Field survey and experimental verification to understand the evolution of mode of sexual reproduction in Closterium peracerosum-strigosum-littorale complex

• Project/Area Number: 15H05237
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 海外学術
• Research Field: Biodiversity/Systematics
• Research Institution: Japan Women's University
• Principal Investigator: SEKIMOTO HIROYUKI 日本女子大学, 理学部, 教授 (20281652)
• Co-Investigator(Kenkyū-buntansha): 土金 勇樹 日本女子大学, 理学部, 助教 (20434152) ; 野崎 久義 東京大学, 大学院理学系研究科(理学部), 准教授 (40250104)
• Co-Investigator(Renkei-kenkyūsha): NISHIYAMA Tomoaki 金沢大学, 学際科学実験センター, 助教 (50390688)
• Research Collaborator: KAWAI Junko ; OOTSUKI Ryo ; KOMIYA Ayumi ; KANDA Naho ; TSUCHIMATSU Takashi ; SHIBATA Aika ; KOBAYASHI Hiroka ; TSUYUKI Natsumi ; HIRATA Yuuka
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000); Fiscal Year 2018: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000); Fiscal Year 2017: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2016: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2015: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
• Keywords: ミカヅキモ / 海外調査 / 生殖様式 / 交配群 / 種分化 / 生殖隔離 / 有性生殖 / 自家交配 / 他家交配

Outline of Final Research Achievements

The Closterium peracerosum-strigosum-littorale complex (C. p-s-l. complex) is a unicellular alga, and is composed of several reproductively isolated mating groups of heterothallic strains and closely related homothallic strains. We carried out field studies in foreign countries and in Japan, to isolate reproductively isolated C. p-s-l. complex. Finally, we established several clonal cultures of new sexually isolated mating groups and the closely related homothallic strains from these fields. We found a gene for mating-type determination in the heterothallic strains and named CpMinus1. In addition, the function of the orthologous gene in the homothallic strain was demonstrated by reverse genetic techniques. We suggest that the CpMinus1 could be a molecular switch between heterothallism and homothallism.

Academic Significance and Societal Importance of the Research Achievements

本研究の研究対象であるヒメミカヅキモは、陸上植物の姉妹群であるホシミドロ目に属する単細胞藻類である。これまでに解析に利用できる系統株が非常に限定されていたが、本研究の結果、解析可能な系統株の確立、系統関係の整理がなされ、さらに生殖様式の切換に関わる候補遺伝子の同定がなされた。自殖（ホモ）、他殖（ヘテロ）の進化は、陸上植物でも頻繁に見られており、特に陸上植物の進化を考える上で非常に有益な情報となり得ることが期待される。

Research Products

• [Int'l Joint Research] Wild Life Watch Group(ネパール)
• [Int'l Joint Research] Wild Life Watch Group(Nepal)
• [Int'l Joint Research] Khon Kaen University(Thailand)
• [Journal Article] Expression of genes from paternal alleles in rice zygotes and involvement of OsASGR-BBML1 in initiation of zygotic development (2019)
  • 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
  • Peer Reviewed / Open Access
• [Journal Article] The genus Closterium, a new model organism to study sexual reproduction in streptophytes (2019)
  • Author(s): Tsuchikane, Y., Sekimoto, H.
  • Journal Title: New Phytol. Volume: 221 Issue: 1 Pages: 99-104
  • DOI: 10.1111/nph.15334
  • Peer Reviewed
• [Journal Article] Identification of a new mating group and reproductive isolation in the Closterium peracerosum-strigosum-littorale complex (2018)
  • Author(s): Tsuchikane, Y., Kobayashi, H., Kato, M., Watanabe, J., Wu, J.-T., Sekimoto, H.
  • Journal Title: J. Plant Res. Volume: 131 Issue: 5 Pages: 735-746
  • DOI: 10.1007/s10265-018-1043-8
  • NAID: 40021652422
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Transcription factor DUO1 generated by neo-functionalization is associated with evolution of sperm differentiation in plants (2018)
  • Author(s): Higo, A., Kawashima, T., Borg, M., Zhao, M., Lopez-Vidriero, I., Sakayama, H., et al. (他18名)
  • Journal Title: Nature Communications Volume: 9 Issue: 1 Pages: 5283-5283
  • DOI: 10.1038/s41467-018-07728-3
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Molecular evolutionary analysis of a gender-limited MID ortholog from the homothallic species Volvox africanus with male and monoecious spheroids (2017)
  • Author(s): Kayoko Yamamoto, Hiroko Kawai-Toyooka, Takashi Hamaji, Yuki Tsuchikane, Toshiyuki Mori, Fumio Takahashi, Hiroyuki Sekimoto, Patrick J. Ferris, Hisayoshi Nozaki
  • Journal Title: PLoS One Volume: 12 Issue: 6 Pages: e0180313-e0180313
  • DOI: 10.1371/journal.pone.0180313
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] CRISPR/Cas9-based knockouts reveal that CpRLP1 is a negative regulator of the sex pheromone PR-IP in the Closterium peracerosum-strigosum-littorale complex. (2017)
  • Author(s): Kanda, N., Ichikawa, M., Ono, A., Toyoda, A., Fujiyama, A., Abe, J., Tsuchikane, Y., Nishiyama, T., Sekimoto, H.
  • Journal Title: Sci. Rep. Volume: 7 Issue: 1 Pages: 17873-17873
  • DOI: 10.1038/s41598-017-18251-8
  • Peer Reviewed / Open Access
• [Journal Article] Sexual reproduction and sex determination in green algae (2017)
  • Author(s): Sekimoto, H.
  • Journal Title: Journal of Plant Research Volume: 印刷中
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] reparation of knockdown transformants of unicellular charophycean alga, Closterium peracerosum-strigosum-littorale complex (2016)
  • Author(s): Abe, J., Hirano, N., Komiya, A., Kanda, N., Fujiwara, A., Hori, S., Tsuchikane, Y., Sekimoto, H.
  • Journal Title: Bio-protocol Volume: 6
  • Peer Reviewed / Open Access
• [Journal Article] Concanavalin A disrupts the release of fibrous material necessary for zygote formation of a unicellular charophycean alga, Closterium peracerosum-strigosum-littorale complex. (2016)
  • Author(s): Abe, J., Hori, S., Sato, M., Sekimoto, H.
  • Journal Title: Frontiers in Plant Science Volume: 7 Pages: 1040-1040
  • DOI: 10.3389/fpls.2016.01040
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Preparation of knockdown transformants of unicellular charophycean alga, Closterium peracerosum-strigosum-littorale complex (2016)
  • Author(s): Abe, J., Hirano, N., Komiya, A., Kanda, N., Fujiwara, A., Hori, S., Tsuchikane, Y., Sekimoto, H.
  • Journal Title: Bio-protocol Volume: 印刷中
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A receptor-like kinase, related with cell wall sensor of higher plants, is required for sexual reproduction in the unicellular charophycean alga, Closterium peracerosum-strigosum-littorale complex. (2015)
  • Author(s): Hirano, N., Marukawa, Y., Abe, J., Hashiba, S., Ichikawa, M., Tanabe, Y., Ito, M., Nishii, I., Tsuchikane, Y., Sekimoto, H.
  • Journal Title: Plant Cell Physiol. Volume: 56 Issue: 7 Pages: 1456-1462
  • DOI: 10.1093/pcp/pcv065
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] The algal ancestor of land plants was pre-adapted for symbiosis. (2015)
  • Author(s): Delaux, P-M., Radhakrishnan, G.V., Jayaraman, D., Cheema, J., Malbreil, M., Volkening, J.D., Sekimoto, H., Nishiyama, T., Melkonian, M., Pokorny, L., Rothfels, C.J., Sederoff, H.W., Stevenson, D.W., Surek, B., Zhang, Y., Sussman, M.R., Dunand, C., Morris, R.J., Roux, C., Wong, G.K-S., Oldroyd, G.E.D., Ané, J-M.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 112 Issue: 43 Pages: 13390-13395
  • DOI: 10.1073/pnas.1515426112
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] ネパール産ミカヅキモ属の多様性と新規交配群Hの発見 (2019)
  • Author(s): 小林弘佳、川口也和子、Mangal Man Shakya、土松隆志、関本弘之、土金勇樹
  • Organizer: 日本植物分類学会18回大会
• [Presentation] CRISPR/Cas9-based knockouts reveal the action mechanism of the sex pheromone PR-IP in unicellular zygnematophycean alga, Closterium peracerosum-strigosum-littorale complex (2018)
  • Author(s): Sekimoto, H.
  • Organizer: EMBO Workshop _New shores in land plant evolution
  • Int'l Joint Research
• [Presentation] Auxin response in conjugating green algae (Zygnematophyceae) (2018)
  • Author(s): Roman Skokan, Stanislav Vosolsob_, Junko Kawai, Hiroyuki Sekimoto, Henrik Buschmann, Jan Petr__ek
  • Organizer: EMBO Workshop _New shores in land plant evolution
  • Int'l Joint Research
• [Presentation] ヒメミカヅキモの有性生殖進行に関わるRWP-RK遺伝子の解析 (2018)
  • Author(s): 平田優香、土金勇樹、西山智明、関本弘之
  • Organizer: 日本植物学会第82回大会
• [Presentation] ヒメミカヅキモのCpMinus1形質転換体の比較トランスクリプトーム解析 (2018)
  • Author(s): 露木奈津美、小宮あゆみ、大槻涼、西山智明、関本弘之
  • Organizer: 日本植物学会第82回大会
• [Presentation] 接合藻ヒメミカヅキモ種内系統間にみられるゲノム構造の変異：生殖様式の進化との関連 (2018)
  • Author(s): 川口也和子、土金勇樹、西山智明、関本弘之、土松隆志
  • Organizer: 日本植物学会第82回大会
• [Presentation] ヒメミカヅキモの性フェロモンPR-IP Inducerによる生殖隔離障壁 (2018)
  • Author(s): 土金勇樹、小林弘佳、吉田睦未、関本弘之
  • Organizer: 日本植物学会第82回大会
• [Presentation] シャジクモ藻類ヒメミカヅキモのモデル植物化 (2018)
  • Author(s): 関本弘之
  • Organizer: 日本植物学会第82回大会
  • Invited
• [Presentation] CpMinus1N遺伝子の過剰発現から明らかにするヒメミカヅキモの姉妹接合 (2018)
  • Author(s): 板野芙美・齋藤真衣・関本弘之・土金勇樹
  • Organizer: 日本植物分類学会第17回大会
• [Presentation] ヒメミカヅキモの失われた交配群とその再発見 (2018)
  • Author(s): 小林弘佳・関本弘之・土金勇樹
  • Organizer: 日本植物分類学会第17回大会
• [Presentation] CRISPR/Cas9 システムを利用したヒメミカヅキモのマイナス型細胞特異的受容体型タンパク質 CpRLP1 の機能解析 (2017)
  • Author(s): 神田奈保、西山智明、土金勇樹、関本弘之
  • Organizer: 日本植物生理学会第58回大会
  • Place of Presentation: 鹿児島大学
  • Year and Date: 2017-03-16
• [Presentation] CRISPR/Cas9 システムを利用したヒメミカヅキモのマイナス型細胞特異的受容体型キナーゼ遺伝子 CpRLK2 の機能解析 (2017)
  • Author(s): 今詩織、神田奈保、藤原安理、市川真知子、関本弘之
  • Organizer: 日本植物生理学会第58回大会
  • Place of Presentation: 鹿児島大学
  • Year and Date: 2017-03-16
• [Presentation] ヒメミカヅキモの CpMinus1 遺伝子の逆遺伝学的解析 (2017)
  • Author(s): 露木奈津美、神田奈保、小宮あゆみ、川井絢子、土金勇樹、西山智明、関本弘之
  • Organizer: 日本植物生理学会第58回大会
  • Place of Presentation: 鹿児島大学
  • Year and Date: 2017-03-16
• [Presentation] 接合藻ヒメミカヅキモの性フェロモンによる生殖隔離と生殖干渉 (2017)
  • Author(s): 小林弘佳、加藤万智、関本弘之、土金勇樹
  • Organizer: 日本植物分類学会第16回大会
  • Place of Presentation: 京都大学
  • Year and Date: 2017-03-09
• [Presentation] ヒメミカヅキモの性決定におけるCpMinus1遺伝子の逆遺伝学的解析 (2017)
  • Author(s): 露木奈津美、神田奈保、小宮あゆみ、川井絢子、土金勇樹、西山智明、関本弘之
  • Organizer: 日本植物学会第81回大会
• [Presentation] CRISPR/Cas9による、ヒメミカヅキモの性特異的受容体型タンパク質の機能解析 (2017)
  • Author(s): 神田奈保、今詩織、藤原安理、西山智明、土金勇樹、関本弘之
  • Organizer: 日本植物学会第81回大会
• [Presentation] 性特異的遺伝子から明らかにするミカヅキモの進化 (2017)
  • Author(s): 土金 勇樹, 土松 隆志, Shakya Mangal Man, 関本 弘之
  • Organizer: 日本植物学会第81回大会
• [Presentation] Functional analyses of sex specific receptor-like proteins in Closterium peracerosum-strigosum-littorale complex, using CRISPR/CAS9 system (2017)
  • Author(s): H. Sekimoto et al.
  • Organizer: 11th International Phycological Congress
  • Int'l Joint Research
• [Presentation] CRISPR/CAS9-induced knockout in unicellular zygnematophycean alga, Closterium peracerosum-strigosum-littorale complex (2017)
  • Author(s): N. Kanda et al.
  • Organizer: 11th International Phycological Congress
  • Int'l Joint Research
• [Presentation] CpMinus1, mt- genome specific gene of heterothallic Closterium peracerosum-strigosum-littorale complex, is involved in the sex determination (2017)
  • Author(s): Tsuyuki
  • Organizer: 11th International Phycological Congress
  • Int'l Joint Research
• [Presentation] Reproductive isolation and reproductive interference by sex pheromones in Closterium peracerosum-strigosum-littorale complex (2017)
  • Author(s): Kabayashi
  • Organizer: 11th International Phycological Congress
  • Int'l Joint Research
• [Presentation] CpMinus1, mating-type minus genome specific gene of Closterium peracerosum-strigosum-littorale complex, promotes mt_ phenotype (2017)
  • Author(s): Tsuyuki
  • Organizer: The 65th NIBB Conference Marchantia Workshop 2017
  • Int'l Joint Research
• [Presentation] New insights into the sexual reproduction in unicellular zygnematophycean alga, Closterium peracerosum-strigosum-littorale complex (2017)
  • Author(s): H Sekimoto
  • Organizer: The 65th NIBB Conference Marchantia Workshop 2017
  • Int'l Joint Research / Invited
• [Presentation] ヒメミカヅキモの受容体型タンパク質CpRLP1の逆遺伝学的解析 (2016)
  • Author(s): 神田奈保、関本弘之
  • Organizer: 日本植物学会第80回大会
  • Place of Presentation: 沖縄コンベンションセンター
  • Year and Date: 2016-09-16
• [Presentation] エレクトロポーレーションによるシャジクモ藻類ヒメミカヅキモの安定形質転換系の確立 (2016)
  • Author(s): 川井絢子、金澤愛樹、早川靖彦、関本弘之
  • Organizer: 日本植物学会第80回大会
  • Place of Presentation: 沖縄コンベンションセンター
  • Year and Date: 2016-09-16
• [Presentation] シャジクモ藻類ヒメミカヅキモの性決定機構の解析 (2016)
  • Author(s): 関本弘之
  • Organizer: 日本植物学会第80回大会
  • Place of Presentation: 沖縄コンベンションセンター
  • Year and Date: 2016-09-16
  • Invited
• [Presentation] The CpMinus1 gene, specifically localized on the mating-type minus genome, is responsible for the sex determination of heterothallic Closterium peracerosum-strigosum-littorale complex (2016)
  • Author(s): Sekimoto, H., Komiya, A., Nishiyama, T.
  • Organizer: EMBO Workshop -New model systems for early land plant evolution
  • Place of Presentation: Vienna, Austria
  • Year and Date: 2016-06-22
  • Int'l Joint Research
• [Presentation] ヒメミカヅキモにおける組換え型性フェロモンを用いた生殖隔離障壁の解析 (2016)
  • Author(s): 土金勇樹、吉田睦未、関本弘之
  • Organizer: 日本藻類学会第40回大会
  • Place of Presentation: 日本歯科大学（東京都千代田区）
  • Year and Date: 2016-03-20
• [Presentation] CpMinus1遺伝子はヒメミカヅキモの性を決定する (2015)
  • Author(s): 小宮 あゆみ,阿部 淳,川井 絢子,鈴木 穣,豊田 敦,藤山 秋佐夫,大槻 涼,土金 勇樹, 西山 智明,関本 弘之
  • Organizer: 日本植物学会第79回大会
  • Place of Presentation: 朱鷺メッセ（新潟市）
  • Year and Date: 2015-09-08
• [Presentation] 接合藻ヒメミカヅキモにおける新規交配群 G の発見とトランスクリプトーム解析 (2015)
  • Author(s): 土金 勇樹,渡邊 樹梨,加藤 万智,鈴木 穣,西山 智明,関本 弘之
  • Organizer: 日本植物学会第79回大会
  • Place of Presentation: 朱鷺メッセ（新潟市）
  • Year and Date: 2015-09-08
• [Presentation] シャジクモ藻類ヒメミカヅキモには、２つのクラスのKNOX遺伝子が存在する (2015)
  • Author(s): 金澤愛樹、坂本季美枝、鈴木穣、豊田敦、藤山秋佐夫、西山智明、関本弘之
  • Organizer: 日本植物学会第79回大会
  • Place of Presentation: 朱鷺メッセ（新潟市）
  • Year and Date: 2015-09-08
• [Presentation] ヒメミカヅキモの受容体型タンパク質CpRLP1の機能解析 (2015)
  • Author(s): 神田奈保,市川真知子,小野彩花,土金勇樹,関本弘之
  • Organizer: 日本植物学会第79回大会
  • Place of Presentation: 朱鷺メッセ（新潟市）
  • Year and Date: 2015-09-07
• [Presentation] Sexual reproduction and sex determination of green algae (2015)
  • Author(s): Hiroyuki Sekimoto
  • Organizer: 日本植物学会第79回大会
  • Place of Presentation: 朱鷺メッセ（新潟市）
  • Year and Date: 2015-09-07
  • Invited
• [Presentation] The CpMinus1 gene, specifically localized on the mating-type minus genome, is responsible for the sex determination of heterothallic Closterium peracerosum-strigosum-littorale complex (2015)
  • Author(s): Komiya, A., Abe, J., Kawai, J., Suzuki, Y., Ootsuki, R., Tsuchikane, Y., Nishiyama, T., Sekimoto, H.
  • Organizer: 6th European Phycological Congress
  • Place of Presentation: London, England
  • Year and Date: 2015-08-25
  • Int'l Joint Research
• [Presentation] Characterization of a sex-specific receptor-like protein, expressing during the conjugation of heterothallic Closterium peracerosum-strigosum-littorale complex (2015)
  • Author(s): Kanda, N., Ichikawa, M., Ono, A., Tsuchikane, Y., Abe, J., Sekimoto, H.
  • Organizer: 6th European Phycological Congress
  • Place of Presentation: London, England
  • Year and Date: 2015-08-25
  • Int'l Joint Research
• [Presentation] A receptor-like kinase, related with cell wall sensor of higher plants, is responsible for sexual reproduction in the unicellular charophycean alga, Closterium peracerosum-strigosum-littorale complex (2015)
  • Author(s): Sekimoto, H.
  • Organizer: 6th European Phycological Congress
  • Place of Presentation: London, England
  • Year and Date: 2015-08-25
  • Int'l Joint Research