甲氧虫酰肼对不同抗性棉铃虫种群谷胱甘肽S-转移酶活性和基因表达量的影响

摘要/Abstract

摘要： 【目的】明确与谷胱甘肽S-转移酶（GST）相关联的棉铃虫 Helicoverpa armigera (Hübner)对甲氧虫酰肼的抗性分子机制, 更有效地开展棉铃虫抗药性的快速监测。【方法】用LC₄₀甲氧虫酰肼处理棉铃虫3龄初幼虫, 测定处理前、后抗性种群GST的活性及5种GST基因的表达量变化, 并比较一种Delta基因GSTd1的编码区序列。【结果】经测序、比对, 抗甲氧虫酰肼种群（R-methoxyfenozide）和同源对照种群（S-methoxyfenozide）GSTd1基因编码区序列相同, 表明其编码的酶结构没有发生改变。甲氧虫酰肼处理前, R-methoxyfenozide种群的GST比活力显著高于S-methoxyfenozide种群；而药剂处理后, 两种群的GST比活力均降低, 但R-methoxyfenozide种群的活性可以快速回升。药剂处理前, R-methoxyfenozide种群的GST基因表达量显著高于S-methoxyfenozide种群。药剂处理对不同抗性种群GST基因表达量的影响差异较大。除了GSTs1, S-methoxyfenozide种群GST基因表达量均降低, 其中GSTd2和GSTe2可以缓慢回升。GSTs1表达量在36 h内没有明显变化, 但在48 h显著升高。R-methoxyfenozide种群的GST基因表达量均先降低, 然后迅速恢复, 除GSTe2基因外, R-methoxyfenozide种群的基因初始表达量和药剂处理后的最终表达量均显著高于S-methoxyfenozide种群。【结论】棉铃虫对甲氧虫酰肼的抗性与GST比活力增强有关, 而GST比活力的增强主要源于多个GST基因的过量表达。

关键词: 棉铃虫, 甲氧虫酰肼, 谷胱甘肽S-转移酶, 酶活性, 基因表达, 实时定量PCR

Abstract: 【Aim】 In order to develop more effective methods for molecular detection of insecticide resistance, the molecular mechanisms of resistance to methoxyfenozide related to glutathione S-transferases (GSTs) in Helicoverpa armigera (Hübner) were studied. 【Methods】 The early 3rd instar larvae of the methoxyfenozide-resistant (R-methoxyfenozide) and the homologous control (S-methoxyfenozide) populations of H. armigera were treated with LC₄₀ of methoxyfenozide, and the specific activities of GST and the expression of five GST genes were measured before and after treatment. The coding sequence of one delta GST gene (GSTd1) was compared in the two populations. 【Results】 The coding sequences of GSTd1 in the R-methoxyfenozide and S-methoxyfenozide populations of H. armigera were identical, suggesting that no structural change occurrs in the isozyme. The GST activity in R-methoxyfenozide population of H. armigera was higher than that in the S-methoxyfenozide population before treatment. After treatment, the GST activity decreased in both populations, but fast recovered in the R-methoxyfenozide population. Except GSTe2, GST genes had a higher expression in the R-methoxyfenozide population than in the S-methoxyfenozide population before treatment. Effects generated by methoxyfenozide treatment on the expression of GST genes were quite different in the two populations. After treatment, the expression of GST genes, except GSTs1, initially decreased in the S-methoxyfenozide population, and then the expression of GSTd2 and GSTe2 recovered gradually. The expression of GSTs1 showed no obvious change within 36 h and increased at 48 h after treatment. But in the R-methoxyfenozide population, the expression of the five GST genes after treatment decreased initially, and then fast recovered. Except GSTe2, the initial and final expression of GST genes in the R-methoxyfenozide population were remarkably higher than that in the S-methoxyfenozide population. 【Conclusion】 The results suggest that resistance to methoxyfenozide in H. armigera is related to enhanced GST activities which are rooted from the over-expression of GST genes.

Key words: Helicoverpa armigera, methoxyfenozide, glutathione S-transferase, enzyme activity, gene expression, qRT-PCR

徐希宝, 张靖, 芮昌辉. 甲氧虫酰肼对不同抗性棉铃虫种群谷胱甘肽S-转移酶活性和基因表达量的影响[J]. , 2014, 57(12): 1381-1388.

XU Xi-Bao, ZHANG Jing, RUI Chang-Hui. Effects of methoxyfenozide on the activities and gene expression levels of glutathione S-transferases (GST) in the methoxyfenozide-resistant and homologous control populations of Helicoverpa armigera (Lepidoptera: Noctuidae)[J]. , 2014, 57(12): 1381-1388.

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甲氧虫酰肼对不同抗性棉铃虫种群谷胱甘肽S-转移酶活性和基因表达量的影响

Effects of methoxyfenozide on the activities and gene expression levels of glutathione S-transferases (GST) in the methoxyfenozide-resistant and homologous control populations of Helicoverpa armigera (Lepidoptera: Noctuidae)

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