Plant-microbe interaction in the rhizosphere is the determinant of soil fertility and plant health. The presence of beneficial bacteria in the vicinity of roots stimulates plant growth. In this way, soil bacteria play very important role in improving plant nutrition and have been utilized for agriculture for long times. However, little has been done at molecular level in Pakistan to explore their potential. The present study was designed with the objectives to isolate bacterial strains from legume rhizospheric soil and nodules, to characterize and identify potential bacterial strains by using molecular tagging of 16S rRNA gene sequencing and to asses rhizobacterial impact on yield, nodulation and N2-fixation of legume crops under controlled and field conditions. Extensive survey was carried out in Pothwar (District Rawalpindi, Attock and Chakwal) for collection of legumes (mash bean and chickpea) nodules and rhizospheric soil. Five samples of each legume crop were collected from each district. Rhizospheric soil bacteria were isolated through dilution plate technique using Phosphate Buffer Saline solution (PBS; 1X) and nutrient media i.e. Tryptic Soya Agar (TSA; Difco). Root nodules for Rhizobium isolation were washed, crushed and directly streaked on yeast extract mannitol (YEM) plates supplemented with Congo red. About 100 bacterial strains of different genera included Rhizobium were isolated and designated as AM-1, AM-2 to AM-100. These isolated bacterial strains were characterized for plant growth promoting (PGP) properties like auxin i.e. indole acetic acid (IAA), P solubilization and production of NH3. On the basis of PGP traits, 10 most potential strains were selected and identified using molecular techniques i.e. 16S rRNA sequencing. The DNA of each strain was amplified using universal primers 9F: (´GAGTTTGATCCTGGCTCAG´) and 1510R: (´GGCTACCTTGTTACGA´).
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