphic bands recorded as 9.5 bands per primer. Almost all primers based on 100% polymorphism and unique band amplification was considered as highly informative primers. Also the virulence of the fungus was observed with a control isolate to check its ability to cause infection of insect pest, cause its death and emerge out of its dead body.