用基因枪法将小麦病程相关蛋白基因TaPR-1导入小麦的研究

Transferring Wheat Pathogenesis-related Proteins 1(TaPR-1) Gene into Wheat via Micro-projectile Bombardment

为验证小麦病程相关蛋白基因TαPR-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TαPR-1基因导入扬麦158幼胚．经3～5mg／L Bialaphos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。

To validate the function of the wheat pathogenesis-related proteinsl gene （TαPR-1）, the transient expression technique of gus gene was used to optimize two bombardment parameters, the amount of gold powder and the bombard distance, in wheat gene transformation by microprojectile bombardment. The results indicated that when the dosage of gold powder per bombardment was 250.0μg and the bombardment distance was 9 number of blue spots per immature embryo were bombardment was gained. Using the parameter immature embryo of Yangmai 158 . Total of 49 tained. Two plants were identified to be positive cm, the ratio of GUS transient expression and the 89.8% and 33.7, respectively, and a perfect effect of optimized,TαPR-1 target gene was transferred into plants resistant to 3-5 mg/ L Bialaphos were obby PCR and Southern hybridization test in To generation. The transformation frequency was 0. 11%. At transgenic plants was evaluated by using detached-leaf mary result showed there was no significant difference the same time, powdery mildew resistance of segments and adult plant inoculation. The pribetween transgenic plants and the wild type.