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首页> 美国卫生研究院文献>Applied and Environmental Microbiology >Development of Bacillus thuringiensis CryIC Resistance by Spodoptera exigua (Hubner) (Lepidoptera: Noctuidae)
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Development of Bacillus thuringiensis CryIC Resistance by Spodoptera exigua (Hubner) (Lepidoptera: Noctuidae)

机译：甜菜夜蛾对苏云金芽孢杆菌CryIC抗性的建立（鳞翅目：夜蛾科）

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摘要

Selection of resistance in Spodoptera exigua (Hubner) to an HD-1 spore-crystal mixture, CryIC (HD-133) inclusion bodies, and trypsinized toxin from Bacillus thuringiensis subsp. aizawai and B. thuringiensis subsp. entomocidus was attempted by using laboratory bioassays. No resistance to the HD-1 spore-crystal mixture could be achieved after 20 generations of selection. Significant levels of resistance (11-fold) to CryIC inclusion bodies expressed in Escherichia coli were observed after seven generations. Subsequent selection of the CryIC-resistant population with trypsinized CryIC toxin resulted, after 21 generations of CryIC selection, in a population of S. exigua that exhibited only 8% mortality at the highest toxin concentration tested (320 (mu)g/g), whereas the 50% lethal concentration was 4.30 (mu)g/g for the susceptible colony. Insects resistant to CryIC toxin from HD-133 also were resistant to trypsinized CryIA(b), CryIC from B. thuringiensis subsp. entomocidus, CryIE-CryIC fusion protein (G27), CryIH, and CryIIA. In vitro binding experiments with brush border membrane vesicles showed a twofold decrease in maximum CryIC binding, a fivefold difference in K(infd), and no difference in the concentration of binding sites for the CryIC-resistant insects compared with those for the susceptible insects. Resistance to CryIC was significantly reduced by the addition of HD-1 spores. Resistance to the CryIC toxin was still observed 12 generations after CryIC selection was removed. These results suggest that, in S. exigua, resistance to a single protein is more likely to occur than resistance to spore-crystal mixtures and that once resistance occurs, insects will be resistant to many other Cry proteins. These results have important implications for devising S. exigua resistance management strategies in the field.
机译：选择甜菜夜蛾对HD-1孢子晶体混合物，CryIC（HD-133）包涵体和苏云金芽孢杆菌亚种的胰蛋白酶解毒的抗性。 aizawai和苏云金芽孢杆菌亚种。昆虫病原体通过实验室生物测定法进行尝试。选择20代后，无法获得对HD-1孢子晶体混合物的抗性。七代后，观察到对在大肠杆菌中表达的CryIC包涵体的显着抗性水平（11倍）。在经过21代CryIC选择后，随后选择了经胰蛋白酶处理的CryIC毒素对CryIC产生抗药性的人群导致了在最高毒素浓度（320μg/ g）下仅表现出8％死亡率的S. exigua种群，而易感菌落的50％致死浓度为4.30μg/ g。对来自HD-133的CryIC毒素具有抗性的昆虫也对受胰蛋白酶消化的CryIA（b）和苏云金芽孢杆菌亚种的CryIC具有抗性。昆虫，CryIE-CryIC融合蛋白（G27），CryIH和CryIIA。与易感性昆虫相比，刷状缘膜囊泡的体外结合实验显示最大CryIC结合力降低了两倍，K（infd）差异提高了五倍，并且抗CryIC的昆虫的结合位点浓度没有差异。通过添加HD-1孢子，对CryIC的抗性大大降低。去除CryIC选择后12代仍可观察到对CryIC毒素的抗性。这些结果表明，在S. exigua中，对单一蛋白质的抗药性比对孢子晶体混合物的抗药性更可能发生，一旦抗药性发生，昆虫将对许多其他Cry蛋白产生抗药性。这些结果对设计该领域的S. exigua抗药性管理策略具有重要意义。

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