CRISPR/Cas9-based functional characterization of Px ABCB1 reveals its roles in the resistance of Plutella xylostella(L.)to Cry1Ac,abamectin and emamectin benzoate  

作　　者：OUYANG Chun-zheng YE Fan WU Qing-jun WANG Shao-li Neil CRICKMORE ZHOU Xu-guo GUO Zhao-jiang ZHANG You-jun 

机构地区：[1]State Key Laboratory of Vegetable Biobreeding,Deapartment of Plant Protection,Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,P.R.China [2]Guangdong Laboratory for Lingnan Modern Agriculture,Guangzhou 510642,P.R.China [3]School of Life Sciences,University of Sussex,Brighton BN19QG,UK [4]Department of Entomology,University of Kentucky,Lexington,KY 40546-0091,USA

出　　处：《Journal of Integrative Agriculture》2023年第10期3090-3102,共13页农业科学学报（英文版）

基　　金：the Laboratory of Lingnan Modern Agriculture Project,China(NT2021003);National Natural Science Foundation of China(32022074,32221004 and 32172458);Beijing Key Laboratory for Pest Control and Sustainable Cultivation of Vegetables,Chinese Academy of Agricultural Sciences,and the Innovation Program of the Chinese Academy of Agricultural Sciences(CAAS-CSCB-202303)。

摘　　要：The identification of functional midgut receptors for pesticidal proteins produced by Bacillus thuringiensis(Bt)is critical for deciphering the molecular mechanism of Bt resistance in insects.Reduced expression of the PxABCB1 gene was previously found to be associated with Cry1Ac resistance in the diamondback moth,Plutella xylostella(L.).To directly validate the potential receptor role of PxABCB1 and its contribution to Bt Cry1Ac toxicity in P.xylostella,we used CRISPR/Cas9 to generate a homozygous knockout ABCB1KO strain with a 5-bp deletion in exon 3 of its gene.The ABCB1KO strain exhibited a 63-fold resistance to Cry1Ac toxin compared to the parental DBM1Ac-S strain.Intriguingly,the ABCB1KO strain also exhibited significant increases in susceptibility to abamectin and emamectin benzoate.No changes in susceptibility to various other Bt Cry proteins or synthetic insecticides were observed.The knockout strain exhibited no significant fitness costs.Overall,our study indicates that PxABCB1 can protect the insect against avermectin insecticides on one hand,while on the other hand it facilitates the toxic effect of the Bt Cry1Ac toxin.The results of this study will help to inform integrated pest management approaches against this destructive pest.

关 键 词：Bacillus thuringiensis Plutella xylostella CRISPR/Cas9 ABCB1 bioinsecticide resistance 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治]