2002 年度実績報告書

植物の可塑的な生長・分化を支える分子機構

研究課題

研究課題/領域番号	13CE2005
研究機関	名古屋大学
研究代表者	松岡信名古屋大学, 生物分子応答研究センター, 教授 (00270992)
研究分担者	前島正義名古屋大学, 大学院・生命農学研究科, 教授 (80181577) 坂神洋次名古屋大学, 大学院・生命農学研究科, 教授 (80107408) 上口智治名古屋大学, 生物分子応答研究センター, 助教授 (20232738) 近藤孝男名古屋大学, 大学院・理学研究科, 教授 (10124223) 魚住信之名古屋大学, 生物分子応答研究センター, 助教授 (40223515)
キーワード	植物分子育種 / 植物生長物質 / 物質輸送機構 / 外環境応答 / 栄養環境応答
研究概要	1.植物生長物質の情報伝達機構の解明とその利用 (1)ジベレリン(GA)の信号伝達において負の分子スイッチとして働いているSLR1タンパク質は、GAシグナル依存的に分解され、その結果、抑制されていた下流の信号伝達が脱抑制されることでGA反応が引き起こされる。本年度は、このGA依存的なSLR1タンパク質の分解機構について解析を行った結果、分解にはSCF複合体を介したタンパク質分解機構が関与していることが判明した。すなわち、SLR1分解が異常になった変異体gibberellin insensitive dwarf2 (gid2)の原因遺伝子を単離してその構造を調べたところ、GID2はF-boxドメインを持つタンパク質をコードしていることが解った。さらに、gid2変異体の中ではSLR1はリン酸化された状態で蓄積されていることから、SRL1はGAシグナル依存的にリン酸化を受け、分解反応が開始されると考えられた。 (2)ペプチド性植物細胞増殖因子ファイトスルフォカイン(PSK)に特異的に結合する膜タンパク質をニンジン培養細胞から精製し、その部分アミノ酸配列を明らかにした。この配列を用いて、PSK結合タンパク質をコードする遺伝子のクローニングに成功した.この遺伝子は、ロイシンリッチリピート、膜貫通領域、セリン・スレオニンキナーゼ領域を含む1021アミノ酸をコードしていた。このタンパク質を過剰発現させた細胞は、PSKに対する結合能が上がり、生長速度も増加した。以上のことから、本結合タンパク質はPSK受容体であると結論した。興味深いことに受容体を過剰発現した細胞は再分化能を失った。これは、PSKの植物における生理作用を反映していると考えられた。 (3)シロイヌナズナサイトカイニン受容体候補遺伝子AHK2,3について、それぞれのノックアウト変異体を分離し、遺伝解析の準備を整えた。また大腸菌内でサイトカイニン受容体を発現させ、サイトカイニンに応答した情報伝達系の再構築に成功した。シロイヌナズナTCP10変異体の表現型解析の結果、TCP10が細胞分裂の正の制御因子であることを強く示唆するデータを得た。またイネTCP遣伝子の一つであるTB1遺伝子の逆遺伝学的解析ならびに当遺伝子の劣性突然変異体の解析から、これがイネ分げつを負に制御する因子であることを明らかにした。 2.植物における物質輸送メカニズムの解明とその利用 (1)イネ生体膜(細胞膜と液胞膜)のCa^<2+>/H^+交換輸送体について焦点をあて、5種のcDNAクローンを得て一次構造の特徴、酵母での異種発現システムによるイオン輸送機能の検定を進め、Ca2+のみでなく、Mn^<2+>、Cd^<2+>の輸送にも関与すること、特定分子種は栽培栄養条件としてのCa^<2+>、Mn^<2+>濃度により遺伝子発現が変動すること等を見出した。 (2)Na^+/K^+トランスポーター(HKT1)のイオン選択性に関与するアミノ酸を同定した。AtHKT1の破壊株について解析を行い、本輸送体は植物地上部から根にNa^+を転流する機能を担っており、本遺伝子破壊により地上部が塩感受性となることを見い出した. 3.外環境応答メカニズムの解明とその利用 (1)光周性の詳しい解析に用いる材料として生理学的実験植物として確立している短日型と長日型の同属2種のウキクサを用い、概日時計、光周性、花芽形成に関わる遺伝子(APRR1/TOC1,GI, CO, FT, AG)のホモログをクローニングした。これらのmRNAレベルの発現を解析した結果、概日リズムや日長条件依存的なパターンを示すものなどを明らかにするとともに、短日型と長日型の発現様式が異なる遺伝子を明らかにした。 (2)シロイヌナズナにおける新規な時計関連遺伝子を5種類同定し、花の開花時期制御や光形態形成との関連を明らかにした。 4.栄養環境応答メカニズムの解明とその利用ヒメツリガネゴケの硝酸イオン能動輸送体、硝酸還元酵素、亜硝酸還元酵素のcDNAを単離して構造を決定した。これらの遺伝子発現は硝酸イオンにより活性化されアンモニアにより抑制された。また、亜硝酸還元酵素のノックアウト株を作成した。シロイヌナズナにおいて個葉の硝酸イオン蓄積量を測定する方法を開発し、またPIIタンパク質のアンチセンス株を作成した。

研究成果
(44件)

すべてその他

すべて文献書誌 (44件)

[文献書誌] Maser, P: "Glycine residues in potassium channel-like selectivity filters determine potassium selectivity in four-loop-per-subunit HKT transporters from plants"Proc.Natl.Acad.Sci. USA. 99. 6428-6433 (2002)
[文献書誌] Imai, K.: "Membrane-bound heparin binding proteins from HL-60 cells purified in a 2-step affinity chromatography differentially eluted with divalent cations"J.Chromatography B. 779. 331-339 (2002)
[文献書誌] Maser, P.: "Altered shoot/root Na^+ distribution and bifurcating salt sensitivity in Arabidopsis by genetic disruption of the Na^+ transporter AtHKT1"FEBS Letters. 2. 157-161 (2002)
[文献書誌] Iwasaki, H.: "KaiA-Stimulated KaiC Phosphorylation in Circadian Timing Loops in Cyanobacteria"Proc.Natl.Acad.Sci.. 99. 15788-15793 (2002)
[文献書誌] Kageyama H: "Circadian Formation of Clock Protein Complexes by KaiA, KaiB, KaiC and SasA in Cyanobacteria"J.Biol.Chem.. 278. 2388-2395 (2003)
[文献書誌] Matsubayashi, Y.: "An LRR receptor kinase involved in perception of a peptide plant hormone, phytosulfokine"Science. 296. 1470-1472 (2002)
[文献書誌] Takayama, S.: "Peptide signaling in plants"Curr.Opin.Plant Biol.. 5. 382-387 (2002)
[文献書誌] Tanigawa, R.: "Transcriptional activation of NtcA-dependent promoters of Synechococcus sp. PCC 7942 by 2-oxoglutarate in vitro"Proc.Natl.Acad.Sci. USA. 99. 4251-4255 (2002)
[文献書誌] Makino, S.: "The APRR1/TOC1 quintet implicated in circadian rhythms of Arabidopsis thaliana : I. Characterization with APRR1-overexpressing plants"Plant Cell Physiol.. 43. 58-69 (2002)
[文献書誌] Matsushika, A.: "The APRR1/TOC1 quintet implicated in circadian rhythms of Arabidopsis thaliana : II. Characterization with CCA1-overexpressing plants"Plant Cell Physiol.. 43. 118-122 (2002)
[文献書誌] Murakami-Kojima, M.: "The APRR3 component of the clock-associated APRR1/TOC1 quintet is phosphorylated by a novel protein kinase belonging to the WNK family, the gene for which is also transcribed rhythmically in Arabidopsis thaliana"Plant Cell Physiol.. 43. 675-683 (2002)
[文献書誌] Hosoda, K.: "Molecular Structure of the GARP Family of Plant Myb-related DNA-binding Motifs of the Arabidopsis Response Regulators"Plant Cell. 14. 2015-2029 (2002)
[文献書誌] Ikeda, M.: "Expression of V-ATPase proteolipid subunit of Acetabularia acetabulum in a VMA3-deficient strain of Saccharomyces cerevisiae and its complementation study"Eur.J.Biochem.. 268. 6097-6104 (2001)
[文献書誌] Ikeda, M.: "Functional expression of Acetabularia acetabulum vacuolar H^+-pyrophosphatase in a yeast VMA3-deficient strain"J.Exp.Botany. 53. 2273-2275 (2002)
[文献書誌] Suga, S.: "Aquaporin isoforms responsive to salt and water stresses and phytohormones in radish seedlings"Plant Cell Physiol.. 43. 1229-1237 (2002)
[文献書誌] Fujiwara, S.: "Molecular properties of a matrix attachment region-binding protein in tobacco cells"Plant Cell Physiol.. 43. 1558-1567 (2002)
[文献書誌] Itoh, H.: "The gibberellin signaling pathway is regulated by the appearance and disappearance of SLENDER RICE1 in nuclei"Plant Cell. 14. 57-70 (2002)
[文献書誌] Sasaki, A.: "A mutant gibberellin-synthesis gene in rice"Nature. 416. 701-702 (2002)
[文献書誌] Ashikari, M.: "Loss-of-function of a rice gibberellin biosynthetic gene, GA20 oxidase(GA20ox-2), led to the rice 'Green Revolution'"Breeding Science. 52. 143-150 (2002)
[文献書誌] Hong, Z.: "Loss-of-function of a rice brassinosteroid biosynthetic enzyme, C-6 oxidase, prevents the organized arrangement and polar elongation of cells in leaves and stem"Plant J.. 32. 495-508 (2002)
[文献書誌] Nishimura, H.: "Mutations in KaiA, a clock protein, extend the period of circadian rhythm of cyanobacterium Synechococcus elogatus PCC7942"Microbiology. 148. 2903-2909 (2002)
[文献書誌] Sasaki, K.: "Effects of phytosulfokine-a on growth and tropane alkaloid production in transformed roots of Atropa belladonna"Plant Growth Regulation. 36. 87-90 (2002)
[文献書誌] Suzuki, T.: "An Arabidopsis histidine-containing phosphotransfer (HPt) factor implicated in phosphorelay signal transduction : Overexpression of AHIP2 in plants results in hypersensitiveness to cytokinin"Plant Cell Physiol.. 43. 123-129 (2002)
[文献書誌] Matsushika, A.: "Aberrant Expression of the Light-Inducible and Circadian-Regulated APRR9 Gene Belonging to the Circadian-Associated APRR1/TOC1 Quintet Results in the Phenotype of Early Flowering in Arabidopsis thaliana"Plant Cell Physiol.. 43. 833-843 (2002)
[文献書誌] Kiba, T.: "Characterization of the ARR15 and ARR16 response regulars with special reference to the cytokinin signaling pathway mediated by the AHK4 histidine kinase in roots of Arabidopsis thalian"Plant Cell Physiol.. 43. 1059-1066 (2002)
[文献書誌] Nakamichi, N.: "Compilation and characterization of the novel WNK family of protein kinases in Arabiodpsis thaliana with special reference to circadian rhythms."Biosci.Biotechno.Biochem.. 66. 2429-2436 (2002)
[文献書誌] Nakamichi, N.: "His-to-Asp phosphorelay circuitry regulates sexual development in Schizosaccharomyces pombe"Biosci.Biotechno.Biochem.. 66. 2663-2672 (2002)
[文献書誌] Oshima, T.: "Transcriptome analysis of all two-component regulatgory system mutants of Escherichia coil K-12"Mol.Microbiol.. 46. 281-291 (2002)
[文献書誌] Mimura T.: "Rapid increase of vacuolar volume in response to salt stress"Planta. 216. 397-402 (2003)
[文献書誌] Yuasa, K.: "Equilibrium dialysis measurements of the Ca^<2+>-binding properties of recombinant radish vacuolar Ca^<2+>-binding protein expressed in E"coil.Biosci.Biotechnol.Biochem.. 66. 2382-2387 (2002)
[文献書誌] Pittman, J.K.: "Distinct N-terminal regulatory domains of Ca^<2+>H^+ antiporters"Plant Physiol.. 130. 1054-1062 (2002)
[文献書誌] Hagiwara, Y.: "Subcellular localization of host and viral proteins associated with tobamovirus RNA replication"EMIBO J.. 22. 344-353 (2003)
[文献書誌] Sato, Y.: "A loss-of-function mutation in the rice KNOX type homeobox gene, OSH3"Plant Cell Physiol.. 43. 44-51 (2002)
[文献書誌] Ashikari, M.: "Application of rice genomics to plant biology and breeding"Bot.Bull.Acad.Sin.. 43. 1-11 (2002)
[文献書誌] Sato, Y.: "Isolation and characterization of a cDNA encoding an orthologue of ROUGH SHEATH2(OsRS2) from rice"Plant Biotech. 19. 11-18 (2002)
[文献書誌] Ito, M.: "Position dependent expression of GL2-type homeobox gene, Roc1: significance for protoderm differentiation and radial pattern formation in early rice embryogenesis"Plant J.. 29. 497-507 (2002)
[文献書誌] Kaneko, M.: "The α-Amylase induction in endosperm during rice seed germination is caused by gibberellin synthesized in epithelium"Plant Physiol.. 128. 1264-1270 (2002)
[文献書誌] Nishimura, A.: "OsPNH1 regulates leaf development and maintenance of the shoot apical meristem in rice"Plant J.. 31. 189-201 (2002)
[文献書誌] Itoh, H.: "Modification of Rice Plant Height by Suppressing the Height-Controlling Gene, D18, in Rice"Breeding Science. 52. 215-218 (2002)
[文献書誌] Iwasaki, H.: ""Histidine kinases in the cyanobacterial circadian system" Histidine Kinases (Eds. M.Inouye and R.Dutta)"Academic. (in press).
[文献書誌] Mizuno, T.: "Role of the Histidine-containing phosphotransfer (HPt)domain in the multistep phosphorelay through the anaerobic hybrid sensor, ArcBs""Histidine Kinases", Inouye, M., ed Academic Press, USA. (in press).
[文献書誌] Mizuno, T.: "Genome-wide analysis of Escherichia coil histidine kinases""Histidine Kinases", Inouye, M., ed Academic Press, USA. (in press).
[文献書誌] Nakanishi, Y.: "Patch clamp analysis of H^+ pump expressed in giant yeast vacuoles"Submitted for publication. (in press).
[文献書誌] Suga, S.: "An abundance of aquaporins in specific cells and osmotic water permeability of individual protoplasts of radish"Plant Cell Physiol.. (in press).

2002 年度 実績報告書

植物の可塑的な生長・分化を支える分子機構

研究代表者

松岡 信 名古屋大学, 生物分子応答研究センター, 教授 (00270992)

研究成果

[文献書誌] Maser, P: "Glycine residues in potassium channel-like selectivity filters determine potassium selectivity in four-loop-per-subunit HKT transporters from plants"Proc.Natl.Acad.Sci. USA. 99. 6428-6433 (2002)

[文献書誌] Imai, K.: "Membrane-bound heparin binding proteins from HL-60 cells purified in a 2-step affinity chromatography differentially eluted with divalent cations"J.Chromatography B. 779. 331-339 (2002)

[文献書誌] Maser, P.: "Altered shoot/root Na^+ distribution and bifurcating salt sensitivity in Arabidopsis by genetic disruption of the Na^+ transporter AtHKT1"FEBS Letters. 2. 157-161 (2002)

[文献書誌] Iwasaki, H.: "KaiA-Stimulated KaiC Phosphorylation in Circadian Timing Loops in Cyanobacteria"Proc.Natl.Acad.Sci.. 99. 15788-15793 (2002)

[文献書誌] Kageyama H: "Circadian Formation of Clock Protein Complexes by KaiA, KaiB, KaiC and SasA in Cyanobacteria"J.Biol.Chem.. 278. 2388-2395 (2003)

[文献書誌] Matsubayashi, Y.: "An LRR receptor kinase involved in perception of a peptide plant hormone, phytosulfokine"Science. 296. 1470-1472 (2002)

[文献書誌] Takayama, S.: "Peptide signaling in plants"Curr.Opin.Plant Biol.. 5. 382-387 (2002)

[文献書誌] Tanigawa, R.: "Transcriptional activation of NtcA-dependent promoters of Synechococcus sp. PCC 7942 by 2-oxoglutarate in vitro"Proc.Natl.Acad.Sci. USA. 99. 4251-4255 (2002)

[文献書誌] Makino, S.: "The APRR1/TOC1 quintet implicated in circadian rhythms of Arabidopsis thaliana : I. Characterization with APRR1-overexpressing plants"Plant Cell Physiol.. 43. 58-69 (2002)

[文献書誌] Matsushika, A.: "The APRR1/TOC1 quintet implicated in circadian rhythms of Arabidopsis thaliana : II. Characterization with CCA1-overexpressing plants"Plant Cell Physiol.. 43. 118-122 (2002)

[文献書誌] Murakami-Kojima, M.: "The APRR3 component of the clock-associated APRR1/TOC1 quintet is phosphorylated by a novel protein kinase belonging to the WNK family, the gene for which is also transcribed rhythmically in Arabidopsis thaliana"Plant Cell Physiol.. 43. 675-683 (2002)

[文献書誌] Hosoda, K.: "Molecular Structure of the GARP Family of Plant Myb-related DNA-binding Motifs of the Arabidopsis Response Regulators"Plant Cell. 14. 2015-2029 (2002)

[文献書誌] Ikeda, M.: "Expression of V-ATPase proteolipid subunit of Acetabularia acetabulum in a VMA3-deficient strain of Saccharomyces cerevisiae and its complementation study"Eur.J.Biochem.. 268. 6097-6104 (2001)

[文献書誌] Ikeda, M.: "Functional expression of Acetabularia acetabulum vacuolar H^+-pyrophosphatase in a yeast VMA3-deficient strain"J.Exp.Botany. 53. 2273-2275 (2002)

[文献書誌] Suga, S.: "Aquaporin isoforms responsive to salt and water stresses and phytohormones in radish seedlings"Plant Cell Physiol.. 43. 1229-1237 (2002)

[文献書誌] Fujiwara, S.: "Molecular properties of a matrix attachment region-binding protein in tobacco cells"Plant Cell Physiol.. 43. 1558-1567 (2002)

[文献書誌] Itoh, H.: "The gibberellin signaling pathway is regulated by the appearance and disappearance of SLENDER RICE1 in nuclei"Plant Cell. 14. 57-70 (2002)

[文献書誌] Sasaki, A.: "A mutant gibberellin-synthesis gene in rice"Nature. 416. 701-702 (2002)

[文献書誌] Ashikari, M.: "Loss-of-function of a rice gibberellin biosynthetic gene, GA20 oxidase(GA20ox-2), led to the rice 'Green Revolution'"Breeding Science. 52. 143-150 (2002)

[文献書誌] Hong, Z.: "Loss-of-function of a rice brassinosteroid biosynthetic enzyme, C-6 oxidase, prevents the organized arrangement and polar elongation of cells in leaves and stem"Plant J.. 32. 495-508 (2002)

[文献書誌] Nishimura, H.: "Mutations in KaiA, a clock protein, extend the period of circadian rhythm of cyanobacterium Synechococcus elogatus PCC7942"Microbiology. 148. 2903-2909 (2002)

[文献書誌] Sasaki, K.: "Effects of phytosulfokine-a on growth and tropane alkaloid production in transformed roots of Atropa belladonna"Plant Growth Regulation. 36. 87-90 (2002)

[文献書誌] Suzuki, T.: "An Arabidopsis histidine-containing phosphotransfer (HPt) factor implicated in phosphorelay signal transduction : Overexpression of AHIP2 in plants results in hypersensitiveness to cytokinin"Plant Cell Physiol.. 43. 123-129 (2002)

[文献書誌] Matsushika, A.: "Aberrant Expression of the Light-Inducible and Circadian-Regulated APRR9 Gene Belonging to the Circadian-Associated APRR1/TOC1 Quintet Results in the Phenotype of Early Flowering in Arabidopsis thaliana"Plant Cell Physiol.. 43. 833-843 (2002)

[文献書誌] Kiba, T.: "Characterization of the ARR15 and ARR16 response regulars with special reference to the cytokinin signaling pathway mediated by the AHK4 histidine kinase in roots of Arabidopsis thalian"Plant Cell Physiol.. 43. 1059-1066 (2002)

[文献書誌] Nakamichi, N.: "Compilation and characterization of the novel WNK family of protein kinases in Arabiodpsis thaliana with special reference to circadian rhythms."Biosci.Biotechno.Biochem.. 66. 2429-2436 (2002)

[文献書誌] Nakamichi, N.: "His-to-Asp phosphorelay circuitry regulates sexual development in Schizosaccharomyces pombe"Biosci.Biotechno.Biochem.. 66. 2663-2672 (2002)

[文献書誌] Oshima, T.: "Transcriptome analysis of all two-component regulatgory system mutants of Escherichia coil K-12"Mol.Microbiol.. 46. 281-291 (2002)

[文献書誌] Mimura T.: "Rapid increase of vacuolar volume in response to salt stress"Planta. 216. 397-402 (2003)

[文献書誌] Yuasa, K.: "Equilibrium dialysis measurements of the Ca^<2+>-binding properties of recombinant radish vacuolar Ca^<2+>-binding protein expressed in E"coil.Biosci.Biotechnol.Biochem.. 66. 2382-2387 (2002)

[文献書誌] Pittman, J.K.: "Distinct N-terminal regulatory domains of Ca^<2+>H^+ antiporters"Plant Physiol.. 130. 1054-1062 (2002)

[文献書誌] Hagiwara, Y.: "Subcellular localization of host and viral proteins associated with tobamovirus RNA replication"EMIBO J.. 22. 344-353 (2003)

[文献書誌] Sato, Y.: "A loss-of-function mutation in the rice KNOX type homeobox gene, OSH3"Plant Cell Physiol.. 43. 44-51 (2002)

[文献書誌] Ashikari, M.: "Application of rice genomics to plant biology and breeding"Bot.Bull.Acad.Sin.. 43. 1-11 (2002)

[文献書誌] Sato, Y.: "Isolation and characterization of a cDNA encoding an orthologue of ROUGH SHEATH2(OsRS2) from rice"Plant Biotech. 19. 11-18 (2002)

[文献書誌] Ito, M.: "Position dependent expression of GL2-type homeobox gene, Roc1: significance for protoderm differentiation and radial pattern formation in early rice embryogenesis"Plant J.. 29. 497-507 (2002)

[文献書誌] Kaneko, M.: "The α-Amylase induction in endosperm during rice seed germination is caused by gibberellin synthesized in epithelium"Plant Physiol.. 128. 1264-1270 (2002)

[文献書誌] Nishimura, A.: "OsPNH1 regulates leaf development and maintenance of the shoot apical meristem in rice"Plant J.. 31. 189-201 (2002)

[文献書誌] Itoh, H.: "Modification of Rice Plant Height by Suppressing the Height-Controlling Gene, D18, in Rice"Breeding Science. 52. 215-218 (2002)

[文献書誌] Iwasaki, H.: ""Histidine kinases in the cyanobacterial circadian system" Histidine Kinases (Eds. M.Inouye and R.Dutta)"Academic. (in press).

[文献書誌] Mizuno, T.: "Role of the Histidine-containing phosphotransfer (HPt)domain in the multistep phosphorelay through the anaerobic hybrid sensor, ArcBs""Histidine Kinases", Inouye, M., ed Academic Press, USA. (in press).

[文献書誌] Mizuno, T.: "Genome-wide analysis of Escherichia coil histidine kinases""Histidine Kinases", Inouye, M., ed Academic Press, USA. (in press).

[文献書誌] Nakanishi, Y.: "Patch clamp analysis of H^+ pump expressed in giant yeast vacuoles"Submitted for publication. (in press).

[文献書誌] Suga, S.: "An abundance of aquaporins in specific cells and osmotic water permeability of individual protoplasts of radish"Plant Cell Physiol.. (in press).

2002 年度実績報告書

松岡信名古屋大学, 生物分子応答研究センター, 教授 (00270992)