Ma Xiaozhi, Li Chunmei, Huang Rui, Zhang Kuan, Wang Qian, Fu Chongyun, Liu Wuge, Sun Changhui, Wang Pingrong, Wang Feng, Deng Xiaojian
State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Rice Research Institute, Sichuan Agricultural University, Chengdu, 611130, China.
Guangdong Provincial Key Laboratory of New Technology in Rice Breeding, Rice Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, 510642, China.
Rice (N Y). 2021 Nov 25;14(1):95. doi: 10.1186/s12284-021-00536-2.
Mechanical strength is a crucial agronomic trait in rice (Oryza sativa), and brittle mutants are thought suitable materials to investigate the mechanism of cell wall formation. So far, almost all brittle mutants are recessive, and most of them are defected in multiple morphologies and/or grain yield, limiting their application in hybrid breeding and in rice straw recycling.
We identified a semi-dominant brittle mutant Brittle culm19 (Bc19) isolated from the japonica variety Nipponbare through chemical mutagenesis. The mutant showed the same apparent morphologies and grain yield to the wild type plant except for its weak mechanical strength. Its development of secondary cell wall in sclerenchyma cells was affected, along with reduced contents of cellulose, hemicellulose, lignin and sugars in culms and leaves. Positional cloning suggested that the Bc19 gene was allelic to OsCESA4, encoding one of the cellulose synthase A (CESA) catalytic subunits. In this mutant, a C-to-T substitution occurred in the coding sequence of BC19, causing the P507S missense mutation in its encoded product, which was located in the second cytoplasmic region of the OsCESA4 protein. Furthermore, introducing mutant gene Bc19 into the wild-type plant resulted in brittle plants, confirming that the P507S point mutation in OsCESA4 protein was responsible for the semi-dominant brittle phenotype of Bc19 mutant. Reverse correlation was revealed between cellulose contents and expression levels of mutant gene Bc19 among the homozygous mutant, the hybrid F plant, and the Bc19 overexpression transgenic plants, implying that gene Bc19 might affect cellulose synthesis in a dosage-dependent manner.
Bc19, a semi-dominant brittle mutant allele of gene OsCESA4, was identified using map-based cloning approach. The mutated protein of Bc19 possessing the P507S missense mutation behaved in a dosage-dependent semi-dominant manner. Unique brittle effect on phenotype and semi-dominant genetic quality of gene Bc19 indicated its potential application in grain-straw dual-purpose hybrid rice breeding.
机械强度是水稻(Oryza sativa)的一个关键农艺性状,脆性突变体被认为是研究细胞壁形成机制的合适材料。到目前为止,几乎所有的脆性突变体都是隐性的,并且它们中的大多数在多种形态和/或谷物产量方面存在缺陷,这限制了它们在杂交育种和稻草回收中的应用。
我们通过化学诱变从粳稻品种日本晴中分离出一个半显性脆性突变体脆茎19(Bc19)。该突变体除了机械强度较弱外,其外观形态和谷物产量与野生型植株相同。其厚壁组织细胞中次生细胞壁的发育受到影响,同时茎和叶中纤维素、半纤维素、木质素和糖类的含量降低。图位克隆表明,Bc19基因与OsCESA4等位,OsCESA4编码纤维素合酶A(CESA)催化亚基之一。在这个突变体中,BC19的编码序列发生了C到T的替换,导致其编码产物中出现P507S错义突变,该突变位于OsCESA4蛋白的第二个细胞质区域。此外,将突变基因Bc19导入野生型植株中导致植株变脆,证实OsCESA4蛋白中的P507S点突变是Bc19突变体半显性脆性表型的原因。在纯合突变体、杂交F1植株和Bc19过表达转基因植株中,纤维素含量与突变基因Bc19的表达水平呈负相关,这意味着基因Bc19可能以剂量依赖的方式影响纤维素合成。
利用图位克隆方法鉴定出基因OsCESA4的一个半显性脆性突变等位基因Bc19。具有P507S错义突变的Bc19突变蛋白表现出剂量依赖的半显性方式。基因Bc19对表型的独特脆性效应和半显性遗传特性表明其在粮饲兼用型杂交水稻育种中的潜在应用价值。