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Genome-edited haploid induction via RNP-CRISPR of activated egg cells in Triticeae

Research Project

Project/Area Number 18K19211
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 39:Agricultural and environmental biology and related fields
Research InstitutionMie University

Principal Investigator

Kakeda Katsuyuki  三重大学, 生物資源学研究科, 教授 (50221867)

Co-Investigator(Kenkyū-buntansha) 岡本 龍史  東京都立大学, 理学研究科, 教授 (50285095)
Project Period (FY) 2018-06-29 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2019: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Fiscal Year 2018: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Keywordsゲノム編集 / in vitro受精 / Triticeae / 半数体
Outline of Final Research Achievements

Wheat egg cells and maize sperm cells were isolated and in vitro-fertilized using electric fusion to produce wheat-maize hybrid zygotes. Plantlets were regenerated from callus cultures derived from these cross-fertilized eggs. Morphological observation, flow cytometry and genome sequencing analysis of the regenerated plants indicated that they were haploid plants of wheat. This study demonstrates that, during the development and regeneration of wheat-maize hybrid zygotes produced in vitro, the sperm cell-derived maize genome is selectively eliminated, as in the case of in planta crosses, resulting in the induction of haploid plants carrying the egg cell-derived wheat genome.

Academic Significance and Societal Importance of the Research Achievements

コムギおよびトウモロコシから単離した雌雄配偶子のin vitro交雑受精卵からコムギ半数体が獲得されたのは世界で初めての成果であり、この実証には挑戦的研究として大きな意義がある。今後、本研究をさらに発展させ、ムギ類の単離卵細胞および受精卵への物質導入系技術の革新を進めることで、形質転換を伴わないDNAフリーゲノム編集体の獲得が可能となり、ゲノム編集を組み入れたムギ類の新たな半数体育種法の開発につながることが期待される。

Report

(5 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Research-status Report
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (51 results)

All 2022 2021 2020 2019 2018

All Journal Article (19 results) (of which Int'l Joint Research: 4 results,  Peer Reviewed: 16 results,  Open Access: 11 results) Presentation (32 results) (of which Int'l Joint Research: 7 results,  Invited: 2 results)

  • [Journal Article] A novel mutant allele at the Cleistogamy 1 locus in barley2021

    • Author(s)
      Wang Ning、Kakeda Katsuyuki、Tomokazu Masahiro、Liu Cheng、Yoshida Megumi、Kawada Naoyuki、Komatsuda Takao
    • Journal Title

      Theoretical and Applied Genetics

      Volume: 134 Issue: 10 Pages: 3183-3193

    • DOI

      10.1007/s00122-021-03884-1

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Development and regeneration of wheat-rice hybrid zygotes produced by in vitro fertilization system2021

    • Author(s)
      Maryenti Tety、Ishii Takayoshi、Okamoto Takashi
    • Journal Title

      New Phytologist

      Volume: 232 Pages: 2369-2383

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Regulatory functions of ROS dynamics via glutathione metabolism and glutathione peroxidase activity in developing rice zygote2021

    • Author(s)
      Rattanawong Kasidit、Koiso Narumi、Toda Erika、Kinoshita Atsuko、Tanaka Mari、Tsuji Hiroyuki、Okamoto Takashi
    • Journal Title

      The Plant Journal

      Volume: 108 Issue: 4 Pages: 1097-1115

    • DOI

      10.1111/tpj.15497

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Dynamics of mitochondrial distribution during development and asymmetric division of rice zygotes2021

    • Author(s)
      Aini Hanifah、Sato Yoshikatsu、Uno Kakishi、Higashiyama Tetsuya、Okamoto Takashi
    • Journal Title

      Plant Reproduction

      Volume: 35 Issue: 1 Pages: 47-60

    • DOI

      10.1007/s00497-021-00430-3

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Effect of Paternal Genome Excess on the Developmental and Gene Expression Profiles of Polyspermic Zygotes in Rice2021

    • Author(s)
      Deushi Ryouya、Toda Erika、Koshimizu Shizuka、Yano Kentaro、Okamoto Takashi
    • Journal Title

      Plants

      Volume: 10 Issue: 2 Pages: 255-255

    • DOI

      10.3390/plants10020255

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] In vitro fertilization system using wheat gametes by electric fusion.2021

    • Author(s)
      Maryenti T., Kato N., Ichikawa M., Okamoto T.
    • Journal Title

      Methods in Mol Biol.

      Volume: -

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Journal Article] CRISPR/Cas9‐Based Genome Editing Using Rice Zygotes2020

    • Author(s)
      Toda Erika、Okamoto Takashi
    • Journal Title

      Current Protocols in Plant Biology

      Volume: 5 Issue: 2

    • DOI

      10.1002/cppb.20111

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Gene Expression and Genome Editing Systems by Direct Delivery of Macromolecules Into Rice Egg Cells and Zygotes2020

    • Author(s)
      Toda Erika、Okamoto Takashi
    • Journal Title

      BIO-PROTOCOL

      Volume: 10 Issue: 14

    • DOI

      10.21769/bioprotoc.3681

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Polyspermy in angiosperms: Its contribution to polyploid formation and speciation2020

    • Author(s)
      Toda Erika、Okamoto Takashi
    • Journal Title

      Molecular Reproduction and Development

      Volume: 87 Issue: 3 Pages: 374-379

    • DOI

      10.1002/mrd.23295

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] In Vitro Production of Zygotes by Electrofusion of Rice Gametes2020

    • Author(s)
      Rahman Md Hassanur、Toda Erika、Okamoto Takashi
    • Journal Title

      Methods Mol. Biol.

      Volume: 2122 Pages: 257-267

    • DOI

      10.1007/978-1-0716-0342-0_18

    • ISBN
      9781071603413, 9781071603420
    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] 植物受精卵を用いたゲノム編集2020

    • Author(s)
      岡本 龍史、戸田絵梨香、加藤 紀夫
    • Journal Title

      植物の化学調節

      Volume: -

    • NAID

      130008060503

    • Related Report
      2019 Research-status Report
  • [Journal Article] イネ受精卵への一過的導入法2020

    • Author(s)
      加藤 紀夫、戸田絵梨香、岡本 龍史
    • Journal Title

      進化するゲノム編集

      Volume: Ⅱ

    • Related Report
      2019 Research-status Report
  • [Journal Article] イネ受精卵発生過程における雌雄配偶子の機能差および父性アリル依存的遺伝子発現による受精卵の発生誘導2019

    • Author(s)
      戸田絵梨香、岡本 龍史
    • Journal Title

      アグリバイオ

      Volume: 4月号 Pages: 73-78

    • NAID

      40021921244

    • Related Report
      2019 Research-status Report
  • [Journal Article] DNA demethylation by ROS1a in rice vegetative cells promotes methylation in sperm2019

    • Author(s)
      Kim M. Yvonne、Ono Akemi、Scholten Stefan、Kinoshita Tetsu、Zilberman Daniel、Okamoto Takashi、Fischer Robert L.
    • Journal Title

      Proceedings of the National Academy of Sciences

      Volume: 印刷中 Issue: 19 Pages: 9652-9657

    • DOI

      10.1073/pnas.1821435116

    • Related Report
      2019 Research-status Report 2018 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Establishment of an In Vitro Fertilization System in Wheat (Triticum aestivum L.)2019

    • Author(s)
      Maryenti Tety、Kato Norio、Ichikawa Masako、Okamoto Takashi
    • Journal Title

      Plant and Cell Physiology

      Volume: 60 Issue: 4 Pages: 835-843

    • DOI

      10.1093/pcp/pcy250

    • Related Report
      2019 Research-status Report 2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Sperm entry into the egg cell induces the progression of karyogamy in rice zygotes.2019

    • Author(s)
      Ohnishi Y*, Kokubu I, Kinoshita T, Okamoto T.
    • Journal Title

      Plant and Cell Physiology

      Volume: 印刷中 Issue: 8 Pages: 1656-1665

    • DOI

      10.1093/pcp/pcz077

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Expression of genes from paternal alleles in rice zygotes and involvement of OsASGR-BBML1 in initiation of zygotic development2019

    • Author(s)
      Rahman, H., Toda, E., Kobayashi, M., Kudo, T., Koshimizu, S., Takahara, M., Iwami, M., Watanabe, Y., Sekimoto, H., Yano, K., Okamoto, T.
    • Journal Title

      Plant Cell Physiol.

      Volume: 60 Issue: 4 Pages: 725-737

    • DOI

      10.1093/pcp/pcz030

    • Related Report
      2019 Research-status Report 2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] An efficient DNA- and selectable-marker-free genome-editing system using zygotes in rice2019

    • Author(s)
      Toda Erika、Koiso Narumi、Takebayashi Arika、Ichikawa Masako、Kiba Takatoshi、Osakabe Keishi、Osakabe Yuriko、Sakakibara Hitoshi、Kato Norio、Okamoto Takashi
    • Journal Title

      Nature Plants

      Volume: 5 Issue: 4 Pages: 363-368

    • DOI

      10.1038/s41477-019-0386-z

    • Related Report
      2019 Research-status Report 2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Effects of an imbalanced parental genome ratio on development of rice zygotes.2018

    • Author(s)
      Toda E, Ohnishi Y, Okamoto T.
    • Journal Title

      Journal of Experimental Botany

      Volume: 印刷中 Issue: 10 Pages: 2609-2619

    • DOI

      10.1093/jxb/ery094

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] Pleiotropic effects of mutation at the miR172 target site in the wheat orthologs of Cly12022

    • Author(s)
      Agetha Nanape, Katsuyuki Kakeda
    • Organizer
      日本育種学会 第141回講演会
    • Related Report
      2021 Annual Research Report
  • [Presentation] Developmental profiles of inter-subfamily polyploid zygotes produced by the fusion of wheat and rice gametes2022

    • Author(s)
      Maryenti T., Ishii T., Okamoto T.
    • Organizer
      日本育種学会 第141回講演会
    • Related Report
      2021 Annual Research Report
  • [Presentation] オオムギ開花性変異遺伝子cly1.b4の遺伝解析2021

    • Author(s)
      掛田 克行, 櫛部 七海, Agetha Nanape
    • Organizer
      日本育種学会 第140回講演会
    • Related Report
      2021 Annual Research Report
  • [Presentation] Characterization of floral/spike traits in the wheat AP2 mutants with reference to FHB resistance2021

    • Author(s)
      A. Nanape, C. Nakashima, K. Kakeda
    • Organizer
      第16回ムギ類研究会
    • Related Report
      2021 Annual Research Report
  • [Presentation] Dynamics of mitochondrial distribution during development and asymmetric division of rice zygotes2021

    • Author(s)
      Aini H., Sato Y., Uno K., Higashiyama T., Okamoto T.
    • Organizer
      日本植物学会 第85回大会
    • Related Report
      2021 Annual Research Report
  • [Presentation] キメラリプレッサーを用いたイネ卵細胞の分裂・発生を誘導する新規転写因子の探索2021

    • Author(s)
      澤本陸、大島良美、貴嶋紗久、光田展隆、池田美穂、高崎寛則、高木優、戸田絵梨香、岡本龍史
    • Organizer
      日本植物学会 第85回大会
    • Related Report
      2021 Annual Research Report
  • [Presentation] イネ受精卵発生を誘導するOsASGR-BBML1の標的遺伝子群の同定2021

    • Author(s)
      増尾心之介、大島良美、貴嶋紗久、光田展隆、池田美穂、高崎寛則、高木優、戸田絵梨香、木下温子、岡本龍史
    • Organizer
      日本植物学会 第85回大会
    • Related Report
      2021 Annual Research Report
  • [Presentation] コムギ in vitro 受精系 : 異質倍数性受精卵の作出とその発生解析2021

    • Author(s)
      岡本龍史
    • Organizer
      麦学オンラインセミナー
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] An efficient DNA- and selectable-marker-free genome-editing system using zygotes in rice and its possible application to other crop species.2021

    • Author(s)
      Toda E, Koiso N, Takebayashi A, Ichikawa M, Kiba T, Osakabe K, Osakabe Y, Sakakibara H, Kato N, Okamoto T.
    • Organizer
      Keystone Symposia “Plant Genome Engineering: From Lab to Field”
    • Related Report
      2020 Research-status Report
    • Int'l Joint Research
  • [Presentation] Developmental profiles of inter-subfamily polyploid zygotes produced by the fusion of wheat and rice gametes2021

    • Author(s)
      Maryenti Tety., Takayoshi Ishii., Okamoto T.
    • Organizer
      日本植物生理学会
    • Related Report
      2020 Research-status Report
  • [Presentation] Expression of Genes from Paternal Alleles in Rice Zygotes and Involvement of OsASGR-BBML1 in Initiation of Zygotic Development2021

    • Author(s)
      ahman MH, Toda E., Kobayashi M., Kudo T., Koshimizu S., Takahara M., Iwami M, Watanabe Y., Sekimoto H., Yano K., Okamoto T.
    • Organizer
      日本植物生理学会
    • Related Report
      2020 Research-status Report
  • [Presentation] Evaluation of the effect of mutations in the wheat AP2 homoeologs on cleistogamous flowering using near-isogenic lines2021

    • Author(s)
      A. Nanape, K. Kakeda
    • Organizer
      日本育種学会
    • Related Report
      2020 Research-status Report
  • [Presentation] Initiation of zygotic development and fertilization-independent egg division in rice. Symposium of Frontiers of research on embryo and endosperm development: Induction of artificial apomixis.2020

    • Author(s)
      Okamoto T.
    • Organizer
      日本植物生理学会
    • Related Report
      2019 Research-status Report
  • [Presentation] ROS-level dynamics during development of zygote in rice2020

    • Author(s)
      Rattanawong K., Koiso N., Okamoto T.
    • Organizer
      日本植物生理学会
    • Related Report
      2019 Research-status Report
  • [Presentation] Accumulation effects of mutations in the wheat AP2 homoeologs on cleistogamous flowering2020

    • Author(s)
      Nanape A., Watanabe K., Haine H.M., Kakeda K.
    • Organizer
      日本育種学会
    • Related Report
      2019 Research-status Report
  • [Presentation] イネ受精卵を用いたDNAおよび選抜マーカーフリーなゲノム編集技術の確立2019

    • Author(s)
      戸田絵梨香
    • Organizer
      プロジェクト横断型公開シンポジウム「植物のゲノム編集基盤技術開発の現状と展望」
    • Related Report
      2019 Research-status Report
  • [Presentation] Establishment of an in vitro fertilization system in wheat2019

    • Author(s)
      Maryenti T., Kato N., Ichikawa M., Okamoto T.
    • Organizer
      ムギ類研究会
    • Related Report
      2019 Research-status Report
  • [Presentation] Accumulation effects of mutant alleles of AP2 homoeologs on cleistogamous flowering in hexaploid wheat2019

    • Author(s)
      Nanape A., Watanabe K., Haine H., Kakeda K.
    • Organizer
      ムギ類研究会
    • Related Report
      2019 Research-status Report
  • [Presentation] コムギ-トウモロコシ交雑受精卵の発生過程における片親ゲノムの選択的な脱落2019

    • Author(s)
      古野真由子, Tety Maryenti, 掛田克行, 岡本龍史
    • Organizer
      日本植物学会
    • Related Report
      2019 Research-status Report
  • [Presentation] 被子植物の受精に伴う遺伝子発現およびpaternal/maternal mRNA degradation の最初の報告2019

    • Author(s)
      大西 由之佑,永田 博基,戸田 絵梨香,國分 巌,岡本 龍史,木下 哲
    • Organizer
      日本植物学会
    • Related Report
      2019 Research-status Report
  • [Presentation] 被子植物の受精に伴うカルシウム応答性遺伝子発現およびmRNA分解2019

    • Author(s)
      永田 博基,大西 由之佑,戸田 絵梨香,國分 巌,岡本 龍史,木下 哲
    • Organizer
      日本植物学会
    • Related Report
      2019 Research-status Report
  • [Presentation] An efficient DNA- and selectable-marker-free genome-editing system using zygotes in rice2019

    • Author(s)
      Toda E, Koiso N, Takebayashi A, Ichikawa M, Kiba T, Osakabe K, Osakabe Y, Sakakibara H, Kato N, Okamoto T.
    • Organizer
      Vienna International Science Conferences and Events Association, Plant Genome Editing & Genome Engineering
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research
  • [Presentation] Gene expression and genome editing systems in rice by direct delivery of macromolecules into egg cell and zygote2019

    • Author(s)
      Toda E, Koiso N, Takebayashi A, Ichikawa M, Kiba T, Osakabe K, Osakabe Y, Sakakibara H, Kato N, Okamoto T.
    • Organizer
      EMBO Practical Course “Functional live imaging of plants”
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research
  • [Presentation] イネ受精卵を用いたDNAおよび選抜マーカーフリーなゲノム編集技術の確立と他作物種への応用.2019

    • Author(s)
      戸田絵梨香、古磯成美、Tety Maryenti、竹林有理佳、市川雅子、木羽隆敏、刑部敬史、刑部祐里子、榊原均、加藤紀夫、岡本龍史
    • Organizer
      第60回日本植物生理学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Establishment of an in vitro fertilization system in wheat (Triticum aestivum L.).2019

    • Author(s)
      Maryenti, T., Kato, N., Ichikawa, M., Okamoto, T.
    • Organizer
      第60回日本植物生理学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Development of gene expression and genome editing systems in rice egg cells and zygotes by direct delivery of macromolecules.2018

    • Author(s)
      Okamoto, T.,Toda E. , Koiso, N., Rahman MH., Takebayashi, A., Ichikawa, M., Kiba, T., Osakabe, Y., Osakabe, T., Sakakibara, H., Kato, N.
    • Organizer
      25th International Congress on Sexual Plant Reproduction
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] Genome editing in rice by direct delivery of preassembled CRISPR-Cas9 vectors or ribonucleoproteins into zygotes.2018

    • Author(s)
      Okamoto, T., Toda E., Koiso, N., Takebayashi, A., Ichikawa, M., Kiba, T., Osakabe, Y., Osakabe, T., Sakakibara, H., Kato, N.
    • Organizer
      International Association for Plant Biotechnology Congress 2018
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] A paternally expressed transcription factor possibly initiates rice early zygotic development.2018

    • Author(s)
      Rahman MH., Toda E., Ohnishi Y., Koiso N., Okamoto T.
    • Organizer
      25th International Congress on Sexual Plant Reproduction
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] Effects of an imbalanced parental genome ratio on development of rice zygotes and possible function of genes expressing in zygotes with paternal allele-specific manner.2018

    • Author(s)
      Toda E., Kobayashi M., Takahara M., Ohnishi Y., Kudo T., Rahman MH., Watanabe Y., Iwami M., Yano K., Okamoto T.
    • Organizer
      25th International Congress on Sexual Plant Reproduction
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] in vitro 受精系:育種への展開.2018

    • Author(s)
      岡本龍史
    • Organizer
      植物科学シンポジウム2018
    • Related Report
      2018 Research-status Report
    • Invited
  • [Presentation] 精細胞侵入に伴う細胞内 Ca2+ レベルの上昇はイネ受精卵内の核合一を促進する.2018

    • Author(s)
      大西由之佑,國分巌,岡本龍史
    • Organizer
      日本植物学会第82回大会
    • Related Report
      2018 Research-status Report
  • [Presentation] 植物受精卵への Cas9タンパク質-gRNA 複合体の直接導入によるゲノム編集技術の確立と応用.2018

    • Author(s)
      戸田絵梨香、古磯成美、竹林有理佳、市川雅子、木羽隆敏、刑部敬史、刑部祐里子、榊原均、岡本龍史、加藤紀夫
    • Organizer
      日本植物学会第82回大会
    • Related Report
      2018 Research-status Report

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Published: 2018-07-25   Modified: 2023-01-30  

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