农杆菌介导的花生遗传转化条件优化

为建立花生快速高效的遗传转化体系，以浸泡吸水后的花生半粒种子为转化受体，利用根癌农杆菌介导法，以除草剂Basta抗性筛选及基因组DNA的Bar基因PCR检测为指标，优化花生遗传转化体系。研究结果表明，在侵染悬浮液中加入1mmol/L二硫苏糖醇（DTT）提高了农杆菌介导的转化效率（最高提升36.5 %）；与YEB培养液相比，AB重悬液的使用显著提高了农杆菌的转化效率（最高提升19.3 %）；农杆菌菌液OD600为0.7时转化效率最高；基因型显著影响转化效率，EXP27-1516的转化效率为69.03%，显著高于14AU01（56.67 %）；以花生半粒种子为转化受体，从种子培养到形成茁壮根系仅需8周，明显短于子叶节转化所需的13周。优化后的农杆菌介导的花生遗传转化体系为花生转基因研究提供了重要的研究工具。

Optimization of Agrobacterium tumefaciens–mediated transformation in peanut

 The aim of the this study was to optimize peanut transformation system to improve the transformation efficiency. Several steps and components were studied for their effect on transformation efficiency mediated by Agrobacterium tumefaciens through detecting the transgenic gene Bar in the genomic DNA of transformed plants. The results showed that the transformation efficiency could be significantly increased by collecting the Agrobacterium at a concentration of OD600 = 0.7, then using an Agrobacterium infection medium AB supplemented with 1 mmol/L dithiothreitol (DTT) to infect the half-seed explants. The Agrobacterium transformation efficiency for peanut variety EXP27-1516 (69.03%) was higher than that of variety 14AU01 (56.67 %). The transformation system described here is an efficient approach that yield vigorous plants within 8 weeks, compared to 13 weeks required in the transformation using cotyledonary nodes, and is the fastest method available for peanut transformation. This study provided an optimized protocol for A. tumefaciens mediated transformation in peanut as a useful reference to perform rapid transgenic research.