甘蓝型油菜花序长发夹ｌｈＲＮＡｉ文库的构建

异源四倍体油菜含有大量多拷贝基因及冗余基因，难以通过T-DNA插入、物理及化学诱变等常规突变体库创制方法发掘功能基因。为开发适应于油菜的简单高效的突变体库创制方法，本研究通过滚环复制方法构建了甘蓝型油菜的花序lhRNAi文库，并对这种新型的干扰文库进行质量评估；然后将该花序lhRNAi文库对甘蓝型油菜进行遗传转化，获得763株T0植株，从中鉴定分离出74株具有可见表型变异的花发育相关突变体，包括花瓣减少、形状异常，柱头卷曲，雄蕊退化萎缩，雄性不育（不能产生花粉）、死蕾或花蕾闭合等。说明通过滚环复制介导的lihRNAi文库的构建方法可以成功应用到油菜中，这将为油菜花序发育相关基因功能的研究提供重要的研究平台。

Construction of a lhRNAi library related to rapeseed flower development mediated by rolling circle replication

Allopolyploid B. napus contains a large number of homologous and redundant genes. It is difficulty to identify novel genes related to important agronomic traits by traditional T-DNA insertional, physical and chemical mutagenesis methods. In this study, a genome-wide inflorescence lhRNAi library of B. napus was constructed by the rolling circle amplification (RCA) - mediated lhRNAi construction technology. Subsequently, the inflorescence lhRNAi library was transformed into B. napus to produce 763 T0 transgetic plants. A total of 74 transgenetic lines exhibited visible mutant phenotypes, including petal reduction in the number, stamen degeneration shrinkage and number reduction, stigma distortion and exposure, dead or closed buds and male sterility. These results demonstrated the construction method of lhRNAi library mediated by rolling circle replication and were successfully applied to rapeseed, which provided an important research platform for the study of gene functions related to rape flower inflorescence development.