发根农杆菌介导药用甘薯西蒙1号的遗传转化

Genetic Transformation of Sweet Potato Simon 1 (Ipomoea batatas)by Agrobacterium rhizogenes

用发根农杆菌A4分别感染药用甘薯西蒙1号的叶片、茎切段、叶柄等外植体,诱导出毛状根,并对毛状根进行了离体培养。采用L9(34)正交设计法优化甘薯西蒙1号的毛状根诱导条件;PCR扩增检测转化毛状根;用高效液相色谱仪检测了毛状根中咖啡酸的含量。结果表明:转化中茎切段是最合适的外植体,最佳感染时间20 min,共培养最佳时间为2天;PCR扩增检测表明发根农杆菌Ri质粒的T-DNA片段已整合进植物的基因组中;经高效液相色谱仪证实毛状根中含有咖啡酸,含量为0.03792 mg/g。

Different types of explants including leaves, stems, petioles obtained from sweet potato Simon 1 were respectively infected by Agrobacterium rhizogenes strain A4. Hairy roots were induced and in vitro cultured. Induction conditions were optimized through orthogonality L9 （34） method ;The transformation of Ri T-DNA was examined by PCR;HPLC detected the caffeic acid in the hairy roots of sweet potato Simon 1. The results showed that the best infected explants is the stem, the best infection time is 20 min, the best co-culture time is 2 d;The transformation of T-DNA from Ri plasmid to the hairy roots was confirmed by PCR analysis; HPLC analysis showed that hairy roots contain caffeic acid, the content of affeic acid in hairy roots is 0. 03792 mg/g.