Genotypic and Phenotypic Profile of Alkalophile Proteolytic Bacillus sp. Associated with Rhizosphere of Apple Trees in Trans Himalayas
AbstractRhizosphere of apple trees of Trans Himalayas is great niche of proteolytic bacteria which may have potential for use in industries. A total of 76 proteolytic bacteria were obtained from rhizosphere soil and root endosphere of apple trees from different sites of four locations viz. Chamba, Shimla, Kinnaur and Kullu of Himachal Pradesh, India, which were screened for plant growth promoting traits including production of indole acetic acid, phosphate solubilization, hydrogen cyanide production and production of antifungal metabolite. Among them, 17 isolates that exhibited maximum alkaline protease activity were selected. The diversity elucidation and differentiation of isolates was achieved through their biochemical characterization and amplified ribosomal DNA restriction analysis with three tetra cutter restriction enzymes (HpaII, HinfI and TaqI) the representative cluster groups were identified by 16S rDNA sequencing. This brings to light the predominance of Bacillus sp. and presence of remarkable functional and genotypic intraspecific diversity in apple rhizosphere. Proteolytic activity among selected isolates ranged between 50 and 2,770 ug/ml/min at pH 8.0 and the maximum was observed for Bacillus amyloliquefaciens (2,770 ug/ml/min) followed by B. subtilis (1,300.97 ug/ml/min) Bacillus amyloliquifaciens in particular showed great potential of alkaline protease production which agrees its exploitation in detergent industry. To the best of authors' knowledge, this is first report for genetic and functional diversity of alkaline protease producing PGPR associated with apple rhizosphere in Trans Himalayan region of Himachal Pradesh. Additionally the generated information may serve as a base line data for developing effective alkaline protease preparation for the industrial use.
Proceedings of the National Academy of Sciences, India Section B: Biological Sciences