| Project/Area Number |
10460006
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Breeding science
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| Research Institution | KOBE UNIVERSITY |
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Principal Investigator |
NAKAMURA C KOBE UNIV., AGRICULTURE, PROFESSOR, 農学部, 教授 (10144601)
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| Co-Investigator(Kenkyū-buntansha) |
TAKUMI S KOBE UNIV., AGRICULTURE, ASSLSTANT PROF., 農学部, 助手 (50249166)
MORI N KOBE UNIV., AGRICULTURE, ASCOIATE PROF., 農学部, 助教授 (60230075)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥11,800,000 (Direct Cost: ¥11,800,000)
Fiscal Year 2000: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1999: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥6,300,000 (Direct Cost: ¥6,300,000)
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| Keywords | cold acclimation / freezing tolerance / cold-responsive genes / Cor-ltr / Lea / Dhn / Rab / chloroplast / mitochondria / common wheat / 低温誘導性遺伝子 / 低温馴化 / ABA / GA_3 / コムギ / 低温誘導性遺伝子(Cor,Rab) / シアン耐性鎖酸化酵素遺伝子 |
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| Research Abstract |
1) Six clones of cold-responsive genes were isolated from a winter-hardy common wheat cultivar 'Mironovska 808', and their structure and expression were studied. a) Wcor14 and Wcs19 : genes gelonging to a Cor (cold-responsive) family. Wcor14 is homologous to cold- and light-responsive barley Bcor14b. Wcs19 has a N-terminal region homologous to that of Wcor14 and Bcor14b. WCOR14, WCS19 and BCOR14b have a 51-amino-acid N-terminal choloroplast leader peptide. b) Wdhn13 : a gene belonging to a hedydrin family. A genomic clone of Wcas19 including a putative promoter was also obtained. c) Wrab 17 : a gene homologous to GA_3-responsive ES2A from a GA_3-insensitive barley mutant. d) Wrab19 : a gene belonging to a group-3 lea gene family. e) Wltr10 : a member of cereal-specific Ltr family. f) Waoxl and Waox2 : genes encoding cyanide-insensitive alternative oxidase.
2) Copy number of each gene was determined by genomic Southern blot analysis. Chromosome assignment was made using nulli-tetrasomic
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lines and D genome chromosome substiotution lines. Waoxl and Waox2 were shown to be nonhomoeologous genes.
3) Bioassy for freezing tolerance was conducted using 'Mironovska 808' and a spring wheat cv.'Chinese Spring'. Enhancement of freezing tolerance by cold acclimation and significant cultivar differences in the level of freezing tolerance were shown.
4) Gene expression was studied by Northern blot analysis. Expression of Wcor14, Wcs19 and Wltr10 was low temperature-specific. Expression of Wrab17 and Wrab19 was induced by low temperature and also modulated by GA_3, and ABA.Expression of Waox1 and Waox2 was enhanced by low temperature and KCN.A positive correlation was observed between the level of expression of these genes and the level of freezing tolerance in the two cultivars.
5) Transportation of WCOR14 and WCS19 to chloroplasts and that of WAOX1 and WAOX2 to mitochondria was proven by the transient expression assay using GFP-fusion constracts.
6) Genomic clones of Waox1 and Waox2 were isolated and their exson-intron structures were determined.
7) WCOR14 and WDHN13 were synthesized in E.coli and their antibodies were raized using mouse and rabbit. Less
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