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ABSTRACTS OF RESEARCH PROJECT,GRANT-IN-AID FOR SCIENTIFIC RESEARCH (1995)

Research Project

Project/Area Number 06660426
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Applied molecular and cellular biology
Research InstitutionNara Institute of Science and Technology

Principal Investigator

HASHIMOTO Takashi  School of Biological Sciences Associate Professor, バイオサイエンス研究科, 助教授 (80180826)

Co-Investigator(Kenkyū-buntansha) YAMADA Yasuyuki  School of Biological Sciences Professor, バイオサイエンス研究科, 教授 (50026415)
Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1994: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywordstobacco / nicotine / mutant / putrescine / molecular biology / 突然変異
Research Abstract

Two semi-dominant loci, Nic1 and Nic2, regulate nicotine contents in tobacco. We previously isolated two classes of cDNA clones that were under-represented in low-nicotine nic1 nic2 mutant tobacco roots. One of the two cDNA classes was found to encode putrescine N-methyltransferase (PMT) , the first enzyme in nicotine biosynthetic pathway. In this study, we isolated the PMT gene as the first step to unravel the regulatory mechanism of nicotine biosynthesis by Nic genes.
From a Nicotiana sylvestris genomic library, we isolated five genomic clones that hybridized to the PMT cDNA probe, which were divided into three groups according to restriction maps. Genomic Southern hybridization analysis also indicated that the genome of N.sylvestris contains three DNA sequences that are homologous to PMT.
3'-Nontranslated region of the PMT cDNA hybridized only one group in the three PMT-related groups. Northen hybridization with the same PMT-specific probe detected a signal only in the root of N.sylvestris. The group was designated as the PMT gene and further analyzed. The nine exons have very high similarity to the PMT cDNA,including the 3'-nontranslated region described above. A TATA-related region was found 113 bp upstream of the first ATG.We also constructed plant vinary vectors in which the 5'-upstream regions were fused translationally to a reporter gene.

Report

(3 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (3 results)

All Other

All Publications (3 results)

  • [Publications] T.Hashimoto: "Regulation of tobacco alkaloid biosyuthesis" Phytochemicals and Health(D.L.Gustine.H.E.Flores,els Am.Soc.Plant Physiol.出版). 130-144 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T.Hashimoto, and Y.Yamada: "Regulation of tobacco alkaloid biosynthesis, In : Phytochemicals and Health, DL Gustine, HE Flores, eds." American Society of Plant Physiologists. 130-144 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Naruhiro Hibi: "Gene expression in tobacco low-nicotine mutants" The Plant Cell. 6. 723-735 (1994)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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