Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto, 611-0011, Japan.
Faculty of Bioscience and Bioindustry, Tokushima University, Kuramoto-cho, Tokushima, 770-8503, Japan.
Plant J. 2019 Jun;98(6):975-987. doi: 10.1111/tpj.14290. Epub 2019 Apr 8.
Breeding approaches to enrich lignins in biomass could be beneficial to improving the biorefinery process because lignins increase biomass heating value and represent a potent source of valuable aromatic chemicals. However, despite the fact that grasses are promising lignocellulose feedstocks, limited information is yet available for molecular-breeding approaches to upregulate lignin biosynthesis in grass species. In this study, we generated lignin-enriched transgenic rice (Oryza sativa), a model grass species, via targeted mutagenesis of the transcriptional repressor OsMYB108 using CRISPR/Cas9-mediated genome editing. The OsMYB108-knockout rice mutants displayed increased expressions of lignin biosynthetic genes and enhanced lignin deposition in culm cell walls. Chemical and two-dimensional nuclear magnetic resonance (NMR) analyses revealed that the mutant cell walls were preferentially enriched in γ-p-coumaroylated and tricin lignin units, both of which are typical and unique components in grass lignins. NMR analysis also showed that the relative abundances of major lignin linkage types were altered in the OsMYB108 mutants.
通过靶向敲除转录抑制因子 OsMYB108 利用 CRISPR/Cas9 介导的基因组编辑,我们在模式草本植物水稻(Oryza sativa)中生成了富含木质素的转基因水稻。OsMYB108 敲除水稻突变体表现出木质素生物合成基因的表达增加和茎细胞壁中木质素沉积增强。化学和二维核磁共振(NMR)分析表明,突变体细胞壁优先富含 γ-对香豆酰基化和松柏醇木质素单元,这两者都是禾本科木质素的典型和独特成分。NMR 分析还表明,OsMYB108 突变体中主要木质素连接类型的相对丰度发生了改变。