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为验证小麦病程相关蛋白基因TaPR-1的功能,利用gus基因瞬间表达技术对基因枪法转化小麦幼胚的两个重要参数金粉用量和轰击距离进行了优化。研究结果表明,金粉用量为250.0μg,轰击距离9cm的轰击参数最为理想,GUS瞬间表达率和平均蓝点数分别可达89.8%和33.7个。采用此轰击参数,将TaPR-1基因导入扬麦158幼胚,经3~5mg/LBialaphos筛选获得49株抗性再生植株,经PCR、Southern杂交分析,其中2株为阳性植株,转化率为0.11%。对转基因小麦植株进行苗期离体叶段和田间成株白粉病接种鉴定,初步结果表明,转基因植株对白粉病抗性与对照相比无明显差别。
In order to verify the function of TaPR-1, a wheat blast disease-related protein gene, gus gene transient expression technique was used to optimize the amount of gold powder and the bombardment distance, which are two important parameters of wheat seedling transformed by particle bombardment. The results showed that the bombardment parameters of 250.0μg gold powder and 9cm bombardment distance were the best, the instantaneous expression rate of GUS and average blue point were 89.8% and 33.7 respectively. Using this bombardment parameter, TaPR-1 gene was introduced into immature embryos of Yangmai 158, and 49 resistant regenerated plants were screened by 3 ~ 5mg / LBialaphos. Two of them were positive plants and the transformation rate was 0.11 %. The transgenic wheat plants were inoculated with in vitro leaf segments and field powdery mildew inoculated at seedling stage. The preliminary results showed that there was no significant difference in the resistance of the transgenic plants to powdery mildew compared with the control.