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Elucidation of the mechanism of cytoplasmic male sterility using mitoTALEN

Research Project

Project/Area Number 23K21187
Project/Area Number (Other) 21H02169 (2021-2023)
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeMulti-year Fund (2024)
Single-year Grants (2021-2023)
Section一般
Review Section Basic Section 39010:Science in plant genetics and breeding-related
Research InstitutionKyushu University

Principal Investigator

Kazama Tomohiko  九州大学, 農学研究院, 准教授 (30431464)

Project Period (FY) 2021-04-01 – 2025-03-31
Project Status Completed (Fiscal Year 2024)
Budget Amount *help
¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000)
Fiscal Year 2024: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2023: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2022: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2021: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Keywords細胞質雄性不稔性 / ミトコンドリアゲノム編集 / mitoTALEN / 細胞質雄性不稔性(CMS) / ミトコンドリア / CMS / 雄性不稔性
Outline of Research at the Start

収量増加の育種において、細胞質雄性不稔性はハイブリッド種子の効率的な採種を支える基盤であり、農業上非常に重要な形質である。しかし、ミトコンドリアへの細胞質雄性不稔性原因遺伝子(CMS原因遺伝子)産物の蓄積が、なぜ雄性不稔性を引き起こすのか?といった細胞質雄性不稔性発生の分子メカニズムは全く明らかになっていない。そこで、ミトコンドリアに存在する遺伝子をノックアウトすることが可能であるミトコンドリア移行シグナル付きTALEN(mitoTALEN)を用いて、CMS原因遺伝子候補の遺伝子破壊株を作成し、CMS原因遺伝子の特定を行うとともに、雄性不稔性を引き起こすメカニズムに関する知見を得る。

Outline of Final Research Achievements

In this research, we have attempted to knock out mitochondrial genes, orf352 and orf307, which have been reported as CMS-associated genes for WA-type and CW-type CMS rice, respectively. Knockout of orf307 led to fertility restoration in CW-type CMS rice. This indicates that orf307 is a cause of CW-type CMS rice. On the other hand, knocking out of orf352 in WA-type CMS rice didn't restore its fertility. However, pollen development proceeds in those orf352-depleted plants rather than in those of WA-type CMS rice. This indicates that orf352 is a part of the CMS-causing gene.
To elucidate the mechanism of male sterility, we performed RNA sequencing analysis. RNA was extracted from anthers at the mature pollen stage from CW-type CMS, T65, and orf307-depleted plants. Comparison among them, genes expressed specifically in CW-type CMS have been found. These genes were supposed to be involved in the retrograde-regulated expression in CW-type CMS.

Academic Significance and Societal Importance of the Research Achievements

核とミトコンドリアの不和合によって引き起こされる細胞質雄性不稔性(CMS)は、自分の花粉によって種子ができないため、効率的なF1採種に重用される表現型である。植物ミトコンドリアへの遺伝子導入が不可能なことより、これまで,CMSの真の原因遺伝子の特定は不可能であったが、本研究ではミトコンドリアゲノム編集技術を用いることで2種類のCMSイネのCMS原因遺伝子を特定することができた。本成果を用いることで、CMSイネとなりうる野生イネのスクリーニングや、F1ハイブリッド育種への応用が可能となり、社会的な意義は大きい。

Report

(3 results)
  • 2024 Final Research Report ( PDF )
  • 2022 Annual Research Report
  • 2021 Annual Research Report
  • Research Products

    (14 results)

All 2023 2022 2021 Other

All Journal Article (3 results) (of which Peer Reviewed: 3 results,  Open Access: 2 results) Presentation (9 results) (of which Invited: 1 results) Remarks (2 results)

  • [Journal Article] A Method for Precisely Identifying Modifications to Plant Mitochondrial Genomes by mitoTALENs2023

    • Author(s)
      Kazama Tomohiko、Arimura Shin-ichi
    • Journal Title

      Methods in Molecular Biology

      Volume: 2615 Pages: 365-378

    • DOI

      10.1007/978-1-0716-2922-2_25

    • ISBN
      9781071629215, 9781071629222
    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] <scp>TALEN</scp> ‐mediated depletion of the mitochondrial gene <i>orf312</i> proves that it is a Tadukan‐type cytoplasmic male sterility‐causative gene in rice2022

    • Author(s)
      Takatsuka Ayumu、Kazama Tomohiko、Arimura Shin‐ichi、Toriyama Kinya
    • Journal Title

      The Plant Journal

      Volume: 未定 Issue: 4 Pages: 1-11

    • DOI

      10.1111/tpj.15715

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Disruption of mitochondrial open reading frame 352 partially restores pollen development in cytoplasmic male sterile rice2021

    • Author(s)
      Shiho Omukai, Shin-ich Arimura, Kinya Toriyama, Tomohiko Kazama
    • Journal Title

      Plant Physiology

      Volume: 187 Issue: 1 Pages: 236-246

    • DOI

      10.1093/plphys/kiab236

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] Tadukanに由来する細胞質雄性不稔性イネの稔性回復候補遺伝子2023

    • Author(s)
      髙塚 歩、風間 智彦、市田 裕之、阿部 知子、鳥山 欽哉
    • Organizer
      第64回 日本植物学会年会
    • Related Report
      2022 Annual Research Report
  • [Presentation] mitoTALENの利用で明らかとなった細胞質雄性不稔イネの原因遺伝2022

    • Author(s)
      風間 智彦
    • Organizer
      中国育種談話会
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] イネミトコンドリアが引き起こす雄性生殖器官の「異常」とそれを「正常」に戻す核の遺伝子2022

    • Author(s)
      髙塚 歩、風間 智彦、有村慎一、市田 裕之、阿部 知子、鳥山 欽哉
    • Organizer
      第45回 日本分子生物学会年会
    • Related Report
      2022 Annual Research Report
  • [Presentation] "Exploration for a restorer of fertility gene for cytoplasmic male sterility derived from Oryza sativa cv. Tadukan"2022

    • Author(s)
      髙塚 歩、風間 智彦、市田 裕之、阿部 知子、鳥山 欽哉
    • Organizer
      第17回 東北育種研究集会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Tadukan型細胞質雄性不稔性イネを稔性回復させる遺伝子の探索2022

    • Author(s)
      髙塚 歩、風間 智彦、市田 裕之、阿部 知子、鳥山 欽哉
    • Organizer
      日本育種学会 第142回講演会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Tadukan型細胞質雄性不稔性イネの花粉発達は正常だが葯の裂開が阻害され、稔性回復は胞子体型的に行われる2022

    • Author(s)
      高塚歩、風間智彦、鳥山欽哉
    • Organizer
      第141回日本育種学会
    • Related Report
      2021 Annual Research Report
  • [Presentation] イネTA-CMSにおいてミトコンドリア移行TALENが引き起こす相同組換え修復およびゲノム構成の変化の解析2021

    • Author(s)
      高塚歩、風間智彦、有村慎一、鳥山欽哉
    • Organizer
      第140回日本育種学会
    • Related Report
      2021 Annual Research Report
  • [Presentation] mitoTALEN導入によるTadukan型細胞質雄性不稔性イネにおけるゲノム再構成の誘発2021

    • Author(s)
      高塚歩、風間智彦、有村慎一、鳥山欽哉
    • Organizer
      第38回日本植物バイオテクノロジー学会
    • Related Report
      2021 Annual Research Report
  • [Presentation] ミトコンドリア移行TALENによるCW型細胞質雄性不稔イネ原因遺伝子の同定2021

    • Author(s)
      木下諄美、有村慎一、鳥山欽哉、風間智彦
    • Organizer
      第38回日本植物バイオテクノロジー学会
    • Related Report
      2021 Annual Research Report
  • [Remarks]

    • URL

      https://kyushu-u.pure.elsevier.com/ja/persons/tomohiko-kazama

    • Related Report
      2022 Annual Research Report
  • [Remarks]

    • URL

      https://kyushu-u.pure.elsevier.com/ja/persons/tomohiko-kazama

    • Related Report
      2021 Annual Research Report

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Published: 2021-04-28   Modified: 2026-01-16  

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