野生天麻DNA导入马铃薯实现遗传转化的研究

采用浸苗法将野生天麻总DNA导入马铃薯试管苗,对筛选得到的转化植株进行蛋白及药用成份的分析。结果显示:(1)在200株转化的马铃薯中有21株的紫外扫描图谱与正常对照组有显著差异,且在220nm有明显吸收峰。(2)5株经PCR扩增出野生天麻抗真菌蛋白(GAFP)基因。(3)转基因马铃薯与正常马铃薯的蛋白表达有明显差异,并且在转基因马铃薯中有一条与野生天麻抗真菌蛋白(GAFP)相同的条带。而正常马铃薯中无此条带。(4)通过薄层层析法检测出3株转基因马铃薯表达野生天麻的有效药用成份天麻素。说明采用浸苗法进行外源总DNA导入是可行的。

A genetic transformation study of introducing wild Gastrodia elata DNA into potato

After the tuber had grown up,the solution of gastrodin was extracted from the potatoes which were transformed by wild Gastrodia elataDNA. SDS-PAGE was used to analyze GAFP gene. TLC was used to analyze the gastrodin of transformed potatoes. Results:(1)21 transformed plants had obvious obsorption peak at 220nm in 200 transformed potatoes. (2)The fragment of wild Gastrodia elataGAFP gene was found by PCR from 5 transformed plants. (3)The protein expressions in transgenic potatoes were obviously different from normal potatoes;A band was detected in transgenic potato and wild Gastrodia elata,but wasn’t detected in normal potatoes. (4)We detected the expression of the gastrodin by TLC from 3 transformed plants. It is feasible to use the soaking seedling method to introduce exogenous DNA into plants.