アブラナおよびペチュニアの自家不和合性の分子機構

• Project/Area Number: 11238205
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Nara Institute of Science and Technology
• Principal Investigator: 磯貝 彰 奈良先端科学技術大学院大学, バイオサイエンス研究科, 教授 (20011992)
• Co-Investigator(Kenkyū-buntansha): 岩野 恵 奈良先端科学技術大学院大学, バイオサイエンス研究科, 教務職員 (50160130) ; 柴 博史 奈良先端科学技術大学院大学, バイオサイエンス研究科, 助手 (20294283) ; 高山 誠司 奈良先端科学技術大学院大学, バイオサイエンス研究科, 助教授 (70273836)
• Project Period (FY): 1999 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥98,700,000 (Direct Cost: ¥98,700,000); Fiscal Year 2003: ¥17,000,000 (Direct Cost: ¥17,000,000); Fiscal Year 2002: ¥21,300,000 (Direct Cost: ¥21,300,000); Fiscal Year 2001: ¥21,200,000 (Direct Cost: ¥21,200,000); Fiscal Year 2000: ¥21,200,000 (Direct Cost: ¥21,200,000); Fiscal Year 1999: ¥18,000,000 (Direct Cost: ¥18,000,000)
• Keywords: 自家不和合性 / アブラナ / ペチュニア / ウメ / S-レセプターキナーゼ / S-RNase / Fボックスタンパク質 / 自他認識 / S遺伝子 / 花粉因子 / 自他識別 / 受容体キナーゼ / リガンド / RNase / F-box蛋白質 / シグナル伝達 / 情報伝達 / S-Rnase

Research Abstract

1.アブラナの自家不和合性
雌ずいのSRK受容体下流の情報伝達系の解明を目的として、自家和合性変異株イエローサルソンの原因遺伝子Mのポジショナルクローングを行った。その結果、M遺伝子は、MLPKと名付けた新規膜結合型蛋白質キナーゼをコードしていることが示された。変異株においては、1塩基非同義置換によりMLPKはキナーゼ活性を失っていることが判明した。この変異株の柱頭に野生型MLPKを一過的に発現させると自家不和合性が回復することが示され、MLPKが自家不和合性の情報を正に伝達する鍵因子であることが明らかとなった。MLPKは、これまでに機能が殆ど解明されていない受容体様細胞質キナーゼ(RLCK)ファミリーに属する蛋白質であったが、細胞膜に局在しており、恐らくSRK近傍で機能していると考えられた。MLPKの活性化機構や基質を明らかにすることで情報伝達系の解明が加速するものと期待される。
2.ペチュニアの自家不和合性
昨年度、ナス科植物ペチュニアと同じ配偶体型自家不和合性を示すバラ科植物ウメのS遺伝子座の解析から、花粉因子の有力な候補遺伝子SLFを発見した。しかし、ウメを用いての形質転換による機能証明は困難であったことから、本年度は、そのオルソログをペチュニアで探索した。その結果、ペチュニアの3つのSハプロタイプからSLFオルソログと推定される遺伝子を単離することができた。現在、花粉因子としての機能の証明を、形質転換の手法を用いて進めている。また、SLFはFボックス蛋白質として、推定標的蛋白質であるS-RNaseの蛋白質分解に関与していると期待される。SLFのS-RNaseとの相互作用の検証と、Fボックス蛋白質としての機能検証が、今後の重要研究課題として残されている。

Research Products

• [Publications] Takayama, S.: "Molecular mechanism of self-recognition in Brassica self-incompatibility."J.Exp.Bot.. 54. 149-156 (2003)
• [Publications] Kakizaki, T.: "Linear dominance relationship among four class-II S haplotypes in pollen is determined by the expression of SP11 in Brassica self-incompatibility."Plant Cell Physiol.. 44. 70-75 (2003)
• [Publications] Shiba, H.: "Genomic organization of the S-locus region of Brassica."Biosci.Biotechnol.Biochem.. 67. 622-626 (2003)
• [Publications] Entani, T.: "Comparative analysis of the self-incompatibility (S-) locus region of Prunus mume : identification of a pollen-expressed F-box gene with allelic diversity."Genes Cells. 8. 203-213 (2003)
• [Publications] Suzuki, G.: "The S haplotypes lacking SLG in the genome of Brassica rapa."Plant Cell Rep.. 21. 911-915 (2003)
• [Publications] Iwano, M.: "Immunohistochemical studies on translocation of pollen S-haplotype determinant in self-incompatibility of Brassica rapa."Plant Cell Physiol.. 44(4). 428-436 (2003)
• [Publications] Watanabe, M.: "Recent progress on self-incompatibility research in Brassica species."Breeding Sci.. 53. 199-208 (2003)
• [Publications] Mishima, M.: "Structure of the male determinant factor for Brarsica self-incompatibility."J.Biol.Chem.. 278(38). 36389-36395 (2003)
• [Publications] Iwano, M.: "Calcium crystals in the anther of Petunia the existence and biological significance in the pollination process."Plant Cell Physiol.. 45. 40-47 (2004)
• [Publications] Murase, K.: "A membrane-anchored protein kinase involved in Brassica self-incompatibility signaling."Science. 303. 1516-1519 (2004)
• [Publications] H.Shiba: "The dominance of alleles controlling self-incompatibility in Brassica pollen is regulated at the RNA level"Plant Cell. 14. 491-504 (2002)
• [Publications] S.Takayama: "Peptide signalling in plants"Curr. Opin. Plant Biol.. 5. 382-387 (2002)
• [Publications] A.Isogai: "Molecular mechanism for the recognition reaction in the self-incompatibility of Brassica species"Porc. Japan Acad.. 78,Ser.B. 241-249 (2002)
• [Publications] S.Takayama: "Molecular mechanism of self-recognition in Brassica self-incompatibility"J. Exp. Bot.. 54. 149-156 (2003)
• [Publications] T.Kakizaki: "Linear dominance relationship among four class-II S haplotypes in pollen is determined by the expression of SP11 in Brassica self-incompatibility"Plnat Cell Physiol.. 44. 70-75 (2003)
• [Publications] T.Entani: "Comparative analysis of the self-incompatibility (S-)locus region of Prunus mume : identification of a pollen-expressed F-box gene with allelic diversity"Genes Cells. 8. 202-213 (2003)
• [Publications] H.Shiba: "Genomic organization of the S-locus region of Brassica"Biosci. Biotechnol. Biochem.. (in press).
• [Publications] G.Suzuki: "The S haplotypes lacking SLG in the genome of Brassica rapa"Plant Cell Rep.. (in press).
• [Publications] M.Iwano: "Immunohistochemical studies on translocation of pollen S-haplotype determinant in self-incompatibility of Brassica rapa"Plant Cell Physiol.. (in press).
• [Publications] 磯貝 彰: "植物は自分の花粉をどうやって見分けるか-アブラナ科植物の自家不和合性の分子機構"蛋白質核酸酵素. 47(6). 700-707 (2002)
• [Publications] H.Shiba: "A pollen coat protein, SP11/SCR, determines the pollen specificity in the self-incompatibility of Brassica"Plant Physiol.. 125(4). 2095-2103 (2001)
• [Publications] S.Takayama: "Direct ligand-receptor complex interaction controls Brassica self-incompatibility"Nature. 413. 534-538 (2001)
• [Publications] H.Shiba: "The dominance of alleles controlling self-incompatibility in Brassica pollen is regulated at the RNA level"Plant Cell. 14(2)(In press). (2002)
• [Publications] 磯貝 彰: "植物は自分の花粉をどうやって見分けるか"蛋白質核酸酵素. (印刷中). (2002)
• [Publications] 柴 博史: "植物の自家不和合性の謎が解けた"化学. 56(8). 18-23 (2001)
• [Publications] Y.Takada: "Characterization of expressed genes in the SLL2 region of self-incompatibile Arabidopsisi thaliana"DNA Research. 8. 215-219 (2001)
• [Publications] K.Hatakeyama: "The S receptor kinase gene determines dominance relationships in stigma expression of self-incompatibility in Brassica"Plant J.. 26(1). 69-76 (2001)
• [Publications] N.Kai: "Sequence comparisons among dispersed members of the Brassica S multigene family in an S9 genome"Mol. Genet. Genomics. 265. 526-534 (2001)
• [Publications] S.Takayama: "The pollen determinant of self-incompatibility in Brassica campestris."Proc.Natl.Acad.Sci.USA. 97(4) . 1920-1925 (2000)
• [Publications] T.Takasaki: "The S receptor kinase determines self-incompatibility in Brassica stigma."Nature. 403. 913-916 (2000)
• [Publications] S.Takayama: "Isolation and characterization of pollen coat proteins of Brassica campestris that interact with S-locus related glycoprotein 1 involved in pollen-stigma adhesion"Proc.Natl Acad.Sci.USA. 97(7). 3765-3770 (2000)
• [Publications] M.Watanabe: "Highly divergent sequences of the pollen self-incompatibility (S) gene in calss-I haplotypes of Brassica campestris (syn.rapa) L."FEBS Letters. 473. 139-144 (2000)
• [Publications] M.Watanabe: "Genomic organization of the SLG/SRK region of the S-locus in Brassica species."Annals Botany. 85 (Supplement). 155-161 (2000)
• [Publications] H.Shiba: "Alteration of the self-incompatibility phenotype in Brassica by transformation of the antisense SLG gene."Biosci.Biotech.Biochem.. 64(5). 1016-1024 (2000)
• [Publications] H.Shiba: "A pollen caot protein, SP11/SCR, determines the pollen specificity in the self-incompatibility of Brassica(in press)"Plant Physiol.. (2001)
• [Publications] T.Entani: "Centromeric localization of an S-Rnase gene in Petunia hybrida Vilm"Theopr.Appl.Genet.. 99. 391-397 (1999)
• [Publications] M.Watanabe: "Molecular biology of self-incompatibility in Brassica species"Plant Biotechnol.. 16(4). 263-272 (1999)
• [Publications] G.Suzuki: "Genomic oraganization of the S locus : Identification and characterization of genes in SLG/SRK region of S9 haplotype of Brassica campestris(svn.rapa)"Genetics. 153. 391-400 (1999)
• [Publications] T.Takasaki: "Introduction of SLG(S locus glycoprotein) alters the phenotype of endogenous S haplotype, but confers no new S haplotype sepcificitv in Brassica rapa L"Plant Mol.Biol.. 40. 659-668 (1999)
• [Publications] M.Watanabe: "Two anther-expressed genes downstream of SLG9 : Identification of a novel S-linked gene specifically expressed in anthers at uninucleate of Brassica campestris(svn.rapa)I"Sex.Plant Reprod.. 12. 127-134 (1999)
• [Publications] T.Entani: "Relationship between polyploydy and pollen self-incompatibility phenotype in Petunis hybrida Vilm"Biosci.Biotechnol.Biochem.. 63(11). 1882-1888 (1999)
• [Publications] T.Takasaki: "The S receptor kinase determines self-incompatibility in Brassica stigma"Nature. (in press).
• [Publications] S.Takayama: "The pollen determinant of self-incompatibility in Brassica campestris"Proc.Natl.Acad.Sci.USA. (in press).
• [Publications] M.Iwano: "X-ray microanalysis of papillar cells and pollen grains in the pollination process in Brassica using variable-pressure scanning electron microscope"J.Electron Microsc.. (In press).
• [Publications] S.Takayama: "Isokation and characterization of coat proteins of Brassica campestris that interact with S-locus realted glvcoprotein 1 involved in pollen-stigma adhesion"Proc.Natl.Acad.Sci.USA. (in press).