不規則な環境変動に応答した気孔開度と花成の制御機構

• Project Area: Multi-layered regulatory system of plant resilience under fluctuating environment
• Project/Area Number: 20H05910
• Research Category: Grant-in-Aid for Transformative Research Areas (A)
• Allocation Type: Single-year Grants
• Review Section: Transformative Research Areas, Section (III)
• Research Institution: Nagoya University
• Principal Investigator: 木下 俊則 名古屋大学, 理学研究科(WPI), 教授 (50271101)
• Co-Investigator(Kenkyū-buntansha): 今泉 貴登 名古屋大学, 遺伝子実験施設, 招へい教員 (60767466) ; 児玉 豊 宇都宮大学, バイオサイエンス教育研究センター, 教授 (00455213)
• Project Period (FY): 2020-11-19 – 2025-03-31
• Project Status: Granted (Fiscal Year 2024)
• Budget Amount: ¥125,190,000 (Direct Cost: ¥96,300,000、Indirect Cost: ¥28,890,000); Fiscal Year 2024: ¥26,260,000 (Direct Cost: ¥20,200,000、Indirect Cost: ¥6,060,000); Fiscal Year 2023: ¥26,260,000 (Direct Cost: ¥20,200,000、Indirect Cost: ¥6,060,000); Fiscal Year 2022: ¥30,420,000 (Direct Cost: ¥23,400,000、Indirect Cost: ¥7,020,000); Fiscal Year 2021: ¥28,860,000 (Direct Cost: ¥22,200,000、Indirect Cost: ¥6,660,000); Fiscal Year 2020: ¥13,390,000 (Direct Cost: ¥10,300,000、Indirect Cost: ¥3,090,000)
• Keywords: 植物 / 環境変動 / 気孔 / 花成 / 光照射 / 光受容体

Outline of Research at the Start

自然界で実際に植物が晒されている不規則な環境変動により引き起こされる気孔開度制御や花成の段階的なステージゲート応答の分子機構について、参画研究者らのこれまでに培ってきた技術・経験を生かした生理・生化学的手法やイメージングを始め、新たな組織、細胞レベルでのオミクスアプローチも駆使して解明する。さらに、これらの知見に基づき気孔開度や花成をより精密に制御することで、植物の成長促進や収量増産の技術の確立を目指す。

Outline of Annual Research Achievements

植物の段階的な乾燥ステージゲート応答の分子機構を明らかにすることを目的として解析を進め、これまでの研究により、bHLH転写因子AKSs/FBHsが重要な役割を 果たすことが明らかとなってきた。そこで、今年度は、AKSs/FBHsのリン酸化部位の経時変化と共同沈降タンパク質について、プロテオミクス解析を進め、AKS1に おける乾燥処理に応答したリン酸化変動や共同沈降タンパク質を同定した。さらに、支援班の協力のもと、aks変異体と野生株の網羅的な発現解析を進め、転写因 子AKSの標的因子について、AKSのプロモーターへの結合実験を行った。 次に、変動光環境における気孔開閉の制御機構について解析を進め、暗転した葉では、葉内[CO2]が急激に高まり、それが孔辺細胞の細胞膜H+-ATPaseの迅速な脱リン酸化を引き起し、気孔閉鎖が誘導されていること、さらに、この葉内[CO2]上昇に応答したH+-ATPaseの急激な脱リン酸化のシグナル伝達に関わる分子実体が明らかとなってきた。また、気孔開口のシグナル伝達についての解析を進め、孔辺細胞の細胞膜H+-ATPaseの新規のリン酸化部位881番目のスレオニン残基が気孔開口に極めて重要な役割を果たしていることを証明し、論文として発表した。さらに気孔開度制御に関わる新規因子を同定し、現在、不均一環境に応答した気孔開度制御への関与 についての解析を進めている。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

大きな研究の柱としている段階的な乾燥ステージゲート応答において鍵となる転写因子を同定し、その標的が次々と明らかとなってきた。今後これら標的の分子機構を解析をすることで、植物における段階的な乾燥ステージゲート応答の分子レベルでの解明につながると考えられる。

Strategy for Future Research Activity

これまでに明らかとなったきた植物の段階的な乾燥ステージゲート応答や変動光環境における気孔開度制御において関わっている因子の孔辺細胞内での具体的な 作用機作の解析をさらに進めることで、本領域の大きな目標である不均一環境における植物の環境応答の分子機構を明らかにしたい。

Research Products

• [Int'l Joint Research] 南京農業大学/北京大学(中国)
• [Int'l Joint Research] ミネソタ大学(米国)
• [Int'l Joint Research] 南京農業大学/北京大学(中国)
• [Int'l Joint Research] IST Austria(オーストリア)
• [Int'l Joint Research] 南京農業大学(中国)
• [Int'l Joint Research] ミネソタ大学(米国)
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  • Journal Title: Plant Molecular Biology Volume: 110 Issue: 3 Pages: 269-285
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  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Ethanol-Mediated Novel Survival Strategy against Drought Stress in Plants (2022)
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• [Journal Article] ABP1-TMK auxin perception for global phosphorylation and auxin canalization (2022)
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• [Journal Article] Elevated CO2 induces rapid dephosphorylation of plasma membrane H+-ATPase in guard cells. (2022)
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• [Journal Article] Tryptophan synthase B subunit 1 affects stomatal phenotypes in Arabidopsis thaliana. (2022)
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• [Journal Article] Photosynthetic-Product?Dependent Activation of Plasma Membrane H+-ATPase and Nitrate Uptake in<i>Arabidopsis</i>Leaves (2022)
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• [Journal Article] Stomatal properties of Arabidopsis cauline and rice flag leaves and their contributions to seed production and grain yield (2022)
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• [Journal Article] The endoplasmic reticulum membrane-bending protein RETICULON facilitates chloroplast relocation movement in Marchantia polymorpha (2022)
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• [Journal Article] Type 2C protein phosphatase clade D family members dephosphorylate guard cell plasma membrane H+-ATPase (2021)
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• [Journal Article] Identification of Abscisic Acid-Dependent Phosphorylated Basic Helix-Loop-Helix Transcription Factors in Guard Cells of Vicia faba by Mass Spectrometry (2021)
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• [Journal Article] Promotion and Upregulation of a Plasma Membrane Proton-ATPase Strategy: Principles and Applications (2021)
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• [Journal Article] TMK-based cell-surface auxin signalling activates cell-wall acidification (2021)
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• [Journal Article] Overexpression of Plasma Membrane H+-ATPase in Guard Cells Enhances Light-Induced Stomatal Opening, Photosynthesis, and Plant Growth in Hybrid Aspen (2021)
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• [Journal Article] Identification of Genes Preferentially Expressed in Stomatal Guard Cells of Arabidopsis thaliana and Involvement of the Aluminum-Activated Malate Transporter 6 Vacuolar Malate Channel in Stomatal Opening (2021)
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• [Journal Article] Molecular basis of plasma membrane H+-ATPase function and potential application in the agricultural production (2021)
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• [Journal Article] Protease Inhibitor-Dependent Inhibition of Light-Induced Stomatal Opening (2021)
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• [Journal Article] An Affordable Image-Analysis Platform to Accelerate Stomatal Phenotyping During Microscopic Observation (2021)
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  • Peer Reviewed / Int'l Joint Research
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  • Peer Reviewed
• [Journal Article] The photo-thermochemical properties and functions of Marchantia phototropin encoded by an unduplicated gene in land plant evolution (2021)
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  • Journal Title: Journal of Photochemistry and Photobiology B: Biology Volume: 224 Pages: 112305-112305
  • DOI: 10.1016/j.jphotobiol.2021.112305
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• [Journal Article] Steric and Electronic Interactions at Gln154 in ZEITLUPE Induce Reorganization of the LOV Domain Dimer Interface (2021)
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  • Journal Title: Biochemistry Volume: 60 Issue: 2 Pages: 95-103
  • DOI: 10.1021/acs.biochem.0c00819
  • Peer Reviewed
• [Journal Article] Flowering Times of Wild Arabidopsis Accessions From Across Norway Correlate With Expression Levels of FT, CO, and FLC Genes (2021)
  • Author(s): Kinmonth-Schultz Hannah、Lewandowska-Sabat Anna、Imaizumi Takato、Ward Joy K.、Rognli Odd Arne、Fjellheim Siri
  • Journal Title: Frontiers in Plant Science Volume: 12
  • DOI: 10.3389/fpls.2021.747740
  • Peer Reviewed
• [Journal Article] Stomatal immunity against fungal invasion comprises not only chitin-induced stomatal closure but also chitosan-induced guard cell death (2020)
  • Author(s): Ye Wenxiu、Munemasa Shintaro、Shinya Tomonori、Wu Wei、Ma Tao、Lu Jiang、Kinoshita Toshinori、Kaku Hanae、Shibuya Naoto、Murata Yoshiyuki
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 117 Issue: 34 Pages: 20932-20942
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  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] DNA methylation is reconfigured at the onset of reproduction in rice shoot apical meristem (2020)
  • Author(s): Higo Asuka、Saihara Noriko、Miura Fumihito、Higashi Yoko、Yamada Megumi、Tamaki Shojiro、Ito Tasuku、Tarutani Yoshiaki、Sakamoto Tomoaki、Fujiwara Masayuki、Kurata Tetsuya、Fukao Yoichiro、Moritoh Satoru、Terada Rie、Kinoshita Toshinori、Ito Takashi、Kakutani Tetsuji、Shimamoto Ko、Tsuji Hiroyuki
  • Journal Title: Nature Communications Volume: 11 Issue: 1 Pages: 1-1
  • DOI: 10.1038/s41467-020-17963-2
  • Peer Reviewed / Open Access
• [Journal Article] CIPK23 regulates blue light-dependent stomatal opening in Arabidopsis thaliana. (2020)
  • Author(s): 1.Inoue S, Kaiserli E, Zhao X, Waksman T, Takemiya A, Okumura M, Takahashi H, Seki M, Shinozaki K, Endo Y, Sawasaki T, Kinoshita T, Zhang X, Christie JM, Shimazaki K
  • Journal Title: The Plant Journal Volume: 104 Issue: 3 Pages: 679-692
  • DOI: 10.1111/tpj.14955
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Role of Proton Motive Force in Photoinduction of Cytoplasmic Streaming in Vallisneria Mesophyll Cells (2020)
  • Author(s): Harada Akiko、Okazaki Yoshiji、Kinoshita Toshinori、Nagai Reiko、Takagi Shingo
  • Journal Title: Plants Volume: 9 Issue: 3 Pages: 376-376
  • DOI: 10.3390/plants9030376
  • Peer Reviewed / Open Access
• [Presentation] CRISPR/Cas9 によるシロイヌナズナ細胞膜H+-ATPase の発現制御と表現型解析 (2024)
  • Author(s): 木下俊則
  • Organizer: 日本植物生理学会
• [Presentation] Light regulation of stomat al movement and plasma membrane H+-ATPase in guard cells (2023)
  • Author(s): Kinoshita T
  • Organizer: the 33rd International Conference on Arabidopsis Research
  • Int'l Joint Research / Invited
• [Presentation] タイプ2CプロテインホスファターゼDクレードによる気孔開度制御 (2023)
  • Author(s): 木下大地，水谷未耶，榊原拓，村上公亮，丹下昭憲，安藤英伍，木下俊則
  • Organizer: 第64回日本植物生理学会年会
• [Presentation] シロイヌナズナ実生での細胞膜 H+-ATPase 活性化が包括的な遺伝子発現に与える影響 (2023)
  • Author(s): 木下(永縄)悟，瀧京美，永縄万由子，長谷川莉子，桐山寛生，大神田淳子，木下俊則
  • Organizer: 第64回日本植物生理学会年会
• [Presentation] CO2 による孔辺細胞の細胞膜 H+-ATPase の急速な脱リン酸化を通じた気孔閉鎖促進 (2023)
  • Author(s): 安藤英伍，Hannes Kollist，深津孝平，木下俊則，寺島一郎
  • Organizer: 第64回日本植物生理学会年会
• [Presentation] ストレプト藻類クレブソルミディウムの細胞膜 H+-ATPase 活性制御 (2023)
  • Author(s): 高橋宏二，堀孝一，太田啓之，木下俊則
  • Organizer: 第64回日本植物生理学会年会
• [Presentation] 気孔開口シグナル伝達経路に作用す る新規化合物の分子機構の解析 (2023)
  • Author(s): 辛洸徹，相原悠介，藤茂雄，佐藤綾人 ，木下俊則
  • Organizer: 第64回日本植物生理学会年会
• [Presentation] キナーゼ阻害剤による気孔開口抑制の分子機構の解析 (2023)
  • Author(s): 桑山翔悟，高橋宏二，林真妃，佐藤綾人，木下俊則
  • Organizer: 第64回日本植物生理学会年会
• [Presentation] 気孔孔辺細胞において青色光によりリン酸化 レベルの変動するタンパク質の機能解析 (2023)
  • Author(s): 深津孝平，林優紀，鈴木孝征，桑田啓子，木下俊則
  • Organizer: 第64回日本植物生理学会年会
• [Presentation] イソチオシアネートを基盤とした気孔開口阻 害剤の機能解析と分子改良 (2023)
  • Author(s): 相原悠介，前田文平，後藤栞奈，野元美佳，高橋宏二，藤茂雄，叶文秀，戸田陽介，多田安臣，佐藤綾人，伊丹健一郎，村上慧，木下俊則
  • Organizer: 第64回日本植物生理学会年会
• [Presentation] アミン合成酵素の機能解析 (2023)
  • Author(s): 祢冝淳太郎，小畑智暉，西村さくら，宋普錫，山柿将，星野奈摘，深津孝平，木下俊則，西 田生郎，射場厚
  • Organizer: 第64回日本植物生理学会年会
• [Presentation] SNS1 を介した長期乾燥ストレス条件への抵抗戦略 (2023)
  • Author(s): 片桐壮太郎，神山佳明，木下俊則，梅澤泰史
  • Organizer: 第64回日本植物生理学会年会
• [Presentation] 孔辺細胞における細胞膜 H+-ATPase の C 末端 のリン酸化を介した新規機能制御の解析 (2023)
  • Author(s): 林優紀，深津孝平，高橋宏二，木下(永縄)悟，加藤恭平 ，桑田啓子 ，鈴木孝征 ，木下俊則
  • Organizer: 第64回日本植物生理学会年会
• [Presentation] TCP転写因子による葉の細胞伸長の制御機構の解析 (2023)
  • Author(s): 小山知嗣，光田展隆，関原明，高橋宏二，木下俊則，別所歩武，國枝正，出村拓，高木優
  • Organizer: 第64回日本植物生理学会年会
• [Presentation] 植物の成長の謎にせまる気孔の科学のはなし (2023)
  • Author(s): 木下俊則
  • Organizer: 名古屋大学オープンレクチャー
  • Invited
• [Presentation] Light regulation of stomatal opening and control of plant growth (2022)
  • Author(s): Toshinori KInoshita
  • Organizer: Stomatal development and function seminar
  • Int'l Joint Research / Invited
• [Presentation] 植物の巧みな環境応答 -植物に欠かせない気孔の働き- (2022)
  • Author(s): 木下俊則
  • Organizer: 2022年度 名大 MIRAI GSC 第1ステージ講義
  • Invited
• [Presentation] How plasma membrane H+-ATPase overexpression increases rice yield (2022)
  • Author(s): Toshinori Kinoshita
  • Organizer: Plant Biology 2022
  • Int'l Joint Research / Invited
• [Presentation] Analysis of phosphoenolpyruvate carboxylase in stomatal guard cells (2022)
  • Author(s): Kohei Fukatsu
  • Organizer: Plant Biology 2022
  • Int'l Joint Research
• [Presentation] Functional characterization of novel compounds that affect signalingpathway in stomatal opening (2022)
  • Author(s): Gwangchol Sin
  • Organizer: Plant Biology 2022
  • Int'l Joint Research
• [Presentation] 植物の巧みな環境応答 (2022)
  • Author(s): 木下俊則
  • Organizer: 長野県立飯山高等学校探求科「名古屋サイエンスツアー・世界を変える研究の最前線！これからの社会と地球の未来を考える」
  • Invited
• [Presentation] タイプ2CプロテインホスファターゼD clade (PP2C.D)による孔辺細胞細胞膜プロトンポンプの脱リン酸化 (2022)
  • Author(s): 木下俊則
  • Organizer: 日本植物学会第86回大会
• [Presentation] イソチオシアネートを基盤とした植物の乾燥耐性付与剤の発見と改良 (2022)
  • Author(s): 相原 悠介，前田 文平，後藤 栞奈，藤 茂雄，叶 文 秀，戸田 陽介，村上 慧，佐藤 綾人，伊丹 健一郎，木下 俊則
  • Organizer: 日本植物学会第86回大会
• [Presentation] ソラマメ孔辺細胞においてABA依存的にリン酸化されるbHLH型転写因子の同定と機能解析 (2022)
  • Author(s): 林 優紀，高橋 洋平，桑田 啓子，鈴木 孝征，木下 俊則
  • Organizer: 日本植物学会第86回大会
• [Presentation] 気孔開度制御による植物の成長と生産量の促進 (2022)
  • Author(s): 木下俊則
  • Organizer: BioJapan2022
• [Presentation] ポンプ植物作出による光合成、養分吸収、収量の促進 (2022)
  • Author(s): 木下俊則
  • Organizer: アグリビジネス創出フェア
• [Presentation] How plasma membrane H+-ATPase overexpression increases rice yield (2022)
  • Author(s): Toshinori Kinoshita
  • Organizer: Plant Biology 2022, Plant Biotechnology Symposium
  • Int'l Joint Research / Invited
• [Presentation] 不規則な環境変動に応答した気孔開度と花成の制御機構 (2022)
  • Author(s): 木下俊則
  • Organizer: 第63回日本植物生理学会年会シンポジウム
  • Invited
• [Presentation] 光合成依存的な細胞膜H+-ATPaseの活性化を介したシロイヌナズナ葉での硝酸取り込みの解析 (2022)
  • Author(s): 木下悟，鈴木孝征，木羽隆敏，榊原均，木下俊則
  • Organizer: 第63回日本植物生理学会年会
• [Presentation] 時計長周期化化合物と標的タンパ クの結合モデル (2022)
  • Author(s): 前田明里，松尾宏美，藤本和宏，木下俊則，山口潤一郎，中道範人
  • Organizer: 第63回日本植物生理学会年会
• [Presentation] 植物概日時計の温度補償性に関わるタンパク質の量的制 御メカニズムの解析 (2022)
  • Author(s): 前田明里，松尾宏美，木下俊則，中道範人
  • Organizer: 第63回日本植物生理学会年会
• [Presentation] 新奇液胞膜型マグネシウム輸送体の気孔開口への関与 (2022)
  • Author(s): 井上晋一郎，林真妃，黄勝，横正健剛，後藤栄治，池松朱夏，奥村将樹，鈴木孝征，木下俊則，馬建鋒
  • Organizer: 第63 回日本植物生理学会年会
• [Presentation] 気孔の光シグナル伝達と光合成や成長の制御機構 (2021)
  • Author(s): 木下俊則
  • Organizer: 日本植物学会第85回大会シンポジウム
  • Invited
• [Presentation] シロイヌナズナ葉における光合成依存的な細胞膜H+-ATPase活性化とのSAUR遺伝子の関与について (2021)
  • Author(s): 木下 悟，鈴木 孝征，木下俊則
  • Organizer: 日本植物学会第85 回大会
• [Presentation] PEPカルボキシラーゼの光による気孔開口の関与と分子機構の解明 (2021)
  • Author(s): 深津 孝平，林 優紀，鈴木 孝征，桑田 啓子，木下俊則
  • Organizer: 日本植物学会第85回大会
• [Presentation] Analysis of mechanism of photosynthesis-dependent plasma membrane H+- ATPase activation in Arabidopsis leaf (2021)
  • Author(s): Kinoshita S, Suzuki T, Kinoshita T
  • Organizer: International Symposium on Plant Photobiology (ISPP) 2021
• [Presentation] 植物の巧みな環境応答 (2021)
  • Author(s): 木下俊則
  • Organizer: 名古屋大学オープンキャンパス・ライブ講義
  • Invited
• [Presentation] 植物を操る- 植物に学び、秘められた能力を引き出す (2021)
  • Author(s): 木下俊則
  • Organizer: 名古屋大学オンラインセミナー “MEBINAR”
  • Invited
• [Presentation] 気孔開口を制御する小分子の作用点解析と分子改良 (2021)
  • Author(s): 相原 悠介、藤 茂雄、戸田 陽介、井上 心平、佐藤 綾人、村上 慧、伊丹 健一郎、木下 俊則
  • Organizer: 日本植物生理学会
• [Presentation] 気孔開口シグナル伝達経路に作用する新規化合物の分子機構の解析 (2021)
  • Author(s): 辛 洸徹、相原 悠介、藤 茂雄、井上 心平、佐藤 綾人、木下 俊則
  • Organizer: 日本植物生理学会
• [Presentation] シロイヌナズナ葉における光合成炭素代謝物と細胞膜 H+-ATPase の関係について (2021)
  • Author(s): 木下 悟、木下 俊則
  • Organizer: 日本植物生理学会
• [Presentation] ソラマメ孔辺細胞を用いた質量分析による AKS1 オーソログの同定 (2021)
  • Author(s): 林 優紀、高橋 洋平、桑田 啓子、鈴木 孝征、木下 俊則
  • Organizer: 日本植物生理学会
• [Presentation] 環境刺激による多様な気孔開度制御機構 (2020)
  • Author(s): 木下俊則
  • Organizer: 日本植物学会
  • Invited
• [Book] ＪＡＴＡＦＦジャーナル１０巻９号「プロトンポンプの機能を高めることによりイネの収量向上と肥料の削減を実現」 (2022)
  • Author(s): 木下俊則
  • Publisher: 公益社団法人農林水産・食品産業技術振興協会
• [Book] 月刊誌「植調」 第55巻第9号 (2021)
  • Author(s): 木下俊則
  • Publisher: 公益財団法人 日本植物調節剤研究協会
• [Book] 光合成 (2021)
  • Author(s): 日本光合成学会
  • Total Pages: 224
  • Publisher: 朝倉書店
  • ISBN: 9784254171761
• [Remarks] 名古屋大学植物生理学グループ
  • URL: http://plantphys.bio.nagoya-u.ac.jp/index.html
• [Remarks]
  • URL: http://plantphys.bio.nagoya-u.ac.jp
• [Patent(Industrial Property Rights)] 植物気孔開口調節剤 (2022)
  • Inventor(s): 木下俊則、相原悠介、佐藤綾人、村上慧
  • Industrial Property Rights Holder: 木下俊則、相原悠介、佐藤綾人、村上慧
  • Industrial Property Rights Type: 特許
  • Filing Date: 2022