Sodium azide induced mutation of Actinomycetes I

Abstract

Sodium azide was employed to generate mutants of actinomycetes from Mount Everest region, viz. Sagarmatha National Park, Jorsale, Kalapatthar and Namche. Individual isolates of actinomycetes were inoculated on Starch Casein Agar plates containing 10, 50 and 100 ppm of sodium azide. Depending on complete inhibition of their growth , actinomycete isolates were categorized as highly sensitive , moderately sensitive and tolerant, respectively. Out of a total of 38 actinomycetes isolates, two (5.3%) were highly sensitive, 23 (60.5%) moderately sensitive and 13 (34.2%) tolerant. A total of 21 mutants were generated by treating 8 of the 23 moderately sensitive actinomycetes with 20, 30 and 40 ppm (sublethal concentrations ) of sodium azide and analyzed morphologically and biochemically. All mutants like wild strains were able to hydrolyse starch, Tween 20 and casein and utilize fructose and lactose. All were urease and catalase positive , but were different from wild strains in colony color or texture or both. Altogether nine (43 % ) mutants having negative mutation in loci of utilization of galactose or mannose (loss-of-function LOF, single mutants ) , two LOF double mutants in utilization of both galactose and arabinose , one gain-of-function (GOF) single mutant having positive mutation in the loci of utilization of galactose, one GOF double mutant in utilization of mannose and galactose and five double mutants (25%) having GOF ( arabinose or sucrose ) as well as LOF (mannose or galactose or arabinose) were obtained. Key words : Mt. Everest, Sagarmatha, actinomycetes, sodium azide, mutation, LOF, GOF DOI: 10.3126/ijls.v3i0.2385