S1基因结构解析与分子进化分析

Structural Analysis and Molecular Evolution of S1 Gene

S1是水稻(Oryza sativa L.)亚非栽培稻种间杂种不育一个最重要的基因座,对杂种的雌雄配子都有选择性杀灭作用。前期我们通过图位克隆获得S1基因座的候选基因后,NCBI查找发现‘日本晴’的S1等位基因序列全长7 kb,局倍区域有高GC分布。为此本研究通过分多段运用有针对性的PCR方法,对所用的材料IRAT216与IRAT216S1进行了扩增并测序,获得了各自的S1全长序列,并进行了比对,发现IRAT216与‘日本晴’的序列完全相同,但IRAT216与IRAT216S1之间存在多处变异。同时针对有效变异区对S1基因进行了分子进化调查,建立了水稻的分子进化树,确定了S1在物种进化中的意义。本研究的S1基因序列测定,将为水稻分子标记辅助育种中分子标记的开发提供序列参照,同时也将为S1基因RNA的表达分析与蛋白功能研究、在各物种中的分化变异和水稻在进化中地位划分提供理论依据,并能为水稻的起源研究提供参考。

S1 is one of the most important loci for interspecific hybrid sterility in Asian-African cultivated rice(Oryza sativa L.), and it has selective killing effect on the male and female gametes of hybrids. In the early stage,we obtained the candidate gene of S1 locus by map cloning. NCBI search found that the S1 allele sequence of‘Nipponbare’had a total length of 7 kb, and high GC distribution was found in partial region. To this end, we used multi-stage targeted PCR methods to amplify and sequence the materials IRAT216 and IRAT216S1. The full-length sequences of S1 were obtained and compared, which found that the sequences of IRAT216 and ‘Nipponbare’ were completely same, but there were multiple variations between IRAT216 and IRAT216S1. At the same time, the molecular evolution of the S1 gene was investigated for the effective variant region, and the molecular phylogenetic tree of rice was established, and the significance of S1 in species evolution was determined. The sequencing of S1 gene in this study would provide a sequence reference for the development of molecular markers in rice molecular marker-assisted breeding. Meanwhile, it would provide a theoretical basis for the analysis of S1 gene RNA expression and protein function, differentiation and variation in various species and the division of rice status in evolution, and provide a reference for the study of rice origin.