Abstract:
In order to establish an Agrobacteriummediated genetic transformation system for Zanthoxylum piperitum DC.var.inerme Makino,using petioles and stems as explants, we examined several influencing factors of transformation efficiency, including precultivation time, optimum bacterium concentration and inflection period, and cocultivation time. The efficient transformation system obtained was 3 days precultivation time, 10 minutes inflection using bacterium at the concentration of OD600 0.5~0.8, followed by 3 days cocultivation. Selection of the transgenic plants was carried out after 7 days regeneration using gradient kanamycin at 30 and 50 mg/L, respectively. Adding 100 μmol/L acetosyringone in the resuspending solution and cocultivation medium increased the transformation efficiency to 24.6%. Transgenic plants were confirmed by GUS histochemical assays and polymerase chain reaction. The results suggest that an efficient Agrobacteriummediated transformation system for stable integration of foreign genes into Z. piperitum DC. var. inerme Makino has been developed, which could be applied in future studies on transferring economically important genes into Z. pipertum DC. var. inerme Makino.