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Insecticidal Activity of a Vip3Ab1 Chimera Is Conferred by Improved Protein Stability in the Midgut of Spodoptera eridania

机译:Vip3Ab1嵌合体的杀虫活性是由改善的斜纹夜蛾中肠的蛋白质稳定性赋予的。

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摘要

Vip3A proteins are important for the control of spodopteran pests in crops, including Spodoptera frugiperda (fall armyworm). Native Vip3Ab1 controls S. frugiperda, but it is ineffective against S. eridania (southern armyworm), a major pest of soybean in South America. Recently, a Vip3Ab1 chimera with a modified C-terminus was described, Vip3Ab1-740, which has increased potency against S. eridania while maintaining activity against S. frugiperda. As S. frugiperda and S. eridania are differentially susceptible to Vip3Ab1, experiments were conducted to identify and understand the mechanism by which this expanded potency is conferred. The role of protein stability, processing, and in vivo effects of Vip3Ab1 and Vip3Ab1-740 in both of these species was investigated. Biochemical characterization of the midgut fluids of these two species indicated no obvious differences in the composition and activity of digestive enzymes, which protease inhibitor studies indicated were likely serine proteases. Histological examination demonstrated that both proteins cause midgut disruption in S. frugiperda, while only Vip3Ab1-740 affects S. eridania. Immunolocalization indicated that both proteins were present in the midgut of S. frugiperda, but only Vip3Ab1-740 was detected in the midgut of S. eridania. We conclude that the gain of toxicity of Vip3Ab1-740 to S. eridania is due to an increase in protein stability in the midgut, which was conferred by C-terminal modification.
机译:Vip3A蛋白对于控制作物中的鳞翅目有害生物非常重要,其中包括Spodoptera frugiperda(秋天夜蛾)。天然Vip3Ab1控制着Srug frugiperda,但它对S. eridania(南方粘虫)无效,后者是南美大豆的主要害虫。最近,描述了具有修饰的C末端的Vip3Ab1嵌合体,Vip3Ab1-740,其具有增强的针对S.eridania的效力,同时保持了针对S.frugiperda的活性。由于S. frugiperda和S. eridania对Vip3Ab1具有不同的敏感性,因此进行了实验以鉴定和了解赋予这种增强的效力的机制。研究了这两个物种中Vip3Ab1和Vip3Ab1-740的蛋白质稳定性,加工和体内作用的作用。这两个物种的中肠液的生化特征表明消化酶的组成和活性没有明显差异,蛋白酶抑制剂研究表明这很可能是丝氨酸蛋白酶。组织学检查表明,这两种蛋白均导致沙门氏菌中肠破裂,而只有Vip3Ab1-740会影响大肠链球菌。免疫定位表明两种蛋白质都存在于弗氏链球菌的中肠中,但是在S. eridania中肠中仅检测到Vip3Ab1-740。我们得出的结论是,Vip3Ab1-740对S. eridania的毒性增加是由于中肠中蛋白质稳定性的提高,这是C末端修饰所致。

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