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采用差异蛋白质组分析和酶活性测定法研究棉铃虫中肠组织对拟除虫菊酯类杀虫剂胁迫的分子适应

Molecular adaptations of Helicoverpa armigera midgut tissue under pyrethroid insecticide stress characterized by differential proteome analysis and enzyme activity assays.

机构信息

Biochemistry Joint Graduate Program, Middle East Technical University, Ankara, Turkey.

出版信息

Comp Biochem Physiol Part D Genomics Proteomics. 2013 Jun;8(2):152-62. doi: 10.1016/j.cbd.2013.04.001. Epub 2013 Apr 24.

DOI:10.1016/j.cbd.2013.04.001
PMID:23685472
Abstract

Helicoverpa armigera is an insect that causes important economic losses in crops. To reduce this loss, pyrethroids have been commonly used against H. armigera in farming areas. However, excess and continuous usage of pyrethroids cause resistance in H. armigera. Therefore, expressions of midgut proteins of two H. armigera field populations were compared to those of a susceptible strain by 2-D PAGE and MALDI-ToF-MS. Our results indicate that H. armigera reacts to pyrethroid-induced stress mainly by increasing the expression of energy metabolism-related proteins, such as ATP synthase and arginine kinase. NADPH cytochrome P450 reductase, also up-regulated, could play a role in detoxification of toxic pyrethroid metabolites, such as 3-phenoxybenzaldehyde. Interestingly, while GSTs were not found up-regulated in the comparative proteome analysis, biochemical assays showed significant increases of enzyme activities in both field populations as compared to the susceptible strain. Similarly, although esterases were not found differentially expressed, biochemical assays showed significant increases of esterase activities in both field populations. Thus, esterases are also proposed to be involved in metabolic responses towards pyrethroid insecticide-induced stress. In conclusion, we suggest increased energy metabolism in the midgut tissue of H. armigera as a general prerequisite for compensating the costs of energy-consuming detoxification processes.

摘要

棉铃虫是一种对农作物造成重大经济损失的昆虫。为了减少这种损失,在农业区通常使用拟除虫菊酯来防治棉铃虫。然而,过量和持续使用拟除虫菊酯会导致棉铃虫产生抗药性。因此,通过 2-DE PAGE 和 MALDI-ToF-MS 比较了两个田间种群和一个敏感品系棉铃虫中肠蛋白的表达。我们的结果表明,棉铃虫对拟除虫菊酯诱导的应激的反应主要是通过增加与能量代谢相关的蛋白质的表达来实现的,如 ATP 合酶和精氨酸激酶。上调的 NADPH 细胞色素 P450 还原酶可能在解毒有毒的拟除虫菊酯代谢物(如 3-苯氧苯甲醛)中发挥作用。有趣的是,虽然 GSTs 在比较蛋白质组分析中没有上调,但生化测定显示两个田间种群的酶活性都明显高于敏感品系。同样,虽然酯酶没有差异表达,但生化测定显示两个田间种群的酯酶活性都显著增加。因此,酯酶也被认为参与了代谢对拟除虫菊酯杀虫剂诱导的应激的反应。总之,我们认为中肠组织中能量代谢的增加是补偿耗能解毒过程成本的一般前提。

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