Christey M C, Sinclair B K, Braun R H, Wyke L
New Zealand Institute for Crop and Food Research Limited, Private Bag 4704, Christchurch, New Zealand.
Plant Cell Rep. 1997 Jun;16(9):587-593. doi: 10.1007/BF01275497.
A procedure for the production of fertile transgenic brassicas via Ri-mediated transformation is reported in this paper. Transgenic hairy root lines were selected for 12 vegetable brassica cultivars and lines representing six varieties: broccoli, Brussels sprouts, cabbage, cauliflower, rapid-cycling (allBrassica oleracea) and Chinese cabbage (B. campestris). Leaf explants or petioles of intact cotyledons were co-cultivated withAgrobacterium strain A4T harbouring various binary vectors. The T-DNA region of all binary vectors contained a neomycin phosphotransferase II gene for kanamycin resistance, in addition to other genes. Hairy root lines grew prolifically on hormone-free medium containing kanamycin. Transgenic shoots were regenerated from all cultivars either spontaneously or after transfer of hairy roots to a hormone-containing medium. Southern analysis confirmed that the plants were transgenic. Plants from all brassica types were successfully transferred to greenhouse conditions. Plants were fertile and segregation analysis confirmed transmission of traits to progeny.
本文报道了一种通过发根农杆菌介导的转化方法生产可育转基因芸苔属植物的过程。从12个蔬菜芸苔属栽培品种和品系中筛选出转基因毛状根系，它们代表了六个品种：西兰花、抱子甘蓝、卷心菜、花椰菜、快生型（均为甘蓝型油菜）和大白菜（白菜型油菜）。完整子叶的叶片外植体或叶柄与携带各种双元载体的发根农杆菌菌株A4T共同培养。所有双元载体的T-DNA区域除了其他基因外，还包含一个用于卡那霉素抗性的新霉素磷酸转移酶II基因。毛状根系在含有卡那霉素的无激素培养基上大量生长。所有品种的转基因芽要么自发再生，要么在将毛状根转移到含激素的培养基后再生。Southern分析证实这些植株是转基因植株。所有芸苔属类型的植株都成功转移到温室条件下。植株可育，分离分析证实性状可传递给后代。
