DNA Synthesis and Fragmentation in Bacteroids during Astragalus sinicus Root Nodule Development.

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  • KOBAYASHI Hajime
    Department of Biotechnology, Graduate School of Engineering, Osaka University
  • SUNAKO Michihiro
    Department of Biotechnology, Graduate School of Engineering, Osaka University
  • HAYASHI Makoto
    Department of Biotechnology, Graduate School of Engineering, Osaka University
  • MUROOKA Yoshikatsu
    Department of Biotechnology, Graduate School of Engineering, Osaka University

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Histo- and cytochemical techniques were used to study the DNA replication and fragmentation patterns in bacteroids formed by Mesorhizobium huakuii subsp. rengei in nodules of Astragalus sinicus. DNA replication was detected by the incorporation of 5-bromo deoxy-uridine. Signals denoting DNA synthesis were observed in plant nuclei within the nodule meristem and in bacteroids near the meristem. The TUNEL (TdT-mediated dUTP nick-end labeling) assay was used to measure DNA fragmentation. In nutrient-depleted 1-mpi (month(s) post inoculation) nodule sections, some bacteroids were in vacuoles, and DNA fragmentation signals were observed only in such bacteroids. In contrast, 1-mpi nodule sections without nutrient depletion showed neither bacteroid localization in vacuoles nor DNA fragmentation signals. The bacteroid translocation into vacuoles upon nutrient starvation might results from autophagy of the plant. In 2-mpi nodule sections, bacteroids with DNA fragmentation signals appeared within the cytoplasm of some nodule cells in the senescence zone.

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