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Positional cloning of fertility restorer genes in sugarbeet and its appliation to molecular breeding

Research Project

Project/Area Number 12556001
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Breeding science
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

MIKAMI Tetsuo  Hokkaido Univ., Grad.School of Agr., Prof., 大学院・農学研究科, 教授 (50133715)

Co-Investigator(Kenkyū-buntansha) TANAKA Masakatsu  National Agri.Res.Center for Hokkaido Region, Head, 北海道農業研究センター, 室長
KISHIMA Yuji  Hokkaido Univ., Grad.School of Agr., Asso.Prof., 大学院・農学研究科, 助教授 (60192556)
KUBO Tomohiko  Hokkaido Univ., Grad.School of Agr., Lec., 大学院・農学研究科, 講師 (40261333)
IKEGUCHI Syoujiro  Hokuren Agr.Res.Institute., Cheaf Res.Sci., 植物工学科, 主任研究員
Project Period (FY) 2000 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥13,400,000 (Direct Cost: ¥13,400,000)
Fiscal Year 2003: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2002: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2001: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2000: ¥6,200,000 (Direct Cost: ¥6,200,000)
Keywordssugarbeet / Owen cytoplasmic male sterility / fertility restorer gene / RfX / positional cloning / BAC library / AFLP marker / BAC contig / 稔性回復核遺伝子 / BACクローン / PPRモチーフ / ミトコンドリア局在性 / コンティグ / RfX遺伝子 / 共優性マーカー / TAIL PCR / X遺伝子 / 細胞質雄性不稔性 / AFLP / RFLP
Research Abstract

Sugarbeet plants containing the Owenmitochondria encoding cytoplasmic male sterility are fertile if they carry two dominant nuclear restorer genes (RfX and RfZ). In order to understand the molecular mechanism of fertility restoration mediated by RfX and RfZ, we decided to clone RfX using a positional cloning strategy. Bulked segregant analysis was utilized to identify RAPD and AFLP markers linked to the RfX locus. Out of 812 RADP markers examined, the closest marker was observed to be located 5.7cM away from RfX. A total of 2721 AFLP primer combinations were further tested for polymorphisms, which allows the screening of approximately 27000 loci for linkage to RfX. We obtained eight markers tightly linked to the RfX locus which was restricted within 3.4cM by these markers. It should also be mentioned that three AFLP markers co-segregated with the RfX locus. A sugarbeet BAC library (3.4 genome equivalents), with an average insert size of 98kb, was constructed and screened with the linked markers. The target region around RfX comprises approximately 300kb and is covered with a minimum of eight BAC clones. By nucleotide suquencing and RT-PCR analysis, we identified some open reading frames within the 300kb-region, which were observed to be actively transcribed in anther tissues and to have mitochondrial targeting sequences. It thus seems reasonable to consider that one of the reading frames identified here is a good candidate for RfX gene.

Report

(5 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (53 results)

All Other

All Publications (53 results)

  • [Publications] S.Nishizawa, T.Kubo, T.Mikami: "Variable number of tandem repeat loci in the mitochondrial genomes of beets."Current Genetics. 37. 34-38 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Kubo, M-P.Yamamoto, T.Mikami: "The nad4L-orf25 gene cluster is conserved and expressed in sugar beet mitochondria."Theoretical and Applied Genetics. 100. 214-220 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Kubo, S.Nishizawa, A.Sugawara, N.Itchoda, A.Estiati, T.Mikami: "The complete nucleotide sequence of the mitochondrial genome of sugar beet (Beta vulgaris L.) reveals a novel gene for tRNACys(GCA)."Nucleic Acids Research. 28. 2571-2576 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] S.Kato, H.Yamaguchi, Y.Shimamoto, T.Mikami: "The chloroplast genomes of azuki bean and its close relatives : a deletion mutation found in weed azuki bean."Hereditas. 132. 43-48 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] S.Kato, T.Koike, T.T.Lei, C.F.Hsieh, K.Ueda, T.Mikami: "Analysis of mitochondrial DNA of an endangered beech species, Fagus hayatae Palibin ex Hayata."New Forests.. 19. 109-114 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Mikami, T.Kubo, M-P.Yamamoto, E.Hagihara: "Molecular analysis of cytoplasmic male sterility in beets."Acta Horticulturae. 560. 117-179 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Kitamura, S.Hashida, T.Mikami, Y.Kishima: "Position effect of the excision frequency of the Antirrhinum transposon Tam3 : implications for the degree of position dependent methylation in the ends of the element."Plant Molecular Biology. 47. 475-490 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Mikami, S.Nishizawa, A.Estiati, T.Kubo: "Gene content and organization in the mitochondrial genome of sugar beet (Beta vulgaris L.)"Proceedings of XVI EUCARPIA Genetic Resources Section Symposium. 237-239 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] N.Itchoda, S.Nishizawa, H.Nagano, T.Kubo, T.Mikami: "The sugar beet mitochondrial nad4 gene : an intron loss and its phylogenetic implication in the Caryophyllales."Theoretical and Applied Genetics.. 104. 209-213 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] N.Itchoda, T.Kubo, A.Estiati, M-P.Yamamoto, H.Handa, T.Mikami: "The rps4 gene in sugar beet mitochondria : insertion/deletion mutations occur within the gene but do not disrupt the reading frame."Journal of Plant Physiology.. 159. 211-215 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] N.Fujii, N.Tomaru, K.Okuyama, T.Koike, T.Mikami, K.Ueda: "Chloroplast DNA phylogeography of Fagus crenata (Fagaceae) in Japan."Plant Systematics and Evolution. 232. 21-23 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] R.Yui, S.Iketani, T.Mikami, T.Kubo: "Antisense inhibition of mitochondrial pyruvate dehydrogenase E1 α subunit in anther tapetum causes male sterility."The Plant Journal. 34. 56-66 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] S.Hashida, K.Kitamura, T.Mikami, Y.Kishima: "Temperature shift coordinately changes the activity and the methylation state of transposon Tam3 in Antirrhinum."Plant Physiology. 132. 1207-1216 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 三上 哲夫: "花-性と生殖の分子生物学"学会出版センター. 258 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 三上 哲夫: "栽培植物の自然史"北海道大学図書刊行会. 241 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 三上 哲夫: "生物学データ大百科事典(上)"朝倉書店. 1459 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 三上 哲夫: "植物遺伝学入門"朝倉書店(印刷中). (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] S.Nishizawa, T.Kubo, T.Mikami: "Variable number of tandem repeat loci in the mitochondrial genomes of beets."Current Genetics. 37. 34-38 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Kubo, M-P.Yamamoto, T.Mikami: "The nad4L-orf25 gene cluster is conserved and expressed in sugar beet mitochondria."Theoretical and Applied Genetics. 100. 214-220 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Kubo, S.Nishizawa, A.Sugawara, N.Itchoda, A.Estiati, T.Mikami: "The complete nucleotide sequence of the mitochondrial genome of sugar beet (Beta vulgaris L.) reveals a novel gene for tRNACys(GCA)."Nucleic Acids Research. 28. 2571-2576 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] S.Kato, H.Yamaguchi, Y.Shimamoto, T.Mikami: "The chloroplast genomes of azuki bean and its close relatives : a deletion mutation found in weed azuki bean."Hereditas. 132. 43-48 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] S.Kato, T.Koike, T.T.Lei, C.F.Hsieh, K.Ueda, T.Mikami: "Analysis of mitochondrial DNA of an endangered beech species, Fagus hayatae Palibin ex Hayata."New Forests. 19. 109-114 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Mikami, T.Kubo, M-P.Yamamoto, E.Hagihara: "Molecular analysis of cytoplasmic male sterility in beets."Acta Horticulturae. 560. 177-179 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Kitamura, S.Hashida, T.Mikami, Y.Kishima: "Position effect of the excision frequency of the Antirrhinum transposon Tam3 : implications for the degree of position dependent methylation in the ends of the element."Plant Molecular Biology. 47. 475-490 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Mikami, S.Nishizawa, A.Estiati, T.Kubo: "Gene content and organization in the mitochondrial genome of sugar beet (Beta vulgaris L.)."Proceedings of XVI EUCARPIA Genetic Resources Section Symposium. 237-239 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] N.Itchoda, S.Nishizawa, H.Nagano, T.Kubo, T.Mikami: "The sugar beet mitochondrial nad4 gene : an intron loss and its phylogenetic implication in the Caryophyllales."Theoretical and Applied Genetics. 104. 209-213 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] N.Itchoda, T.Kubo, A.Estiati, M-P.Yamamoto, H.Handa, T.Mikami: "The rps4 gene in sugar beet mitochondria : insertion/deletion mutations occur within the gene but do not disrupt the reading frame."Journal of Plant Physiology. 159. 211-215 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] N.Fujii, N.Tomaru, K.Okuyama, T.Koike, T.Mikami, K.Ueda: "Chloroplast DNA phylogeography of Fagus crenate (Fagaceae) in Japan."Plant Systematics and Evolution. 232. 21-33 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] R.Yui, S.Iketani, T.Mikami, T.Kubo: "Antisense inhibition of mitochondrial pyruvate dehydrogenase E1 α subunit in anther tapetum causes male sterility."Plant Journal. 34. 57-66 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] S.Hashida, K.Kitamura, T.Mikami, Y.Kishima: "Temperature shift coordinately changes the activity and the methylation state of transposon Tam3 in Antirrhinum majus."Plant Physiology. 132. 1207-1216 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] R.Yui: "Antisense inhibition of mitochondrial pyruvate dehydrogenase E1 α subunit in anther tapetum causes male sterility"The Plant Journal. 34. 56-66 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] S-N.Hashida: "Temperature shift coordinately changes the activity and the methylation state of transposon Tam3 in Antirrhinum"Plant Physiology. 132. 1207-1216 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 三上 哲夫: "植物遺伝学入門(三上哲夫編著)"朝倉書店(印刷中). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] N.Itchoda: "The sugar beet mitochondrial nad4 gene : an intron loss and its phylogenetic implication in the Caryophyllales"Theoretical and Applied Genetics. 104. 209-213 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] N.Itchoda: "The rps4 gene in sugar beet mitochondria : insertion/deletion mutations occur within the gene but do not disrupt the reading frame"Journal of Plant Physiology. 159. 211-215 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] N.Fujii: "Chloroplast DNA phylogeography of Fagus crenata(Fagaceae)in Japan"Plant Systematics and Evolution. 232. 21-33 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] R.Yui: "Antisense inhibition of mitochondrial pyruvate dehydrogenase E1 α subunit in anther tapetum causes male sterility"The Plant Journal. (in press).

    • Related Report
      2002 Annual Research Report
  • [Publications] S-N.Hashida: "Temperature shift coordinately changes the activity and the methylation state of transposon Tam3 in Antirrhinum"Plant Physiology. (in press).

    • Related Report
      2002 Annual Research Report
  • [Publications] T. Mikami: "Molecular analysis of cytoplasmic male sterility in beets"Acta Horticulturae. 560. 177-179 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] K. Kitamura: "Position effect of the excision frequency of the Antirrhinum transposon Tam3 : implications for the degree of position-dependent methylation in the ends of the element"Plant Molecular Biology. 47. 475-490 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] N. ltchoda: "The sugar beet mitochondrial nad4 gene : an intron loss and its phylogenetic implication in the Caryophyllales"Theoretical and Applied Genetics. (in press).

    • Related Report
      2001 Annual Research Report
  • [Publications] N. ltchoda: "The rps4 gene in sugar beet mitochondria : insertion/deletion mutations occur within the gene but do not disrupt the reading frame"Journal of Plant Physiology. (in press).

    • Related Report
      2001 Annual Research Report
  • [Publications] N. Fujii: "Chloroplast DNA phylogeography of Fagus crenata(Fagaceae)in Japan"Plant Systematics and Evolution. (in press).

    • Related Report
      2001 Annual Research Report
  • [Publications] T. Mikami: "Gene content and organization in the mitochondrial genome of sugar beet(Beta vulgaris L.)"Proceedings of XVI EUCARPIA Genetic Resources Section Symposium. (in press).

    • Related Report
      2001 Annual Research Report
  • [Publications] 三上 哲夫: "花 - 性と生殖の分子生物学(日向 康吉 編)"学会出版センター. 258 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] S.Nishizawa: "Variable number of tandem repeat loci in the mitochondrial genomes of beets."Current Genetics. 37. 34-38 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] T.Kubo: "The nad4L-orf25 gene cluster is conserved and expressed in sugar beet mitochondria."Theoretical and Applied Genetics. 100. 214-220 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] T.Kubo: "The complete nucleotide sequence of the mitochondrial genome of sugar beet (Beta vulgaris L) reveals a novel gene for tRNACys (GCA)."Nucleic Acids Research. 28. 2571-2576 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] S.Kato: "The chloroplast genomes of azuki bean and its close relatives : a deletion mutation found in weed azuki bean."Hereditas. 132. 43-48 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] S.Kato: "Analysis of mitochondrial DNA of an endangered beech species, Fagus hayatae Palibin ex Hayata."New Forests. 19. 109-114 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] T.Mikami: "Molecular analysis of cytoplasmic male sterility in beets."Acta Horticulturae. ( in press ) . (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 三上哲夫: "花-性と生殖の分子生物学(日向康吉 編)"学会出版センター(印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] 三上哲夫: "栽培植物の自然史(山口裕文 編)"北海道大学図書刊行会(印刷中). (2001)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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