---

期 刊 ：植物保护学报 2007年

关键词：益母草种子；抗菌蛋白；转基因；抗病性；

Keywords:motherwort seed, antimicrobial protein, transgene, disease resistance,

摘 要 ：为了验证来自益母草Leonums japonicus Houtt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24h,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。

Abstract: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.

---