Sex determining mechanism in streptophytes approached by the sex determining gene of Closterium peracerosum-strigosum-littorale complex

• Project/Area Number: 26650147
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• 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): 坂山 英俊 神戸大学, 理学研究科, 准教授 (60391108)
• Co-Investigator(Renkei-kenkyūsha): NISHIYAMA Tomoaki 金沢大学, 学際科学実験センター, 助教 (50390688) ; TSUCHIKANE Yuki 日本女子大学, 理学部, 助教 (20434152) ; ABE Jun 日本女子大学, 理学部, 学術研究員 (10424764)
• Research Collaborator: KAWAI Junko 日本女子大学, 理学部, 学術研究員 (30727673) ; KOMIYA Ayumi 日本女子大学, 理学研究科, 大学院生 ; FUJIWARA Anri 日本女子大学, 理学研究科, 大学院生 ; KANDA Naho 日本女子大学, 理学研究科, 大学院生 ; TSUYUKI Natsumi 日本女子大学, 理学研究科, 大学院生
• Project Period (FY): 2014-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
• Keywords: ミカヅキモ / 性決定 / ゲノム解読 / シャジクモ / ストレプトファイツ

Outline of Final Research Achievements

The CpMinus1 gene is a mt- genome-specific gene from the Closterium peracerosum-strigosum-littorale complex (C. psl. complex).
We developed the CpMinus1-expressing mt+ transformants. They showed a weak sexual reaction without mating partners, like as the case of homothallic strains, but tended to form zygotes with mt+ strain but not with mt- strain. The gene expression profile was drastically changed to that of wild mt- strains. We also obtained CpMinus1-knockout mutants from mt- background. The gene expression profile was quite similar to that of wild mt+ cells. We concluded that the CpMinus1 gene was a master gene for the sex determination in C. psl. complex. On the other hand, we could not find the CpMinus2 gene, which would be tightly linked to the CpMinus1 in the genome.
Through the Iso-seq data of sexual dimorphic Chara australis, two gene showing weak similarity to the CpMinus1 was found. One of them expressed only in the male strain, although the expression level was very low.

Research Products

• [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] Preparation of knockdown transformants of unicellular charophycean alga, Closterium peracerosum-strigosum-littorale complex (2016)
  • Author(s): be, 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
• [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): 1. 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: 印刷中
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Detailed analyses on the parthenospore formation in Closterium moniliferum (Zygnematophyceae, Charophyta). (2014)
  • Author(s): Yuki Tsuchikane, Ayaka Nakai, and Hiroyuki Sekimoto
  • Journal Title: Phycologia. Volume: 53 Issue: 6 Pages: 571-578
  • DOI: 10.2216/14-35r1.1
  • Peer Reviewed / Acknowledgement Compliant
• [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): Sekimoto, H., Kanda, N., Kon, S., Nishiyama, T., Tsuchikane, Y.
  • 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): Kanda, N., Tsuyuki, N., Kon, S., Nishiyama, T., Tsuchikane, Y., Sekimoto, H.
  • 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, N., Komiya, A., Kanda, N., Tsuchikane, Y., Nishiyama, T., Sekimoto, H.
  • 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): Kobayashi, H., Kato, M., Sekimoto, H., Tsuchikane, Y.
  • 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, N., Komiya, A., Kanda, N., Tsuchikane, Y., Nishiyama, T., Sekimoto, H.
  • 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): Sekimoto, H.
  • 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（新潟県新潟市）
  • 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（新潟県新潟市）
  • 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（新潟県新潟市）
  • 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（新潟県新潟市）
  • 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（新潟県新潟市）
  • 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
• [Presentation] ホモタリックなヒメミカヅキモにおけるCpMinus1相同遺伝子の機能解析 (2015)
  • Author(s): 土金勇樹、横山佳奈、鈴木穣、西山智明、関本弘之
  • Organizer: 日本藻類学会
  • Place of Presentation: 九州大学（福岡県福岡市）
  • Year and Date: 2015-03-21 – 2015-03-22
• [Presentation] 自殖するミカヅキモにおける性決定遺伝子CpMinus1の機能解析 (2015)
  • Author(s): 横山佳奈、関本弘之、鈴木穣、西山智明、土金勇樹
  • Organizer: 日本植物分類学会
  • Place of Presentation: 福島大学（福島県福島市）
  • Year and Date: 2015-03-06 – 2015-03-08
• [Presentation] ヒメミカヅキモの－型細胞ゲノム特異的遺伝子CpMinus1の機能解析 (2014)
  • 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
  • Place of Presentation: 明治大学（神奈川県川崎市）
  • Year and Date: 2014-09-12 – 2014-09-14
• [Presentation] ヒメミカヅキモの性フェロモンと結合するファシクリンIタンパク質の機能解析 (2014)
  • Author(s): 市原健介、滝口若菜、原真由美、土金勇樹、阿部淳、関本弘之
  • Organizer: 日本植物学会
  • Place of Presentation: 明治大学（神奈川県川崎市）
  • Year and Date: 2014-09-12 – 2014-09-14
• [Presentation] 接合藻ヒメミカヅキモにおけるホモタリズムの進化 (2014)
  • Author(s): 土金勇樹、西山智明、関本弘之
  • Organizer: 日本植物学会
  • Place of Presentation: 明治大学（神奈川県川崎市）
  • Year and Date: 2014-09-12 – 2014-09-14
• [Presentation] ヒメミカヅキモの－型細胞特異的な受容体型タンパク質の特性 (2014)
  • Author(s): 藤原安理、市川真知子、関本弘之
  • Organizer: 日本植物学会
  • Place of Presentation: 明治大学（神奈川県川崎市）
  • Year and Date: 2014-09-12 – 2014-09-14
• [Presentation] シャジクモ藻類ヒメミカヅキモにおける陸上植物特異的な転写因子の探索と発現解析 (2014)
  • Author(s): 川井絢子、阿部淳、鈴木穣、豊田敦、藤山秋佐夫、土金勇樹、西山智明、関本弘之
  • Organizer: 日本植物学会
  • Place of Presentation: 明治大学（神奈川県川崎市）
  • Year and Date: 2014-09-12 – 2014-09-14