利用类甜蛋白基因诱导表达提高马铃薯对晚疫病的抗性研究

Studies on Enhancement of Transgenic Potato's Resistance to Late blight by Inducing the Expression of Thaumatin like Protein Gene

将类甜蛋白基因 (TLP)及其紧密连锁的抗除草剂bar基因以农杆菌介导法导入脱毒微型种薯“津引薯 8号” ,经除草剂筛选后的转基因单株无性繁殖用于分子检测 ,标记基因bar基因的PCR反应和含TLP基因特异探针的Southern检测结果 ,转基因植株分别显示出 0 54kb和 3 0kb的特异性条带 ,证明TLP基因已成功地整合到转基因马铃薯基因组中。利用TLP基因特异性抗血清对转基因植株进行Western反应检测 ,结果表明 ,转基因植株中TLP基因表达量显著高于对照组 ,经晚疫病菌游离孢子接种离体抗性分析 。

Thaumatin like protein (TLP)gene and bar gene which is closely combined with TLP gene were transferred into virus free micropotato variety Jing Yinshu 8 by agrobacterium mediated transformation.Transgenic plants selected by bialaphos were amplified for detection.The results of PCR of maker bar gene and Southern blot of specific probe with TLP gene showed that transgenic plants displayed 0.54 kb and 3.0 kb bands,respectively,indicating TLP gene has been successfully integrated into the chromosome of transgenic potatoes.The western reaction of transgenic plants using TLP antiserum showed the expression of TLP gene in transgenic plants was much higher than the control.Through the vitro assay of leaves inoculated by alive spores of phytophthora,leaves of transgenic potato plants exhibited significant inhibition on late blight disease and delayed development of disease symptoms.