通过PCR法扩增出陆地棉GhRDL1基因ATG上游1.2 kb的片段,将该片段融合GUS报告基因进行棉花、拟南芥及烟草转化.分析显示,GhRDL1基因启动子是一个棉纤维或表皮毛特异的启动子,能够驱动靶基因在单细胞的表皮毛(如棉纤维和拟南芥表皮毛)表达;在烟草表皮毛中,GUS基因在多细胞的腺毛(包括顶端的腺体细胞和柄细胞)和非腺毛中皆有表达.表明GhRDL1基因启动子是一个多功能的表皮毛特异启动,可以用于棉花基因工程改良和植物次生代谢工程的研究.
A 1.2 kb upstream fragment of GhRDL1 was isolated form Gossypium hirsutum and fused to a β-glucuronidase(GUS) reporter gene,and was transferred into cotton,Arabidopsis and tobacco.In transgenic cotton plants,GhRDL1 promoter exhibited activities predominantly in developing fiber cellsand 0-DPA ovule and to a less extent in trichomes of other aerial organs,including leaf,stem,bract,sepal and petal.In transgenic Arabidopsis plants the promoter was also specific to trichomes.Different from Arabidopsis and cotton fiber that have unicellular simple trichomes,tobacco has glandular secreting trichomes(GSTs) and simple trichomes,both are multicellular.In tobacco the GhRDL1 promoter directed GUS gene expression exclusively in the GSTs(Including the head cells and stalk cell) and multicellular simple trichomes.This promoter provides a tool not only for genetic engineering of cotton fiber improvement,but also for the GSTs-based plant secondary metabolism engineering.
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