Kang Xin-Le, Zhang Meng, Wang Kang, Qiao Xian-Feng, Chen Mao-Hua
Key Laboratory of Crop Pest Integrated Pest Management on the Loess Plateau of Ministry of Agriculture, College of Plant Protection, Northwest A&F University, Yangling, Shaanxi, China.
Arch Insect Biochem Physiol. 2016 May;92(1):65-84. doi: 10.1002/arch.21334.
The ATP-binding cassette (ABC) transporters are important transmembrane proteins encoded by a supergene family. The majority of ABC proteins are primary active transporters that bind and hydrolyze ATP to mediate the efflux of a diverse range of substrates across lipid membranes. In this study, we cloned and characterized a putative multidrug resistance associated protein 1 (MRP1) from Rhopalosiphum padi encoded by ABCC1. Structural analysis showed that this protein has structural features typical of the ABC transporter family. Phylogenetic analysis indicated that the amino acid sequence was highly similar that of the corresponding protein from Acyrthosiphon pisum. Real-time quantitative polymerase chain reaction (PCR) analysis showed that ABCC1 was expressed throughout all R. padi developmental stages, with the highest level of expression in the fourth larval instar. We also examined ABCC1 expression in four different tissue types and found that it was most highly expressed in the midgut. Exposing R. padi to imidacloprid and chlorpyrifos increased ABCC1 expression. Furthermore, ABCC1 expression was higher in the imidacloprid-resistant (IR) and chlorpyrifos-resistant (CR) strains than in an insecticide-susceptible strain (SS) of R. padi. Exposing R. padi to verapamil in combination with insecticides significantly increased the toxicity of the insecticides. The respective synergy factor of CR and IR R. padi strain was 1.33 and 1.26, which was lower than that (2.72 and 1.64, respectively) of the SS. Our results clarify the biological function of ABCC1 in R. padi, particularly its role in insecticide resistance, and suggest novel strategies for pest management that use ABC transporter inhibitors to increase the effectiveness of insecticides.
ATP结合盒（ABC）转运蛋白是由一个超基因家族编码的重要跨膜蛋白。大多数ABC蛋白是初级主动转运蛋白，它们结合并水解ATP，以介导多种底物跨脂质膜的外排。在本研究中，我们克隆并鉴定了由ABCC1编码的来自禾谷缢管蚜的一个假定的多药耐药相关蛋白1（MRP1）。结构分析表明，该蛋白具有ABC转运蛋白家族的典型结构特征。系统发育分析表明，其氨基酸序列与来自豌豆蚜的相应蛋白高度相似。实时定量聚合酶链反应（PCR）分析表明，ABCC1在禾谷缢管蚜的所有发育阶段均有表达，在四龄幼虫期表达水平最高。我们还检测了ABCC1在四种不同组织类型中的表达，发现其在中肠中表达最高。将禾谷缢管蚜暴露于吡虫啉和毒死蜱中会增加ABCC1的表达。此外，ABCC1在禾谷缢管蚜的抗吡虫啉（IR）和抗毒死蜱（CR）品系中的表达高于杀虫剂敏感品系（SS）。将禾谷缢管蚜暴露于维拉帕米与杀虫剂的组合中会显著增加杀虫剂的毒性。CR和IR禾谷缢管蚜品系各自的协同因子分别为1.33和1.26，低于SS品系（分别为2.72和1.64）。我们的结果阐明了ABCC1在禾谷缢管蚜中的生物学功能，特别是其在抗药性中的作用，并提出了利用ABC转运蛋白抑制剂提高杀虫剂有效性的害虫管理新策略。
