Functionally analysis of legume symbiotic specific genes in transgenic Lotus japonicus

• Project/Area Number: 16580047
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Plant nutrition/Soil science
• Research Institution: Kagawa University
• Principal Investigator: TAJIMA Shigeyuki Kagawa University, Faculty of Agriculture, Professor, 農学部, 教授 (50116894)
• Co-Investigator(Kenkyū-buntansha): AGARIE-NOMURA Mika Kagawa University, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (50294749)
• Project Period (FY): 2004 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥3,700,000 (Direct Cost: ¥3,700,000); Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000); Fiscal Year 2004: ¥2,700,000 (Direct Cost: ¥2,700,000)
• Keywords: Lotus japonicus / transgenic plant / nodule / SNARE / promoter analysis / root cortex / PEPC / C / N metabolism / 二次元電気泳動 / 共生窒素固定

Research Abstract

(1)For surveying symbiotic differentiation of key organelle, mitochondria, soybean nodule and root mitochondria were compared after two dimensional electrophoresis and the proteins were characterized in combination with MALDI-TOF MS. We also surveyed the soybean nodule bacteroid by the methods of proteome analysis.
(2)We identified a Sed5-like clone LjSYP32-1 which contributes to nodule tissue formation and plant growth in Lotus japonicus. Promoter analysis and in situ hybridization revealed that LjSYP32-1 expressed significantly in inner cortex cell layer surrounding the infected zone of young nodules and at meristem area of developing lateral root. The anti-sense plants showed the nodules were much smaller and some nodules fused each other by sharing the vascular bundles. Such irregular nodules were observed in the rate of two times more comparing to those of wild type plant. The data suggest that LjSYP32-1 is contributing to support plant growth and normal nodule tissue differentiation.
(3)Using antisense technique, we diminished the expression levels of the nodule enhanced PEPC gene (Ljpepc1) in a determinate legume plant (Lotus japonicus) for surveying the influence to symbiotic phenotype and biochemical parameters. In nodules of Asppc plants, PEPC activity reduced to ca. 10% of non-transformants and the plants showed a typical nitrogen deficient symptom without nitrogen nutrient supply, and returned to normal growth with supply of 2.5 mM nitrate. Significant reduction of sucrose, succinate, asparagine, asparatate and glutamate contents also decreased in Asppc nodules. The data was discussed on a role of PEPC for the carbon/nitrogen metabolic flux in nodules.
(4)We isolated the some novel protein kinase possessing the subdomain VIB sequences characteristic of Ser/Thr kinase. To determine the function of the gene, we have made transgenic Lotus japonicus by RNAi methods.

Research Products

• [Journal Article] Expression cloning of a variety of novel protein kinase in Lotus japonicus. (2005)
  • Author(s): Kameshita, I, Nishida, T, Nakamura, S, Sugiyama, Y, Sueyoshi, N, Umehara, Y, Nomura, M, Tajima,S.
  • Journal Title: J.Biochemistry 137 Pages: 33-39
  • NAID: 10017342482
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Expression cloning of a variety of novel protein kinase in Lotus japonicus. (2005)
  • Author(s): Kameshita, I., Nishida, T., Nakamura, S., Sugiyama, Y., Sueyoshi, N., Umehara, Y., Nomura, M., Tajima, S.
  • Journal Title: J.Biochemistry 137 Pages: 33-39
  • NAID: 10017342482
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Expression cloning of a variety of novel protein kinase in Lotus japonicus. (2005)
  • Author(s): Kameshita, I., Nishida, T., Nakamura, S., Sugiyama, Y., Sueyoshi, N., Umehara, Y., Nomura, M., Tajima, S.
  • Journal Title: J. Biochemistry 137 Pages: 33-39
  • NAID: 10017342482
• [Journal Article] Expression cloning of a variety of novel protein kinase in Lotus japonicus. (2005)
  • Author(s): Nomura, M., Tajima, S 他6名
  • Journal Title: J.Biochemistry 137 Pages: 33-39
  • NAID: 10017342482
• [Journal Article] The promoter for C_4-type mitochondrial aspartate aminotransferase does not direct bundle sheath-specific expression in transgenic rice plants. (2005)
  • Author(s): Nomura, M., Higuchi, T., Katayama, K., Taniguchi, M., Tajima, S.他2名
  • Journal Title: Plant and Cell Physiology 46.5(In press)
  • NAID: 10015661862
• [Journal Article] Proteomic analysis on symbiotic differentiation of mitochondria in soybean nobules. (2004)
  • Author(s): Le Thi-Phong Hoa, Nomura, M, Tajima, S, 他2名
  • Journal Title: Plant and Cell Physiology 45 Pages: 300-308
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Characterization of bacteroid proteins in soybean nodules formed with Bradyrihizobium japonicum USDA110. (2004)
  • Author(s): Le Thi-PhongHoa, Mika Nomura, Shigeyuki Tajima
  • Journal Title: Microbes and Environments 19 Pages: 71-75
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Expression islands clustered on symbiosis island of Mesorhizobium loti genome. (2004)
  • Author(s): S.Tajima, M.nomura, A.Suzuki, Y.Shinoda K.Shioya, M.Abe, K.Mi namisawa 他16名
  • Journal Title: J. Bacteriology 186 Pages: 2439-2446
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Large scale analysis of gene expression profiles during early stages of root nobule formation in model legume, Lotus japonicus (2004)
  • Author(s): Kouchi, H, Tajima, S, 他15名
  • Journal Title: DNA research 11 Pages: 263-274
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Ureide biosynthesis in legume nodules (2004)
  • Author(s): Tajima, S, Nomura, M, Kouchi, H.
  • Journal Title: Frontiers in bioscience 9 Pages: 1374-1381
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Proteomic analysis on symbiotic differentiation of mitochondria in soybean nodules. (2004)
  • Author(s): Hoa, L.T.-P., Nomura, M., Kajiwara, H., Day, D.A., Tajima, S.
  • Journal Title: Plant and Cell Physiol. 45,3 Pages: 300-308
  • NAID: 10012724055
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Characterization of bacteroid proteins in soybean nodules formed with Bradyrihizobium japonicum USDA110. (2004)
  • Author(s): Hoa, L.T-P., Nomura, M., Tajima, S.
  • Journal Title: Microbes and Environments. 19,1 Pages: 71-75
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Expression islands clustered on symbiosis island of Mesorhizobium loti genome. (2004)
  • Author(s): Uchiumi, T., Owada, T., Itakura, M., Mitsui, Nukui, N., Dawadi, P., Kaneko, T., Yokoyama, T., Tejima, K., Saeki, K., Oomori, H., Hayashi, M., Maekawa, T., Sriprang, R., Murooka, Y., Tajima, S., Shimomura, K., Nomura, M., Suzuki, A., Shimoda, Y., Shioya, K., Abe, M., Minamisawa, K.
  • Journal Title: J.Bacteriology 186,8 Pages: 2439-2448
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Large scale analysis of gene expression profiles during early stages of root nodule formation in model legume, Lotus japonicus (2004)
  • Author(s): Kouchi, H., Tajima, S.etc
  • Journal Title: DNA research 11 Pages: 263-274
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Ureide biosynthesis in legume nodules. (2004)
  • Author(s): Shigeyuki Tajima, Mika Nomura, Hiroshi Kouchi
  • Journal Title: Frontiers in Bioscience 9 Pages: 1374-1381
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Proteomic analysis on symbiotic differentiation of mitochondria in soybean nodules. (2004)
  • Author(s): Le Thi-Phong Hoa, Nomura, M., Tajima, S. 他2名
  • Journal Title: Plant and Cell Physiology 45 Pages: 300-308
  • NAID: 10012724055
• [Journal Article] Expression islands clustered on symbiosis island of Mesorhizobium loti genome. (2004)
  • Author(s): S.Tajima, M., Nomura, A., Suzuki, Y., Shimoda, K., Shioya, M.Abe, K., Minamisawa 他16名
  • Journal Title: J.Bacteriology 186 Pages: 2439-2446
• [Journal Article] Large scale analysis of gene expression profiles during early stages of root nodule formation in model legume, Lotus japonicus (2004)
  • Author(s): Kouchi, H., Tajima, S.他15名
  • Journal Title: DNA research 11 Pages: 263-274