Physiological and molecular characterization of locally adapted Rhizobium strains of lentil (Lens culinaris Medik.) having restricted phage sensitivity
Tóm tắt
Indigenous rhizobia were isolated from root nodules of lentil plants collected from various agro-climatic regions of India. These isolates together with four standard lentil Rhizobium strains were screened for sensitivity against eight phages. Four strains, USDA 2431, BHULR 104, BHULR 113 and BHULR 115 having restricted sensitivity to lytic phages LRP-1, LRP-4, LRP-13 and LRP-15 respectively, were characterized for both physiological and molecular characters. All strains had a generation time of between 3.8 and 5.6 h in yeast extract–mannitol (YM) broth, and colonies on YM agar plates showed an acidic reaction. Compared to other strains, strain USDA 2431 grew poorly when sucrose was the sole carbon source and showed maximum growth in arabinose-containing medium. The intrinsic antibiotic resistance level in all strains was tested against seven antibiotics and found to be high with ampicillin and kanamycin (50–60 μg ml-1) but very low with neomycin (0.03 μg ml-1). With the exception of strain BHULR 113, all strains expressed ex planta nitrogenase activity, with strain USDA 2431 showing the maximum activity (26.8 nmol C2H4 h-1 mg-1 protein) after 6 h of incubation. Genomic and phylogenetic relationships among the strains were examined by randomly amplified polymorphic DNA and 16S rRNA sequence analysis. Genetic distance varied from 0.09 to 0.23 among the strains, and the primer OPL-11 was found to be suitable for the discrimination of these strains. The 16S rRNA sequence analysis revealed 99–100% similarity with Rhizobium leguminosarum bv. viciae. These results clearly indicate that phage sensitivity is a useful marker for discriminating locally soil-adapted rhizobial strains forming effective nodules in lentil.
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