苎麻雄性不育相关atp6和atp9基因RNA干扰载体的构建

植物线粒体基因缺陷是细胞质雄性不育的主要原因。为了获得苎麻atp6和atp9基因RNA干扰(RNAi)表达载体，根据已报道的苎麻atp6和atp9基因序列设计引物，利用RT-PCR克隆了atp6和atp9基因的部分cDNA片段，将目的基因正反向片段连接入RNAi载体pHANNIBAL，再将其表达框连入表达载体pCAMBIA 1300。结果表明，所克隆的cDNA片段经序列比对后确认为目的基因片段，经酶切和测序验证确认完成了atp6和atp9基因RNAi载体pCAM-6SR和pCAM-9SR的构建。atp6和atp9基因RNAi载体是验证苎麻雄性不育的基础，也为苎麻遗传工程改良奠定了技术基础。

Construction of male sterility related atp6 and atp9 RNAi vectors in Ramie

Defects in mitochondrial genes are the main sources of cytoplasmic male sterility in plants. Arabidopsis and rice have exactly the same gene sequence copies of atp6 and atp9 in nuclear genomes as compared to their mitochondrial genomes, but weather the nuclear copy is expressed has less been understood. On the other hand, Ecto-F1-ATPase is found on the surface of cell membrane, but the origin of its subunit genes is still unknown. This article cloned atp6 and atp9 gene sense and reverse fragments into pHANNIBAL, and then the expression frame was cloned into pCAMBIA 1300 to complete the construction of atp6 and atp9 RNAi vectors. The purpose is to study if defects of atp6 and atp9 can resulted in male sterility and to lay the technical foundation of genetic improvement in ramie