论文部分内容阅读
为建立简单高效的黄瓜转基因技术体系,本研究以携带有抗除草剂Bar基因的农杆菌工程菌浸泡处理开花期黄瓜嫩梢,进行活体植株转基因方法研究。研究结果表明最佳转化体系为:表面活性剂silwet-77适宜浓度0.01%~0.05%;菌液浸泡最佳位置为顶部以下10 cm嫩梢,最适子房发育时期为开花前5~12 d子房;农杆菌菌株EHA105浸染效果优于LBA4404;3次重复浸泡处理有利于提高转化频率。黄瓜苗期除草剂抗性筛选适宜浓度为17%草胺膦1100倍稀释液。研究获得除草剂抗性植株1320株,PCR阳性植株36株,其中30株经Southern斑点杂交后有阳性信号,最高转化效率为9.5%。
In order to establish a simple and efficient system of cucumber transgenic technology, transgenic plants of transgenic cucumber plants were treated with Agrobacterium tumefaciens carrying anti-herbicide Bar gene soaking the tender shoots of cucumber at flowering stage. The results showed that the best transformation system was as follows: the suitable concentration of surfactant silwet-77 was 0.01% ~ 0.05%; the best place for soaking was 10 cm fresh shoots below the top, and the optimum developmental period was 5 ~ 12 d Ovary; Agrobacterium tumefaciens strain EHA105 is better than LBA4404; 3 times repeated immersion treatment is conducive to improve the frequency of transformation. Cucumber seedling herbicide resistance screening appropriate concentration of 17% phosphinothricin 1100 times dilution. 1320 herbicide-resistant plants and 36 PCR-positive plants were obtained. Among them, 30 were positive after Southern blotting and the highest transformation efficiency was 9.5%.