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利用转座子mini Tn5将δ 毒素基因cryIA(c)整合到生防细菌荧光假单胞菌P30 3 1的染色体上 ,构建成荧光假单胞工程菌。以棉铃虫二龄幼虫作供试昆虫的离体生测表明 :在同一剂量水平上 ,PT4 5、PT51、PT6 1、PT71等工程菌株对棉铃虫幼虫的杀虫活性均高于野生菌株HD 73;浓度最高时PT4 5、PT51、PT6 1、PT71、HD 73处理棉铃虫的死亡率分别为 86 7%、76 7%、80 0 %、86 7%、70 0 % ,死亡率与浓度呈正相关 ,体重与浓度呈负相关。用上述工程菌接种棉花活体 ,喷雾处理 3周后 ,用棉叶饲喂的棉铃虫死亡率分别达 70 0 %、6 0 0 %、6 0 0 %、6 6 7%、33 3% ;用棉顶尖饲喂的棉铃虫死亡率分别达 70 0 %、6 3 3%、6 0 0 %、73 3%、30 0 % ;用棉蕾饲喂的棉铃虫死亡率分别为 6 3 3%、53 3%、50 0 %、6 3 3%、2 6 7%。
The delta toxin gene cryIA (c) was integrated into the biocontrol bacterial Pseudomonas fluorescens P30 3 1 chromosome using the transposon mini Tn5 to construct a fluorescent pseudomonas engineering bacterium. The in vitro bioassay of the second-instar larvae of Helicoverpa armigera showed that the insecticidal activities of PT4 5, PT51, PT6 1, PT71 and other engineering strains on H. armigera larvae at the same dose level were higher than those of the wild strain HD 73 The highest mortality rates were 86.7%, 76.7%, 80.0%, 86.7%, 70.0% for PT4 5, PT51, PT6 1, PT71 and HD 73, respectively. The mortality was positively correlated with the concentration , Body weight and concentration was negatively correlated. Inoculation of the cotton in vivo with the above-mentioned engineered bacteria showed that the death rates of cotton bollworm fed with cotton leaf were 70%, 60%, 60%, 66% and 33% respectively after spraying for 3 weeks; The death rates of cotton bollworm fed with top cotton were 70%, 63%, 60%, 73% and 30% respectively. The mortality of cotton bollworm fed with cotton buds was respectively 63.3% 53 3%, 50 0%, 63 3%, 267%.