影响农杆菌介导的花生遗传转化条件的研究

Studies on Genetic Transformation Conditions of Peanut Mediated by Agrobacterium tumefaciens

利用花生幼叶高效再生体系,对影响农杆菌介导的花生遗传转化条件,抗生素(Km)筛选压、预培养时间、共培养时间进行了研究。最后确立的转化条件为:Km筛选压为100mg/L,预培养时间为1～2d,共培养时间为3d。并对得到的再生植株进行了PCR检测,初步证实了目的基因转到了花生基因组中。

Applying to young leaflets regeneration system of peanut, genetic transformation of peanut using Agrobacterium-mediated approach were carrying on the research. The factors affecting the transformation, such as the selective pressure of antibiotic, pre-cuhured, co-cultivation were examined. The optimized procedure was as following： after pre-cuhured for 1-2 days, the young leaflets were dipped into an Agrobacterium suspension for 10-15 min, then co-cultivation for three days, the explants were transferred to selective media with 100mg/L Km and500mg/L Cef. These conditions benefited to improve the transformation efficiency. PCR analysis showed that transgenic peanut plants were obtained.