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      • KCI등재

        Assessment of nifH Diversity in Rhizobial Isolates of Different Origin and the Role of Antioxidant in Respiratory Protection

        Bhaganagare, Govindraj Ramakantrao,Kesawat, Mahipal Singh,Das, Basanta Kumar,Suresh, Arumuganainar,Surwase, Babasaheb Shivmurti,Manorama, Manorama 한국작물학회 2013 Journal of crop science and biotechnology Vol.16 No.1

        Rhizobia diversity is considered as one of the most useful resources for bioprospecting due to their symbiotic nitrogen-fixing ability with members of Leguminosae. The highly conserved nature of the nitrogenase reductase gene (nifH) makes it an ideal molecular tool to determine the potential for biological nitrogen fixation in any environment. In the present investigation, 250 rhizobial strains were isolated from legumes belonging to different geographical locations of Chhattisgarh, India. Genetic diversity of the nitrogen-fixing bacterial community was analyzed using the nifH gene-specific primer. The polymorphism was found among the nitrogen-fixing population of different sources and origin but not in same source of rhizobia. Further, the symbiotic plasmid DNA was characterized on the basis of size and copy number of plasmids. The plasmid number varying from one to three in different rhizobial isolates had a size greater than 23 kb, while in some rhizobial isolates plasmids were absent. In addition, to examine the role of ascorbate in respiratory protection, the clear black spot margin of ascorbate was observed in the endodermis region of the nodule whereas scarcely dispersed in the infected region. Therefore, our findings demonstrated that knowing the rhizobial nifH gene diversity along with copy number of the plasmid is important for strain identification, deciding its fertility, productivity standards, and potential of biological nitrogen fixation across the geographical region.

      • KCI등재

        Assessment of nifH Diversity in Rhizobial Isolates of Different Origin and the Role of Antioxidant in Respiratory Protection

        Govindraj Ramakantrao Bhaganagare,Mahipal Singh Kesawat,Basanta Kumar Das,Arumuganainar Suresh,Babasaheb Shivmurti Surwase,Manorama 한국작물학회 2013 Journal of crop science and biotechnology Vol.16 No.1

        Rhizobia diversity is considered as one of the most useful resources for bioprospecting due to their symbiotic nitrogen-fixing ability with members of Leguminosae. The highly conserved nature of the nitrogenase reductase gene (nifH) makes it an ideal molecular tool to determine the potential for biological nitrogen fixation in any environment. In the present investigation, 250 rhizobial strains were isolated from legumes belonging to different geographical locations of Chhattisgarh, India. Genetic diversity of the nitrogenfixing bacterial community was analyzed using the nifH gene-specific primer. The polymorphism was found among the nitrogen-fixing population of different sources and origin but not in same source of rhizobia. Further, the symbiotic plasmid DNA was characterized on the basis of size and copy number of plasmids. The plasmid number varying from one to three in different rhizobial isolates had a size greater than 23 kb, while in some rhizobial isolates plasmids were absent. In addition, to examine the role of ascorbate in respiratory protection, the clear black spot margin of ascorbate was observed in the endodermis region of the nodule whereas scarcely dispersed in the infected region. Therefore, our findings demonstrated that knowing the rhizobial nifH gene diversity along with copy number of the plasmid is important for strain identification, deciding its fertility, productivity standards, and potential of biological nitrogen fixation across the geographical region.

      • KCI등재후보

        Isolation and Characterization of Lipopolysaccharides from Different Rhizobial Isolates

        Mahipal Singh Kesawat,Basanta Kumar Das,GR Bhaganagare,Vinay Sharma,Manorama 한국작물학회 2009 Journal of crop science and biotechnology Vol.12 No.3

        Formation of nodules on roots or in stems (in some cases) of leguminous plants is the unique ability of gram-negative bacteria, Rhizobia, which converts atmospheric nitrogen into usable forms by the host plant. Lipopolysaccharide (LPS) is the outer membrane component of the gram-negative bacteria, known to be an essential factor in host recognition, specificity, and initial infection processes. In the present study, we extracted lipopolysaccharides from different rhizobial isolates by a modified phenol-water method and partially characterized by polyacrylamide gel electrophoresis with silver staining. The results showed two separate banding regions, LPS-I and LPS-II. The high molecular weight and electrophoretic mobility of LPS-I region resembles that of lysozyme, used as a standard marker. The LPS-II region has a low molecular weight and electrophoretic mobility greater than that of lysozyme. The LPS-II region was due to incomplete LPS, which either lacks the entire O-antigen repeating unit or contains only one or two repeating units. The banding patterns of LPS vary among the different rhizobial isolates. Results revealed that the type of LPS structure and banding regions greatly facilitate the further characterization of the LPS modifications required for symbiosis. Formation of nodules on roots or in stems (in some cases) of leguminous plants is the unique ability of gram-negative bacteria, Rhizobia, which converts atmospheric nitrogen into usable forms by the host plant. Lipopolysaccharide (LPS) is the outer membrane component of the gram-negative bacteria, known to be an essential factor in host recognition, specificity, and initial infection processes. In the present study, we extracted lipopolysaccharides from different rhizobial isolates by a modified phenol-water method and partially characterized by polyacrylamide gel electrophoresis with silver staining. The results showed two separate banding regions, LPS-I and LPS-II. The high molecular weight and electrophoretic mobility of LPS-I region resembles that of lysozyme, used as a standard marker. The LPS-II region has a low molecular weight and electrophoretic mobility greater than that of lysozyme. The LPS-II region was due to incomplete LPS, which either lacks the entire O-antigen repeating unit or contains only one or two repeating units. The banding patterns of LPS vary among the different rhizobial isolates. Results revealed that the type of LPS structure and banding regions greatly facilitate the further characterization of the LPS modifications required for symbiosis.

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