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转群体感应淬灭基因attM烟草抗青枯病的研究

DOI: 10.7685/j.issn.1000-2030.2012.03.012, PP. 64-70

Keywords: 转attM基因烟草,青枯病,抗病性

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Abstract:

自根癌农杆菌(Agrobacteriumtumefaciens)克隆获得attM基因。采用叶盘转化法,农杆菌介导转化烟草,经过卡那霉素抗性筛选获得抗性愈伤组织,对其分化产生的植物株系进行PCR、PCR-Southern和荧光定量PCR及GUS染色检测分析。结果表明attM基因被成功插入烟草基因组DNA中。在转基因株系中均能够表达,相对表达量最大可达125.8,最小为31.0。以野生型烟草和转pBI121空载体的烟草为对照,分别进行了离体及活体检测试验,结果表明转attM基因烟草植株对茄科雷尔氏菌引起的青枯病抗性显著。1×107~1×108CFU?mL-1的青枯菌接种离体叶片,野生型烟草和转pBI121空载体的烟草的中毒面积与接种面积比值为15~25,转基因植株的比值低于3;青枯菌3×106CFU?mL-1伤根接种盆栽烟草植株14d,野生株和转空载体烟草均青枯死亡,病情指数为100,转基因植株病情指数为31.25。

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