Global analysis of symbiotic tissue specific gene expressions of nitrogen and carbon metabolisms in legume nodules

豆科植物根瘤中氮和碳代谢共生组织特异性基因表达的整体分析

• 批准号: 14560054
• 负责人: TAJIMA Shigeyuki
• 金额: $ 2.37万
• 依托单位: Kagawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14560054/
• 关键词: Symbiotic Nitrogen Fixation; Macroarray; Proteome; RNAi; Transgenic plants; Lotus Japonicus; SNARE; Antisense; ダイズ; マメ科植物; 根粒; ミトコンドリア; PMF(peptide mass fingerprint)解析; 自動エドマン法

1)Proteome analysis of nodule mitochondria and nodule bacteroids of soybean plants was surveyed, In. the case of nodule mitochondria 2D-EP following Protein Mass Finger print and LCMSMS analysis revealed that 465 spots (nodule mitochondria) and 383 spots (root mitochondria) were tissue specific. Using Edman degradation methods, Protein Mass Finger print and LCMSMS analysis identification of these protein spots were performed. Although there are many spots which the function were not registered in the database, the data showed 7 up-regulating and 8 down-regulating proteins were identified. Six suppressed proteins in nodule mitochondria were identified too. The role in nodule metabolism of these proteins were discussed. Similar approach was employed in the case of nodule bacteroids too. In this bacteroid proteome experiment, we identified 62 proteins which were up-regulated in nodule bacteroids comparing to cultured bacteria.2)For preparing genome sequencing data we joined to the project team of Kazusa DNA Research Institute and Dr. Kouchi' laboratory. In this experiment we sequenced more than 9,000 cDNA clones from 5'site of the clone and the data is going to supplied to the public through Kazusa DNA Research Institute.

1)对大豆根瘤线粒体和根瘤类细菌的蛋白质组进行了研究。蛋白质质量指纹和LCMSMS分析后的结节线粒体2D-EP的情况显示465个点（结节线粒体）和383个点（根线粒体）是组织特异性的。采用Edman降解法、蛋白质指纹图谱和LCMSMS分析对这些蛋白质点进行鉴定。虽然有许多点的功能在数据库中没有登记，数据显示，7个上调和8个下调蛋白质的鉴定。在根瘤线粒体中还发现了6个抑制蛋白。讨论了这些蛋白在根瘤代谢中的作用。类似的方法也用于结节类杆菌的情况。在这个类杆菌蛋白质组实验中，我们鉴定了62个蛋白质，与培养的细菌相比，这些蛋白质在结核类杆菌中表达上调。2）为了准备基因组测序数据，我们加入了Kazusa DNA研究所的项目团队和Kouchi博士的实验室。在本实验中，我们对9,000多个克隆的5 '位点的cDNA进行了测序，数据将通过Kazusa DNA研究所提供给公众。

项目成果

Hoa, L., Mika Nomura, Hideyuki Kajiwara, David Alexander Day, Shigeyuki Tajima: "Proteomic analysis on symbiotic differentiation of mitochondria in soybean nodules."Plant Cell Physiol. 45. 300-308 (2004)

Hoa，L.，Mika Nomura，Hideyuki Kajiwara，David Alexander Day，Shigeyuki Tajima：“大豆根瘤中线粒体共生分化的蛋白质组学分析。”植物细胞生理学。

Hoa, L., Mika Nomura, Shigeyuki Tajima: "Characterization of bacteroid proteins in soybean nodules formed with Bradyrhizobium japonicum USDA110."Microbes & Environment. 19. 71-75 (2004)

Hoa, L.、Mika Nomura、Shigeyuki Tajima：“用日本慢生根瘤菌 USDA110 形成的大豆根瘤中类菌蛋白的特征。”微生物

Uchiumi T, Ohwada T, Itakura M, Mitsui H, Nukui N, Dawadi P, Kaneko T, Tabata S, Yokoyama T, Tejima K, Saeki K, Omori H, Hayashi M, Maekawa T, Sriprang R, Murooka Y, Tajima S, Simomura K, Nomura M, Suzuki A, Shimoda Y, Sioya K, Abe M, Minamisawa: "Express

Uchiumi T、Ohwada T、Itakura M、Mitsui H、Nukui N、Dawadi P、Kaneko T、Tabata S、Yokoyama T、Tejima K、Saeki K、Omori H、Hayashi M、Maekawa T、Sriprang R、Murooka Y、Tajima S

Uchiumi T, Ohwada T, Itakura M, Mitsui H, Nukui N, Dawadi P, Kaneko T, Tabata S, Yokoyama T, Tejima K, Saeki K, Omori H, Hayashi M, Maekawa T, Sriprang R, Murooka Y, Tajima S, Simomura K, Nomura M, Suzuki A, Shimoda Y, Sioya K, Abe M: "Minamisaw, Expressi

Uchiumi T、Ohwada T、Itakura M、Mitsui H、Nukui N、Dawadi P、Kaneko T、Tabata S、Yokoyama T、Tejima K、Saeki K、Omori H、Hayashi M、Maekawa T、Sriprang R、Murooka Y、Tajima S

Hoa, L., Mika Nomura, Hideyuki Kajiwara, David Alexander Day, Shigeyuki Tajima: "Proteomic analysis on symbiotic differentiation of mitochondria in soybean nodules"Plant Cell Physiol. 45. 300-308 (2004)

Hoa，L.，Mika Nomura，Hideyuki Kajiwara，David Alexander Day，Shigeyuki Tajima：“大豆根瘤中线粒体共生分化的蛋白质组学分析”植物细胞生理学。