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Studies of the cell autonomy and tissue dependency of clock-gene functions in plants

Research Project

Project/Area Number 19H03245
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 44030:Plant molecular biology and physiology-related
Research InstitutionKyoto University

Principal Investigator

Oyama Tokitaka  京都大学, 理学研究科, 准教授 (30324396)

Project Period (FY) 2019-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥15,860,000 (Direct Cost: ¥12,200,000、Indirect Cost: ¥3,660,000)
Fiscal Year 2021: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
Fiscal Year 2020: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2019: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
Keywords概日時計 / 植物細胞 / 時計遺伝子 / 発光レポーター / 細胞間相互作用 / 細胞自律性 / 植物 / 細胞 / 遺伝子機能
Outline of Research at the Start

本研究課題は、細胞自律性を基本としながら、個体内部環境の影響を受ける組織依存性も示す植物概日時計システムの特性を単一細胞レベルの概日リズム観測をもとに明らかにすることを目的とする。多彩な変異体があるシロイヌナズナを材料に、パーティクルガンを用いた概日発光レポーター導入/細胞遺伝子改変操作および微弱な細胞発光連続自動モニタリング系を主な実験手法として研究を進めていく。時計遺伝子機能(周期長制御や位相決定)が細胞自律性(周囲の組織(細胞)依存性)、さらに、個体レベルの同期状態依存性(非依存性)を明らかにする。これらの成果をもとに、最も基本的な細胞概日振動子を構成する時計遺伝子セットを明確化する。

Outline of Final Research Achievements

The plant circadian system is based on self-sustained cellular oscillations and these cellular clocks are coordinated in the plant body. In order to understand the plant circadian system from the aspect of cellular clocks, the bioluminescence reporter system was utilized for Arabidopsis and duckweed plants at a single cell level. Characterization of circadian rhythms of isolated cells, cells in the plant, and proliferating plants was done and the spontaneous order formation of circadian rhythms in the plant was analyzed. A Tool for gene disruption by the CRISPR/cas9 system and a dual-color luminescence monitoring system were developed and evaluated. A cellular circadian rhythm that was unaffected by perturbation of clock gene expression was revealed. The existence of a cellular physiological rhythm that is uncoupled with the cellular clock provides a new insight into our understanding of the plant circadian system.

Academic Significance and Societal Importance of the Research Achievements

植物個体内で細胞時計と細胞の概日リズムが分離可能であることを本研究で明確に示したことで、植物の概日時計システムの基本的な概念を新たに提案することができた。この点は、時間生物学分野に加えて一般的な植物生理学分野にも広く新たな展開を引き起こすと期待される。また、生物発光測定技術や遺伝子改変技術についての革新的な成果は、植物に限らず細胞レベルの遺伝子発現解析・機能解析に広く応用されていくと期待される。

Report

(4 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Annual Research Report
  • 2019 Annual Research Report
  • Research Products

    (37 results)

All 2022 2021 2020 2019

All Journal Article (10 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 6 results,  Open Access: 3 results) Presentation (24 results) (of which Int'l Joint Research: 5 results,  Invited: 2 results) Book (3 results)

  • [Journal Article] Adaptive Diversification in the Cellular Circadian Behavior of <i>Arabidopsis</i> Leaf- and Root-Derived Cells2022

    • Author(s)
      Nakamura Shunji、Oyama Tokitaka
    • Journal Title

      Plant and Cell Physiology

      Volume: 63 Issue: 3 Pages: 421-432

    • DOI

      10.1093/pcp/pcac008

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Interspecific divergence of circadian properties in duckweed plants2022

    • Author(s)
      Isoda Minako、Ito Shogo、Oyama Tokitaka
    • Journal Title

      Plant, Cell & Environment

      Volume: - Issue: 6 Pages: 1942-1953

    • DOI

      10.1111/pce.14297

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] An endogenous basis for synchronisation characteristics of the circadian rhythm in proliferating <i>Lemna minor</i> plants2021

    • Author(s)
      Ueno Kenya、Ito Shogo、Oyama Tokitaka
    • Journal Title

      New Phytologist

      Volume: 233 Issue: 5 Pages: 2203-2215

    • DOI

      10.1111/nph.17925

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Return of the Lemnaceae: duckweed as a model plant system in the genomics and postgenomics era2021

    • Author(s)
      Acosta Kenneth、Appenroth Klaus J、Borisjuk Ljudmilla、Edelman Marvin、Heinig Uwe、Jansen Marcel A K、Oyama Tokitaka、Pasaribu Buntora、Schubert Ingo、Sorrels Shawn、Sree K Sowjanya、Xu Shuqing、Michael Todd P、Lam Eric
    • Journal Title

      The Plant Cell

      Volume: 33 Issue: 10 Pages: 3207-3234

    • DOI

      10.1093/plcell/koab189

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Detection of uncoupled circadian rhythms in individual cells of Lemna minor using a dual-color bioluminescence monitoring system2021

    • Author(s)
      Watanabe Emiri、Isoda Minako、Muranaka Tomoaki、Ito Shogo、Oyama Tokitaka
    • Journal Title

      Plant and Cell Physiology

      Volume: - Issue: 5 Pages: 815-826

    • DOI

      10.1093/pcp/pcab037

    • Related Report
      2021 Annual Research Report 2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Bioluminescent monitoring of circadian rhythms in isolated mesophyll cells of Arabidopsis at single-cell level.2021

    • Author(s)
      Nakamura Shunji, Oyama Tokitaka
    • Journal Title

      Bioluminescennce: Methods and Protocols. 4th edition. Edited by Kim, S.-B. in Springer Protocols series

      Volume: -

    • Related Report
      2021 Annual Research Report
  • [Journal Article] An endogenous basis for synchronization manners of the circadian rhythm in proliferating Lemna minor plants2021

    • Author(s)
      Ueno Kenya、Ito Shogo、Oyama Tokitaka
    • Journal Title

      bioRxiv

      Volume: -

    • DOI

      10.1101/2021.02.09.430421

    • Related Report
      2020 Annual Research Report
    • Open Access
  • [Journal Article] Application of Single-Cell Bioluminescent Imaging to Monitor Circadian Rhythms of Individual Plant Cells2020

    • Author(s)
      Muranaka Tomoaki、Oyama Tokitaka
    • Journal Title

      Bioluminescent Imaging, Methods in Molecular Biology

      Volume: 1098 Pages: 231-242

    • DOI

      10.1007/978-1-4939-9940-8_17

    • ISBN
      9781493999392, 9781493999408
    • Related Report
      2020 Annual Research Report
  • [Journal Article] Monitoring single-cell bioluminescence of <i>Arabidopsis</i> leaves to quantitatively evaluate the efficiency of a transiently introduced CRISPR/Cas9 system targeting the circadian clock gene <i>ELF3</i>2019

    • Author(s)
      Kanesaka Yuki、Okada Masaaki、Ito Shogo、Oyama Tokitaka
    • Journal Title

      Plant Biotechnology

      Volume: 36 Issue: 3 Pages: 187-193

    • DOI

      10.5511/plantbiotechnology.19.0531a

    • NAID

      130007724760

    • ISSN
      1342-4580, 1347-6114
    • Year and Date
      2019-09-25
    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] A solvable model of entrainment ranges for the circadian rhythm under light/dark cycles2019

    • Author(s)
      Takakura Ren、Ichikawa Masatoshi、Oyama Tokitaka
    • Journal Title

      BioRxiv

      Volume: -

    • DOI

      10.1101/683615

    • Related Report
      2019 Annual Research Report
    • Open Access
  • [Presentation] シロイヌナズナ葉を用いた低温における概日リズムの維持機構の解明に向けて2022

    • Author(s)
      中村 駿志、小山 時隆
    • Organizer
      第63回日本植物生理学会年会
    • Related Report
      2021 Annual Research Report
  • [Presentation] Construction of a CRISPR/Cas9-Induced One-Cell Bioluminescence Reporter System in Individual Plants2022

    • Author(s)
      上野 稜平、伊藤 照悟、 小山 時隆
    • Organizer
      第63回日本植物生理学会年会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 原形質分離がシロイヌナズナ葉内組織の細胞概日リズムに与える影響の解析2021

    • Author(s)
      中村 駿志、小山 時隆
    • Organizer
      第28回時間生物学会学術大会
    • Related Report
      2021 Annual Research Report
  • [Presentation] Evaluation of plant circadian rhythms based on the cellular circadian behaviour2021

    • Author(s)
      Tokitaka Oyama
    • Organizer
      19th Congress of the European Society for Photobiology
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Robustness of the circadian periodicity of Arabidopsis mesophyll cells against influences of cell density on their rhythms2021

    • Author(s)
      Shunji Nakamura、Tokitaka Oyama
    • Organizer
      85th Cold Spring Harbor Laboratory Symposium on Quantitative Biology Biological Time Keeping,
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Uncoupled cellular circadian rhythms of a duckweed plant detected by a dual-color bioluminescence monitoring system2021

    • Author(s)
      Emiri Watanabe、Shogo Ito、Tokitaka Oyama
    • Organizer
      85th Cold Spring Harbor Laboratory Symposium on Quantitative Biology Biological Time Keeping,
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ウキクサ植物Wolffiella hyalinaのサリチル酸/安息香酸誘導性花成経路の解析2021

    • Author(s)
      磯田 珠奈子、北山七海、伊藤 照悟、小山 時隆
    • Organizer
      第62回日本植物生理学会年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 概日周期の多様性がもたらす限界日長の多様性2021

    • Author(s)
      村中 智明、工藤 洋、小山 時隆
    • Organizer
      第62回日本植物生理学会年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] シロイヌナズナの単離細胞における細胞間コミュニケーションが与える細胞概日リズムの安定性への影響2021

    • Author(s)
      中村 駿志、小山 時隆
    • Organizer
      第62回日本植物生理学会年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 2 波長発光レポーター測定系を用いたウキクサの細胞内非連結性リズムの挙動の解析2021

    • Author(s)
      渡邊 絵美理、伊藤 照悟、小山 時隆
    • Organizer
      第62回日本植物生理学会年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 温度条件がシロイヌナズナの単離細胞の概日時計に与える影響の解析2020

    • Author(s)
      中村 駿志、小山 時隆
    • Organizer
      日本植物学会第84回大会
    • Related Report
      2020 Annual Research Report
  • [Presentation] ウキクサ植物Wolffiella属の概日リズムにおける温度/光条件の依存性2020

    • Author(s)
      磯田 珠奈子、小山 時隆
    • Organizer
      第27回時間生物学会学術大会
    • Related Report
      2020 Annual Research Report
  • [Presentation] コウキクサにみられる細胞内非連結性概日リズムの解析2020

    • Author(s)
      渡邊 絵美理、伊藤 照悟、小山 時隆
    • Organizer
      第27回時間生物学会学術大会
    • Related Report
      2020 Annual Research Report
  • [Presentation] シロイヌナズナの単離細胞を用いた温度補償性の細胞基盤の解析2020

    • Author(s)
      中村 駿志、小山 時隆
    • Organizer
      第27回時間生物学会学術大会
    • Related Report
      2020 Annual Research Report
  • [Presentation] シロイヌナズナの単離した細胞を用いた細胞概日リズムの安定性の解析2020

    • Author(s)
      中村 駿志、伊藤 照悟、小山 時隆
    • Organizer
      第61回日本植物生理学会年会(大阪大学)
    • Related Report
      2019 Annual Research Report
  • [Presentation] 波長の異なる概日発光レポーターの共導入によるウキクサの細胞内非連結性リズムの観測2020

    • Author(s)
      渡邊絵美理,伊藤照悟,小山時隆
    • Organizer
      第61回日本植物生理学会年会(大阪大学)
    • Related Report
      2019 Annual Research Report
  • [Presentation] Circadian rhythms and single-cell analysis in duckweed2019

    • Author(s)
      Tokitaka Oyama
    • Organizer
      The 5th International Conference on Duckweed Research and Application (Rehovot, Israel)
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Analysis of floral induction by salicylic acid in a duckweed, Wolffiella hyalina2019

    • Author(s)
      Minako Isoda, Shogo Ito, Tokitaka Oyama
    • Organizer
      The 5th International Conference on Duckweed Research and Applications, Weizmann Institute of Science. Rehovot, Israel(国際 学会)
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 細胞から見る植物概日リズム2019

    • Author(s)
      小山時隆
    • Organizer
      第26回日本時間生物学会学術大会・シンポジウム「細胞・組織・個体に表出されるリズムの自律性と非自律性の分界」
    • Related Report
      2019 Annual Research Report
  • [Presentation] 植物における明暗同調の有無による恒明条件下での概日周期・振幅の時空間特性の違い2019

    • Author(s)
      上野賢也、磯田珠奈子、伊藤照悟、小山時隆
    • Organizer
      第26回日本時間生物学会学術大会(金沢市文化ホール)
    • Related Report
      2019 Annual Research Report
  • [Presentation] コウキクサの細胞内非連結性リズムの2波長発光レポーター測定系による検出2019

    • Author(s)
      渡邊絵美理、伊藤照悟、小山時隆
    • Organizer
      第26回日本時間生物学会学術大会(金沢市文化ホール)
    • Related Report
      2019 Annual Research Report
  • [Presentation] 弱光および局所的な明暗刺激を与えたウキクサ個体の概日リズムの応答2019

    • Author(s)
      吉永彩夏、四方純、伊藤照悟、小山時隆
    • Organizer
      第26回日本時間生物学会学術大会(金沢市文化ホール)
    • Related Report
      2019 Annual Research Report
  • [Presentation] 異なる温度環境下でのウキクサ植物 Lemna 属と Wolffiella 属の概日リズム特性の比較2019

    • Author(s)
      磯田珠奈子、小山時隆
    • Organizer
      第26回日本時間生物学会学術大会(金沢市文化ホール)
    • Related Report
      2019 Annual Research Report
  • [Presentation] シロイヌナズナの葉と根における光応答に対する細胞概日リズム特性の解析2019

    • Author(s)
      中村駿志、伊藤照悟、小山時隆
    • Organizer
      第26回日本時間生物学会学術大会(金沢市文化ホール)
    • Related Report
      2019 Annual Research Report
  • [Book] "The application of single cell bioluminescent imaging to monitor circadian rhythms of individual plant cells. pp 231- 242" in "Methods in Molecular Biology (Ripp, S. ed.)"2020

    • Author(s)
      T. Muranaka, T. Oyama.
    • Total Pages
      246
    • Publisher
      Humana Press (Springer Nature)
    • ISBN
      9781493999392
    • Related Report
      2019 Annual Research Report
  • [Book] "植物個体内の単一細胞発光モニタリング" 実験医学別冊 『発光イメージング実験ガイド(永井健治、小澤岳昌編),145- 158頁』2019

    • Author(s)
      村中智明、小山時隆
    • Total Pages
      222
    • Publisher
      羊土社
    • Related Report
      2019 Annual Research Report
  • [Book] "生き物の時計 生き物の時間", 『星雲, 第61号 58-66頁』2019

    • Author(s)
      小山時隆
    • Total Pages
      555
    • Publisher
      中山報恩会
    • Related Report
      2019 Annual Research Report

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Published: 2019-04-18   Modified: 2023-01-30  

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