Maruyama Ryo, Mayuzumi Yasuyoshi, Morisawa Jun, Kawai Shinya
United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, Fuchu, Tokyo 183-8509, Japan.
Graduate School of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Tokyo 183-8509, Japan.
Plant Biotechnol (Tokyo). 2021 Sep 25;38(3):379-386. doi: 10.5511/plantbiotechnology.21.0616a.
Lignocellulosic materials are potential renewable sources of fermentable sugars for bioethanol production. In this study, we used the gene encoding CcAbf62A, a putative extracellular α-L-arabinofuranosidase, cloned from the mycotrophic basidiomycete . CcAbf62A acts on arabinoxylan, the major hemicellulose of grasses, releasing arabinose. was introduced into rice with the aim of enhancing delignification efficiency and the availability of lignocellulosic materials without reducing lignin content. Among the 32 lines of regenerated transgenic rice, 13 exhibited markedly disrupted elongation growth and excessive tillering (dwarf), seven showed delayed elongation growth (retarded-growth), and 12 showed phenotypes similar to those of control plants (normal). Additionally, the dwarf lines showed reduced acclimation. RT-PCR analysis revealed that dwarf lines had higher levels of expression than retarded-growth and normal lines. Although the lignin content of transgenic rice plants expressing did not differ significantly from that of control rice plants, dwarf lines were characterized by delayed deposition of lignin in the culms compared with the controls. The reduced acclimation ability of dwarf lines is believed to be associated with increased water loss and reduced water conductivity concomitant with delayed lignin deposition. Contrary to expectations, the alkaline delignification rates of dwarf and retarded-growth Abf lines were slightly lower than those of control rice plants. Our findings indicate that CcAbf62A reduces ferulate-lignin cross-links by detaching arabinose side chains from arabinoxylan and increases the relative abundance of alkaline-resistant benzyl ether cross-links. CcAbf62A is anticipated to provide new approaches for breeding plants containing altered lignocellulosic materials or lodging-resistant crops.
木质纤维素材料是生物乙醇生产中潜在的可发酵糖可再生来源。在本研究中,我们使用了从菌根担子菌中克隆的编码假定的细胞外α-L-阿拉伯呋喃糖苷酶CcAbf62A的基因。CcAbf62A作用于阿拉伯木聚糖(禾本科植物的主要半纤维素),释放阿拉伯糖。将其导入水稻以提高脱木质素效率和木质纤维素材料的可利用性,同时不降低木质素含量。在32株再生转基因水稻株系中,13株表现出明显的伸长生长受阻和分蘖过多(矮化),7株表现出伸长生长延迟(生长迟缓),12株表现出与对照植株相似的表型(正常)。此外,矮化株系的适应性降低。RT-PCR分析表明,矮化株系中该基因的表达水平高于生长迟缓和正常株系。虽然表达该基因的转基因水稻植株的木质素含量与对照水稻植株没有显著差异,但与对照相比,矮化株系茎中木质素的沉积延迟。矮化株系适应性降低的能力被认为与水分流失增加和木质素沉积延迟伴随的水分传导率降低有关。与预期相反,矮化和生长迟缓的Abf株系的碱性脱木质素率略低于对照水稻植株。我们的研究结果表明,CcAbf62A通过从阿拉伯木聚糖上分离阿拉伯糖侧链来减少阿魏酸-木质素交联,并增加耐碱性苄基醚交联的相对丰度。预计CcAbf62A将为培育含有改变的木质纤维素材料的植物或抗倒伏作物提供新方法。