棉花胞质不育育性恢复候选基因载体构建及其功能初步分析

棉花细胞质雄性不育是研究杂种优势利用和质核互作机理的理想材料。本实验对棉花细胞质雄性不育育性恢复基因ZH46-3437利用pBI121载体构建了过表达载体。同时构建了amiRNA干扰载体即amiR3437,利用病毒诱导基因沉默体系pCLCrV浸染F1植株的子叶。转基因植株的相对荧光定量数据表明,amiR3437的相对表达量上调,而ZH46-3437的相对表达量下调,利用碘-碘化钾染色法鉴定转基因出现半不育性状,表明ZH46-3437基因可能与棉花花粉育性有关。

Vector Construction and Preliminarily Functional Analysis of a Candidate Gene for Cytoplasmic Male Sterility in Cotton

Cytoplasmic male sterility (CMS) in cotton is an ideal system to use heterosis to study nucleo-cytoplasmic interactions. In this study, an over-expression vector of the candidate fertility restoration gene ZH46-3437 was constructed using a pBI121 vector. An amiRNA interference vector, amiR3437, was also constructed and inoculated into the cotyledon of an F1 hybrid plants using the CLCrV (Cotton leaf crumple virus)-induced gene silencing system. qRT-PCR analysis of transgenic plants containing the transgene showed that the expression level of amiR3437 increased, while the expression level of ZH46-3437 decreased. The semi-sterility of the pollen of the transgenic plants could be detected by I₂-KI staining method. This research implied that ZH46-3437 may play a role in cotton fertility.