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2003 Fiscal Year Final Research Report Summary

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
Keywordssugarbeet / Owen cytoplasmic male sterility / fertility restorer gene / RfX / positional cloning / BAC library / AFLP marker / BAC contig
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.

  • Research Products

    (30 results)

All Other

All Publications (30 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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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

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Published: 2005-04-19  

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