发根土壤杆菌介导的大豆发根转化体系改进

Improvement of Soybean Hairy-root System Mediated by Agrobacterium Rhizogenes

鉴于大豆难以获得转基因植株的现实,借鉴Kereszt等建立的发根土壤杆菌(Agrobacterium rhizogenes)K599介导的大豆转化体系,并针对实验中遇到的具体问题,对原体系进行了适当改进。结果表明:改进的体系操作更简单,对发根培养条件、人员操作技能要求更低,所形成的植株更具生命力。同时,应用改进体系以及RNAi技术,构建了表达线虫特异基因dsRNA的转基因发根植株,可在60 d内在转基因大豆发根上对植物线虫基因的RNAi效果完成验证。改进的体系为培育抗大豆胞囊线虫品种奠定了基础。

In consideration of the fact that it is difficult to acquire transgenic soybean plants, we established a transformation system of soybean mediated by Agrobacterium rhizogenes K599 according to the method of Kereszt et el. , and improved it be-cause of some specific questions in experiment. The improved transformation system and the original one were compared. Take advantage of the improved system and RNAi technology, transgenic hairy-root-plants that expressing nematode dsRNA of specif-ic genes were constructed and the RNAi effects of these genes could be tested within 60 days on the transgenic hairy roots. The results indicate that the improved system is easier to manipulate,have less demands to environment conditions and personnel skilling levels, and improve the vitality of acquired plants. Our results established the foundation for breeding new crop cultivars with highly resistant to soybean cyst nematode.