Molecular identification of dominant microflora associated with ‘Hawaijar’ — A traditional fermented soybean ( Glycine max (L.)) food of Manipur, India

Abstract

The dominant microorganisms in ‘Hawaijar’, a traditional non-salted fermented soybean ( Glycine max (L.)) food of Manipur, India, were isolated and identified by molecular techniques. Bacillus spp. were the predominant microorganisms in ‘Hawaijar’. A total of 274 isolates were obtained from forty-one ‘Hawaijar’ samples collected from household preparations and markets of valley districts of Manipur. Phenotypic grouping followed by Amplified Ribosomal DNA Restriction Digestion Analysis (ARDRA), PCR amplification of 16S–23S rDNA region, RFLP by Hae III and Hind III double digest (by comparing MTCC type strains) and sequencing of 9-1514 region of 16S rDNA resulted in three major phylogenic groups. Bacillus subtilis group comprising B. subtilis and B. licheniformis representing 112 isolates, B. cereus group representing of 128 isolates and Staphylococcus spp. group comprising S. aureus and S. sciuri representing 23 isolates. A few bacterial isolates belongs to Alkaligenes spp. and Providencia rettgeri were also found. Genetic diversity of B. subtilis phylogenic group was investigated by RAPD-PCR. Eighty-two strains of B. subtilis phylogenic group were identified using RAPD-PCR using OPA-18 and OPA-20 primers. These strains will be investigated for potential starter culture selection.