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The Lotus japonicus Sen1 gene controls rhizobial differentiation into nitrogen-fixing bacteroids in nodules

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Abstract

A Lotus japonicus mutant, Ljsym75, which forms ineffective symbiotic nodules and defines a new locus involved in the process of nitrogen fixation, was characterized in detail in order to identify the stage of developmental arrest of the nodules. No nitrogen-fixing activity was detectable in Ljsym75 nodules at any stage during plant development, and plant growth was markedly retarded. Ljsym75 plants formed twice as many nodules as the wild-type Gifu, and this phenotype was not influenced by the application of low concentrations of nitrate. Although the ineffective nodules formed on Ljsym75 were anatomically similar to effective Gifu nodules, Ljsym75 nodules senesced prematurely. Microscopic examination revealed that bacteria endocytosed into Ljsym75 nodules failed to differentiate into bacteroids. Moreover, the bacteria contained no nitrogenase proteins, whereas leghemoglobin was detected in the cytosol of the nodules. These results indicate that Ljsym75 is required for bacterial differentiation into nitrogen-fixing bacteroids in nodules, and thus the Ljsym75 gene was renamed sen1 (for stationary endosymbiont nodule). Linkage analysis using DNA markers showed that Sen1 is located on chromosome 4.

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Acknowledgements

The authors would like to thank Dr. F.C. Guinel (Wilfrid Laurier University, Laval, Canada) for critical reading of the manuscript and Dr. T. Bisseling (Agricultural University, Wageningen, Netherlands) for providing antisera against nitrogenase components I and II from pea bacteroids. They would also like to acknowledge Dr. S. Sato (Kazusa DNA Research Institute) for providing information about DNA markers, Dr. H. Kouchi (National Institute of Agrobiological Resources) and Dr. H. Kitano (Nagoya University) for technical suggestions. This work was supported by special coordination funds for promoting science and technology from the Japanese Ministry of Education, Culture, Sports, Science, and Technology.

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  1. S. Akao

    Present address: Faculty of Agriculture, Miyazaki University, Miyazaki, Miyazaki 889-2192, Japan

Authors and Affiliations

  1. Department of Life Science, Aichi University of Education, Kariya, Aichi 448-8542, Japan

    N. Suganuma, Y. Nakamura, M. Yamamoto & T. Ohta

  2. Iwate Biotechnology Research Center, Kitakami, Iwate 024-0003, Japan

    H. Koiwa

  3. National Institute of Agrobiological Resources, Tsukuba, Ibaraki 305-8602, Japan

    S. Akao

  4. Department of Environmental Sciences, Faculty of Science, Niigata University, Niigata, Niigata 950-2181, Japan

    M. Kawaguchi

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  1. N. Suganuma
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  2. Y. Nakamura
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Correspondence to N. Suganuma.

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Communicated by A. Kondorosi

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Suganuma, N., Nakamura, Y., Yamamoto, M. et al. The Lotus japonicus Sen1 gene controls rhizobial differentiation into nitrogen-fixing bacteroids in nodules. Mol Gen Genomics 269, 312–320 (2003). https://doi.org/10.1007/s00438-003-0840-4

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