多聚半乳糖醛酸酶反义基因转化加工番茄  

Polygalacturonase Antisense Gene into Processing Tomato

作　　者：陈丽萍 程智慧[2] 赵根 韩明丽 

机构地区：[1]湖州市农业科学研究院,浙江湖州313000 [2]西北农林科技大学园艺学院,陕西杨凌712100

出　　处：《中国农学通报》2014年第7期70-76,共7页Chinese Agricultural Science Bulletin

基　　金：新疆建设兵团博士资金项目"加工番茄果实硬度栽培生理和生物育种技术研究";国家自然科学基金项目"番茄转化酶在逆境生态下的表达及生理调节研究"(30370977)

摘　　要：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.

关 键 词：加工番茄 多聚半乳糖醛酸酶(PG) 反义基因 遗传转化 

分 类 号：S641.2[农业科学—蔬菜学]