抗菌蛋白AP1编码基因对马铃薯的转化及其介导的青枯病抗性研究  

Transformation of anti-microbial protein encoded by gene apl and the mediated resistance to bacterial wilt of the transgenic potato

作　　者：梁成罡[1] 何礼远[1] 

机构地区：[1]中国农业科学院植物保护研究所,北京100094

出　　处：《植物保护学报》2002年第1期51-56,共6页Journal of Plant Protection

基　　金：国家自然科学基金(39630220);国家"863"计划课题(101-04-01-04)资助

摘　　要：根据已知马铃薯抗青枯菌蛋白AP1编码基因序列,合成了特异性引物,经PCR扩增、酶切以及连接、转化等分子操作,成功地获得了AP1编码基因植物表达载体pHap1,该载体具有-2E-P35S-12-ap1-结构。以电激法将pHap1导入根癌农杆菌LBA4404,用叶盘法转化马铃薯青枯病感病品种Mira、中-5-1及金冠试管苗,分别获得了卡那霉素(kan)抗性再生苗。经PCR、Southern、Northern检测,表明ap1基因已成功整合到转基因植株染色体上并正确表达。对Mira品种的转基因植株进行了青枯病抗病性鉴定,结果表明,转基因植株的抗病性比非转基因植株对照明显提高,发病或出现萎蔫症状的时间延迟、病情指数降低。APl is a plant endogenic anti-microbial protein that was isolated from a bacterial wilt-resistant potato variety MS42.3. Based on the nucleotide sequence of its encoded gene apl, a pair of specific primers were designed and synthesized. The apl fragment was obtained by PCR and cloned into an intermediate plasmid pG Ω 4A to obtain the plant expression box -2E-P35S-Ω-apl- . After inserted '-2E-P35S-Ω-apl- into plasmid pBI121, the plant expression vector of apl was obtained and named as pHapl. Mediated by Agrobacterium tumefacients strain LBA4404, pHapl was introduced into potato cultivars Mira, Jinguan and Zhong-5-1 with leaf disc transformation. Some kanamycin resistant seedlings were obtained consequently. After molecular analysis of transgenic plants by PCR, Southern and Northern-blotting, apl was proved to be integrated into the potato chromosome DNA and efficiently expressed. Resistance testing of transgenic plants of the cultivar Mira indicated that apl transformation had led to an increased resistance of transgenic potato plants to bacterial wilt caused by Ralstonia solanacearum .

关 键 词：抗菌蛋白AP1 apl基因 马铃薯 转化 青枯病 抗病性 介导 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]