Harnessing synthetic chemistry to elucidate novel mechanisms of stomatal patterning

• Project/Area Number: 19H00990
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Medium-sized Section 44:Biology at cellular to organismal levels, and related fields
• Research Institution: Nagoya University
• Principal Investigator: 鳥居 啓子 名古屋大学, トランスフォーマティブ生命分子研究所, 客員教授 (60506103)
• Co-Investigator(Kenkyū-buntansha): 下遠野 明恵 名古屋大学, トランスフォーマティブ生命分子研究所, 特任講師 (70647544) ; 村上 慧 関西学院大学, 理学部, 准教授 (90732058) ; 望田 啓子 (桑田啓子) 名古屋大学, トランスフォーマティブ生命分子研究所, 特任講師 (70624352)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Project Status: Declined (Fiscal Year 2022)
• Budget Amount: ¥44,980,000 (Direct Cost: ¥34,600,000、Indirect Cost: ¥10,380,000); Fiscal Year 2022: ¥12,610,000 (Direct Cost: ¥9,700,000、Indirect Cost: ¥2,910,000); Fiscal Year 2021: ¥11,960,000 (Direct Cost: ¥9,200,000、Indirect Cost: ¥2,760,000); Fiscal Year 2020: ¥10,010,000 (Direct Cost: ¥7,700,000、Indirect Cost: ¥2,310,000); Fiscal Year 2019: ¥10,400,000 (Direct Cost: ¥8,000,000、Indirect Cost: ¥2,400,000)
• Keywords: 植物発生 / 有機合成化学 / 気孔 / シグナル伝達 / ケミカルバイオロジー / bHLH型転写因子 / ステロイド / 有機合成 / 低分子化合物 / 植物の発生 / 生理活性 / 構造活性相関 / 非対称分裂 / 植物成長 / 合成化学 / 低分子生化合物 / ストレス応答 / ラベルフリープロテオミクス / 低分子性化合物

Outline of Research at the Start

陸上植物の表皮に散在する気孔は、植物と大気とのガス交換や植物体の水分調節を担っている 。気孔の存在は、バイオマス生産と生存に必要不可欠である。本申請研究は、植物発生学、有機合成化学、分析化学の専門家の綿密なコラボレーションを通して、化合物をキーワードに、従来のアプローチでは捉えられなかった気孔の発生を制御する新奇メカニズムの解明を目指すものである。本研究は、動植物を問わず、細胞の分化やパターン形成の普遍的な仕組みに迫るものである。また、将来的には、低分子化合物を用いて気孔の数を人為的に操作することにより、植物の成長制御につながる可能性がある。

Outline of Annual Research Achievements

本申請研究は、植物発生学、有機合成化学、分析化学の専門家の綿密なコラボレーションを通して、化学の力で気孔の数と分布を制御する未知のメカニズムに斬り込むとともに、化合物による有用植物のバイオマス促進に繋がるものである。本年度は、気孔の数を増やす化合物kC9とChatty、気孔幹細胞であるメリステモイド細胞の発生停止を引き起こすAYSJ929(Stomatazoline)に関する生理機能解析と標的候補因子の探索を主に継続した。
(1) Stomatazoline - 気孔の発生に関わる様々な突然変異体に対する生理作用の詳細な解析から、Stomatazolineはメリステモイド細胞から孔辺母細胞への分化を指令するMUTEの活性を阻害することが示唆された。Stomatazolineは、気孔系譜の指令因子であるｂHLHタンパク質のC末端に存在するACTL領域に結合すること、それにより、MUTEーSCREAMヘテロ二量体形成が阻害されることが明らかになった。
(2) kC9 - kC9の気孔のパターン形成に対する効果は、免疫シグナル伝達を活性化させることにより無効化する。そのため、kC9は気孔形成と免疫シグナル伝達経路に共通する因子に対して作用すると考え、両経路の共通因子であるMAPキナーゼに着目した。その結果、ｋC９はMPK6に結合すること、一過的にin vivoにおけるMAPK活性を阻害することがわかった。
(3) Chatty -アフィニティー担体を用いたプロテオミクスの結果、一連のステロイドホルモン合成酵素が標的因子候補とし同定された。実際に、Chattyを添加した暗所植物芽生えは、ブラシノステロイド欠損変異体と同様の光形態形成反応を示していた。これらの結果をもとにChattyの作用と、気孔形成とブラシノステロイドの作用に関して、既存ドグマと異なる知見を得た。

Research Progress Status

翌年度、交付申請を辞退するため、記入しない。

Strategy for Future Research Activity

翌年度、交付申請を辞退するため、記入しない。

Research Products

• [Int'l Joint Research] University of Washington/Texas A&M University(米国)
• [Int'l Joint Research] Swedish Univ. Agricultural Sciences(スウェーデン)
• [Int'l Joint Research] Spanish National Research Council(スペイン)
• [Int'l Joint Research] University of Cambridge/John Innes Centre(英国)
• [Int'l Joint Research] University of Zurich/University of Lausanne(スイス)
• [Int'l Joint Research]
• [Int'l Joint Research] テキサス大学/テキサスA&M大学/ワシントン大学(米国)
• [Int'l Joint Research] イーストアングリア大学/セインズベリー研究所(英国)
• [Int'l Joint Research] デュッセルドルフ大学(ドイツ)
• [Int'l Joint Research] スイス連邦工科大学（ETHチューリッヒ）(スイス)
• [Journal Article] Deceleration of the cell cycle underpins a switch from proliferative to terminal divisions in plant stomatal lineage (2022)
  • Author(s): Han Soon-Ki、Herrmann Arvid、Yang Jiyuan、Iwasaki Rie、Sakamoto Tomoaki、Desvoyes Benedicte、Kimura Seisuke、Gutierrez Crisanto、Kim Eun-Deok、Torii Keiko U.
  • Journal Title: Developmental Cell Volume: 57 Issue: 5 Pages: 569-582.e6
  • DOI: 10.1016/j.devcel.2022.01.014
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• [Journal Article] Intragenic suppressors unravel the role of the SCREAM ACT-like domain for bHLH partner selectivity in stomatal development (2022)
  • Author(s): Seo Hyemin、Sepuru Krishna Mohan、Putarjunan Aarthi、Aguirre Lyndsey、Burrows Benjamin A.、Torii Keiko U.
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  • DOI: 10.5511/plantbiotechnology.21.0508a
  • NAID: 130008091603
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  • Year and Date: 2021-09-25
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• [Journal Article] Effective range of non-cell autonomous activator and inhibitor peptides specifying plant stomatal patterning (2020)
  • Author(s): Zeng Scott M.、Lo Emily K. W.、Hazelton Bryna J.、Morales Miguel F.、Torii Keiko U.
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  • DOI: 10.1093/plphys/kiaa061
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  • Author(s): Qi Xingyun、Yoshinari Akira、Bai Pengfei、Maes Michal、Zeng Scott M、Torii Keiko U
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• [Journal Article] SCREAMing Twist on the Role of ICE1 in Freezing Tolerance (2020)
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  • DOI: 10.1105/tpc.20.00124
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• [Journal Article] Heat Shocking the Jedi Master: HSP90's Role in?Regulating Stomatal Cell Fate (2020)
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  • DOI: 10.1016/j.molp.2020.03.001
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• [Journal Article] Designer palmitoylation motif-based self-localizing ligand for sustained control of protein localization in living cells and Caenorhabditis elegans (2020)
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  • Journal Title: ACS Chemical Biology Volume: 15 Issue: 4 Pages: 837-843
  • DOI: 10.1021/acschembio.0c00014
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• [Journal Article] Chemogenetic control of protein anchoring to endomembranes in living cells with lipid-tethered small molecules (2020)
  • Author(s): Akinobu Nakamura, Rika Katahira, Shunsuke Sawada, Eri Shinoda, Keiko Kuwata, Tatsuyuki Yoshii, Shinya Tsukiji
  • Journal Title: Biochemistry Volume: 59 Issue: 2 Pages: 205-211
  • DOI: 10.1021/acs.biochem.9b00807
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• [Journal Article] Kleptoprotein bioluminescence: Parapriacanthus fish obtain luciferase from ostracod prey (2020)
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• [Journal Article] Dissecting plant hormone signaling with synthetic molecules: perspective from the chemists (2019)
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  • Journal Title: Current Opinion in Plant Biology Volume: 47 Pages: 32-37
  • DOI: 10.1016/j.pbi.2018.09.002
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• [Journal Article] Stomatal Development and Perspectives toward Agricultural Improvement (2019)
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  • Journal Title: Cold Spring Harbor Perspectives in Biology Volume: 11 Issue: 5 Pages: a034660-a034660
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• [Journal Article] Bipartite anchoring of SCREAM enforces stomatal initiation by coupling MAP kinases to SPEECHLESS (2019)
  • Author(s): Putarjunan Aarthi、Ruble Jim、Srivastava Ashutosh、Zhao Chunzhao、Rychel Amanda L.、Hofstetter Alex K.、Tang Xiaobo、Zhu Jian-Kang、Tama Florence、Zheng Ning、Torii Keiko U.
  • Journal Title: Nature Plants Volume: 5 Issue: 7 Pages: 742-754
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• [Journal Article] Chemical Synthesis of Atomically Tailored SUMO E2 Conjugating Enzymes for the Formation of Covalently Linked SUMO?E2?E3 Ligase Ternary Complexes (2019)
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• [Journal Article] Casein kinase 1 family regulates PRR5 and TOC1 in the Arabidopsis circadian clock (2019)
  • Author(s): Uehara TN.、Mizutani Y、Kuwata K、Hirota T、Sato A、Mizoi J、Takao S、Matsuo H、Suzuki T、Ito S、Saito AN.、Nishiwaki-Ohkawa T、Yamaguchi-Shinozaki K、Yoshimura T、Kay SA.、Itami K、Kinoshita T、Yamaguchi J、Nakamichi N.
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 116 Issue: 23 Pages: 11528-11536
  • DOI: 10.1073/pnas.1903357116
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• [Journal Article] Molecular Mechanism of Cellular Oxidative Stress Sensing by Keap1 (2019)
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  • Journal Title: Cell Reports Volume: 28 Issue: 3 Pages: 746-758
  • DOI: 10.1016/j.celrep.2019.06.047
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 気孔を増やす低分子化合物Chattyの標的探索 (2022)
  • Author(s): 中川彩美, 桑田啓子, 山田柊哉, Gregory J.P. Perry, 廣田毅, 佐藤綾人, 打田直行, 下遠野明恵, 伊丹健一郎, 村上慧, 鳥居啓子
  • Organizer: 第63回日本植物生理学会年会
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• [Presentation] Slowing Down for Perfection: Transcription factor-orchestrated cell cycle switch from proliferative- to-terminal division in plants (2021)
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  • Organizer: Southwest Society for Developmental Biology
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• [Presentation] 小胞ストレス応答因子の表皮細胞分化への役割の解析 (2021)
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  • Organizer: 第62回日本植物生理学会年会
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  • Author(s): 村上慧
  • Organizer: 第6回有機若手ワークショップ
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  • Organizer: GTR Annual Meeting 2020 (Online)
• [Presentation] Photoredox-catalyzed direct aminomethylation towards aryl bromides (2021)
  • Author(s): Jaehyun Jung, Yota Sakakibara, Kei Murakami, Kenichiro Itami
  • Organizer: 3rd ICReDD International Symposium
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• [Presentation] Photoredox-catalyzed decarboxylative direct aminomethylation towards aryl bromides: Straightforward synthesis of primary benzylamine (2021)
  • Author(s): Jaehyun Jung, Yota Sakakibara, Kei Murakami, Kenichiro Itami
  • Organizer: IRCCS 第４回国際シンポジウム “Multidimensional control over material structure and function”
  • Int'l Joint Research
• [Presentation] 光レドックス触媒が駆動するハロゲン化アリールの脱炭酸型アミノメチル化反応 (2021)
  • Author(s): Jaehyun Jung, Yota Sakakibara, Kei Murakami, Kenichiro Itami
  • Organizer: 日本化学会第101春季年会
• [Presentation] Decarboxylative transformation modulated by photocatalyst (2021)
  • Author(s): 榊原陽太,村上慧,伊丹健一郎
  • Organizer: 日本化学会第101春季年会
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  • Author(s): Keiko U. Torii
  • Organizer: Sainsbury Lab Symposium 2020 "Pluriopotency in Plant Development" 9.22-25.2020
  • Int'l Joint Research / Invited
• [Presentation] 窒素に着目した反応開発と生物活性分子の創出 (2020)
  • Author(s): 村上慧
  • Organizer: ACE meeting online II
  • Invited
• [Presentation] Photoredox-catalyzed Decarboxylative Cross-coupling of Aryl Halides and Glycine (2020)
  • Author(s): Jaehyun Jung, Yota Sakakibara, Kei Murakami, Kenichiro Itami
  • Organizer: The 9th JACI/GSC Symposium (Online)
• [Presentation] 光レドックス触媒が駆動するアリール酢酸の脱炭酸型変換 (2020)
  • Author(s): 榊原陽太,村上慧,伊丹健一郎
  • Organizer: ACE meeting online I
• [Presentation] Harnessing synthetic chemistry to dissect plant hormone signaling in development (2020)
  • Author(s): Keiko U. Torii
  • Organizer: International Symposium on Chemical Biology, Geneva, Switzerland, January 22-24, 2020
  • Int'l Joint Research / Invited
• [Presentation] Harnessing synthetic chemistry to understand and manipulate plant development (2019)
  • Author(s): Keiko U. Torii
  • Organizer: Walter E and Helen Parke Loomis Lecture and Symposium
  • Int'l Joint Research / Invited
• [Presentation] Cellular decision making during stomatal patterning and differentiation (2019)
  • Author(s): Keiko U. Torii
  • Organizer: EMBO Live Imaging Workshop, ITbM Nagoya, Nagoya, Japan, May 22, 2019
  • Int'l Joint Research / Invited
• [Presentation] Make decisions and keep moving forward (2019)
  • Author(s): Keiko U. Torii
  • Organizer: Nature Symposium “Plants of the Future”. NYU, New York, NY, USA, June 14, 2019
  • Int'l Joint Research / Invited
• [Presentation] Synthesis of imidazolone-cored heterocycles via sequential amidation/oxazole rearrangement and the discovery of its bioactivity in reducing the number of stomata (2019)
  • Author(s): Yip Shu Jan、中川 彩美、打田 直行、村上 慧、鳥居 啓子、伊丹 健一郎
  • Organizer: ITbMサイトビジット,P-18, 名古屋大学豊田講堂 東山キャンパス, 愛知, 2019年8月5, 6日
• [Presentation] Small compounds that increase the number of stomata: toward target identification (2019)
  • Author(s): Ayami Nakagawa, Shuya Yamada, Keiko Kuwata, Gregory J. P. Perry, Ayato Sato, Kei Murakami, Naoyuki Uchida, Kenichiro Itami, Keiko Torii
  • Organizer: ITbMサイトビジット, 名古屋大学豊田講堂 東山キャンパス, 愛知, 2019年8月5, 6日
• [Presentation] Synthetic study of arylated heteroaromatics: catalyst development and molecular probe synthesis for target identification of the stomata-increasing protein (2019)
  • Author(s): 山田 柊哉, 中川 彩美, Gregory J. P. Perry, Kaylin Nicole Flesch, 佐藤 綾人, 打田 直行, 鳥居 啓子, 村上 慧, 伊丹 健一郎
  • Organizer: ITbMサイトビジット,P-17, 名古屋大学豊田講堂 東山キャンパス, 愛知, 2019年8月5, 6日
• [Presentation] クロロフルオロアセタミドを利用した不可逆的キナーセ&阻害剤の開発 (2019)
  • Author(s): 進藤直哉、渕田大和、佐藤磨美、渡公佑、桑田啓子、小野眞弓、王子田彰夫
  • Organizer: 日本ケミカルバイオロジー学会第14回年会 2019年6月10-12日（名古屋）
• [Presentation] プロテオミクスとコア施設機能拡充とワークライフバランスと (2019)
  • Author(s): 桑田啓子
  • Organizer: 理研シンポジウム 「分子構造解析2019 : MSとNMRの基礎と実践」 2019年6月18日
  • Invited
• [Presentation] 日本の現状の紹介・男女共同参画の取り組み紹介・フリーディスカッション (2019)
  • Author(s): 桑田啓子
  • Organizer: 第15回質量分析技術者研究会 2019年8月27日（大阪）
• [Presentation] HIGH STEROL ESTER 1 is a key regulator in plant sterol biosynthesis (2019)
  • Author(s): Shimada, T. L., Shimada, T., Okazaki, Y., Higashi, Y., Saito, K., Kuwata, K., Oyama, K., Kato, M., Ueda, H., Nakano, A., Ueda, T., Takano, Y., & Hara-Nishimura, I
  • Organizer: Asian-Oceanian Symposium on Plant Lipids19-22 Nov 2019Australia
  • Int'l Joint Research / Invited
• [Presentation] 7-Aminomethyl-7-deazaguanine をもつPNA の合成 (2019)
  • Author(s): 森谷俊介、柴崎初音、桑田啓子、今村保忠、出水庸介、栗原正明、橘高敦史、杉山亨
  • Organizer: 第45回反応と合成の進歩シンポジウム2019年10月28日～29日
• [Book] 私の科学者ライフ (2021)
  • Author(s): 女性科学者に明るい未来をの会
  • Total Pages: 288
  • Publisher: 日本評論社
  • ISBN: 9784535789302
• [Remarks] Unraveling how one of the most important..
  • URL: https://research.utexas.edu/showcase/articles/view/unraveling-how-one-of-the-most-important-cell-types-form
• [Remarks] 植物の気孔はじっくり時間をかけてつくられる（名古屋大学 研究フロントライン）
  • URL: https://note.com/nagoya_ura/m/m1e09136b6f58
• [Patent(Industrial Property Rights)] US Provisional patent application. A super sensitive Auxin Induced Degradation (AID) system in mammalian cells using Synthetic, Engineered auxin-TIR1 pair (2020)
  • Inventor(s): KU Torii, K Nishimura 他５名
  • Industrial Property Rights Holder: KU Torii, K Nishimura 他５名
  • Industrial Property Rights Type: 特許
  • Filing Date: 2020
  • Overseas