1995 年度実績報告書

自家不和合性関連遺伝子の遺伝子導入試験

研究課題

研究課題/領域番号	05556002
研究機関	東北大学
研究代表者	日向康吉東北大学, 農学部, 教授 (00005589)
研究分担者	渡辺正夫東北大学, 農学部, 助手 (90240522) 鳥山欽哉東北大学, 農学部, 助教授 (20183882) 中西テツ神戸大学, 大学院・自然科学研究科, 教授 (80031227) 崎山文夫大阪大学, 蛋白質研究所, 教授 (40029947) 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)
キーワード	自家不和合性 / BRASSICA CAMPESTRIS / PYRUS PILIFOLIA / SLG / SRK / S-RNASE / 遺伝子操作 / トランスジェニック植物
研究概要	胞子体型の自家不和合性を示すアブラナ科植物および配偶体型に分類されるバラ科植物に関する自家不和合性関連遺伝子を単離し、これらの遺伝子を同じ植物種あるいは他の植物種に導入して、自家不和合性の制御の可能性を検討することが本研究の狙いである。日向・磯貝・渡辺・鳥山らはアブラナ科植物を材料として、既報の自家不和合性関連遺伝子の同義・非同義塩基置換の程度から、進化的にはSRAがまず分化し、それに続いて、SRB, SLG/SRKが分化したことを示唆した。さらに、SLG8,SLG9,SLG12,SRK8,SRK9,SRK12など、関連遺伝子としてSRA1,SRA2,SRA3,SRA4、およびSRBなどのcDNAを単離し,その塩基配列を決定し,S遺伝子間の相同性と特異性を明らかにした。そして、SLGのセンスおよびアンチセンス遺伝子を導入した形質転換植物を作成し、アンチセンスSLGを導入すると自家不和合性が失われること、センスSLGを導入するとコサプレッションが起きることを示した。また、タバコにSLG, SRKを導入し、SRKの発現について検討した。崎山はニホンナシの自家不和合性種「二十世紀」(S2S4)花柱より2種類のS-RNase(S2-RNase,S4-RNase)と自家不和合性に関与しないnon-S-RNaseをコードするcDNAをクローニングした。さらに、二十世紀の変異種で自家和合性の「おさ二十世紀」について検討し、この和合性はS4-RNaseの欠損によることを明らかにした。中西はニホンナシへの遺伝子導入法を検討し、Agrobacteriumの感染能を確認し、茎頂培養における不定芽の誘導と種子外植体からのシュートの再生にThidiazuronが効果的であることを明らかにした。しかし、残念ながらこの研究は阪神の災害で中断している。一方、ニホンナシS2-,S4-RNaseの塩基配列をペチュニアのS3プロモーターにつなぎ、トランスジェニック植物を得て、現在その遺伝子発現を検討中である。

研究成果

(19件)

すべてその他

すべて文献書誌 (19件)

[文献書誌] Yamakawa, S,, Watanabe, M., Isogai A., Takayama, S., Satoh, S., Hinata, K. and Suzuki, A.: "The cDNA sequence of NS3-glycoprotein from Brassica campestris and its homology to related proteins." Plant Cell Physiol.34. 173-175 (1993)
[文献書誌] Nou, I. S., Watanabe, M., Isuzugawa, K., Isogai, A., and Hinata, K.: "Isolation of S-allele from a wild population of Brassica campestris L. at Balcesme, Turkey and their characterization by S-glycoprotein." Sex. Plant Reprod.6. 71-78 (1993)
[文献書誌] Nou, I. S., Watanabe, M., Isogai, A., and Hinata, K.: "Comparison of S-alleles and S-glycoproteins between wild populations of Brassica cam pestris in Turkey and Japan." Sex. Plant Reprod. c. 6. 79-86 (1993)
[文献書誌] Hinata, K., Watanabe, M., Toriyama, K,, and Isogai, A.: "A review of recent studies on homomorphic self-incompatibility." Inter. Rev. Cytol.143. 257-296 (1993)
[文献書誌] Wong, K. C., Watanabe, M. and Hinata, K.: "Protein profiles in pin and thrum floral organs of distylous Averrhoa carambola L." Sex. Plant Reprod. 7. 7. 107-115 (1994)
[文献書誌] Wong, K. C., Watanabe, M. and Hinata, K.: "Fluorescnese and scanning electron microscopic studies on self-incompatibility in disty lous Averrhoa carambola L." Sex. Plant Reprod.7. 116-121 (1994)
[文献書誌] Yamakawa, S., Shiba, H., Watanabe, M., Shiozawa, H., Takayama, S., Hinata, K., Isogai, A. and Suzuki, A.: "The sequence of S-glycoproteins involved in self-incompatibility of Brassica campestris and their distribution among Brassicaceae." Biosci. Biotech. Biochem.58. 921-925 (1994)
[文献書誌] Watanabe, M., Takasaki, T., Toriyama, K., Yamakawa, S., Isogai, A., Suzuki, A. and Hinata, K.: "A high degree of homology exists between the protein encoded by SLG and the S recept or domain encoded by SRK in self-incompatible Brassica campestris L." Plant Cell Physiol.35. 1221-1229 (1994)
[文献書誌] Hinata, K., Watanabe, M., Isogai, A., Yamakawa, S., Satta, Y. and Takahata, N.: "Genetic studies of self-incompatible Brassica campestris." “Pollen-pistil interactions and pollen tube growth (Current Topics in Plant Physiology: An American Society of Plant Physiologists Series. ). 12. 103-110 (1994)
[文献書誌] Matsushita, M., Watanabe, M., Yamakawa, S., Isogai, A. and Hinata, K.: "Base sequence variation of the S-glycoprotein within the same S12-allele in Brassica campestris." “Pollen-pistil interactions and pollen tube growth (Current Topics in Plant Physiology: An American Society of Plant Physiologists Series. ). 12. 313-315 (1994)
[文献書誌] Yamakawa, S., Watanabe, M., Hinata, K., Suzuki, A. and Isogai, A.: "The sequences of S-receptor kinase(SRK)involved in self-incompatibility and their homologies to S-locus glycoprotein of Brassica campestris." Biosci. Biotech. Biochem.59. 161-162 (1995)
[文献書誌] Hinata, K., Watanabe, M., Yamakawa, S., Satta, Y. and Isogai, A.: "Evolutionary aspects of the S-related genes of the Brassica self-incompatibility system:synonymous and nonsynonyomous base substitutions." Genetics. 140. 1099-1104 (1995)
[文献書誌] Suzuki,G.,Watanabe, M., Toriyama, K., Isogai, A. and Hinata, K.: "Molecular cloning of members of the S-multigene family in self-incompatible Brassica campestris L." Plant Cell Physiol.36. 1273-1280 (1995)
[文献書誌] Shinba, H., Hinata, K., Suzuki, A. and Isogai. A.: "Breakdown of self-incompatibility in Brassica by the antisense RNA of SLG gene." Proc. Japan Academy,. 71B. 81-83 (1995)
[文献書誌] Kuroda, S., Norioka, S., Mitta, M., Kato, I., and Sakiyama, F.: "Primary structure of a novel stylar RNaseunassociated with self-incompatibility in tobacco plant, Nicotiana alata." J. Protein Chem.13. 438-439 (1994)
[文献書誌] Ishimizu, T., Miyagi, M., Norioka, S., Ying-Hong Liu, Clarke, A. E. and Sakiyama, F.: "Identification of histidine 31 and cysteine 95 in the active site of self-incompatibility a ssociated S6-RNase in Nicotiana alata." J. Biochem.118. 1007-1013 (1995)
[文献書誌] Norioka, N., Ohnishi, Y., Norioka, S., Ishimizu, T., Nakanishi, T. & Sakiyama F.: "Nucleotide Sequences of cDNAs Encodinf S2-and S4-RNases from Japanese Pear(Pyrus pynfolia Nakai)" Plant Physiol.108. 1343- (1995)
[文献書誌] 渡辺正夫・鳥山欽哉: "花粉学事典(日本花粉学会編,分担執筆)" 朝倉書店, (1994)
[文献書誌] Hinata, K., Isogai, A. and Isuzugawa, K.,: "Genetic control of self-incompatibility and reproductive development in flowering plants" eds. by Williams, E. G., Clarke, A. E. and Knox, R. B.," Kluwer Academic Publishers,102-115 (1994)

1995 年度 実績報告書

自家不和合性関連遺伝子の遺伝子導入試験

研究代表者

日向 康吉 東北大学, 農学部, 教授 (00005589)

研究成果

[文献書誌] Yamakawa, S,, Watanabe, M., Isogai A., Takayama, S., Satoh, S., Hinata, K. and Suzuki, A.: "The cDNA sequence of NS3-glycoprotein from Brassica campestris and its homology to related proteins." Plant Cell Physiol.34. 173-175 (1993)

[文献書誌] Nou, I. S., Watanabe, M., Isuzugawa, K., Isogai, A., and Hinata, K.: "Isolation of S-allele from a wild population of Brassica campestris L. at Balcesme, Turkey and their characterization by S-glycoprotein." Sex. Plant Reprod.6. 71-78 (1993)

[文献書誌] Nou, I. S., Watanabe, M., Isogai, A., and Hinata, K.: "Comparison of S-alleles and S-glycoproteins between wild populations of Brassica cam pestris in Turkey and Japan." Sex. Plant Reprod. c. 6. 79-86 (1993)

[文献書誌] Hinata, K., Watanabe, M., Toriyama, K,, and Isogai, A.: "A review of recent studies on homomorphic self-incompatibility." Inter. Rev. Cytol.143. 257-296 (1993)

[文献書誌] Wong, K. C., Watanabe, M. and Hinata, K.: "Protein profiles in pin and thrum floral organs of distylous Averrhoa carambola L." Sex. Plant Reprod. 7. 7. 107-115 (1994)

[文献書誌] Wong, K. C., Watanabe, M. and Hinata, K.: "Fluorescnese and scanning electron microscopic studies on self-incompatibility in disty lous Averrhoa carambola L." Sex. Plant Reprod.7. 116-121 (1994)

[文献書誌] Yamakawa, S., Shiba, H., Watanabe, M., Shiozawa, H., Takayama, S., Hinata, K., Isogai, A. and Suzuki, A.: "The sequence of S-glycoproteins involved in self-incompatibility of Brassica campestris and their distribution among Brassicaceae." Biosci. Biotech. Biochem.58. 921-925 (1994)

[文献書誌] Yamakawa, S., Watanabe, M., Hinata, K., Suzuki, A. and Isogai, A.: "The sequences of S-receptor kinase(SRK)involved in self-incompatibility and their homologies to S-locus glycoprotein of Brassica campestris." Biosci. Biotech. Biochem.59. 161-162 (1995)

[文献書誌] Hinata, K., Watanabe, M., Yamakawa, S., Satta, Y. and Isogai, A.: "Evolutionary aspects of the S-related genes of the Brassica self-incompatibility system:synonymous and nonsynonyomous base substitutions." Genetics. 140. 1099-1104 (1995)

[文献書誌] Suzuki,G.,Watanabe, M., Toriyama, K., Isogai, A. and Hinata, K.: "Molecular cloning of members of the S-multigene family in self-incompatible Brassica campestris L." Plant Cell Physiol.36. 1273-1280 (1995)

[文献書誌] Shinba, H., Hinata, K., Suzuki, A. and Isogai. A.: "Breakdown of self-incompatibility in Brassica by the antisense RNA of SLG gene." Proc. Japan Academy,. 71B. 81-83 (1995)

[文献書誌] Kuroda, S., Norioka, S., Mitta, M., Kato, I., and Sakiyama, F.: "Primary structure of a novel stylar RNaseunassociated with self-incompatibility in tobacco plant, Nicotiana alata." J. Protein Chem.13. 438-439 (1994)

[文献書誌] Ishimizu, T., Miyagi, M., Norioka, S., Ying-Hong Liu, Clarke, A. E. and Sakiyama, F.: "Identification of histidine 31 and cysteine 95 in the active site of self-incompatibility a ssociated S6-RNase in Nicotiana alata." J. Biochem.118. 1007-1013 (1995)

[文献書誌] Norioka, N., Ohnishi, Y., Norioka, S., Ishimizu, T., Nakanishi, T. & Sakiyama F.: "Nucleotide Sequences of cDNAs Encodinf S2-and S4-RNases from Japanese Pear(Pyrus pynfolia Nakai)" Plant Physiol.108. 1343- (1995)

[文献書誌] 渡辺正夫・鳥山欽哉: "花粉学事典(日本花粉学会編,分担執筆)" 朝倉書店, (1994)

[文献書誌] Hinata, K., Isogai, A. and Isuzugawa, K.,: "Genetic control of self-incompatibility and reproductive development in flowering plants" eds. by Williams, E. G., Clarke, A. E. and Knox, R. B.," Kluwer Academic Publishers,102-115 (1994)

1995 年度実績報告書

日向康吉東北大学, 農学部, 教授 (00005589)