Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
Plant Physiol. 2012 Aug;159(4):1440-52. doi: 10.1104/pp.112.195529. Epub 2012 Jun 4.
Plant chitinases, a class of glycosyl hydrolases, participate in various aspects of normal plant growth and development, including cell wall metabolism and disease resistance. The rice (Oryza sativa) genome encodes 37 putative chitinases and chitinase-like proteins. However, none of them has been characterized at the genetic level. In this study, we report the isolation of a brittle culm mutant, bc15, and the map-based cloning of the BC15/OsCTL1 (for chitinase-like1) gene affected in the mutant. The gene encodes the rice chitinase-like protein BC15/OsCTL1. Mutation of BC15/OsCTL1 causes reduced cellulose content and mechanical strength without obvious alterations in plant growth. Bioinformatic analyses indicated that BC15/OsCTL1 is a class II chitinase-like protein that is devoid of both an amino-terminal cysteine-rich domain and the chitinase activity motif H-E-T-T but possesses an amino-terminal transmembrane domain. Biochemical assays demonstrated that BC15/OsCTL1 is a Golgi-localized type II membrane protein that lacks classical chitinase activity. Quantitative real-time polymerase chain reaction and β-glucuronidase activity analyses indicated that BC15/OsCTL1 is ubiquitously expressed. Investigation of the global expression profile of wild-type and bc15 plants, using Illumina RNA sequencing, further suggested a possible mechanism by which BC15/OsCTL1 mediates cellulose biosynthesis and cell wall remodeling. Our findings provide genetic evidence of a role for plant chitinases in cellulose biosynthesis in rice, which appears to differ from their roles as revealed by analysis of Arabidopsis (Arabidopsis thaliana).
植物几丁质酶是糖苷水解酶的一类,参与正常植物生长和发育的各个方面,包括细胞壁代谢和抗病性。水稻(Oryza sativa)基因组编码 37 种假定的几丁质酶和几丁质酶样蛋白。然而,它们都没有在遗传水平上进行过特征描述。在本研究中,我们报告了脆性穗突变体 bc15 的分离和受突变影响的 BC15/OsCTL1(几丁质酶样 1)基因的图谱克隆。该基因编码水稻几丁质酶样蛋白 BC15/OsCTL1。BC15/OsCTL1 的突变导致纤维素含量和机械强度降低,而植物生长无明显改变。生物信息学分析表明,BC15/OsCTL1 是一种 II 类几丁质酶样蛋白,缺乏氨基端富含半胱氨酸的结构域和几丁质酶活性基序 H-E-T-T,但具有氨基端跨膜结构域。生化分析表明,BC15/OsCTL1 是一种定位于高尔基体的 II 型膜蛋白,缺乏经典的几丁质酶活性。定量实时聚合酶链反应和β-葡萄糖醛酸酶活性分析表明,BC15/OsCTL1 广泛表达。使用 Illumina RNA 测序对野生型和 bc15 植物的全球表达谱进行研究,进一步提出了 BC15/OsCTL1 介导纤维素生物合成和细胞壁重塑的可能机制。我们的研究结果为植物几丁质酶在水稻纤维素生物合成中的作用提供了遗传证据,这似乎与通过拟南芥(Arabidopsis thaliana)分析揭示的作用不同。