EXPLORING NATURAL COMBINATION FOR IDENTIFICATION OF UPREGULATED NITROGEN FIXING BACTERIA SPECIFIC TO CHICKPEA IN TARGETED GEOGRAPHY: A PHYSICAL, BIOCHEMICAL, AND In Silico APPROACH

The object of the study is to understand the concept of symbiosis in a more localized natural selective way to optimize the crop yield of chickpea in a targeted geography. Specifically the aim is to isolate and identify the specific Rhizobium strains for Cicer arietinum from Bhadrachalam forest lands using physical, biochemical and in silico analysis. Soils were collected randomly from 45 locations of Bhadrachalam forest and sown with chickpea seeds in triplets. All the samples are found to contain similar levels of NPK and micronutrients. Top five soil samples which supported the best plant growth were considered for further investigation. Pure cultures of isolated Rhizobial species obtained from the root nodules were maintained. Lag phase cultures in broth form were inoculated on the seeds sown in sterile soil samples maintaining a control. Tremendous growth has been identified in the inoculated soils when compared with the control proving the efficacy of Rhizobia. The organisms which are contributing for plant growth were identified by phylogenetic analysis were found to be Mesorhizobium Ciceri, Rhizobium meliloti, Rhizobium fredii, Bradyrhizobium japonicum, and Mesorhizobium loti sp. These species has shown high rate of plant growth not only at green house conditions but also in low vegetative agriculture soils of the same geography. The 16S rRNA gene sequencing revealed the fact that soil sample S 4 contains abundant M. Ciceri sp., with 99% similarity while R. fredii, M. loti, R. meliloti, and B. japonicum were found in sample 1, 2, 3 and 5 respectively. Furthermore in silico analysis of Nif A protein revealed the underlining precursors of IAA production and nitrogenase activity. Therefore this method could be adopted for easy identification of species specific rhizobium in targeted geographies.