Sodium azide induced mutaion of Actinomycetes II: Biochemical and Genetic Characterization

Pradeep Subedi; Simon Shrestha; Rubin Thapa Magar; Bharat Pokhrel; Kiran Babu Tiwari

Authors

Pradeep Subedi Universal Science College, Pokhara Universty, Chakupat, Kathmandu
Simon Shrestha Universal Science College, Pokhara Universty, Chakupat, Kathmandu
Rubin Thapa Magar Universal Science College, Pokhara Universty, Chakupat, Kathmandu and Padmashree Institute of Management and sciences, Bangalore, India
Bharat Pokhrel Universal Science College, Pokhara Universty, Chakupat, Kathmandu and Padmashree Institute of Management and sciences, Bangalore, India
Kiran Babu Tiwari Universal Science College, Pokhara Universty, Chakupat, Kathmandu and Research Laboratory for Biotechnology and Biochemistry (RLABB), Sanepa, Kathmandu, Nepal

Keywords:

Actinomycetes, biochemical and genetic polymorphisms, Mt. Everest, mutation, sodium azide

Abstract

Sodium azide was employed to generate mutants of an actinomycete isolate (Snp1e) from Sagarmatha National Park of Mount Everest region to establish biochemical and genetic changes among them. Nine mutants were selected from Starch Casein Agar plates containing 50, 75 and 100 ppm sodium azide based on observed morphological differences of individual colonies and purified subsequently. Individual isolates were tested for carbon assimilation tests, Nitrate reduction, Catalase production and hydrolysis tests. Five decamer primers were used to establish genomic polymorphisms among the isolates by Randomly Amplified Polymorphic DNA – PCR method. Most of the mutants grew faster than the wild type isolate in Starch-Casein broth. Many of the mutants were able to hydrolyze urea and Tween-20 and produce Nitrate reductase. All mutants were able to show one or more of either GOF mutation or LOF mutation or both. The primers allowed scoring of total 173 bands of which 53 (30.64%) were polymorphic. Primer P1 demonstrated DNA polymorphisms above 50% among the isolates and large numbers of bands were observed with primers P3 and P5. Ample biochemical and genetic variations were observed among mutants derived from an actinomycete using Sodium azide.