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Related Experiment Videos

Polypeptide synthesis by Rhizobium bacteroids and bacteria.

B D Shaw, W D Sutton

    Biochimica Et Biophysica Acta
    |June 20, 1979
    PubMed
    Summary

    Rhizobium bacteroids synthesize specific proteins, including nitrogenase components, identified using [35S]-methionine labeling. Protein patterns vary with nodule maturity and Rhizobium strain, differing from cultured bacteria.

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    Area of Science:

    • Microbiology
    • Plant-Bacterial Interactions
    • Molecular Biology

    Background:

    • Rhizobium bacteroids are key symbionts in legume nitrogen fixation.
    • Understanding bacteroid protein synthesis is crucial for optimizing symbiosis.
    • Previous studies identified nitrogenase polypeptides but lacked detailed protein synthesis profiles.

    Purpose of the Study:

    • To characterize the de novo synthesized polypeptides in Rhizobium bacteroids.
    • To investigate how environmental conditions and developmental stages affect bacteroid protein profiles.
    • To compare protein synthesis patterns between bacteroids and cultured bacteria.

    Main Methods:

    • Isolation and aerobic incubation of Rhizobium bacteroids (strain NZP 2257) from lupin nodules.
    • Incorporation of [35S]-methionine to label newly synthesized polypeptides.
    • Analysis of labeled polypeptides using SDS-polyacrylamide gel electrophoresis.
    • Pulse-chase experiments to assess polypeptide stability.
    • In situ labeling of bacteroids within detached nodules.

    Main Results:

    • A distinct set of polypeptides was synthesized by isolated bacteroids, with many matching Coomassie blue-stained bands.
    • Labeled polypeptides were stable for several hours.
    • Ion concentrations (H+, K+, Mg2+) affected overall labeling but not relative patterns.
    • Distinct labeling patterns were observed in mature vs. immature bacteroids, correlating with nitrogenase appearance.
    • Two major labeled components in mature bacteroids (60 and 34 kDa) likely represent nitrogenase subunits.
    • Bacteroids from the same strain but different hosts showed similar patterns; different strains showed distinct patterns.
    • Bacteroid labeling patterns differed significantly from those of broth-cultured Rhizobium.

    Conclusions:

    • Rhizobium bacteroids exhibit specific protein synthesis profiles, distinct from cultured bacteria.
    • Bacteroid polypeptide synthesis is regulated by plant developmental stage and Rhizobium strain.
    • The identified labeled polypeptides, particularly the 60 and 34 kDa components, are strong candidates for nitrogenase subunits, providing insights into symbiosis regulation.

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