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分别在北京和江苏省连云港市对携带Btcry基因的荧光假单胞菌工程菌进行了田间残留与扩散的追踪检测 .对越冬前后的试验地和保护地土壤样品进行抗生素抗性平板分离 ,能检测到极少量的具有与出发菌株相同抗性的荧光假单胞菌菌落 ,但没有发现工程菌株的残留与扩散 .对江苏试验地样品还进行了工程菌株质粒卡那霉素和壮观霉素抗性标记基因的抗性菌落分离 ,绝大多数样品中都能分离到包括荧光假单胞菌在内的抗性菌落 ,土样中菌密度n(cfu) =10 4 ～ 10 5g-1,但进行Btcry基因PCR -RFLP检测时没有从样品中得到特异扩增产物 .研究结果表明 :工程菌株抗性标记基因在自然界广泛存在 ,工程菌在株环境中没有残留和扩散 ,具有良好的生物安全性 .表 4参 8
Traces of Pseudomonas fluorescens-carrying bacteria carrying Btcry gene were tracked in the field in Beijing and Lianyungang, Jiangsu Province, respectively. Antibiotic resistance plates were isolated from soil samples before and after overwintering and soil samples from protected areas To a very small number of Pseudomonas fluorescens colonies that had the same resistance as the original strain but no residual and spread of the engineered strains were found.The engineered strains were also tested for kanamycin and spectinomycin resistance Resistant colonies of the marker genes were isolated, resistant colonies including Pseudomonas fluorescens were isolated in the vast majority of samples, and bacterial density n (cfu) = 104 to 105g-1 in soil samples Btcry gene PCR-RFLP did not detect the specific amplification products from the samples.The results showed that: the engineered strains of resistance markers exist widely in nature, engineering bacteria in the plant environment without residual and proliferation, with good biosecurity. Table 4 Reference 8