葉の縦・横の決定機構

• Project/Area Number: 14036228
• 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 (2005-2006); National Institutes of Natural Sciences Okazaki Research Facilities (2004); Okazaki National Research Institutes (2002-2003)
• Principal Investigator: 塚谷 裕一 東京大学, 大学院・理学系研究科, 教授 (90260512)
• Project Period (FY): 2002 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥88,800,000 (Direct Cost: ¥88,800,000); Fiscal Year 2006: ¥17,700,000 (Direct Cost: ¥17,700,000); Fiscal Year 2005: ¥17,700,000 (Direct Cost: ¥17,700,000); Fiscal Year 2004: ¥17,700,000 (Direct Cost: ¥17,700,000); Fiscal Year 2003: ¥17,700,000 (Direct Cost: ¥17,700,000); Fiscal Year 2002: ¥18,000,000 (Direct Cost: ¥18,000,000)
• Keywords: 葉形態形成 / 極性 / 発生・分化 / 遺伝子 / 植物 / 葉 / 遺伝子制御 / 発生遺伝 / 分子遺伝学 / シロイヌナズナ / 細胞分裂 / 細胞伸長 / 発生遺伝学

Research Abstract

まず、葉の横への極性伸長制御因子・ANの細胞内局在を精密に再解析した結果、ER/ゴルジを中心とすることが判明した。そこでCtBPファミリーとの間で、分子機能が保存されているかどうかを調べるため、二歩博士との共同研究により、キイロショウジョウバエのAN安定形質転換体を作成し解析した結果、ANはdCtBPの機能の一部のみを代替することが判明した。さらに高野博士との共同研究により、裸子植物のカラマツからホモログを単離し、解析を行なった結果、カラマツのホモログは、シロイヌナズナan-1の表現型を完全に回復させることが確認された。
一方、葉の細胞観察の新方法を2種、開発し(Horiguchi et al.2006)これを用いて、葉の細胞数と細胞サイズを指標とした、シロイヌナズナ変異体の大規模スクリーニングを行ない、多くの葉変異体を新たに取得した(Horiguchi et al.2006)。そのうち、葉の細胞サイズに特異的な欠損を示すxs変異体群の解析から、葉の細胞伸長経路は少なくとも独立の2経路からなることが判明した(Fujikura et al.2007)。また補償作用を示す新たな変異体の解析からは、葉原基における細胞伸長と細胞分裂の間の共役を明らかにした(Ferjani et al.,in press)。
一方、横幅方向への細胞増殖を正に制御するAN3の下流遺伝子の候補を探索した結果、AN3による葉原基における細胞増殖制御系は、興味深いことに、葉の極性決定機構との問に遺伝的連関を持つことが判明した。
さらに葉の縦への細胞分裂の負の制御因子群・RTFLファミリーに関しては、イネよりホモログを単離し、植物で構成的に発現させてみたところ、ROT4同様、シロイヌナズナ、イネ共に葉を短くすることを確認した(Yamaguchi and Tsukaya 2006)。また35Sp::ROT4::GFPを導入した系統を親株とした変異原処理をおこない、エンハンサー系統を十数系統単離した。さらにROT4のC端・N端それぞれから部分的に削ったコンストラクトを作成し、解析した結果、ROT4は、RTFLドメインに含まれる小さなペプチド断片として機能する可能性を示唆する結果を得た。

Research Products

• [Journal Article] Dissection of enhanced cell expansion processes in leaves triggered by defect in cell proliferation, with reference to roles of endoreduplication. (2007)
  • Author(s): Fujikura U, Horiguchi G, Tsukaya H
  • Journal Title: Plant and Cell Physiology 48 Pages: 278-286
  • NAID: 10018519516
• [Journal Article] Analysis of leaf development in fugu mutants of Arabidopsis thaliana reveals three compensation modes that modulate cell expansion in determinate organs. (2007)
  • Author(s): Ferjani A, Horiguchi G., Tsukaya H
  • Journal Title: Plant Physiology (in press)
• [Journal Article] Large-scale histological analysis of leaf mutants using two simple leaf observation methods : identification of novel genetic pathways governing the size and shape of leaves. (2006)
  • Author(s): Horiguchi G., Fujikura U, Ferjani A, Ishikawa N, Tsukaya H
  • Journal Title: Plant Journal 48 Pages: 638-644
• [Journal Article] Gravitropism in Leaves of Arabidopsis thaliana (L.) Heynh. (2006)
  • Author(s): Mano E., Horiguchi G, Tsukaya H
  • Journal Title: Plant and Cell Physiology 47 Pages: 217-223
  • NAID: 10017272479
• [Journal Article] Leaf-shape variation of Paederia foetida L. in Japan : reexamination of the small, narrow leaf form from Miyajima Island. (2006)
  • Author(s): Tsukaya H., Imaichi R, Yokoyama J
  • Journal Title: Journal of Plant Research 119 Pages: 303-308
• [Journal Article] Coordination of cell proliferation and cell expansion in the control of leaf size in Arabidopsis thaliana (2006)
  • Author(s): Horiguchi G, Ferjani A, Fujikura U, Tsukaya H
  • Journal Title: Journal of Plant Research 119 Pages: 37-42
  • NAID: 40007130208
• [Journal Article] Mechanism of leaf shape determination. (2006)
  • Author(s): Tsukaya H.
  • Journal Title: Ann. Rev. Plant Biol 57 Pages: 477-496
• [Journal Article] A new member of the CtBP/BARS family from plants : Angustifolia. (2006)
  • Author(s): Tsukaya H.
  • Journal Title: G. Landes Bioscience, Georgetown, Texas, USA. (CtBP Family Proteins, G. Chinnadurai (ed.)) 12 Pages: 112-118
• [Journal Article] ROTUNDIFOLIA4, a plant-specific small peptide involved in the polar cell proliferation of leaves in Arabidopsis thaliana. (2006)
  • Author(s): Yamaguchi T, Tsukaya H
  • Journal Title: Elsevier, San Diego, USA (Handbook of Biologically Active Peptides, A J Kastin (ed.)) Pages: 37-40
• [Journal Article] 日本国内におけるネジバナ(Spiranthes sinensis var. australis)の分子多型と種内変異. (2006)
  • Author(s): 塚谷裕一
  • Journal Title: Plant Morphology 17 Pages: 31-34
  • NAID: 130004303797
• [Journal Article] Genetics, cell cycle and cell expansion in organogenesis in plants. (2006)
  • Author(s): Tsukaya H., G.Beemster
  • Journal Title: Journal of Plant Research 119 Pages: 1-4
  • NAID: 10017270429
• [Journal Article] 葉の形態形成と植物ホルモン. (2006)
  • Author(s): 山口貴大, 堀口吾朗, 塚谷裕一
  • Journal Title: 植物の生長調節 41 Pages: 122-130
  • NAID: 110005716847
• [Journal Article] Coordination of cell proliferation and cell expansion in the control of leaf size in Arabidopsis thaliana. (2006)
  • Author(s): Horiguchi G, Ferjani A, Fujikura U, Tsukaya H.
  • Journal Title: J.Plant Res. 119 Pages: 37-42
  • NAID: 40007130208
• [Journal Article] Gravitropism in leaves of Arabidopsis thaliana (L.) Heynh. (2006)
  • Author(s): Mano E, Horiguchi G, Tsukaya H.
  • Journal Title: Plant Cell Physiol 47 Pages: 217-223
  • NAID: 10017272479
• [Journal Article] Genetics, cell cycle and cell expansion in organogenesis in plants. (2006)
  • Author(s): Tsukaya H., G.Beemster
  • Journal Title: J.Pant Res. 119 Pages: 1-4
  • NAID: 10017270429
• [Journal Article] The biosynthesis of active forms of brassinosteroids in Arabidopsis. (2005)
  • Author(s): Kim, G.-T., Fujioka, S., Kozuka, T., Tax, F.E., Takatsuto, S., Yoshida, S., Tsukaya, H.
  • Journal Title: Plant J. 44 Pages: 710-721
• [Journal Article] The different growth responses of the Arabidopsis thaliana leaf blade and the petiole during shade avoidance are regulated by photoreceptors and sugar. (2005)
  • Author(s): Kozuka, T., Horiguchi, G., Kim, G.-T., Ohgishi, M., Sakai, T., Tsukaya, H.
  • Journal Title: Plant Cell Physiol 46 Pages: 213-223
  • NAID: 10014311153
• [Journal Article] Characterization of a member of the AN subfamily, IAN, from Ipomoea nil. (2005)
  • Author(s): Cho, K.-H., Shindo, T., Kim, G T., Nitasaka, E., Tsukaya, H.
  • Journal Title: Plant Cell Physiol 46 Pages: 250-255
  • NAID: 10014311308
• [Journal Article] The transcription factor AtGRF5 and the transcription coactivator AN3 regulate cell proliferation in leaf primordia of Arabidopsis thaliana. (2005)
  • Author(s): Horiguchi, G., Kim, G.-T., Tsukaya, H.
  • Journal Title: Plant J. 43 Pages: 68-78
• [Journal Article] Leaf shape : genetic controls and environmental factors. (2005)
  • Author(s): Tsukaya, H.
  • Journal Title: Int.J.Dev.Biol. 49 Pages: 547-555
• [Journal Article] Photomorphogenesis of leaves : Shade-avoidance syndrome and differentiation of sun/shade leaves. (2005)
  • Author(s): Kim, G.-T., Yano, S., Kozuka, T., Tsukaya, H.
  • Journal Title: Photochem.Photobiol.Sci. 4 Pages: 770-774
• [Journal Article] 葉の形態形成 (2005)
  • Author(s): 塚谷裕一, 堀口吾朗
  • Journal Title: 蛋白質核酸酵素 50 Pages: 627-632
• [Journal Article] The biosynthesis of active forms of brassinosteroids in Arabidopsis. (2005)
  • Author(s): Kim, G.-T., et al.
  • Journal Title: Plant Journal 41 Pages: 710-721
• [Journal Article] The different growth responses of the Arabidopsis thaliana leaf blade and petiole during shade avoidance are regulated by photoreceptors and sugar. (2005)
  • Author(s): Kozuka et al.
  • Journal Title: Plant & Cell Physiology 46 Pages: 213-223
• [Journal Article] Characterization of a member of the AN subfamily, IAN, from Ipomoea nil. (2005)
  • Author(s): Cho et al.
  • Journal Title: Plant & Cell Physiology 46 Pages: 250-255
• [Journal Article] Over-expression of a novel small peptide ROTUNDIFOLIA4 decreases cell proliferation and alters leaf shape in Arabidopsis. (2004)
  • Author(s): Narita et al.
  • Journal Title: Plant Journal 38 Pages: 699-713
• [Book] 植物の進化(植物の表現型可塑性とその進化.植物細胞工学)(長谷部光泰・清水健太郎編、分担執筆) (2007)
  • Author(s): 塚谷裕一
  • Publisher: 秀潤社(in press)
• [Book] プラント・ミメティックス(分担執筆) (2006)
  • Author(s): 柘植知彦, 塚谷裕一
  • Total Pages: 664
  • Publisher: エヌ・ティー・エス
• [Book] 変わる植物学 広がる植物学 (2006)
  • Author(s): 塚谷裕一
  • Total Pages: 227
  • Publisher: 東京大学出版会
• [Book] CtBP Family Proteins (分担) (2006)
  • Author(s): Tsukaya H.
  • Publisher: Landes Bioscience
• [Book] モデル植物の実験プロトコール(分担) (2005)
  • Author(s): 堀口吾朗, 塚谷裕一
  • Publisher: 秀順社
• [Publications] Ha, C.-H., Kim, G.-T., Kim, B.-C., Jun, J.-H., Soh, M.-S., Ueno, Y., Machida, Y., Tsukaya, H., Nam, H.-G.: "The BLADE-ON-PETIOLE gene control leaf pattern formation through regulation of meristematic activity."Development. 130. 161-172 (2003)
• [Publications] Sulpice, R., Tsukaya, H., Nonaka, H., Mustardy, R., Chen, T., Murata, N.: "Enhanced formation of flowers in salt-stressed Arabidopsis after genetic engineering of the accumulation of glycine betaine."Plant J.. 36. 165-176 (2003)
• [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 of cell proliferation and alters in Arabidopsis."Plant J.. (in press). (2004)
• [Publications] Tsukaya, H.: "Organ shape and size : a lesson from studies of leaf morphogenesis."Curr.Opin.Plant Biol.. 6. 57-62 (2003)
• [Publications] 塚谷裕一: "葉の形とサイズは環境に応じて変化する--葉形の可塑性と進化--"遺伝. 57. 54-59 (2003)
• [Publications] Machida, C., Iwakawa, H., Ueno, Y., Semiarti, E., Tsukaya, H., Hasebe, H., Kojima, S., Machida, Y(分担): "Sekimura, T., Noji, S., Ueno, N. and maini, P.K. eds., Morphogenesis and Pattern Formation in Biological Systems : Experiments and Models."Springer-Verlag Tokyo.. 398 (2003)
• [Publications] 塚谷裕一(分担執筆): "『生物の形の多様性と進化』関村利朗・野地澄晴・森田利仁編"裳華房. 352 (2003)
• [Publications] Ha, C.-H., Kim, G.-T., Kim, B.-C., Jun, J.-H., Soh, M.-S., Ueno, Y., Machida, Y., Tsukaya, H., Nam, H.-G.: "The BLADE-ON-PETIOLE gene controls leaf pattern formation through regulation of meristematic activity"Development. 130. 161-172 (2003)
• [Publications] Nakaya, M., Tsukaya, H., Murakami, N., Kato, M.: "Brassinosteroids control the proliferation of the leaf cells in Arabidopsis thaliana"Plant Cel Physiol.. 43. 239-244 (2002)
• [Publications] Kim, G.-T., Shoda, K., Tsuge, T., Cho, K.-H., Uchimiya, H., Yokoyama, R., Nishitani, K., Tsukaya, H.: "The ANGUSTIFOLIA gene of Arabidopsis, a plant CtBP gene, regulates leaf-cell expansion, the arrangement of corticalmicrotubules in leaf cells and expression of a gene involved in cell-wall formation"EMBO J.. 21. 1267-1279 (2002)
• [Publications] Iwakawa, H., Ueno, Y., Semiarti, E., Onouchi, H., Kojima, S., Tsukaya, H., Hasebe, M., Soma, T., Ikezaki, M., Machida, C., Machida, Y.: "The ASYMMETRIC LEAVES2 gene of Arabidopsis thaliana, required for formation of a symmetric flat leaf lamina, encodes a member of a novel family of protein characterized by cysteine repeats and a leucine zipper"Plant Cell Physiol. 43. 467-478 (2002)
• [Publications] Tsukaya, H.: "Leaf anatomy of a rheophyte, Dendranthema yoshinaganthum (Asteraceae), and of hybrids between D. yoshinaganthum and a closely related non-rheophyte species, D. indicum"J. Plant Res. 115. 329-333 (2002)
• [Publications] Stoynova-Bakalova, E., Petrov, P., Tsukaya, H.: "Cell division and cell enlargement in isolated Cucurbita cotyledons grown in darkness and in light"J. Plant Res. 115. 375-380 (2002)
• [Publications] Tsukaya, H., Kozuka, T., Kim, G.-T: "Genetic control of petiole length in Arabidopsis thaliana"Plant Cell Physiol. 43. 1221-1228 (2002)
• [Publications] Tsukaya, H.: "Organ shape and size"Curr. Opin. Plant Biol.. 6. 57-62 (2003)
• [Publications] Tsukaya, H.: "Interpretation of mutants in leaf morphology : genetic evidence for a compensator system in leaf morphogenesis that provides a new link between Cell and Organismal theory"Int. Rev. Cytol. 217. 1-39 (2002)
• [Publications] Tsukaya, H.: "Leaf Development"The Arabidopsis Book, eds. C.R. Somerville and E.M. Meyerowitz, American Society of Plant Biologists, Rockville, MD http://www.aspb.org/downloads/arabidopsis/tsukaya.pdf. (web.). (2002)
• [Publications] Tsukaya, H.: "The Leaf Index : Heteroblasty, Natural Variation, and the Genetic Control of Polar Processes of Leaf Expansion"Plant Cell Physiol. 43. 372-378 (2002)
• [Publications] 塚谷裕一: "葉の形成-葉の長さ、幅、大きさを決める遺伝子制御"蛋白質核酸酵素. 47. 1576-1580 (2002)
• [Publications] 塚谷裕一: "横幅を決める形態形成遺伝子・AnとCtBP"Medical Science Digest. 28. 4-5 (2002)
• [Publications] Tsukaya, H.: "Himalayan Botany in the Twentieth and Twenty-first Centuries"Molecular mechanisms related to the evolution of leaf shape, with a special emphasis on leaf index. In Himalayan Botany in the Twentieth and Twenty-first Centuries. (2002)
• [Publications] 塚谷裕一: "植物工学の基礎"第5章「モデル植物としてのシロイヌナズナ」(pp.152-179)、第6章「イネゲノムプロジェクト」(pp.180-190). 208 (2002)