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Differential expression of the detoxification enzyme genes in different developmental stages of the whitefly,Bemisia tabaci( Hemiptera: Aleyrodidae)
Pages: 1166-1173
Year: Issue:  11
Journal: Acta Entomologica Sinica

Keyword:  Bemisia tabaciresistancethiamethoxamdetoxification enzymemicroarraydevelopmental stage specificity;
Abstract: 【Aim】Based on the transcriptome data of the whitefly,Bemisia tabaci,this study aims to systematically analyze the expression profiles of genes encoding detoxification enzymes in thiamethoxam resistant strain,and to focus on discussing the biological significance of developmental stage specific differential expression of these genes. 【Methods】Thiamethoxam resistant( TH-R) and susceptible( THS) strains of B. tabaci were maintained in a greenhouse,and individuals at three developmental stages( egg,the 4th instar nymph and 1 day-old unmated female adult) were collected from the TH-R and THS strains. The array design was based on 8 394 ESTs from transcriptome of B. tabaci with putative association with insecticide resistance. By using a custom made microarray,the gene expression in eggs,nymphs and female adults of the TH-R strain of B. tabaci was compared. Nine ESTs were randomly selected for quantitative real-time PCR( qRT-PCR) analysis to validate the microarray response between the TH-R and TH-S strains at three different developmental stages. 【Results】In the TH-R strain,a total of 3 424 ESTs were differentially expressed between the egg and 4th instar nymphal stages. Among them,489 ESTs encode detoxification enzymes( carboxylesterase,glutathione S-transferases,cytochrome P450monooxygenases). Fourteen ESTs( 10 ESTs encoding P450 s, and 4 ESTs encoding GSTs) were overexpressed in the 4th instar nymphal stage. There were 1 273 ESTs which were differentially expressed between the 4th instar nymphal and female adult stages. Among them,193 genes encoding detoxification enzymes and nine ESTs encoding P450 s showed higher expression level( 10-fold) in the female adult stage rather than the 4th instar nymphal stage. The qRT-PCR results revealed that nine ESTs were upregulated in the TH-R strain at three developmental stages compared to those in the TH-S strain. The EST p06027 and the EST p06013 encoding P450 s were overexpressed in egg and the 4th instar nymphal stage of the TH-R strain. The ESTs p05885,p07806 and the EST p00988 encoding P450 s were highly overexpressed in the 4th instar nymphal stage of the TH-R strain. The ESTs p05916 and p00478had low expression levels in the egg and 4th instar nymphal stages but overexpressed in the female adult stage of the TH-R strain. The ESTs p00059 and p00428 were significantly up-regulated in the female adult stage but had low expression level in the other two immature stages of TH-R strain,and the EST p00059 encoding CYP4C1 with the highest expression level( 15. 15-fold) was seen in the female adult stage. 【Conclusion 】 The results suggest that the overexpression of CYP4C1 and CYP6 B may underlie the resistance to thiamethoxam in B. tabaci. According to the microarray data analyzed,the detoxification enzyme genes are differentially expressed in different developmental stages of B. tabaci,which may be associated with the distribution and effective utilization of energy under the stress of insecticides and also an adaptive mechanism under the environmental selection pressures.
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