拟南芥WRKY13基因表达载体构建及鉴定分析

Construction and identification of Arabidopsis WRKY13 gene expression vector

铁是植物生长所必需的微量元素,目前植物受到缺铁胁迫的现象广泛存在,严重影响了植物的生长,减少了作物的产量。因此筛选和克隆植物缺铁胁迫耐受基因对解决农业生产问题具有极大的科学价值和现实意义。文章以拟南芥为实验材料,通过聚合酶链式反应(polymerase chain reaction,PCR)技术克隆WRKY13基因的启动子区域,成功构建了ProWRKY13∶GUS融合表达载体;利用浸花法将ProWRKY13∶GUS载体转化拟南芥,经鉴定分析后可知,获得了阳性转基因植株。对转基因植株进行缺铁诱导的GUS组织染色,实验结果为研究缺铁胁迫下WRKY13基因的功能及转录水平变化提供了重要依据。

Iron(Fe)is an essential micronutrient for plant growth.Nowadays plants are widely affected by Fe deficiency stress,which seriously affects plant growth and reduces crop yield.Therefore,screening and cloning of Fe deficiency stress tolerance genes are of great scientific value and practical significance to solve agricultural production problems.In this study,Arabidopsis thaliana was used as experimental material and the promoter region of the WRKY13 gene was cloned by polymerase chain reaction(PCR).The ProWRKY13∶GUS fusion expression vector was successfully constructed.The ProWRKY13∶GUS vector was transformed into Arabidopsis by the floral dip method,and positive transgenic plants were obtained after identification and analysis.The transgenic plants were subjected to GUS tissue staining induced by Fe deficiency.The results provide an important basis for studying the function and transcriptional changes of WRKY13 gene under Fe deficiency stress.