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RNA interference revealed the roles of two carboxylesterase genes in insecticide detoxification in Locusta migratoria

机译:RNA干扰揭示了两个羧酸酯酶基因在南方蝗中的杀虫剂解毒中的作用

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摘要

Carboxylesterases (CarEs) play key roles in metabolism of specific hormones and detoxification of dietary and environmental xenobiotics in insects. We sequenced and characterized CarE cDNAs putatively derived from two different genes named LmCesA1 and LmCesA2 from the migratory locust, Locusta migratoria, one of the most important agricultural pests in the world. The full-length cDNAs of LmCesA1 (1892 bp) and LmCesA2 (1643 bp) encode 543 and 501 amino acid residues, respectively. The two deduced CarEs share a characteristic α/β-hydrolase structure, including a catalytic triad composed of Ser-Glu (Asp)-His and a consensus sequence GQSAG, which suggests that both CarEs are biologically active. Phylogenetic analysis grouped both LmCesA1 and LmCesA2 into clade A which has been suggested to be involved in dietary detoxification. Both transcripts were highly expressed in all the nymphal and adult stages, but only slightly expressed in eggs. Analyses of tissue-dependent expression and in situ hybridization revealed that both transcripts were primarily expressed in gastric caeca. RNA interference (RNAi) of LmCesA1 and LmCesA2 followed by a topical application of carbaryl or deltamethrin did not lead to a significantly increased mortality with either insecticide. However, RNAi of LmCesA1 and LmCesA2 increased insect mortalities by 20.9% and 14.5%, respectively, when chlorpyrifos was applied. These results suggest that these genes might not play a significant role in detoxification of carbaryl and deltamethrin but are most likely to be involved in detoxification of chlorpyrifos in L. migratoria.
机译:羧酸酯酶(CarEs)在昆虫体内特定激素的代谢以及饮食和环境异生物的排毒中起关键作用。我们测序和表征了CarE cDNA,推测它们来自两个不同的基因,分别来自蝗虫Locusta migratoria,这是世界上最重要的农业害虫之一,分别命名为LmCesA1和LmCesA2。 LmCesA1(1892 bp)和LmCesA2(1643 bp)的全长cDNA分别编码543和501个氨基酸残基。这两个推论的CarEs具有特征性的α/β水解酶结构,包括由Ser-Glu(Asp)-His和共有序列GQSAG组成的催化三联体,这表明这两个CarEs都具有生物活性。系统发育分析将LmCesA1和LmCesA2归类为进化枝A,该进化枝A被认为与饮食排毒有关。两种转录本在所有若虫和成虫阶段均高表达,但在卵中仅少量表达。组织依赖性表达和原位杂交的分析表明,两种转录本主要在胃盲肠中表达。 LmCesA1和LmCesA2的RNA干扰(RNAi),然后局部应用西维因或溴氰菊酯不会导致任何一种杀虫剂的死亡率显着增加。然而,使用毒死rif时,LmCesA1和LmCesA2的RNAi分别使昆虫死亡率增加20.9%和14.5%。这些结果表明,这些基因可能在西维因和溴氰菊酯的解毒中不发挥重要作用,但最有可能参与了对L. migratoria中毒死rif的解毒。

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