TA29-barnase基因转化甘蓝产生雄性不育植株

用PCR技术从烟草革新1号品种的总DNA中扩增了TA29基因的启动子和从解淀粉芽孢杆菌的总DNA中扩增了核糖核酸水解酶基因(barnse)，将其构建成融合基因，并克隆于pCAMBIA2301载体上。通过根癌农杆茵介导转化甘蓝下胚轴，经Km选择压下连续选择、扩繁和进行生根培养，获得了甘蓝转基因植株。经GUS、PCR和Southernblot检测，证明TA29-bar-nase融合基因已经整合至转基因植株的染色体中。经花器官观察，转基因植株中有雄蕊退化的雄性不育和半不育植株出现。用正常花粉对不育株进行人工授粉，不育株能正常结实，这表明转基因不育植株的雌性器官发育正常，其不育性与TA29-barnse融合基因在转基因植株中的表达有关。

Male Sterile TransgenicCabbage Plants with TA29-barnase Gene

RNase gene (barnase) was cloned from Bacillus amyloliquefaciensgenomic DNA,and the upstream regulatory region (-1300～+51) of a tapetum-specific gene TA29 was cloned from tobacco (Nicotiana tabacum L.)total DNA by PCR and sequenced. Then a chimeric gene containing the regulatory region of TA29 gene and coding region of barnase gene was constructed (Fig. 1 ) and introduced into cabbage with Agrobacterium tumefaciens (Plate Ⅰ-1). Completely sterile, and half-sterile were observed among some transformed cabbages carring TA29-barnase chimeric gene (Plate -2～5 ), and other traits were basically identical to untransformed control plants, but filaments of stamens of transformed plant were shorter than normal ones, anthers were shriveled and without pollen. Self pollination of transformed plants failed to produce fruit capsules and seeds, but hybrid seeds were obtained when these plants were cross-pollinated artificially with normal pollen from untransformed cabbage plants. The results showed that these transformed plants were male sterile ( Plate Ⅰ-6 ).