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Studies on inhibition of apple ripening using antisense technique

Research Project

Project/Area Number 07660003
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Breeding science
Research InstitutionHirosaki University

Principal Investigator

HARADA Takeo  Hirosaki University, Department of Agriculture, Assosiate Professor, 農学部, 助教授 (10228645)

Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1996: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1995: ¥1,700,000 (Direct Cost: ¥1,700,000)
KeywordsApple / ACC shynthase gene / Ethylene / Fruit storage / Antisense / アニチセンス / 再分化 / 組換え植物 / 遺伝子導入
Research Abstract

Apple is also a typical climacteric fruit. Therefore, it is appeared to be possible to extend the storage life of apple fruit by inhibiting ethylene biosynthesis using antisense technique. Furthermore, we are interested in molecular mechanism of the differences of the storagelongevity among apple cultivars. Lay-Yee and Knighton (1995) reported afull-length cDNA (MdACS-1) encoding ACS from ripening apple. Recently, two more cDNAs of ACS (MdACS-2, MdACS-3) from ripening apple were isolated (Rosenfield et al., 1997). Thus, it is considered that apple has also a multigene family of ACS to control ripening process like tomato. However, their genomic sequences are unknown yet. As a first step for the elucidation of the mechanism controlling the storage longevity, the sequence of gene encoding the MdACS-1 gene was isolated by screening a genomic library from Malus domestica L.Borkh cv Golden Delicious. A probe DNA was made from PCR using primers designed from MdACS-1 cDNA sequence and 'Golden Delicious' genomic DNA as a template. Three of 13 positive phage clones were appeared to contain the full length of the gene coding regions. One of them (1-6) contained partialy a restriction map which was identical to that of the cDNA.The sequencing of the subcloned fragment (5.6kb) revealed the presence of MdACS-1 gene which consists of exons and three introns. The number and size of four exons and location of introns are similar to other ACS genes isolated from tomato, rice, and Arabidopsis (Lincoln et al., 1993 ; Zarembinski and Theologis, 1993 ; Abel et al., 1995). The sequence also included 2,111b and 1,011b of the 5'-and 3'-flanking regions, respectively. We constructed a chimeric gene containing the full-length promotor region fuged to the coding sequence for the GUS gene. By using this characterization of the promotor is currently in progress.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (7 results)

All Other

All Publications (7 results)

  • [Publications] Chiba T. et al.: "Transcription of tRNA genes from a large-scale plastid DNA deletion celary reveals the action of nuclear-encoded RNA polymerase in the plastid." J. Plant Physiol.148. 652-656 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Harada, T. et al.: "Genomic nucleotide sequence of a ripening-related 1-aminocyclopropane-1-carboxylate synthase gene (MdACS-1) in apple (Accession No. U89156)." Plant Physiol.(in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Chiba T.et al.: "Transcription of tRNA genes from a large-scale plastid DNA deletion celary reveals the action of nuclear-encoded RNA polymerase in the plastid." J.Plant Physiol.148. 652-656 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Harada, T.et al.: "Genomic nucleotide sequence of a ripening-related 1-aminocyclopropane-1-carboxylate synthase gene (MdACS-1) in apple (Accession No.U89156)." Plant Physiol.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Chiba T et al.: "Transcription of tRNA genes from a large-scale plastrd DNA deletion clearly reveais the action of unclear-encoded RNA polymerase in the plastid." J. Plant Physiol. 148. 652-656 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Harada T: "A timeric form of the bean phaseolins produced by a baculovirus expression system." Plant Physiol.108. 146 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Chiba T et al.: "Transcription of tRNA genes from a large-scale plastid DNA deletion celarly reveals the action of nuclear-encoded RNA polymerase in the plastid." J.Plant Physiol.(in press). (1996)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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