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Common developmental mechanisms between somatic cells and zygotes

Research Project

Project/Area Number 16K14742
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Developmental biology
Research InstitutionTokyo Metropolitan University

Principal Investigator

Takashi Okamoto  首都大学東京, 理学研究科, 教授 (50285095)

Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2018: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywords細胞分化 / 分化全能性 / 体細胞 / 受精卵 / イネ / 物質導入 / 微量オーム解析 / 植物
Outline of Final Research Achievements

In the study, the identification of the proliferative protoplasts among somatic protoplasts isolated from rice callus was conducted. Through the establishment of modified methods for protoplast isolation and culture, we successfully traced the initial developmental profiles of isolated somatic protoplasts, and it was shown that approximate 3.5% of isolated/cultured protoplasts possess proliferative ability. These suggest that the single cell level analysis for proliferative and non-proliferative protoplasts is possible to elucidate the molecular mechanisms of totipotency in plant somatic cells. Moreover, comparison of transcriptome between proliferating protoplasts and zygotes, the natural totipotent cell, is also available to estimate the common cellular mechanisms in somatic and zygotic cellular development. To broaden the scope of investigations using these cells, a procedure for delivery of macromolecules, such as DNA and proteins, into these cells was also established.

Academic Significance and Societal Importance of the Research Achievements

一般に被子植物の体細胞は分化全能性をもつとされているが、体細胞組織を構成する細胞群から分化全能性を保持している細胞を特定し、解析することは困難であった。本研究では、イネカルスから調製したプロトプラストから増殖性プロトプラストを特定する手法を確立した。さらに、それら増殖性プロトプラストや受精卵といった全能性をもつ細胞の超微量オーム解析やそれら細胞への物質導入系を可能にし、植物細胞の分化全能性機構の解明に向けた基盤を確立した。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (47 results)

All 2019 2018 2017 2016 Other

All Int'l Joint Research (1 results) Journal Article (17 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 16 results,  Open Access: 12 results,  Acknowledgement Compliant: 3 results) Presentation (25 results) (of which Int'l Joint Research: 8 results,  Invited: 3 results) Remarks (3 results) Patent(Industrial Property Rights) (1 results)

  • [Int'l Joint Research] UC Berkeley(米国)

    • Related Report
      2018 Annual Research Report
  • [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
      2018 Annual Research 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
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] In vitro production of zygotes by electro-fusion of rice gametes.2019

    • Author(s)
      Rahman MH, Toda E., Okamoto T.
    • Journal Title

      Methods Mol. Biol.

      Volume: 0

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [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
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [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
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [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 Annual Research Report 2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Cell cycle in egg cell and its progression during zygotic development in rice2017

    • Author(s)
      Sukawa Yumiko、Okamoto Takashi
    • Journal Title

      Plant Reproduction

      Volume: 31 Issue: 1 Pages: 107-116

    • DOI

      10.1007/s00497-017-0318-x

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Development of gene expression system in egg cells and zygotes isolated from rice and maize2017

    • Author(s)
      Koiso Narumi、Toda Erika、Ichikawa Masako、Kato Norio、Okamoto Takashi
    • Journal Title

      Plant Direct

      Volume: 1 Issue: 3

    • DOI

      10.1002/pld3.10

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Insights into Land Plant Evolution Garnered from the Marchantia polymorpha Genome.2017

    • Author(s)
      Bowman J, Kouchi T,..(106名)..Yokoyama R, ..(4名)
    • Journal Title

      Cell

      Volume: 171 Issue: 2 Pages: 287-304

    • DOI

      10.1016/j.cell.2017.09.030

    • NAID

      120006353839

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Development of polyspermic zygote and possible contribution of polyspermy to polyploid formation in angiosperms.2017

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

      Journal of Plant Research

      Volume: - Issue: 3 Pages: 485-490

    • DOI

      10.1007/s10265-017-0913-9

    • Related Report
      2017 Research-status Report 2016 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Nuclear migration during karyogamy in rice zygotes is mediated by continuous convergence of actin meshwork toward the egg nucleus.2017

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

      Journal of Plant Research

      Volume: 130 Issue: 2 Pages: 339-348

    • DOI

      10.1007/s10265-016-0892-2

    • Related Report
      2017 Research-status Report 2016 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Analysis of Proteins Enriched in Rice Gamete2017

    • Author(s)
      Okamoto Takashi
    • Journal Title

      Methods Mol. Biol.

      Volume: 1669 Pages: 251-263

    • DOI

      10.1007/978-1-4939-7286-9_20

    • ISBN
      9781493972852, 9781493972869
    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Analysis of proteins enriched in rice gamete.2017

    • Author(s)
      Okamoto T.
    • Journal Title

      Methods Mol. Biol.

      Volume: -

    • Related Report
      2016 Research-status Report
    • Peer Reviewed
  • [Journal Article] in vitro 受精系と育種2017

    • Author(s)
      岡本龍史
    • Journal Title

      育種学研究

      Volume: 19 Pages: 34-35

    • Related Report
      2016 Research-status Report
  • [Journal Article] Electro-fusion of gametes and subsequent culture of zygotes.2016

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

      Bio-protocol

      Volume: 6 Issue: 24

    • DOI

      10.21769/bioprotoc.2074

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Development of polyspermic rice zygotes.2016

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

      Plant Physiol.

      Volume: 171 Issue: 1 Pages: 206-214

    • DOI

      10.1104/pp.15.01953

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] DNA demethylation is initiated in the central cells of Arabidopsis and rice.2016

    • Author(s)
      Park K., Kim Y., Vickers M., Park JS., Hyun Y., Okamoto T., Zilberman D., Fischer R., Feng X., Choi Y., Scholten S.
    • Journal Title

      Proc Natl Acad Sci USA

      Volume: 113 Issue: 52 Pages: 15138-15143

    • DOI

      10.1073/pnas.1619047114

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / 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 Annual Research Report
  • [Presentation] Development of gene expression and genome editing systems in rice egg cells and zygotes by directdelivery of macromolecules2018

    • Author(s)
      Okamoto, T., Toda E., T., 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 Annual Research 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 Annual Research Report
    • Int'l Joint Research
  • [Presentation] A paternally expressed transcription factor possibly initiates rice early zygotic development2018

    • Author(s)
      Rahman MH., Toda E., Ohnishi Y., Koiso N., Okamoto T.
    • Organizer
      25th International Congress on Sexual Plant Reproduction
    • Related Report
      2018 Annual Research 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 manner2018

    • 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 Annual Research 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 Annual Research 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 Annual Research 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 Annual Research Report
  • [Presentation] イネ初期受精卵発生における活性酸素種レベルの変動と発生に及ぼす影響2018

    • Author(s)
      古磯成美,戸田絵梨香,加藤紀夫,岡本龍史
    • Organizer
      第59回日本植物生理学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] イネ受精卵へのCRISPR/Cas9ベクターおよび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、刑部祐里子、岡本龍史、加藤紀夫
    • Organizer
      日本育種学会第133回講演会
    • Related Report
      2017 Research-status Report
  • [Presentation] ネ受精卵への 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、刑部祐里子、岡本龍史、加藤紀夫
    • Organizer
      第59回日本植物生理学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] 単離イネ卵細胞の受精非依存的な分裂および発生2018

    • Author(s)
      戸塚香里、須川友実子、岡本龍史
    • Organizer
      第59回日本植物生理学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] A Paternally Expressed AP2-Type Transcription Factor, OsASGR-BBML1, Possibly Contribute to Early Zygotic Development in Rice2018

    • Author(s)
      Rahman MdH, Toda E, Kobayashi M, Kudo T, Ohnishi Y, Yano K, Okamoto T.
    • Organizer
      第59回日本植物生理学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] 植物受精卵を用いた一過的遺伝子発現系およひ植物体誘導法の開発2017

    • Author(s)
      戸田絵梨香、古磯成美、綱島雅子、加藤紀夫、岡本龍史
    • Organizer
      日本育種学会第131回講演会
    • Place of Presentation
      名古屋
    • Year and Date
      2017-03-29
    • Related Report
      2016 Research-status Report
  • [Presentation] セタリア配偶子およひ受精卵の単離と受精卵の培養2017

    • Author(s)
      戸田絵梨香、岡本龍史、加藤紀夫
    • Organizer
      第58回日本植物生理学会年会
    • Place of Presentation
      鹿児島
    • Year and Date
      2017-03-16
    • Related Report
      2016 Research-status Report
  • [Presentation] In vitro fertilization system with isolated gametes: For basic and applied plant sciences2017

    • Author(s)
      Okamoto T. Toda E., Kakeda K., Ohnishi Y.
    • Organizer
      VISCEA Conference on Plant Cells In Vitro: Fundamentals and Applications II
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Possible contribution of a paternally expressed AP2-type transcription factor to early zygotic development in rice2017

    • Author(s)
      Rahman MdH, Toda E, Kobayashi M, Kudo T, Ohnishi Y, Yano K, Okamoto T.
    • Organizer
      Taiwan-Japan Plant Biology
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Effects of imbalanced parental genome ratio on zygotic development and possible function of genes expressing in zygotes with paternal allele-specific manner2017

    • Author(s)
      Toda, E., Kobayashi, M., Takahara, M., Ohnishi Y., Rahman, H., Watanabe, Y., Iwami M., Sekimoto, H., Yano, K., Okamoto, T.
    • Organizer
      Cold Spring Harbor Asia Conference on Plant Cell and Developmental Biology
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Development of gene expression system in egg cell and zygote isolated from rice and maize2017

    • Author(s)
      Toda E, Koiso N, Tunashima M, Kato N, Okamoto T.
    • Organizer
      Vienna International Science Conferences and Events Association, Plant Transformation & Biotechnology IV
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] イネ受精卵初期発生における父性鍵因子の探索および解析2017

    • Author(s)
      岩見百華、戸田絵梨香、小林正明、高原美嶺、大西由之佑、関本弘之、矢野健太郎、岡本龍史
    • Organizer
      日本植物学会第81回大会
    • Related Report
      2017 Research-status Report
  • [Presentation] トウモロコシ受精卵への物質導入と植物体再生2017

    • Author(s)
      加藤紀夫、戸田絵梨香、市川雅子、岡本龍史
    • Organizer
      第35回日本植物細胞分子生物学会大会
    • Related Report
      2017 Research-status Report
  • [Presentation] 植物卵細胞・受精卵を用いた一過的遺伝子発現系の確立および受精卵発生機構の解析へ向けた利用2017

    • Author(s)
      古磯成美、戸田絵梨香、綱島雅子、加藤紀夫、岡本龍史
    • Organizer
      日本植物学会第81回大会
    • Related Report
      2017 Research-status Report
  • [Presentation] イネ受精卵発生カスケードに与えるCa2+イオンの影響2017

    • Author(s)
      國分巌、大西由之佑、岡本龍史
    • Organizer
      日本植物学会第81回大会
    • Related Report
      2017 Research-status Report
  • [Presentation] in vitro 受精系と育種2016

    • Author(s)
      岡本龍史
    • Organizer
      日本育種学会第130回講演会ワークショップ「受精後雑種障壁研究の育種利用に向けて」
    • Place of Presentation
      鳥取
    • Year and Date
      2016-09-24
    • Related Report
      2016 Research-status Report
    • Invited
  • [Presentation] イネ受精卵における核合一機構の解析:卵核方向へのアクチンメッシュの集約に依存した核移動2016

    • Author(s)
      大西由之佑、岡本龍史
    • Organizer
      日本植物学会第80回大会
    • Place of Presentation
      沖縄
    • Year and Date
      2016-09-16
    • Related Report
      2016 Research-status Report
  • [Remarks] 首都大学東京 植物発生生理学研究室ホームページ

    • URL

      http://www.biol.se.tmu.ac.jp/labo.asp?ID=horcel

    • Related Report
      2018 Annual Research Report
  • [Remarks] 首都大学東京 植物発生生理学研究室

    • URL

      http://www.biol.se.tmu.ac.jp/labo.asp?ID=horcel

    • Related Report
      2017 Research-status Report
  • [Remarks] 研究室ホームページ

    • URL

      http://www.biol.se.tmu.ac.jp/labo.asp?ID=horcel

    • Related Report
      2016 Research-status Report
  • [Patent(Industrial Property Rights)] 植物に物質を導入する方法2017

    • Inventor(s)
      加藤紀夫、岡本龍史、古磯成美、木羽隆敏、戸田絵梨香
    • Industrial Property Rights Holder
      加藤紀夫、岡本龍史、古磯成美、木羽隆敏、戸田絵梨香
    • Industrial Property Rights Type
      特許
    • Filing Date
      2017-01-31
    • Related Report
      2016 Research-status Report

URL: 

Published: 2016-04-21   Modified: 2020-03-30  

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