超量表达益母草种子抗菌蛋白提高番茄的抗病性

为了验证来自益母草Leonums japonicusHoutt种子的抗菌蛋白基因LjAMP1和LjAMP2对植物病害的广谱抗性,用根癌农杆菌Agrobacterium tumefaciens介导法,将其分别转入台湾圣女番茄品种。结果显示,利用黄萎病菌毒素浸泡番茄离体枝条,处理24h,空载转基因对照和非转基因再生植株枝条全部萎蔫,而LjAMP1和LjAMP2转基因番茄T0代枝条未出现萎蔫的株系比率分别为11.11%和6.25%;用离体叶片接种菌块,分别对T0代抗或耐黄萎病菌毒素的T1代株系接种早疫病菌,接种10天,空载转基因对照和非转基因再生植株的病情指数达到100,而LjAMP1和LjAMP2转基因番茄病情指数最低的株系分别为17.5和10.0,表明转基因番茄能同时提高对黄萎病菌毒素和早疫病的抗性;进而用叶盘法检测转基因植株对番茄青枯病菌的抑制作用,结果显示对真菌病害抗性强的转基因植株叶片对青枯病菌的抑菌圈更大。转基因番茄抗病鉴定结果表明,来自益母草种子的LjAMP1和LjAMP2基因对植物病害具有广谱抗性。

Enhancing disease resistance in transgenic tomato over-expressing antimicrobial proteins, LjAMP1 and LjAMP2 from motherwort seeds

To confirm the antimicrobial protein function of LjAMP1 and LjAMP2 genes cloned from motherwort,these two genes were introduced into tomato(cv.Taiwan shenlu)by Agrobacterium tumefaciens transformation method.In vitro assessment of the resistance against Verticillium wilt by dipping plant branches into the toxin preparation of Verticillium dahliae,indicated that the rate of showing no wilting symptom in transgenic tomato lines over-expressing LjAMP1 and LjAMP2 is 11.11% and 6.25% respectively,while the wild type and the transgenic plants holding empty construction branches were 100% severely wilted.To further investigate the resistance against Alternaria solani Jones et Grout,detached leaves were inoculated with the pathogen.In T1 progeny transgenic tomato lines over-expressing LjAMP1 and LjAMP2,those showing resistance or tolerance to Verticillium wilt toxin preparation,also exhibited resistance to A.solani.The lowest disease index of LjAMP1 and LjAMP2 T1 progeny transgenic lines was 17.5 and 10.0 respectively,while the disease index of wild type and transgenic plants holding empty construction are 100.Analysis of leaf discs inhibition zone to Ralstonia solanacearum revealed that in transgenic lines those exhibiting resistance to fungal pathogens also showed resistance to the bacterial pathogen,confirming the broad-spectrum resistances of LjAMP1 and LjAMP2.