利用amiRNA技术沉默矮牵牛查尔酮合成酶基因  

Silencing of Chalcone Synthase Genes by Artificial microRNA in Petunia

作　　者：王会平[1] 遇玲[1] 邹世慧[1] 陈璟[1] 郭余龙[1] 李名扬[1] 

机构地区：[1]西南大学园艺园林学院,南方山地园艺学教育部重点实验室,重庆市花卉工程技术中心,重庆400716

出　　处：《园艺学报》2012年第12期2491-2498,共8页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(30671481)

摘　　要：转基因介导的基因沉默技术是进行基因功能分析和利用基因工程改良作物的重要手段。在植物中,amiRNA(artificial microRNA)是具有高度特异性的基因沉默技术。根据amiRNA设计的原则设计了用于沉默矮牵牛查尔酮合成酶(chalcone synthase,CHS)基因的amiRNA:amiRchs1,用拟南芥miR319a的前体作为表达amiRchs1的骨架,重叠PCR扩增,将天然miR319a序列替换成amiRchs1,合成的DNA片段置于35S启动子之后,转化导入矮牵牛。转基因植株花色变淡,花冠出现弥散的白色斑点或不规则的白色斑块,严重的几乎呈纯白色,花色素苷含量明显下降,CHS的mRNA含量明显降低,表明拟南芥miR319a的前体作为表达amiRNA的骨架在矮牵牛中可行,且能有效沉默结构基因。Transgene-mediated gene silencing is a useful technique for elucidating biological function of genes and modifying crops by genetic engineering. In plants, amiRNAs make an effective tool for specific gene silencing. We designed the amiRchsl for silencing the petunia chalcone synthase genes according to the amiRNA design principles refined from the structure characters of plant microRNA. We used the precursor for Arabidopsis miR319a as backbone for the amiRchs 1 expression under the control of 35S promoter, exchanging the natural miR319a sequence with the sequence of amiRchsl by overlapping PCR. The transgenic petunia plants expressing the amiRchsl precursor produced flowers with varying amounts of diffused white dots or irregular white patches, and nearly white flowers. Both the contents of anthocyanin and CHS mRNA in corollas of the transgenic plants were reduced significantly. These results showed that the precursor for Arabidopsis miR319a can be used to express amiRNA in petunia, and structure genes can be knocked down effectively by the amiRNA technology.

关 键 词：矮牵牛 基因沉默 amiRNA 查尔酮合成酶基因 

分 类 号：S685.11[农业科学—观赏园艺]