Xu Jing, Deng Yiwen, Li Qun, Zhu Xudong, He Zuhua
National Key Laboratory of Plant Molecular Genetics and National Center of Plant Gene Research, Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, China.
China National Rice Research Institute, Hangzhou 31006, China.
J Genet Genomics. 2014 Oct 20;41(10):539-48. doi: 10.1016/j.jgg.2014.05.008. Epub 2014 Jun 19.
Mutants with abnormal leaf coloration are good genetic materials for understanding the mechanism of chloroplast development and chlorophyll biosynthesis. In this study, a rice mutant st2 (stripe2) with stripe leaves was identified from the γ-ray irradiated mutant pool. The st2 mutant exhibited decreased accumulation of chlorophyll and aberrant chloroplasts. Genetic analysis indicated that the st2 mutant was controlled by a single recessive locus. The ST2 gene was finely confined to a 27-kb region on chromosome 1 by the map-based cloning strategy and a 5-bp deletion in Os01g0765000 was identified by sequence analysis. The deletion happened in the joint of exon 3 and intron 3 and led to new spliced products of mRNA. Genetic complementation confirmed that Os01g0765000 is the ST2 gene. We found that the ST2 gene was expressed ubiquitously. Subcellular localization assay showed that the ST2 protein was located in mitochondria. ST2 belongs to the cytidine deaminase-like family and possibly functions as the dCMP deaminase, which catalyzes the formation of dUMP from dCMP by deamination. Additionally, exogenous application of dUMP could partially rescue the st2 phenotype. Therefore, our study identified a putative dCMP deaminase as a novel regulator in chloroplast development for the first time.
叶色异常的突变体是研究叶绿体发育和叶绿素生物合成机制的良好遗传材料。本研究从γ射线辐照突变体库中鉴定出一个具有条纹叶的水稻突变体st2(stripe2)。st2突变体表现出叶绿素积累减少和叶绿体异常。遗传分析表明,st2突变体受一个隐性单基因座控制。通过图位克隆策略,将ST2基因精细定位到第1染色体上一个27 kb的区域,并通过序列分析鉴定出Os01g0765000存在一个5 bp的缺失。该缺失发生在外显子3和内含子3的连接处,导致mRNA产生新的剪接产物。遗传互补实验证实Os01g0765000就是ST2基因。我们发现ST2基因在各处均有表达。亚细胞定位分析表明,ST2蛋白定位于线粒体。ST2属于胞苷脱氨酶样家族,可能作为dCMP脱氨酶发挥作用,通过脱氨作用催化dCMP形成dUMP。此外,外源施加dUMP可部分挽救st2的表型。因此,我们的研究首次鉴定出一种假定的dCMP脱氨酶是叶绿体发育中的一种新型调节因子。