多聚半乳糖醛酸酶反义基因转化加工番茄

Polygalacturonase Antisense Gene into Processing Tomato

利用农杆菌EHA105介导转化,将PG基因转入加工番茄B04植株。通过对影响外植体转化的因素(预培养时间、侵染时间、农杆菌浓度、共培养时间)进行了深入探讨,建立了加工番茄品种B04子叶外植体的高频遗传转化体系,并通过农杆菌介导法将PG反义基因导入加工番茄品种B04,获得PG抗性植株。其最佳预培养时间为2天;当农杆菌的菌液浓度为OD600=0.6、侵染时间为10 min时转化率最高;其最佳的共培养时间为2天。对获得的41株抗卡那霉素转基因植株进行PCR检测,其中9株为阳性植株,其转化率为22%。初步证实了部分加工番茄B04植株中已导入PG反义基因。

Agrobacterium mediated transformation using EHA105, PG gene was transferred into processing tomato B04 plants. Through the influence of explants conversion factor（pre-cultural time, invading time, concentration of bacteria, total co-cultural time） were studied, to establish a high frequency of genetic transformation system for processing tomato cotyledon explants of B04. Agrobacterium was used for transformation, and then transgenic plantlets with polygalacturonase gene of processing tomato B04 were obtained. The best pre-culturing time was 2 d. The suit pre-cultural time for processing tomato was 48 hours; OD600 =0.6 was the fitful bacteria concentration for invading; and the time for invading was 10 minutes; the explants and the bacteria had been best co-cultured for 48 hours. By PCR analysis, 9 plants were positive in 41 plants, and the rate of transformation was 22%. The results proved that the foreign genes had been integrated into B04 processing tomato.