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Transcript and metabolite analysis of activation-tagging lines of a model legume

Research Project

Project/Area Number 14540604
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 植物生理
Research InstitutionNihon University

Principal Investigator

AOKI Toshio  Nihon University, College of bioresource sciences, Associate professor, 生物資源科学部, 助教授 (80287606)

Co-Investigator(Kenkyū-buntansha) AYABE Shin-ichi  Nihon University, College of bioresource sciences, Professor, 生物資源科学部, 教授 (40050679)
AKASHI Tomoyoshi  Nihon University, College of bioresource sciences, Research associate, 生物資源科学部, 助手 (80328707)
Project Period (FY) 2002 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2003: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2002: ¥1,700,000 (Direct Cost: ¥1,700,000)
KeywordsLotus japonicus / T-DNA / tagging / mutant / model plant / transformation / T-DNAタギング / Lotus japonicus / アクティベーションタギング / Agrobacterium
Research Abstract

We constructed T-DNA insertional lines of a model legume, Lotus japonicus, using a multifunctional vector for gene and exon activation tagging. The vector had the CaMV 35S promoter together with two additional enhancer elements, the start codon, and splice donor and acceptor sites facing to the left border, in anticipation of the activation of T-DNA flanking genes and forced expression of flanking exons. The improved transformation technique yielded more than 3,500 lines, including 45 dominant mutant candidates with abnormal phenotypes in aerial parts, nodules and roots. Among selected 44 lines, one copy of T-DNA was inserted into the genome of 37 lines (84%). The T-DNA flanking regions of seven lines were isolated by thermal asymmetric interlaced(TAIL)-PCR or reverse transcription(RT)-PCR, and the corresponding genomic clones were analyzed. The transcripts of four genes adjacent to T-DNA out of eleven genes tested were increased in the T_1 generation, demonstrating that gene and exon activation effects by the newly developed tagging vector are heritable. This is the first report on the activation tagging of a legume, and the T-DNA insertional population of L.japonicus will provide legume-specific dominant mutants.

Report

(4 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (17 results)

All 2005 2004 2003 2002 Other

All Journal Article (15 results) Book (1 results) Publications (1 results)

  • [Journal Article] Molecular and biochemical characterization of 2-hydroxyisoflavanone dehydratase. Involvement of carboxylesterase-like proteins in leguminous isoflavone biosynthesis2005

    • Author(s)
      Akashi et al.
    • Journal Title

      Plant Physiology 137

      Pages: 882-891

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Multiple mutagenesis of P450 isoflavonoid synthase reveals a key active-site residue2005

    • Author(s)
      Sawada et al.
    • Journal Title

      Biochemical and Biophysical Research Communications 330

      Pages: 907-913

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Molecular and biochemical characterization of 2-hydroxyisoflavanone dehydratase. Involvement of carboxylesterase-like proteins in leguminous isoflavone biosynthesis.2005

    • Author(s)
      Akashi, T., Aoki, T., Ayabe, S.
    • Journal Title

      Plant Physiology 137(3)

      Pages: 882-891

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Multiple mutagenesis of P450 isoflavonoid synthase reveals a key active-site residue.2005

    • Author(s)
      Sawada, Y., Ayabe, S.
    • Journal Title

      Biochemical and Biophysical Research Communications 330(3)

      Pages: 907-913

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Large-Scale Analysis of Gene Expression Profiles during Early Stages of Root Nodule Formation in a Model Legume, Lotus japonicus2004

    • Author(s)
      Kouchi et al.
    • Journal Title

      DNA Research 11

      Pages: 263-274

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Large-Scale Analysis of Gene Expression Profiles during Early Stages of Root Nodule Formation in a Model Legume, Lotus japonicus2004

    • Author(s)
      Kouchi, H., Shimomura, K., Hata, S., Hirota, A., Wu, G., Kumagai, H., Tajima, S., Suganuma, N., Suzuki, A., Aoki, T., Hayashi, M., Yokoyama, T., Ohyama, T., Asamizu, E., Kuwata, C., Shibata, D., Tabata, S.
    • Journal Title

      DNA Research 11(4)

      Pages: 263-274

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] cDNA cloning and biochemical characterization of S-adenosyl-L-methionine : 2,7 4'-trihydroxyisoflavanone 4'-O-methyltransferase, a critical enzyme of the legume isoflavonoid phytoalexin pathway2003

    • Author(s)
      Akashi et al.
    • Journal Title

      Plant and Cell Physiology 44

      Pages: 103-112

    • NAID

      10010272519

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A cluster of genes encodes the two types of chalcone isomerase involved in the biosynthesis of general flavonoids and legume-specific 5-deoxy(iso)flavonoids in Lotus japonicus2003

    • Author(s)
      Shimada et al.
    • Journal Title

      Plant Physiology 131

      Pages: 941-951

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] cDNA cloning and biochemical characterization of S-adenosyl-L-methionine : 2,7,4'-trihydroxyisoflavanone 4'-O-methyltransferase, a critical enzyme of the legume isoflavonoid phytoalexin pathway.2003

    • Author(s)
      Akashi, T., Sawada, Y., Shimada, N., Sakurai, N., Aoki, T., Ayabe, S.
    • Journal Title

      Plant and Cell Physiology 44(2)

      Pages: 103-112

    • NAID

      10010272519

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A cluster of genes encodes the two types of chalcone isomerase involved in the biosynthesis of general flavonoids and legume-specific 5-deoxy(iso)flavonoids in Lotus japonicus.2003

    • Author(s)
      Shimada, N., Aoki, T., Sato, S., Nakamura, Y., Tabata, S., Ayabe, S.
    • Journal Title

      Plant Physiology 131(3)

      Pages: 941-951

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Efficient Agrobacterium-mediated transformation of Lotus japonicus with reliable antibioticselection2002

    • Author(s)
      Aoki et al.
    • Journal Title

      Plant Cell Reports 21

      Pages: 238-243

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Key amino acid residues required for aryl migration catalyzed by the cytochrome P450 2-hydroxyisoflavanone synthase2002

    • Author(s)
      Sawada et al.
    • Journal Title

      Plant Journal 31

      Pages: 555-564

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Efficient Agrobacterium-mediated transformation of Lotus japonicus with reliable antibioticselection.2002

    • Author(s)
      Aoki, T., Kamizawa, A., Ayabe, S.
    • Journal Title

      Plant Cell Reports 21

      Pages: 238-243

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Key amino acid residues required for aryl migration catalyzed by the cytochrome P450 2-hydroxyisoflavanone synthase.2002

    • Author(s)
      Sawada, Y., Kinoshita, K., Akashi, T., Aoki, T., Ayabe, S.
    • Journal Title

      Plant Journal 31(5)

      Pages: 555-564

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Efficient Agrobacterium-mediated transformation of Lotus japonicus with reliable antibiotic selection2002

    • Author(s)
      Aoki, T., Kamizawa, A., Ayabe, S.
    • Journal Title

      Plant Cell Reports 21(3)

      Pages: 238-243

    • Related Report
      2004 Annual Research Report
  • [Book] Methods in Enzymology, Cytochrome P450, Part C (Eds, Johnson, E.F., Waterman, M.R.)2002

    • Author(s)
      Ayabe et al.
    • Publisher
      Academic Press
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] Aoki, T., Kamizawa, A., Ayabe, S.: "Efficient Agrobacterium-mediated transformation of Lotus japonicus with reliable antibiotic selection"Plant Cell Report. 21. 238-243 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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