【目的】为探讨小菜蛾Plutella xylostella ABC转运蛋白(ATP-binding cassette transporter)与氯虫苯甲酰胺代谢的关系【方法】采用实时荧光定量RT-PCR技术,测定了氯虫苯甲酰胺敏感和抗性小菜蛾及用LC50剂量的氯虫苯甲酰胺处理敏感种群不同时间后ABCC1~ABCC5 mRNA的表达量;并检测了其中过表达的ABCC3~ABCC5在小菜蛾不同发育阶段和4龄幼虫不同组织中的表达量。【结果】所检测的5个ABCC基因在氯虫苯甲酰胺抗性品系中均上调表达,最高为敏感品系的2倍;LC50的氯虫苯甲酰胺处理后ABCC3、ABCC4和ABCC5 mRNA的表达量分别上调2.16倍、2.81倍和1.85倍,ABCC1和ABCC2则分别下调为对照组的65.8%和37.2%。ABCC3的mRNA在幼虫期和雄性成虫中表达量显著高于其它时期;ABCC4的mRNA在各发育阶段表达量差异不大,其中在预蛹期最高;ABCC5的mRNA在3龄幼虫中表达量显著高于其它龄期。在4龄幼虫各组织中,ABCC3、ABCC4和ABCC5在中肠中的表达量均显著高于其它组织。【结论】小菜蛾ABCC3、ABCC4和ABCC5可能在其对氯虫苯甲酰胺的代谢中起作用,该研究结果为进一步探究小菜蛾对氯虫苯甲酰胺的抗性与ABCC1~ABCC5的关系奠定了基础。
[Objectives] To elucidate the relationship between chlorantraniliprole and the ABC transporter(ATP-binding cassette transporter) in the diamondback moth, Plutella xylostella. [Methods] ABCC1-ABCC5 mRNA expression in different strains of the diamondback moth, and the LC50 of a susceptible strain to chlorantraniliprole, were determined using real-time quantitative RT-PCR, after which the expression of ABCC3, ABCC4 and ABCC5 in different developmental stages and tissues of 4th instar larvae was measured. [Results] All 5 genes were up-regulated in the resistant strain(CHR) where their expression was significantly higher than in the susceptible strain(CHS). Expression of ABCC3, ABCC4 and ABCC5 were 2.16, 2.81, and 1.85 times higher in the CHS following treatment with the LC50 dose of chlorantraniliprole than in the untreated control, but ABCC1 and ABCC2 were down-regulated and their expression was only 65.8% and 37.2% that of the control, respectively. The expression of ABCC3 mRNA in the larval stage and in male adults was significantly higher than in other developmental stages. There was no obvious difference in the expression of ABCC4 mRNA among the developmental stages examined. The expression of ABCC5 mRNA in third instar larvae was significantly higher than in other developmental stages. In 4th instar larvae, the expression of ABCC3, ABCC4 and ABCC5 mRNA in the midgut was significantly higher than in other tissues, and the expression of ABCC4 mRNA was significantly higher than that of ABCC3 and ABCC5 in the same tissue. [Conclusion] The results indicate that ABCC3, ABCC4 and ABCC5 may be involved in the detoxification of chlorantraniliprole in the diamondback moth. Among three genes, ABCC4 gene may be the most important in detoxification. These results provide a basis for further study of the relationship between ABCC1-ABCC5 and the metabolic mechanisms involved in the detoxification of chlorantraniliprole in the diamondback moth.