茎頂根端軸の形成・維持を支配する細胞間相互作用の解析

• Project/Area Number: 14036205
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokyo
• Principal Investigator: 福田 裕穂 東京大学, 大学院理学系研究科, 教授 (10165293)
• Co-Investigator(Kenkyū-buntansha): 出村 拓 理化学研究所, 植物科学研究センター, チームリーダー(研究職) (40272009) ; 澤 進一郎 東京大学, 助教授 (00315748)
• Project Period (FY): 2002 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥172,000,000 (Direct Cost: ¥172,000,000); Fiscal Year 2006: ¥32,500,000 (Direct Cost: ¥32,500,000); Fiscal Year 2005: ¥32,500,000 (Direct Cost: ¥32,500,000); Fiscal Year 2004: ¥35,500,000 (Direct Cost: ¥35,500,000); Fiscal Year 2003: ¥35,500,000 (Direct Cost: ¥35,500,000); Fiscal Year 2002: ¥36,000,000 (Direct Cost: ¥36,000,000)
• Keywords: シロイヌナズナ / 細胞間相互作用 / 脂質 / CLEペプチド / 小胞輸送 / 維管束 / レセプターキナーゼ / 突然変異体 / メリステム / 前形成層 / 葉酸 / ザイロジェン / 管状要素 / ダイナミン / ジーントラップ / 細胞内極性 / ARF / ヒャクニチソウ / 軸 / ファイトスルフォカイン / 二次細胞壁

Research Abstract

1)シロイヌナズナ変異体を用いた維管束の連続性に関する研究
VAN3およびその3つのシロイヌナズナホモローグの機能解析を進めた。VAN3のホモローグそれぞれの、ダブルミュータントおよび、トリプルミュータントは維管束の連続性に顕著な影響を及ぼさなかった。しかし、van3との4重変異体では、主脈の断片化を含むvan3より重篤な表現型が現れた。これにより、ACAPタイプのARF-GAPが維管束の連続性に極めて重要な働きをすることが、その中でもVAN3が中心的な役割を担うことが明らかになった。また、脂質との関連について、脂質合成の突然変異体との掛け合わせ、さらには、各ドメインを単独で導入した変異体を用いた解析などから、PHドメインでのptdins(4)Pへの結合が、VAN3の機能発現に必要なことが明らかになった。また、van3による多様なPINの局在に対する影響を調べたところ、1種のPINのみの局在に影響を与えることが分かり、このPINはVAN3による輸送系の制御を受けている可能性が示唆された。
2)発生における局所的細胞間相互作用の解析:
維管束阻害因子、TDIF(Tracheary elements Differentiation Inhibitory Factor)の同定に成功した。この原因遺伝子を単離したところ、シロイヌナズナのCLE41、CLE44に相当する遺伝子であることが分かった。また、CLE41、CLE44タンパク質は12個のアミノ酸として切り出され、さらに水酸化されることにより、活性ペプチドとなることが分かった。シロイヌナズナに存在する全CLEペプチド26種を化学合成し、その活性をシロイヌナズナ茎頂の成長阻害、シロイヌナズナ・イネの根の成長阻害、シロイヌナズナ維管束形成阻害、コケの伸長成長阻害について調べたところ、それぞれに特異性と共通性のあることが明らかとなった。
3)イネの横走維管束形成に関する研究
イネ葉の横走維管束の間隔が短くなる突然変異体を単離し、その解析を行った。その結果、この原因遺伝子がレセプターキナーゼをコードしていることが明らかになった。

Research Products

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• [Journal Article] VAN3 ARF-GAP-mediated vesicle transport is involved in leaf vascular network formation. (2005)
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  • Journal Title: Development (In press)
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• [Journal Article] Identification and molecular characterization of novel anther-specific genes in Diyza sativa, L.by using cDNA microarray. (2004)
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• [Journal Article] 維管束形成を制御するシグナル分子 (2004)
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• [Book] Glycosylphosphatidylinositol-anchored proteins in plants. In Frontiers in Life Sciences, 2006 (ed. M. Fujiwara, N Sato, and S. Ishiura) (2006)
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• [Book] Vascular cell differentiation. In Cell Separation and Adhesion, (eds. J. Roberts and Z. Gonzalez-Carranza) (2006)
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• [Publications] Ito-Ohashi, K., Fukuda, H.: "HD-Zip III homeobox genes that include a novel member, ZeHB-13(Zinnia)/ATH 15 (Arabidopsis), are involved in procambium and xylem cell differentiation"Plant Cell Physiol.. 44. 1350-1358 (2003)
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• [Publications] Nara-Sato, K., Demura, T., Fukuda, H.: "Expression of photosynthesis-related genes and their regulation by light during somatic embryogenesis in Daucuscarota"Planta. (in press). (2004)
• [Publications] Minami, A., Fukuda, H.: "Trypsin-like endopeptidase activity is induced in the last stage of trache element differentiation of Zinnia cells"Plant Biotech.. (in press). (2004)
• [Publications] Narita, N.N., Moore, S., Horiguchi, G., Kubo, M., Demura, T., Fukuda, H., Goodrich, J., Tsukaya, H.: "Over-expression of a novel small peptide ROTUNDIFOLIA4 decreases cell proliferation and alters leaf shape in Arabidopsis"Plant J.. (in press). (2004)
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• [Publications] Demura, T., Tashiro, G.Horiguchi, G., Kishimoto, N., Kubo, M., Matsuoka, N., Minami, A., Nagata-Hiwatashi, M., Nakamura, K., Okamura, Y., Sassa, N., Suzuki, S., Yazaki, J., Kikuchi, S., Fukuda, H.: "Visualization by comprehensive microarray analysis of gene expression programs during transdifferentition of mesophyll cells into xylem cells"Proc. Natl. Acad. Sci. USA. 99. 15794-15799 (2002)
• [Publications] Kuriyama, H., Fukuda, H.: "Developmental Programmed Cell Death in Plants"Curr Opin Plant Biol.. 5. 568-573 (2002)
• [Publications] Ito-Ohashi, K., Demura, T., Fukuda, H.: "Promotion of transcript accumulation of novel Zinnia immature xylem-specific HD-Zip III homeobox genes by brassinosteroids"Plant Cell Physiol. 43. 1146-1153 (2002)
• [Publications] Nakanomyo, I., Kost, B., Chua, N.-H., Fukuda, H.: "Preferential and asymmetrical accumulation of a Rac small GTPase mRNA in differentiating xylem cells of Zinnia elegans"Plant Cell Physiol.. (in press). (2002)
• [Publications] Shinohara, N., Fukuda, H.: "Isolation of monoclonal antibodies recognizing rare and dominant epitopes in plant vascular cell walls by phage display subtraction"J. Immunological Methods. 264. 187-194 (2002)
• [Publications] Endo, M., Matsubara, H., Kokubun, T., Masuko, H., Takahata, Y., Tsuchiya, T., Fukuda, H., Demura, T., Watanabe, M.: "The advantages of cDNA microarray as an effective tool for identification of reproductive organ-specific genes in a model legume, Lotus japonicus"FEBS Lett.. 514. 229-237 (2002)
• [Publications] Ohdaira, Y., Kakegawa K., Amino, S., Sugiyama, M., Fukuda, H.: "Activity of cell-wall autolysis associated with differentiation of isolated mesophyll cells of Zinnia elegans into tracheary elements"Planta. 215. 177-184 (2002)
• [Publications] Nishitani, C., Demura, T., Fukuda, H.: "Analysis of the early stages of wound vascular differentiation processes using two molecular marker genes, TED3 and ZeHB3"Plant Cell Physiol.. 43. 79-90 (2002)
• [Publications] 福田裕穂, 出村拓, 松岡 健: "高等植物の形態形成メカニズムの解明を目指して"日本農芸化学会誌. 74(1). 42-44 (2002)
• [Publications] 福田裕穂: "液胞による細胞死の演出"遺伝別刷、細胞のミクロコスモス-進化とゲノムからその素顔に迫る-. 14. 87-95 (2002)
• [Publications] 福田裕穂: "維管束システムの構築"タンパク質核酸酵素増刊、植物の形づくり-遺伝子からみた分子メカニズム. 47(12). 1593-1598 (2002)