基于原芯片杂交结果,油菜杂合双显性雄性核不育系差异表达基因Bnl9070与拟南芥At2g19070基因高度同源,在油菜可育株雄蕊中高量表达,不育株雄蕊中表达量极低。从雄性不育基因At2g19070入手,利用amiR—NAi(人工微RNA干扰)技术对其两个RNA区段分别设计靶标,转化拟南芥。在多个转基因拟南芥植株和后代中普遍观察到小孢子数量减少、成熟花粉萎缩失去活力、不结实或者少结实的不育表型。两个靶标的干扰使转基因拟南芥出现了类似的表型,基因表达抑制率都在90%以上,干扰效果好,暗示amiRNAi技术在靶标选择上具有一定的灵活性。At2g19070基因的干扰对Bnl9070的功能研究有借鉴作用。
Based on previous microarray result of Brassica napus (heterozygous pairs of dominant male sterile line) and Arabidopsis thaliana genome, a differentially expressed B. napus gene Bn19070 which demonstrated high homology with At2gl9070 emerged. It was well expressed in fertile stem with low expression level in male sterile stem. To understand Bnl9070g function, amiRNA (artificial mieroRNA) gene silencing technique was used to dis- able the function of homologous gene At2g19070. Two interference targets from At2g19070 mRNA were selected and transformed into Arabidopsis. amiRNAi (amiRNA interference ) phenotypes related to male sterility were ob- served in transgenic Arabidopsis, including decreased mierosporc number, deteriorated anther and fruitless. Expres- sions of both targets decreased remarkably ( more than 90% ) in T2 lines with similar phenomena. Results indicated the flexibility of amiRNAi in target site selection. The interference research of At2g19070 would contribute to Bn19070 gene study.