以萱草‘香宝’愈伤组织为外植体，以10 d为继代周期，将携有 PSN1301-CHS 的农杆菌 EHA105菌株侵染受体材料，使 CHS 基因导入到萱草‘香宝’中。建立萱草遗传转化的适宜条件：抑菌剂以浓度为350 mg/L 的羧苄青霉素为宜；潮霉素为转化子的筛选剂，25 mg/L 为适宜的作用浓度；转化中宜选择预培养时间为2 d；农杆菌菌液浓度 OD600在0.6左右，选用重悬液为 MS+3%糖；侵染时间为15 min；在菌液和共培养基中同加100μmol/L 的乙酰丁香酮；共培养基时间为2 d。%With the callus of Hemerocallis hybrid as explant, 10 d as one subculture period, Agrobacterium EHA105 carried with PSN1301-CHS was used to infect explant to tranfer CHS gene into Hemerocallis hybrid. The main factors influencing the gene transformation of Hemerocallis hybrid were as follows: the best carbenicillin concentration was 350 mg/L as bacteriostatic agent;hygromycin was suitable to select transformed tissue, and appropriate screening concentration was 25 mg/L; the time of preculture were 2 days for explant; the OD600 value of appropriate concentration of Agrobacterium was 0.6; MS+3% sucrose was chose as new medium to suspension bacteria; the most appropriate infection time was 15 min. It was beneficial to add 100 μmol/L acetosyringone into cocultivation medium for improving the transfer rate. 2 days were the best co-culture time for explant.
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