百合遗传转化体系的建立

Establishment of genetic transformation system of lily

通过根癌农杆菌介导的遗传转化方法,建立了百合金黄精灵(ButterPixie)鳞茎的遗传转化体系。确定了适宜百合转化的筛选剂草甘磷的浓度:芽分化阶段为1.6mg.L-1,根分化阶段为0.8mg.L-1。除菌剂头孢的使用浓度为500mg.L-1。并且确定OD600=0.5的菌液浓度侵染20min,共培养pH5.2培养3d时对百合有最高转化效率。研究确定了预培养时间为4d,在共培养培养基中去除NH4NO3或加入阿魏酸均可提高转化效率。得到抗性植株12株,PCR初步检测发现其中7株为PCR阳性植株,转化效率为58.3%。

The paper established genetic transformation system of lily （Butter Pixie） bulbs mediated by Agrobacterium tumefacien. PPT is an effective selection substance in lily transformation, and the concentration in the period of buds differentiation was 1.6 mg·L^-1, the roots differentiation period was 0.8 mg·L^-1. The concentration of antibiotic Cef for restraining the propagation of excess A grobacterium was 500 mg·L^-1. Activated bacterial concentration was OD600=0.5, infection time was 20 min. Co-cultivation time was 3 days and the pH of co-cultivation medium was 5.2. Relatively high transformation ratio could be obtained in the above-mentioned conditions. At the same time, the 4 d of pre-culture time and co-cultivation medium MS added trans-Rerulic acid and subtracted NH4NO3 could improve the transformation ratio. Total 12 transformated plants were gained, and 7 plants were positive, the percentage of PCR positive plant was 58.3%.