Regulation mechanisms of the momilactone gene cluster and its evolution in momilactone producing plantss

• Project/Area Number: 17H03811
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bioorganic chemistry
• Research Institution: The University of Tokyo
• Principal Investigator: Okada Kazunori 東京大学, 生物生産工学研究センター, 准教授 (20312241)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥17,290,000 (Direct Cost: ¥13,300,000、Indirect Cost: ¥3,990,000); Fiscal Year 2019: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2018: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2017: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
• Keywords: 二次代謝物質 / 生合成遺伝子クラスター / 転写制御 / 病害抵抗性 / テルペノイド / モミラクトンＡ遺伝子クラスター / 蘚類ハイゴケ / 転写因子 / ゲラニルゲラニル２リン酸 / ファイトアレキシン / 遺伝子クラスター / モミラクトン / オキシリピン / 転写制御因子 / ハイゴケ / 野生イネ / アレロパシー / 植物成長調節物質

Outline of Final Research Achievements

The cultivated rice Oryza sativa has shown to hold two biosynthetic gene clusters on their genome for the production of antimicrobial diterpenoid phytoalexins. In this study, we first showed in planta functions of P450 oxygenases that comprise the biosynthetic pathway of ditepenoid phytoalexins in cultivated rice by analyzing metabolic flows in the pathway using a feeding approach with stable isotopes. We also analyzed two major transcriptional factors involved in the regulation of the genes in the diterpenoid phytoalexin biosynthetic gene clusters and demonstrated that OsTAGP1 has an important role on an allelopathic activity in roots and DPF is both genetically and functionally conserved in wild-rice. Finally, we discovered a functional biosynthetic gene cluster for momilactone A in bryophyte Calohypnum plumiforme, and suggested a genomic evidence for convergent evolution of the momilactone biosynthetic gene cluster in rice, barnyardgrass, and the moss.

Academic Significance and Societal Importance of the Research Achievements

植物に競争力を与えるモミラクトンはこれまでイネだけが利用していると考えられてきたが、イヌビエやハイゴケなど複数の進化的に離れた植物においても、ゲノム上の遺伝子クラスターの存在と共にモミラクトン合成能を維持していることが明らかになり、陸上植物が利用可能な生理活性物質としてのポテンシャルが高いことが示された。さらに、ハイゴケのモミラクトン生合成遺伝子クラスター内の遺伝子を用い、異種植物におけるモミラクトン生産にも成功した。これらの成果を利用することで、モミラクトンを生産し病害抵抗性を付与した新たな作物の開発につながる期待が大きく、本研究成果は学術的に重要なだけでなく社会的意義を持つものである。

Research Products

• [Int'l Joint Research] Universitat Autonoma de Barcelona (UAB)(スペイン)
• [Int'l Joint Research] Zhejiang University/Sichuan Agricultural University(中国)
• [Int'l Joint Research] Universitat Autonoma de Barcelona (UAB)(スペイン)
• [Int'l Joint Research] 浙江大学(中国)
• [Journal Article] Genomic evidence for convergent evolution of gene clusters for momilactone biosynthesis in land plants (2020)
  • Author(s): Mao L, Kawaide H, Higuchi T, Chen M, Miyamoto K, Hirata Y, Kimura H, Miyazaki S, Teruya M, Fujiwara K, Tomita K, Yamane H, Hayashi K, Nojiri H, Jia L, Qiu J, Ye C, Timko MP, Fan L, and Okada K
  • Journal Title: Proc Natl Acad Sci USA Volume: Online ahead of print Issue: 22 Pages: 12472-12480
  • DOI: 10.1073/pnas.1914373117
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Osa-miR7695 enhances transcriptional priming in defense responses against the rice blast fungus (2019)
  • Author(s): Sanchez-Sanuy F, Peris-Peris C, Tomiyama S, Okada K, Hsing YI, San Segundo B, Campo S.
  • Journal Title: BMC Plant Biol. Volume: 19 Issue: 1 Pages: 563-563
  • DOI: 10.1186/s12870-019-2156-5
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Characterization of diterpene synthase genes in the wild rice species Oryza brachyatha provides evolutionary insight into rice phytoalexin biosynthesis (2018)
  • Author(s): Toyomasu Tomonobu、Goda Chisato、Sakai Arisa、Miyamoto Koji、Shenton Matthew R.、Tomiyama Shiho、Mitsuhashi Wataru、Yamane Hisakazu、Kurata Nori、Okada Kazunori
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 503 Issue: 3 Pages: 1221-1227
  • DOI: 10.1016/j.bbrc.2018.07.028
  • Peer Reviewed
• [Journal Article] Expression of RSOsPR10 in rice roots is antagonistically regulated by jasmonate/ethylene and salicylic acid via the activator OsERF87 and the repressor OsWRKY76, respectively (2018)
  • Author(s): Yamamoto, T., Yoshida, Y., Nakajima, K., Tominaga, M., Gyohda, A., Suzuki, H., Okamoto, T., Nishimura, T., Yokotani, N., Minami, E., Nishizawa, Y., Miyamoto, K., Yamane, H., Okada, K., Koshib, T.
  • Journal Title: Plant Direct Volume: ３ Issue: 3 Pages: 1-17
  • DOI: 10.1002/pld3.49
  • Peer Reviewed / Open Access
• [Journal Article] OsDCL1a activation impairs phytoalexin biosynthesis and compromises disease resistance in rice. (2018)
  • Author(s): Salvador-Guirao R, Baldrich P, Tomiyama S, Hsing YI, Okada K, San Segundo B.
  • Journal Title: Annals of Botany Volume: 123 Issue: 1 Pages: 79-93
  • DOI: 10.1093/aob/mcy141
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] In planta functions of cytochrome P450 monooxygenase genes in the phytocassane biosynthetic gene cluster on rice chromosome 2 (2018)
  • Author(s): Zhongfeng Ye, Kohei Yamazaki, Hiromi Minoda, Koji Miyamoto, Sho Miyazaki, Hiroshi Kawaide, Arata Yajima, Hideaki Nojiri, Hisakazu Yamane and Kazunori Okada
  • Journal Title: Bioscience, Biotechnology and Biochemistry Volume: 印刷中 Issue: 6 Pages: 1-10
  • DOI: 10.1080/09168451.2017.1398067
  • Peer Reviewed
• [Journal Article] Biochemical synthesis of uniformly 13C-labeled diterpene hydrocarbons and their bioconversion to diterpenoid phytoalexins in planta (2017)
  • Author(s): Ye Z, Nakagawa K, Natsume M, Nojiri H, Kawaide H, Okada K.
  • Journal Title: Biosci. Biotech. Biochem. Volume: 81(6) Issue: 6 Pages: 1176-1184
  • DOI: 10.1080/09168451.2017.1285689
  • Peer Reviewed
• [Journal Article] OsMYC2, an essential factor for JA-inductive sakuranetin production in rice, interacts with MYC2-like proteins that enhance its transactivation ability. (2017)
  • Author(s): Ogawa S, Miyamoto K, Nemoto K, Sawasaki T, Yamane H, Nojiri H, Okada K.
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 40175-40175
  • DOI: 10.1038/srep40175
  • Peer Reviewed / Open Access
• [Journal Article] OsMYC2 mediates numerous defence-related transcriptional changes via jasmonic acid signalling in rice. (2017)
  • Author(s): Ogawa S, Kawahara-Miki R, Miyamoto K, Yamane H, Nojiri H, Tsujii Y, Okada K.
  • Journal Title: Biochem Biophys Res Commun. Volume: 486(3) Issue: 3 Pages: 796-803
  • DOI: 10.1016/j.bbrc.2017.03.125
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Barnyardgrass (Echinochloa crus-galli) genome analysis provides insight into its adaptation and invasiveness as a weed. (2017)
  • Author(s): Guo L, Qiu J, Ye C, Jin G, Mao L, Zhang H, Yang X, Peng Q, Wang Y, Jia L, Lin Z, Li G, Fu F, Liu C, Chen L, Aoki D, Yumoto E, Yokota T, Miyamoto M, Okada K, Kim DC, Cai D, Zhang C, Lou Y, Qian Q, Yamaguchi H, Yamane H, Kong CH,Timko MP, Bai L, Fan L et al.,
  • Journal Title: Nature Communications Volume: 8(1) Issue: 1
  • DOI: 10.1038/s41467-017-01067-5
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] OsTGAP1 is responsible for JA-inducible diterpenoid phytoalexin biosynthesis in rice roots with biological impacts on allelopathic interaction. (2017)
  • Author(s): Yoshida Y, Miyamoto K, Yamane H, Nishizawa Y, Minami E, Nojiri H, Okada K
  • Journal Title: Physiol Plant. Volume: 161(4) Issue: 4 Pages: 532-544
  • DOI: 10.1111/ppl.12638
  • Peer Reviewed
• [Presentation] 蘚類ハイゴケの染色体上でクラスター化したモミラクトン生合成遺伝子群の機能解析 (2019)
  • Author(s): 樋口俊哉, 照屋美優, 藤原薫, 宮本皓司, 山根久和, 林謙一郎, 川出洋, Longjiang Fan, 野尻秀昭, 岡田憲典
  • Organizer: 第54回 植物化学調節学会 2019/11/15-17、鳥取
• [Presentation] イネのジテルペン系ファイトアレキシン生合成遺伝子のノックアウトイネの作出と解析 (2019)
  • Author(s): 石川一輝, 山村千紘, 和田美樹, 冨山詩歩, 前田哲, 岡田憲典, 鎌倉高志, ○森昌樹
  • Organizer: 第54回 植物化学調節学会 2019/11/15-17、鳥取
• [Presentation] 新規殺菌剤トルプロカルブに関する研究（第13報） －イネにおけるジテルペン型ファイトアレキシン類の蓄積－ (2019)
  • Author(s): 髙橋明里，○鳥邉翔，冨山詩歩，富田啓介，有村源一郎，萩原寛之，小原敏明，岡田憲典
  • Organizer: 農薬学会
• [Presentation] Regulatory machinery for the coordinated expression on the phytoalexin biosynthetic gene clusters conserved in wild rice genome (2019)
  • Author(s): Shiho Tomiyama, Ryouka Kawahara-Miki, Koji Miyamoto, Hisakazu Yamane, Hideaki Nojiri, Naoki Yamamoto, ○Kazunori Okada.
  • Organizer: 17th International Symposium on Rice Functional Genomics (2019/910/4～6, @taiwan)
  • Int'l Joint Research
• [Presentation] Chemical genomics approach to dissect the mode of action of momilactone B, a major contributor to chemical defense in rice (2019)
  • Author(s): Keisuke Tomita, Yasuhiro Matsuo, Makoto Kawamukai, Yoko Yashiroda, Minoru Yoshida, Hideaki Nojiri, Kazunori Okada
  • Organizer: TERPNET 2019 - The 14th International Meeting on Biosynthesis, Function and Synthetic Biology of Isoprenoids (2019/8/26～30 @Halle, Germany)
  • Int'l Joint Research
• [Presentation] Regulators of stress-inducible prenyldiphosphate synthases that define types of terpenoids production in rice (2019)
  • Author(s): Miki Wada, Tomonori Shinya, Ivan Galis, Rika Ozawa, Gen-ichiro Arimura, Masaki Mori, Hideaki Nojiri, Kazunori Okada
  • Organizer: TERPNET 2019 - The 14th International Meeting on Biosynthesis, Function and Synthetic Biology of Isoprenoids (2019/8/26～30 @Halle, Germany)
  • Int'l Joint Research
• [Presentation] Evolutionary history of gene clustering for diterpenoid phytoalexins in the tribe Oryzeae (2019)
  • Author(s): Kazunori Okada
  • Organizer: 2nd China-Japan Joint Symposium on Natural Product Biosynthesis, January 14-15, 2019, Guangzhou, China (Jinan University)
  • Int'l Joint Research / Invited
• [Presentation] 植物種を超えて存在するモミラクトン生合成遺伝子クラスター (2019)
  • Author(s): 岡田憲典
  • Organizer: 日本農芸化学会2019年度大会（シンポジウム）
  • Invited
• [Presentation] イネの化学防御を担うモミラクトンBの作用機序に関する研究 (2019)
  • Author(s): 富田 啓介、松尾 安浩、川向 誠、八代田 陽子、松本 健、吉田 稔、野尻 秀昭、岡田 憲典
  • Organizer: 日本農芸化学会2019年度大会
• [Presentation] 野生イネにも保存されているファイトアレキシン生産制御因子DPFの機能解析 (2019)
  • Author(s): 冨山 詩歩、川原 玲香、宮本 皓司、山根 久和、豊増 知伸、野尻 秀昭、岡田 憲典
  • Organizer: 日本農芸化学会2019年度大会
• [Presentation] Conservation of inductive expression mechanism of diterpenoid phytoalexin biosynthetic gene clusters among the tribe Oryzeae (2018)
  • Author(s): Shiho Tomiyama, Ryouka Kawahara-Miki, Koji Miyamoto, Hisakazu Yamane, Hideaki Nojiri, Kazunori Okada
  • Organizer: 16th International Symposium on Rice Functional Genomics, September 5-7, 2018, Tokyo, Japan
  • Int'l Joint Research
• [Presentation] イネのストレス誘導型プレニル2リン酸合成酵素遺伝子の発現制御機構の解明 (2018)
  • Author(s): 和田 美樹, 田渕 雄夢, 森 昌樹, 野尻 秀昭, 岡田 憲典
  • Organizer: 植物化学調節学会 2018年11月2-4日 (北海道大学)
• [Presentation] ハイゴケにおけるモミラクトン生合成遺伝子クラスターの存在 (2018)
  • Author(s): 樋口 俊哉, 照屋 美優, 藤原 薫, 宮本 皓司, 山根 久和, 林 謙一郎, 川出 洋, Longjiang Fan, 野尻 秀昭, 岡田 憲典
  • Organizer: 植物化学調節学会 2018年11月2-4日 (北海道大学)
• [Presentation] イネ属に保存されている遺伝子クラスターの同調的発現制御機構 (2018)
  • Author(s): 冨山 詩歩, 川原 玲香, 宮本 皓司, 山根 久和, 野尻 秀昭, 岡田 憲典
  • Organizer: ■イネ遺伝学・分子生物学ワークショップ 2018年7月5-6日（国立遺伝学研究所）
• [Presentation] Disrupting cis-region by CRISPR/Cas9 affects momilactones biosynthetic cluster genes regulation (2017)
  • Author(s): Yuri Yoshida, Koji Miyamoto, Hisakazu Yamane, Hideaki Nojiri, Kazunori Okada
  • Organizer: ゲノム編集学会
• [Presentation] Evolutionary insights into regulatory mechanisms of the phytoalexin biosynthetic gene cluster in rice (2017)
  • Author(s): Shiho Tomiyama, Ryouka Kawahara-Miki, Kaihei Koshio, Koji Miyamoto, Hisakazu Yamane, Longjiang Fan, Hideaki Nojiri, Kazunori Okada
  • Organizer: TERPNET2017
  • Int'l Joint Research
• [Presentation] Conservation of inductive expression pattern of phytoalexins biosynthetic gene clusters in wild Oryza species (2017)
  • Author(s): Shiho Tomiyama, Ryouka Kawahara-Miki, Kaihei Koshio, Koji Miyamoto, Hisakazu Yamane, Hideaki Nojiri, Kazunori Okada
  • Organizer: ISRFG 2017
  • Int'l Joint Research
• [Presentation] An evolutionary point of view of the biosynthetic gene clusters formation for phytoalexins in rice and related species. (2017)
  • Author(s): Shiho Tomiyama, Ryouka Kawahara-Miki, Kaihei Koshio, Koji Miyamoto, Hisakazu Yamane, Jie Qiu, Longjiang Fan, Tomonobu Toyomasu, Nori Kurata, Hideaki Nojiri, Kazunori Okada
  • Organizer: ISRFG 2017
  • Int'l Joint Research
• [Presentation] 植物がつくりだす化学防御物質モミラクトンの分裂酵母に対する生育抑制効果 (2017)
  • Author(s): 富田 啓介，松尾 安浩，川向 誠，八代田 陽子，吉田 稔，三橋 渉，野尻 秀昭，岡田 憲典
  • Organizer: 植物化学調節学会
• [Presentation] 様々な生物種の生育を抑制するジテルペノイド化合物モミラクトンの作用機序解析 (2017)
  • Author(s): 富田 啓介，松尾 安浩，川向 誠，八代田 陽子，吉田 稔，三橋 渉，野尻 秀昭，岡田 憲典
  • Organizer: 日本農芸化学会
• [Book] 基礎から学ぶ植物代謝生化学 (2019)
  • Author(s): 水谷正治、士反伸和、杉山曉史（編）、岡田憲典・髙橋征司（分担執筆）
  • Total Pages: 325
  • Publisher: 羊土社
  • ISBN: 9784758120906