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晚疫病菌 (Phytophthorainfestans)引起的马铃薯晚疫病是危害全球马铃薯生产的严重病害。通过基因工程方法把外源基因导入植物体内以增强抗病性被证明是一条行之有效的途径。应用植物基因工程技术将具有能激活植物自身防御系统的无毒基因与适合于植物背景、非专一性的病原物诱导启动子组合成嵌合基因构建到植物表达载体中。通过农杆菌或基因枪的介导转化植物 ,可筛选出高效广谱的抗真菌和细菌病害的转基因植株。本研究从病原细菌PseudomonassyringaePV .tomato中获得的无毒基因avrD (0·93kb)和从病原真菌Phytophthoraparasitica中获得的无毒基因Elicitin(0 2 94kb)分别与非专一性病原物诱导启动子Pill和BG组成含 2个嵌合基因 (Pill -avrD ,BG -Elicitin)的植物表达载体pYH144和pYHEt。通过农杆菌LBA4 4 0 4介导转化马铃薯 ,其中用pYH144载体转化 2个品种 (克新 1号 ,2号 ) ,用pYHEt载体转化 3个品种 (Desiree ,克新 2号 ,4号 ) ,通过组织培养分别获得潮霉素 (HygromycinB)标记的转基因马铃薯试管苗。将转基因试管苗扩繁 ,应用马铃薯脱毒微型种薯生产技术获得转无毒基因微型薯 ,在温室 (15～ 2 5℃和湿度高 )条件下 ,观察转无毒基因马铃薯植株中对晚疫病菌自然感染的抗性。 1998年和 1999年 (每年的 3-
Late blight of potato caused by Phytophthora infestans is a serious disease that affects global potato production. The introduction of exogenous genes into plants by genetic engineering to enhance disease resistance has proven to be an effective way. The application of plant genetic engineering will have non-toxic genes can activate the plant’s own defense system with plant-specific, non-specific pathogen-induced promoter into a chimeric gene into the plant expression vector. Through Agrobacterium or gene gun-mediated transformation of plants, can be screened efficient broad-spectrum anti-fungal and bacterial disease transgenic plants. In this study, the non-toxic gene avrD (0.93kb) obtained from the pathogenic bacterium Pseudomonas salivarius PVTtomato and the non-toxic gene Elicitin (0 2 94kb) obtained from the pathogenic fungus Phytophthora parasitica were respectively compared with the non-specific pathogen-inducible promoter Pill and BG composed of plant expression vectors pYH144 and pYHEt containing two chimeric genes (Pill-avrD, BG -Elicitin). The potato was transformed with Agrobacterium tumefaciens LBA4404, in which two varieties (Kexin No.1 and No.2) were transformed with pYH144 vector and three varieties (Desiree, Kexin No.2 and No.4) were transformed with pYHEt vector, Tissue culture Hygromycin B-labeled transgenic potato plantlets were obtained respectively. The transgenic plantlets were expanded and multiplied. The transgenic potato seedlings were obtained by the technique of potato detoxification miniature seed potato. The transgenic potato seedlings were observed under the conditions of 15 ~ 25 ℃ and high humidity in the greenhouse. Resistance of bacteria naturally infected. 1998 and 1999 (annual 3-