芥菜hau胞质雄性不育源导致近缘种缺绿性状的核质互作分子机制

细胞质雄性不育（CMS）是十字花科蔬菜杂种优势利用的主要途径。但是新的雄性不育源在利用中容易出现蜜腺不发达、叶片缺绿等问题，极大地影响蔬菜商品性和杂种产量。项目申请人前期报道了芥菜hau CMS不育基因的克隆并创制了白菜叶片缺绿hau细胞质雄性不育系1409A和叶色正常的保持系1409B。本项目拟以此为基础，在基因组和蛋白质组水平上对1409A和1409B的叶绿体进行分析，获得具有差异表达的基因与蛋白；同时对1409A和1409B进行总RNA转录组差异表达分析，筛选与叶片缺绿性状相关的核基因，并通过转基因功能验证确定候选基因；然后进行酵母文库互作蛋白的筛选和GST pull-down、BiFC蛋白互作的验证，确定叶片缺绿基因的互作蛋白。该项目的实施不仅可以挖掘芥菜hau CMS引起近缘种缺绿性状的关键基因和蛋白，而且将进一步揭示其内在的核质互作分子机制，具有重要的理论和育种实践价值。

Cytoplasmic male sterile (CMS) is the main way to be used in the heterotic utility of cruciferous vegetables. However, defective nectaries and etiolation in newly CMS lines would affect the quality of vegetable and hybrids producing. The discovery of a mustard hau CMS associated gene has been reported. In addition, by back-crossing for multiple generations in cabbage, the hau CMS line 1409A with yellow leaves and its isonuclear maintainer line 1409B with green leaves were developed. Based on these previous studies, the mitochondria and chloroplast of the male sterile line 1409A and maintainer 1409B will be analyzed on the genomic and proteomic levels, to obtain the differentially expressed genes and proteins;. Meanwhile, expression profile analysis in 1409A and 1409B will also be carried out to screen differential expressed genes related to chlorophyll synthesis. Furthermore, candidate genes associated with leaf etiolation will be verified by transformation. Then, combined with screening of yeast library interaction proteins, GST pull-down and BiFC, the interacting protein of leaf etiolation gene would be further idenified. The implementation of this project will not only explore the key genes and proteins causing leaf etiolation, but also further reveal its intrinsic molecular mechanism of nucleo-cytoplasmic interaction. This research has important theoretical and practical value in breeding.