Longping Branch, Graduate School of Hunan University, Changsha, China.
Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China.
Pest Manag Sci. 2020 Feb;76(2):712-720. doi: 10.1002/ps.5569. Epub 2019 Aug 29.
Rapid evolution of pest resistance has seriously threatened the sustainable use of Bacillus thuringiensis (Bt). The diamondback moth, Plutella xylostella (L.), is the first pest to develop resistance to Bt biopesticides in the open field, which renders it an excellent model to explore the molecular basis of Bt resistance in insects. Our previous midgut transcriptome and RNA-Seq profiles showed that the P-glycoprotein gene PxABCB1 was down-regulated in two Cry1Ac-resistant P. xylostella strains, suggesting its potential involvement in Cry1Ac resistance in P. xylostella.
In this study, the bona fide full-length cDNA sequence of the PxABCB1 gene was cloned and analyzed, and the expression of the PxABCB1 gene was detected in all tissues and developmental stages, with the highest expression in midgut tissue and the female adult stage. Although no consistent non-synonymous mutations were identified between the susceptible and resistant strains, PxABCB1 gene expression was remarkably decreased in all resistant strains, and the association was further validated by Cry1Ac selection in the moderately resistant SZ-R strain. Moreover, knockdown of the PxABCB1 gene expression resulted in significantly reduced larval susceptibility to Cry1Ac toxin in the DBM1Ac-S strain, and decreased expression of the PxABCB1 gene was tightly linked to Cry1Ac resistance in P. xylostella.
Our results demonstrated that down-regulation of the PxABCB1 gene is associated with both laboratory-selected and field-evolved Cry1Ac resistance in P. xylostella. This knowledge will be conducive to further elucidating the complicated molecular basis of Bt resistance and developing new insect resistance management tactics. © 2019 Society of Chemical Industry.
害虫抗药性的快速进化严重威胁到苏云金芽孢杆菌(Bt)的可持续使用。小菜蛾是田间第一个对 Bt 生物农药产生抗性的害虫,它是研究昆虫对 Bt 抗性分子基础的理想模型。我们之前的中肠转录组和 RNA-Seq 图谱显示,两个 Cry1Ac 抗性小菜蛾品系中 P-糖蛋白基因 PxABCB1 下调,表明其可能参与了小菜蛾对 Cry1Ac 的抗性。
本研究克隆并分析了 PxABCB1 基因的全长 cDNA 序列,并检测了 PxABCB1 基因在所有组织和发育阶段的表达情况,其中在中肠组织和雌性成虫阶段表达量最高。虽然在敏感和抗性品系之间没有发现一致的非同义突变,但所有抗性品系中 PxABCB1 基因的表达均显著降低,并且在中度抗性 SZ-R 品系中通过 Cry1Ac 选择进一步验证了这种关联。此外,PxABCB1 基因表达的敲低导致 DBM1Ac-S 品系幼虫对 Cry1Ac 毒素的敏感性显著降低,并且 PxABCB1 基因的低表达与小菜蛾对 Cry1Ac 的抗性紧密相关。
我们的研究结果表明,PxABCB1 基因的下调与小菜蛾实验室选择和田间进化的 Cry1Ac 抗性均有关。这些知识将有助于进一步阐明 Bt 抗性的复杂分子基础,并开发新的昆虫抗性管理策略。 © 2019 英国化学学会。