Metabolic engineering of flavonoid-incorporated lignins for improving grass biomass utilization

• Project/Area Number: 17F17103
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 外国
• Research Field: Wood science
• Research Institution: Kyoto University
• Host Researcher: 飛松 裕基 京都大学, 生存圏研究所, 准教授 (20734221)
• Foreign Research Fellow: LAM PUI YING 京都大学, 生存圏研究所, 外国人特別研究員
• Project Period (FY): 2017-11-10 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥2,200,000 (Direct Cost: ¥2,200,000); Fiscal Year 2019: ¥500,000 (Direct Cost: ¥500,000); Fiscal Year 2018: ¥1,100,000 (Direct Cost: ¥1,100,000); Fiscal Year 2017: ¥600,000 (Direct Cost: ¥600,000)
• Keywords: リグニン / 植物細胞壁 / バイオマス / フラボノリグニン / イネ科植物 / 細胞壁 / フラボノイド / ケイ皮酸エステル

Outline of Annual Research Achievements

イネ科植物は、樹木を凌ぐ高いバイオマス生産性と優れた環境適応性を併せ持つ有用植物種を多数含み、経済性の優れた木質バイオリファイナリーシステムを担うバイオマス供給源として世界的に期待されている。本研究では、イネ科バイオマスの主要構成成分として最近見出されたフラボノイド修飾型リグニンの生合成と代謝工学に関わる研究を進め、樹木とは一線を画す、イネ科植物特有の木質形成機構の解明を目指すとともに、イネ科バイオマスの利用性向上を目指した究明を行う。具体的には、フラボノイド修飾型リグニンの形成に関わるイネ遺伝子群の同定を進め、その量や構造を様々に改変した形質転換イネを作出する。さらに、作成する一連の形質転換イネにつき、植物体の成長特性やバイオマスの各種利用性について比較解析を行い、フラボノイド修飾型リグニンの生理学的役割やそのイネ科バイオマスの各種利用特性に及ぼす寄与の解明を目指す。

これまで、フラボノイド修飾型リグニンを欠失した形質転換イネの更なる特性解析を進めるとともに、アシル修飾型リグニンの形成に関わるとリグニン前駆体アシル化酵素の解析を進めた。前者については、特にフラボノイド芳香核水酸化酵素及び芳香核メチル化酵素の機能欠損変異体の解析を行い、両者においてバイオマス構造の特徴的な変化とともに、バイオマス糖化性の向上が顕著に見られることを明らかにし、論文発表を行った。また、フラボノイド生合成経路の基点で作用する幾つかの新たな酵素遺伝子の解析も進め、CRISPR/Cas9を用いたゲノム編集により得た変異体の性状解析を完了し、論文発表の準備を行っている。一方、リグニン前駆体アシル化酵素の解析についても、CRISPR/Cas9を用いたゲノム編集により、アシル修飾型リグニンの形成に関わるリグニン前駆体アシル化酵素の機能欠損変異体の作出に成功し、引き続き、その性状解析を進めている。

Research Progress Status

令和元年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和元年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] 香港大学/中国科学院植物生理生態研究所(中国)
• [Int'l Joint Research] ウィスコンシン大学/オクラホマ大学(米国)
• [Int'l Joint Research] 香港大学(中国)
• [Int'l Joint Research] ウィスコンシン大学(米国)
• [Journal Article] Flavonoids are indispensable for complete male fertility in rice (2020)
  • Author(s): Lanxiang Wang, Pui Ying Lam , Andy CW Lui, Fu‐Yuan Zhu, Mo‐Xian Chen, Hongjia Liu, Jianhua Zhang, Clive Lo
  • Journal Title: Journal of Experimental Botany Volume: in press
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Convergent recruitment of 5´‐hydroxylase activities by CYP75B flavonoid B‐ring hydroxylases for tricin biosynthesis in Medicago legumes (2020)
  • Author(s): Andy CW Lui, Pui Ying Lam, Chan Kwun-Ho, Lanxiang Wang, Yuki Tobimatsu, Clive Lo
  • Journal Title: New Phytologist Volume: in press
  • DOI: 10.1111/nph.16498
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Fiber‐specific regulation of lignin biosynthesis improves biomass quality in Populus (2020)
  • Author(s): Jinshan Gui, Pui Ying Lam, Yuki Tobimatsu, Jiayan Sun, Cheng Huang, Shumin Cao, Yu Zhong, Toshiaki Umezawa, Laigeng Li
  • Journal Title: New Phytologist Volume: 226 Pages: 1074-1087
  • DOI: 10.1111/nph.16411
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Lignin-Inspired Surface Modification of Nanocellulose by Enzyme-Catalyzed Radical Coupling of Coniferyl Alcohol in Pickering Emulsion (2020)
  • Author(s): Kyohei Kanomata, Naoya Fukuda, Takuma Miyata, Pui Ying Lam, Toshiyuki Takano, Yuki Tobimatsu, Takuya Kitaoka
  • Journal Title: ACS Sustainable Chemistry and Engineering Volume: 8 Pages: 1185-1194
  • DOI: 10.1021/acssuschemeng.9b06291
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] OsCAldOMT1 is a bifunctional O-methyltransferase involved in the biosynthesis of tricin-lignins in rice cell walls (2019)
  • Author(s): Pui Ying Lam, Yuki Tobimatsu, Naoyuki Matsumoto, Shiro Suzuki, Wu Lan, Yuri Takeda, Masaomi Yamamura, Masahiro Sakamoto, John Ralph, Clive Lo & Toshiaki Umezawa
  • Journal Title: Scientific Reports Volume: 9 Pages: 11597-11597
  • DOI: 10.1038/s41598-019-47957-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Recruitment of specific flavonoid B‐ring hydroxylases for two independent biosynthesis pathways of flavone‐derived metabolites in grasses (2019)
  • Author(s): Pui Ying Lam, Andy Lui, Masaomi Yamamura, Lanxiang Wang, Yuri Takeda, Shiro Suzuki, Hongjia Liu, Fu-Yuan Zhu, Mo-Xian Chen, Jian-Hua Zhang, Toshiaki Umezawa, Yuki Tobimatsu, Clive Lo
  • Journal Title: New Phytologist Volume: 印刷中 Pages: 2014-2019
  • DOI: 10.1111/nph.15795
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Altered lignocellulose chemical structure and molecular assembly in CINNAMYL ALCOHOL DEHYDROGENASE-deficient rice (2019)
  • Author(s): Martin, A.F., Tobimatsu, Y., Kusumi, R., Matsumoto, N., Miyamoto, T., Lam, P.Y., Yamamura, M., Koshiba, T., Sakamoto, M., Umezawa, T.
  • Journal Title: Scientific Reports Volume: 9 Pages: 17153-17153
  • DOI: 10.1038/s41598-019-53156-8
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Recruitment of pathway-specific flavonoid B-ring hydroxylases for the independent generation of soluble flavone C-glycosides and cell-wall-bound tricin-lignin (2020)
  • Author(s): Pui Ying Lam, Yuki Tobimatsu, Toshiaki Umezawa, Clive Lo
  • Organizer: 第61回日本植物生理学会年会
• [Presentation] イネ科植物の細胞壁結合型フェルラ酸の生合成に関与するアルデヒドデヒドロゲナーゼの探索 (2020)
  • Author(s): 山本千莉、飛松裕基、Lam Pui Ying、武田ゆり、刑部祐里子、刑部敬史、Laura E. Bartley、梅澤俊明
  • Organizer: 第70回日本木材学会大会
• [Presentation] Comparative analysis of lignocellulose deconstruction properties in lignin-altered CAldOMT and CAD rice mutants (2020)
  • Author(s): Andri Fadillah Martin, Yuki Tobimatsu, Yuri Takeda, Pui Ying Lam, Toshiaki Umezawa
  • Organizer: 第70回日本木材学会大会
• [Presentation] 樹木の木化に学ぶ Pickering エマルションを用いた ナノセルロースの表面修飾 (2020)
  • Author(s): 福田直弥、鹿又喬平、宮田拓摩、Pui Ying Lam、高野俊幸、飛松裕基、北岡卓也
  • Organizer: 第70回日本木材学会大会
• [Presentation] Biosynthesis and Bioengineering of cell wall cross-linking ferulates in grasses (2019)
  • Author(s): Senri Yamamoto, Lam Pui Ying, Yuri Takeda, Yuriko Osakabe, Keishi Osakabe, Yuki Tobimatsu, Toshiaki Umezawa
  • Organizer: The 4th Asia Research Node Symposium on Humanosphere Science -Present and Future of Humanosphere Science-
  • Int'l Joint Research
• [Presentation] Recritment of specific flavonoid B-ring hydroxylases for two independent biosynthesis pathways of flavone-derived metabolites including tricin-lignin in grasses (2019)
  • Author(s): Pui Ying Lam, Andy C. W. Lui, Masaomi Yamamura, Lanxiang Wang, Yuri Takeda, Shiro Suzuki, Hongjia Liu, Fu-Yuan Zhu,Mo-xian Chen, Jianhua Zhang, Toshiaki Umezawa, Yuki Tobimatsu, Clive Lo
  • Organizer: 細胞壁研究者ネットワーク第13回定例研究会
• [Presentation] Altered cell wall chemical structure and molecular assembly in lignin-modified rice mutants (2019)
  • Author(s): Andri Fadillah Martin, Yuki Tobimatsu, Pui Ying Lam, Naoyuki Matsumoto, Shiro Suzuki, Ryosuke Kusumi, Takuto Tanaka, Takuji Miyamoto, Yuri Takeda, Masaomi Yamamura, Taichi Koshiba, Keishi Osakabe, Yuriko Osakabe, Masahiro Sakamoto, Toshiaki Umezawa
  • Organizer: 細胞壁研究者ネットワーク第13回定例研究会
• [Presentation] Altered lignin structures and improved biomass properties of rice mutants deficient in tricin biosynthetic genes (2019)
  • Author(s): Pui Ying Lam, Yuki Tobimatsu, Toshiaki Umezawa, Clive Lo
  • Organizer: 1st International Lignin Symposium
  • Int'l Joint Research
• [Presentation] Pathway-specific flavonoid B-ring hydroxylases involved in the biosynthesis of flavone C-glycosides and tricin-lignins in rice (2019)
  • Author(s): Pui Ying Lam, Andy C. W. Lui, Masaomi Yamamura, Lanxiang Wang, Yuri Takeda, Shiro Suzuki, Hongjia Liu, Fu-Yuan Zhu, Mo-Xian Chen, Jianhua Zhang, Toshiaki Umezawa, Yuki Tobimatsu, Clive Lo
  • Organizer: 第37回日本植物細胞分子生物学会
• [Presentation] Positive transcriptional feedback loop in secondary cell wall (SCW) formation in poplar (2019)
  • Author(s): Naoki Takata, Pui Ying Lam, Shiro Suzuki, Yuki Tobimatsu, Natsumaro Kutsuna, Toru Taniguchi
  • Organizer: IUFRO Tree Biotechnology 2019 Meeting
  • Int'l Joint Research
• [Presentation] Structurally-regulated dehydrogenative polymerization of coniferyl alcohol on Pickering emulsion stabilized by nanocellulose (2019)
  • Author(s): Naoya Fukuda, Kyohei Kanomata, Takuma Miyata, Pui Ying Lam, Yuki Tobimatsu, Takuya Kitaoka
  • Organizer: 2019 International Conference on Nanotechnology for Renewable Materials (TAPPI NANO)
• [Presentation] Altered lignin structures and improved cell wall digestibity of rice mutants deficient in tricin biosynthesis genes (2018)
  • Author(s): Pui Ying Lam, Yuki Tobimatsu, Toshiaki Umezawa, Clive Lo
  • Organizer: 第63回リグニン討論会
• [Presentation] Altered cell wall structure and properties of rice mutants deficient in flavonolignin (2018)
  • Author(s): Pui Ying Lam, Yuki Tobimatsu, Toshiaki Umezawa, Clive Lo
  • Organizer: 第12回細胞壁ネットワーク定例研究会
• [Presentation] Improved biomass digestibility of rice mutants deficient in tricin-lignins (2018)
  • Author(s): Pui Ying Lam, Yuki Tobimatsu, Toshiaki Umezawa, and Clive Lo
  • Organizer: The 3rd Asia Research Node Symposium on Humanosphere Science
  • Int'l Joint Research
• [Presentation] A bifunctional O-methyltransferase involved in the biosynthesis of flavonolignin in rice (2018)
  • Author(s): Pui Ying Lam, Naoyuki Matsumoto, Shiro Suzuki, Wu Lan, Yuri Takeda, Masaomi Yamamura, Masahiro Sakamoto, Clive Lo, John Ralph, Yuki Tobimatsu, Toshiaki Umezawa
  • Organizer: 第36回日本植物細胞分子生物学会
• [Presentation] Altered cell wall structures and improved biomass digestibility of tricin-truncated rice mutants (2018)
  • Author(s): Pui Ying Lam, Yuki Tobimatsu, Toshiaki Umezawa, and Clive Lo
  • Organizer: Lignin Gordon Research Conference
  • Int'l Joint Research
• [Presentation] OsCAldOMT1 is a bifunctional O-methyltransferase involved in the biosynthesis of tricin-lignins in rice cell walls (2018)
  • Author(s): Pui Ying Lam，Naoyuki Matsumoto，Shiro Suzuki，Wu Lan，Yuri Takeda，Masaomi Yamamura，Masahiro Sakamoto，Clive Lo，John Ralph，Yuki Tobimatsu，Toshiaki Umezawa
  • Organizer: 第68回日本木材学会大会
• [Presentation] Altered cell wall structure and improved biomass digestibility in tricin-truncated rice mutant deficient in flavone synthase II (2017)
  • Author(s): Pui Ying Lam, Yuki Tobimatsu, Yuri Takeda, Shiro Suzuki, Masaomi Yamamura, Toshiaki Umezawa, Clive Lo
  • Organizer: 第62回リグニン討論会
• [Presentation] Tricin‐depleted flavone synthase II mutant of rice displays altered lignin and improved cell wall digestibility (2017)
  • Author(s): Pui Ying Lam, Yuki Tobimatsu, Yuri Takeda, Shiro Suzuki, Masaomi Yamamura, Toshiaki Umezawa, Clive Lo
  • Organizer: 第11回細胞壁ネットワーク定例研究会
• [Book] 月刊 アグリバイオ 2019年6月号 (2019)
  • Author(s): 飛松裕基, Pui Ying Lam, 梅澤俊明, Clive Lo
  • Total Pages: 100
  • Publisher: 北隆館