利用花粉管通道法将BcWRKY2抗旱基因导入玉米的研究

Transferring BcWRKY2 Drought Resistance Gene into Maize Inbred Lines by Pollen Tube Pathway

利用花粉管通道法将含有PPT抗性筛选标记的BcWRKY2转录因子基因导入优良玉米自交系。采用正交设计对玉米品种、导入时间及DNA浓度等条件进行了优化。对T0代种子进行了除草剂抗性筛选,结果表明玉米品种龙抗11授粉后16h进行导入及DNA导入浓度为200ng/μL时效果最佳。对获得的除草剂抗性植株进行PCR及PCR-Southern检测,得到PCR阳性植株71株,初步证明了外源目的基因已整合到玉米基因组中。

In this study, Be WRKY2 gene was transformed into four elite maize inbred lines by pollen tube pathway. Varieties, transforming period, concentration of exogenous DNA were optimized by orthogonal tests. Herbicide resistante test was used to screen the seeds of TO generation. The result showed that the combination of variety Longkang 11, transforming 16 hours after pollination and with a concentration of 200ng/μL DNA was optimal. The herbicideresistant plants were further detected by PCR and PCR-Southern. 71 PCR positive plants were obtained. It showed that BcWRKY2 gene was inserted into the genome of the transgenie plants preliminarily.