农杆菌介导AVP1基因转化甜菜

Agrobacterium-mediated Transformation of Sugar Beet with Vacuolar H+-PPase (AVP1) Gene

甜菜是我国重要的糖料作物之一。为了提高甜菜含糖量和产量,增强其抗逆性,本研究应用植物基因工程手段对甜菜进行遗传改良。实验以甜菜DF1031品系为受体材料,利用农杆菌介导法将AVP1基因转入甜菜子叶节,获得转基因植株。研究了菌液侵染浓度、侵染时间、共培养时间、抗生素羧苄青霉素(Carb)、头孢霉素(Cef)以及筛选剂卡那霉素(Kan)等因素对遗传转化效率的影响,优化了甜菜的遗传转化体系。结果表明:菌液浓度0.4～0.5,侵染时间6～8min,共培养时间36～48h,Cef浓度500mg/L,卡那霉素(Kan)浓度100mg/L时得到较高转化率(11.3%),获抗性植株72株,检测到阳性植物37株,阳性率51.4%。

Sugar beet is one of the important sugar crops. In order to improve the sugar content and yield of sugar beet, enhance its resistance, this study applies plant gene engineering means of sugar beet on genetic improvement. AVP1 gene was successfully transformed into the explants of cotyledon node. The study on effects of Agrobacterium concentration, immersed time, co-culture time, Carb, Cef and Kan on genetic transformation frequencies. An efficient genetic transformation system was established. The results showed that bacterium concentration OD600=0.4-0.5, bacterium infection 6-8 min, 36-48 h of co-culture, Cef concentration 500 mg/L, Kan concentration 100 mg/L has obtain the lightest transformation rate （11.3%）. 72 resistant plants and 37 transgenic plants were obtained.