ゼニゴケ油体をモデルとしたオルガネラ周期の証明と中心因子の同定

• Project Area: Intrinsic periodicity of cellular systems and its modulation as the driving force behind plant development
• Project/Area Number: 19H05675
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: National Institute for Basic Biology
• Principal Investigator: 上田 貴志 基礎生物学研究所, 細胞動態研究部門, 教授 (10311333)
• Co-Investigator(Kenkyū-buntansha): 近藤 洋平 大学共同利用機関法人自然科学研究機構(新分野創成センター、アストロバイオロジーセンター、生命創成探究, 生命創成探究センター, 助教 (00724444) ; 河内 孝之 京都大学, 生命科学研究科, 教授 (40202056) ; 金澤 建彦 基礎生物学研究所, 細胞動態研究部門, 助教 (60802783)
• Project Period (FY): 2019-06-28 – 2024-03-31
• Project Status: Granted (Fiscal Year 2021)
• Budget Amount: ¥169,910,000 (Direct Cost: ¥130,700,000、Indirect Cost: ¥39,210,000); Fiscal Year 2021: ¥32,760,000 (Direct Cost: ¥25,200,000、Indirect Cost: ¥7,560,000); Fiscal Year 2020: ¥32,760,000 (Direct Cost: ¥25,200,000、Indirect Cost: ¥7,560,000); Fiscal Year 2019: ¥34,970,000 (Direct Cost: ¥26,900,000、Indirect Cost: ¥8,070,000)
• Keywords: ゼニゴケ / 油体周期 / イメージング / トランスクリプトーム / 油体 / 分泌経路 / オルガネラ周期

Outline of Research at the Start

油体周期をオルガネラ周期のモデルとし，油体形成過程の高精細ライブイメージングと画像解析に一細胞トランスクリプトーム解析を組み合わせ，油体周期の各相を分離することで油体周期の実態を明らかにする．また，生体分子ダイナミクス解析により油体周期の中心振動子を推定するとともに，その分子機能を実験的に検証する．続いて環境変動を与えた際の油体周期の変調が油体の分布パターンの変化として表出する仕組みを解明する．

Outline of Annual Research Achievements

油体周期の存在をライブイメージングで実証するべく準備を進めた。油体周期仮説では、分泌経路の方向が細胞膜方向に向いている細胞膜期と油体方向に向いている油体期が相互に転換することを想定している。我々はこれまでに，相同な2種の細胞膜局在タンパク質（MpSYP12BおよびMpSYP13B）がそれぞれ油体膜と細胞膜に局在し，この局在が転写レベルで制御されていることを見いだしている.このことから，MpSYP12Bが油体期に，MpSYP13Bが細胞膜器に発現しているとの仮説を立て，これをプロモーターレポーターアッセイにより検証した．MpSYP12BプロモーターにPEST配列を連結したELucを発現し発光を検出したところ，発光が周期的に増強と減弱を繰り返す様子が観察された．これにより，油体周期の実在が強く支持された．現在MpSYP13Bのプロモータ活性をLucRを用いて検出する系の確立を行っており，今後2色の発光を用いて油体周期を可視化する予定である．
油体周期が発振するタイミングについても重要な知見を得た．mCitrine-MpSYP12BとmRFP-MpSYP13Bを発現する植物を用いて無性芽の長時間タイムラプス観察を行ったところ，MpSYP12Bにより油体膜が認識できるようになる約12時間前に油体母細胞が不等分裂し，娘細胞のうちの小さい細胞に油体が形成される様子が複数回観察された．このことから，油体細胞への運命決定が不等分裂にともなって起こり，この不等分裂が油体周期発振のキューとなっている可能性が考えられる．現在この不等分裂に関わる因子の探索を進めるとともに，近藤と協働して将来油体細胞へと分化する細胞の特徴抽出に取り組んでいる．

Current Status of Research Progress

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

Reason

植物育成装置の不具合のため当初の研究計画よりもバイオイメージング解析に時間を要したが、今年度の研究活動により上述の通り順調に成果があがっている。

Strategy for Future Research Activity

今後は引き続きバイオイメージング解析を中心とした油体周期仮説の検証を進めるのに加え、単一油体細胞核トランスクリプトーム解析に向けた条件検討を進める。

Research Products

• [Int'l Joint Research] Monash University/The University of Melbourne(オーストラリア)
• [Int'l Joint Research] Heidelberg University/Max Planck Institute/Osnabruck University(ドイツ)
• [Int'l Joint Research] Universidad Nacional del Litoral(アルゼンチン)
• [Journal Article] Fungal-Type Terpene Synthases in Marchantia polymorpha Are Involved in Sesquiterpene Biosynthesis in Oil Body Cells (2021)
  • Author(s): Takizawa Ryosuke、Hatada Miki、Moriwaki Yuta、Abe Sachika、Yamashita Yuko、Arimitsu Ryoma、Yamato Katsuyuki T、Nishihama Ryuichi、Kohchi Takayuki、Koeduka Takao、Chen Feng、Matsui Kenji
  • Journal Title: Plant and Cell Physiology Volume: in press
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  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Hierarchical modeling of mechano-chemical dynamics of epithelial sheets across cells and tissue (2021)
  • Author(s): Asakura Yoshifumi、Kondo Yohei、Aoki Kazuhiro、Naoki Honda
  • Journal Title: Scientific Reports Volume: 11 Pages: 1-15
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  • Peer Reviewed / Open Access
• [Journal Article] Temperature-dependent fasciation mutants provide a link between mitochondrial RNA processing and lateral root morphogenesis (2021)
  • Author(s): Otsuka Kurataka、Mamiya Akihito、Konishi Mineko、Nozaki Mamoru、Kinoshita Atsuko、Tamaki Hiroaki、Arita Masaki、Saito Masato、Yamamoto Kayoko、Hachiya Takushi、Noguchi Ko、Ueda Takashi、Yagi Yusuke、Kobayashi Takehito、Nakamura Takahiro、Sato Yasushi、Hirayama Takashi、Sugiyama Munetaka
  • Journal Title: eLife Volume: 10
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  • Peer Reviewed / Open Access
• [Journal Article] Cargo sorting zones in the trans-Golgi network visualized by super-resolution confocal live imaging microscopy in plants (2021)
  • Author(s): Shimizu Yutaro、Takagi Junpei、Ito Emi、Ito Yoko、Ebine Kazuo、Komatsu Yamato、Goto Yumi、Sato Mayuko、Toyooka Kiminori、Ueda Takashi、Kurokawa Kazuo、Uemura Tomohiro、Nakano Akihiko
  • Journal Title: Nature Communications Volume: 12 Pages: 1901-1901
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  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identification of genes involved in <i>Meloidogyne incognita</i>-induced gall formation processes in <i>Arabidopsis thaliana</i> (2021)
  • Author(s): Suzuki Reira、Ueda Takashi、Wada Takuji、Ito Masaki、Ishida Takashi、Sawa Shinichiro
  • Journal Title: Plant Biotechnology Volume: 38 Issue: 1 Pages: 1-1
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  • ISSN: 1342-4580, 1347-6114
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Chromatin organization in early land plants reveals an ancestral association between H3K27me3, transposons, and constitutive heterochromatin. (2020)
  • Author(s): Montgomery SA, Tanizawa Y, Galik B, Wang N, Ito T, Mochizuki T, Akimcheva S, Bowman JL, Cognat V, Drouard L, Ekker H, Houng SF, Kohchi T, Lin SS, Liu LYD, Nakamura Y, Valeeva LR, Shakirov EV, Shippen DE, Wei WL, Yagura M, Yamaoka S, Yamato KT, Liu C, Berger F.
  • Journal Title: Curr. Biol. Volume: 30 Pages: 573-588
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  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Regulation of the Poly(A) Status of Mitochondrial mRNA by Poly(A)-Specific Ribonuclease Is Conserved among Land Plants (2020)
  • Author(s): Kanazawa Mai、Ikeda Yoko、Nishihama Ryuichi、Yamaoka Shohei、Lee Nam-Hee、Yamato Katsuyuki T、Kohchi Takayuki、Hirayama Takashi
  • Journal Title: Plant Cell Physiol. Volume: 61 Pages: 470-480
  • DOI: 10.1093/pcp/pcz212
  • Peer Reviewed
• [Journal Article] Kinase in the liverwort Marchantia polymorpha; A photosynthesis-dependent kinase for carbon metabolism (2020)
  • Author(s): Koide E, Suetsugu N, Iwano M, Gotoh E, Nomura Y, Stolze SC, Nakagami H, Kohchi T, Nishihama R
  • Journal Title: Plant and Cell Physiology Volume: 61 Pages: 631-643
  • DOI: 10.1093/pcp/pcz232
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Positional cues regulate dorsal organ formation in the liverwort Marchantia polymorpha (2020)
  • Author(s): Suzuki H, Harrison CJ, Shimamura M, Kohchi T, Nishihama R
  • Journal Title: Journal of Plant Research Volume: 133 Pages: 311-321
  • DOI: 10.1007/s10265-020-01180-5
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Design principles of a minimal auxin response system (2020)
  • Author(s): Kato Hirotaka、Mutte Sumanth K.、Suzuki Hidemasa、Crespo Isidro、Das Shubhajit、Radoeva Tatyana、Fontana Mattia、Yoshitake Yoshihiro、Hainiwa Emi、van den Berg Willy、Lindhoud Simon、Ishizaki Kimitsune、Hohlbein Johannes、Borst Jan Willem、Boer D. Roeland、Nishihama Ryuichi、Kohchi Takayuki、Weijers Dolf
  • Journal Title: Nature Plants Volume: 6 Pages: 473-482
  • DOI: 10.1038/s41477-020-0662-y
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Deep Imaging Analysis in VISUAL Reveals the Role of YABBY Genes in Vascular Stem Cell Fate Determination (2020)
  • Author(s): Nurani Alif Meem、Ozawa Yasuko、Furuya Tomoyuki、Sakamoto Yuki、Ebine Kazuo、Matsunaga Sachihiro、Ueda Takashi、Fukuda Hiroo、Kondo Yuki
  • Journal Title: Plant Cell Physiol. Volume: 61 Pages: 255-264
  • DOI: 10.1093/pcp/pcaa002
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Induction of Multichotomous Branching by CLAVATA Peptide in Marchantia polymorpha (2020)
  • Author(s): Hirakawa Yuki、Fujimoto Toko、Ishida Sakiko、Uchida Naoyuki、Sawa Shinichiro、Kiyosue Tomohiro、Ishizaki Kimitsune、Nishihama Ryuichi、Kohchi Takayuki、Bowman John L.
  • Journal Title: Curr. Biol Volume: 30 Pages: 3833-3840
  • DOI: 10.1016/j.cub.2020.07.016
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The Arabidopsis V-ATPase is localized to the TGN/EE via a seed plant-specific motif (2020)
  • Author(s): Lupanga U., Rohrich R., Askani J., Hilmer S., Kiefer C., Krebs M., Kanazawa T., Ueda T., Schumacher K.
  • Journal Title: eLife Volume: 9
  • DOI: 10.7554/elife.60568
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Improvement of Phycocyanobilin Synthesis for Genetically Encoded Phytochrome-Based Optogenetics (2020)
  • Author(s): Uda Youichi、Miura Haruko、Goto Yuhei、Yamamoto Kei、Mii Yusuke、Kondo Yohei、Takada Shinji、Aoki Kazuhiro
  • Journal Title: ACS Chemical Biology Volume: 15 Pages: 2896-2906
  • DOI: 10.1021/acschembio.0c00477
  • Peer Reviewed / Open Access
• [Journal Article] The liverwort oil body is formed by redirection of the secretory pathway (2020)
  • Author(s): Kanazawa T., Morinaka H., Ebine K., Shimada T.L., Ishida S., Minamino N., Yamaguchi K., Shigenobu S., Kohchi T., Nakano A., Ueda T.
  • Journal Title: Nature Communications Volume: 11 Pages: 6152-6152
  • DOI: 10.1038/s41467-020-19978-1
  • Peer Reviewed / Open Access
• [Journal Article] Oil Body Formation in Marchantia polymorpha Is Controlled by MpC1HDZ and Serves as a Defense against Arthropod Herbivores (2020)
  • Author(s): Romani F., Banic E., Florent S.N., Kanazawa T., Goodger J.Q.D., Mentink R.A., Dierschke T., Zachgo S., Ueda T., Bowman J.L., Tsiantis M., Moreno J.E.
  • Journal Title: Current Biology Volume: 30
  • DOI: 10.1016/j.cub.2020.05.081
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Characterization of ancestral myosin XI from Marchantia polymorpha by heterologous expression in Arabidopsis thaliana (2020)
  • Author(s): Duan Zhongrui、Tanaka Misato、Kanazawa Takehiko、Haraguchi Takeshi、Takyu Akiko、Era Atsuko、Ueda Takashi、Ito Kohji、Tominaga Motoki
  • Journal Title: The Plant Journal Volume: 104 Pages: 460-473
  • DOI: 10.1111/tpj.14937
  • Peer Reviewed
• [Journal Article] Longin R-SNARE is retrieved from the plasma membrane by ANTH domain-containing proteins in Arabidopsis (2020)
  • Author(s): Fujimoto Masaru、Ebine Kazuo、Nishimura Kohji、Tsutsumi Nobuhiro、Ueda Takashi
  • Journal Title: Proc. Natl. Acad. Sci. USA. Volume: 117 Pages: 25150-25158
  • DOI: 10.1073/pnas.2011152117
  • Peer Reviewed / Open Access
• [Journal Article] Purification and Interaction Analysis of a Plant-Specific RAB5 Effector by In Vitro Pull-Down Assay (2020)
  • Author(s): Ito Emi、Choi Seung-won、Ueda Takashi
  • Journal Title: Methods in Molecular Biology Volume: 2177 Pages: 183-197
  • DOI: 10.1007/978-1-0716-0767-1_15
  • Peer Reviewed
• [Journal Article] Role of Autophagy in Male Reproductive Processes in Land Plants (2020)
  • Author(s): Norizuki Takuya、Minamino Naoki、Ueda Takashi
  • Journal Title: Frontiers in Plant Science Volume: 11 Pages: 756-756
  • DOI: 10.3389/fpls.2020.00756
  • Peer Reviewed / Open Access
• [Journal Article] Cytokinin signaling is essential for organ formation in Marchantia polymorpha (2019)
  • Author(s): Aki Shiori S、Mikami Tatsuya、Naramoto Satoshi、Nishihama Ryuichi、Ishizaki Kimitsune、Kojima Mikiko、Takebayashi Yumiko、Sakakibara Hitoshi、Kyozuka Junko、Kohchi Takayuki、Umeda Masaaki
  • Journal Title: Plant Cell Physiol. Volume: 60 Pages: 1842-1854
  • DOI: 10.1093/pcp/pcz100
  • NAID: 120006711216
  • Peer Reviewed
• [Journal Article] Cytokinin signaling coordinates development of diverse organs in Marchantia polymorpha (2019)
  • Author(s): Aki Shiori S.、Nishihama Ryuichi、Kohchi Takayuki、Umeda Masaaki
  • Journal Title: Plant Signal. Behav. Volume: 14 Pages: 1668232-1668232
  • DOI: 10.1080/15592324.2019.1668232
  • Peer Reviewed
• [Journal Article] The RopGEF KARAPPO Is Essential for the Initiation of Vegetative Reproduction in Marchantia polymorpha (2019)
  • Author(s): Hiwatashi T., Goh H., Yasui Y., Koh L.Q., Takami H., Kajikawa M., Kirita H., Kanazawa T., Minamino N., Togawa T., Sato M., Wakazaki M., Yamaguchi K., Shigenobu S., Fukaki H., Mimura T., Toyooka K., Sawa S., Yamato K.T.,Ueda T., Urano D., Kohchi T., Ishizaki K.
  • Journal Title: Curr. Biol. Volume: 29 Pages: 3525-3531
  • DOI: 10.1016/j.cub.2019.08.071
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] An Early Arising Role of the MicroRNA156/529-SPL Module in Reproductive Development Revealed by the Liverwort Marchantia polymorpha (2019)
  • Author(s): Tsuzuki Masayuki、Futagami Kazutaka、Shimamura Masaki、Inoue Chikako、Kunimoto Kan、Oogami Takashi、Tomita Yuki、Inoue Keisuke、Kohchi Takayuki、Yamaoka Shohei、Araki Takashi、Hamada Takahiro、Watanabe Yuichiro
  • Journal Title: Current Biology Volume: 29 Pages: 3307-3314
  • DOI: 10.1016/j.cub.2019.07.084
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Protamines from liverwort are produced by posttranslational cleavage and C-terminal di-aminopropanelation of several male germ-specific H1 histones (2019)
  • Author(s): D'Ippolito, RA., Minamino, N, Rivera-Casas, C., Cheema, MS, Bai, DL., Kasinsky, HE., Shabanowitz, J., Eirin-Lopez, JM., Ueda, T., Hunt, DF., and Ausio, J.
  • Journal Title: Journal of Biological Chemistry Volume: 未定 Pages: 16364-16373
  • DOI: 10.1074/jbc.ra119.010316
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] HIGH STEROL ESTER 1 is a key factor in plant sterol homeostasis (2019)
  • Author(s): Shimada Takashi L.、Shimada Tomoo、Okazaki Yozo、Higashi Yasuhiro、Saito Kazuki、Kuwata Keiko、Oyama Kaori、Kato Misako、Ueda Haruko、Nakano Akihiko、Ueda Takashi、Takano Yoshitaka、Hara-Nishimura Ikuko
  • Journal Title: Nature Plants Volume: 5 Pages: 1154-1166
  • DOI: 10.1038/s41477-019-0537-2
  • Peer Reviewed
• [Journal Article] GEMMA CUP-ASSOCIATED MYB1, an ortholog of axillary meristem regulators, is essential in vegetative reproduction in Marchantia polymorpha. (2019)
  • Author(s): Yasui Y, Tsukamoto S, Sugaya T, Nishihama R, Wang Q, Kato H, Yamato KT, Fukaki H, Mimura T, Kubo H, Theres K, Kohchi T, Ishizaki K.
  • Journal Title: Curr. Biol. Volume: 29 Pages: 3987-3995
  • DOI: 10.1016/j.cub.2019.10.004
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A conserved regulatory mechanism mediates the convergent evolution of plant shoot lateral organs. (2019)
  • Author(s): Naramoto S, Jones VAS, Nicola, N, Sato M,Toyooka K, Shimamura M, Ishida S, Nishitani K,Ishizaki K, Nishihama R, Kohchi T, Dolan L, Kyozuka J.
  • Journal Title: PLOS Biol. Volume: 17 Pages: 12-12
  • DOI: 10.1371/journal.pbio.3000560
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Enrichment of Phosphatidylinositol 4,5-Bisphosphate in the Extra-Invasive Hyphal Membrane Promotes Colletotrichum Infection of Arabidopsis thaliana (2019)
  • Author(s): Shimada Takashi L、Betsuyaku Shigeyuki、Inada Noriko、Ebine Kazuo、Fujimoto Masaru、Uemura Tomohiro、Takano Yoshitaka、Fukuda Hiroo、Nakano Akihiko、Ueda Takashi
  • Journal Title: Plant Cell Physiol. Volume: 60 Pages: 1514-1524
  • DOI: 10.1093/pcp/pcz058
  • Peer Reviewed
• [Journal Article] Marchantia polymorpha, a New Model Plant for Autophagy Studies (2019)
  • Author(s): Norizuki T., Kanazawa T., Minamino N., Tsukaya H., Ueda T.
  • Journal Title: Front. Plant Sci. Volume: 10 Pages: 935-935
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  • Peer Reviewed / Open Access
• [Presentation] How Plants acquired new organelles -Lessons from Marchantia (2020)
  • Author(s): Takashi Ueda
  • Organizer: International Webinar Series: From Cellular dynamics to Morphology
• [Presentation] Organelle functions and membrane trafficking in plant cells (2019)
  • Author(s): Takashi Ueda
  • Organizer: EMBO practical course