Regeneration of transgenic vegetable brassicas (Brassica oleracea and B-campestris) via Ri-mediated transformation
Regeneration of transgenic vegetable brassicas (Brassica oleracea and B-campestris) via Ri-mediated transformation
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DOI：
10.1007/bf01275497
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发表时间：
1997-06-01
影响因子：
6.2
通讯作者：
Wyke, L
中科院分区：
文献类型：
--
作者：
Christey, MC;Sinclair, BK;Wyke, L
本文报道了利用Pi介导法获得可育转基因甘蓝的方法。以青花菜、布鲁塞尔芽苗菜、甘蓝、花椰菜、快周期（均为甘蓝）和大白菜（B）6个品种的12个甘蓝品种和品系为材料，筛选出转基因毛状根系。campestris）。将完整子叶的叶外植体或叶柄与携带各种二元载体的农杆菌菌株A4 T共培养。所有双元载体的T-DNA区域除了包含其他基因之外，还包含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菌培养基上大量生长。转基因芽再生从所有品种自发或转移后的毛状根到一个含琼脂糖培养基。Southern分析证实这些植物是转基因的。将来自所有芸苔属类型的植物成功地转移到温室条件下。植株可育，分离分析证实性状传递给后代。
A procedure for the production of fertile transgenic brassicas via Pi-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 (all Brassica oleracea) and Chinese cabbage (B. campestris). Leaf explants or petioles of intact cotyledons were co-cultivated with Agrobacterium 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.