論文

査読有り 国際誌
2019年2月

Lignin characterization of rice CONIFERALDEHYDE 5-HYDROXYLASE loss-of-function mutants generated with the CRISPR/Cas9 system.

The Plant journal : for cell and molecular biology
  • Yuri Takeda
  • ,
  • Shiro Suzuki
  • ,
  • Yuki Tobimatsu
  • ,
  • Keishi Osakabe
  • ,
  • Yuriko Osakabe
  • ,
  • Safendrri K Ragamustari
  • ,
  • Masahiro Sakamoto
  • ,
  • Toshiaki Umezawa

97
3
開始ページ
543
終了ページ
554
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1111/tpj.14141

The aromatic composition of lignin is an important trait that greatly affects the usability of lignocellulosic biomass. We previously identified a rice (Oryza sativa) gene encoding coniferaldehyde 5-hydroxylase (OsCAld5H1), which was effective in modulating syringyl (S)/guaiacyl (G) lignin composition ratio in rice, a model grass species. Previously characterized OsCAld5H1-knockdown rice lines, which were produced via an RNA-interference approach, showed augmented G lignin units yet contained considerable amounts of residual S lignin units. In this study, to further investigate the effect of suppression of OsCAld5H1 on rice lignin structure, we generated loss-of-function mutants of OsCAld5H1 using the CRISPR/Cas9-mediated genome editing system. Homozygous OsCAld5H1-knockout lines harboring anticipated frame-shift mutations in OsCAld5H1 were successfully obtained. A series of wet-chemical and two-dimensional NMR analyses on cell walls demonstrated that although lignins in the mutant were predictably enriched in G units all the tested mutant lines produced considerable numbers of S units. Intriguingly, lignin γ-p-coumaroylation analysis by the derivatization followed by reductive cleavage method revealed that enrichment of G units in lignins of the mutants was limited to the non-γ-p-coumaroylated units, whereas grass-specific γ-p-coumaroylated lignin units were almost unaffected. Gene expression analysis indicated that no homologous genes of OsCAld5H1 were overexpressed in the mutants. These data suggested that CAld5H is mainly involved in the production of non-γ-p-coumaroylated S lignin units, common in both eudicots and grasses, but not in the production of grass-specific γ-p-coumaroylated S units in rice.

リンク情報
DOI
https://doi.org/10.1111/tpj.14141
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/30375064

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