甘蓝型油菜花序长发夹lhRNAi文库的构建  

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

作　　者：曾新华[1] 赵圣博 李可琪[1] 袁荣[1] 吴刚[1] 闫晓红[1] ZENG Xin-hua;ZHAO Sheng-bo;LI Ke-qi;YUAN Rong;WU Gang;YAN Xiao-hong(Oil Crops Research Institute of Chinese Academy of Agricultural Sciences / Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, 430062, China)

机构地区：[1]中国农业科学院油料作物研究所/农业部油料作物生物学与遗传育种重点实验室

出　　处：《中国油料作物学报》2019年第3期340-348,共9页Chinese Journal of Oil Crop Sciences

基　　金：国家自然科学基金面上项目(31671733);转基因生物新品种培育重大专项(2018ZX0801104B)

摘　　要：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、死蕾或花蕾闭合等。说明通过滚环复制介导的lhRNAi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的功能研究提供重要的研究平台。Allopolyploid Brassica 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 transgenic plants.A total of 74 transgenic lines exhibited visible mutant phenotypes,including petal reduction in the number,stamen degeneration shrinkage,stigma distortion and exposure,dead or closed buds and male sterility.These results demonstrated the construction method of lhRNAi library which were successfully applied to rapeseed and provided an important research platform for gene functions related to rape flower inflorescence development.

关 键 词：油菜 滚环复制 长发夹RNAi文库 花发育 

分 类 号：S435.654[农业科学—农业昆虫与害虫防治]