Genome editing of the SfABCC2 gene confers resistance to Cry1F toxin from Bacillus thuringiensis in Spodoptera frugiperda

doi:10.1016/S2095-3119(19)62772-3

Journal of Integrative Agriculture ›› 2021, Vol. 20 ›› Issue (3): 815-820.DOI: 10.1016/S2095-3119(19)62772-3

收稿日期:2019-06-26 出版日期:2021-03-01 发布日期:2021-02-18

Genome editing of the SfABCC2 gene confers resistance to Cry1F toxin from Bacillus thuringiensis in Spodoptera frugiperda

JIN Ming-hui^{1, 2}, TAO Jia-hui¹, LI Qi¹, CHENG Ying¹, SUN Xiao-xu², WU Kong-ming², XIAO Yu-tao¹

¹ Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518120, P.R.China
² State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China

Received:2019-06-26 Online:2021-03-01 Published:2021-02-18
Contact: Correspondence XIAO Yu-tao, E-mail: xiaoyutao@caas.cn
About author:JIN Ming-hui, E-mail: jinminghui722@163.com;
Supported by:
This study was financially supported by the Key Project for Breeding Genetic Modified Organisms of China (2016ZX08012004003).

摘要/Abstract

摘要： ABCC2基因是多种鳞翅目昆虫的Bt受体，有研究表明对Cry1F具有抗性的草地贪夜蛾田间种群与ABCC2突变遗传连锁，为探明SfABCC2突变是否介导Cry1F抗性，我们采用CRISPR/Cas9基因编辑技术对草地贪夜蛾ABCC2基因进行敲除。对筛选得到的纯合敲除品系进行生物测定，结果显示Cry1F毒素对敲除品系的LC50为39.2 (29.2–54.2) μg cm^–2，是野生品系的118倍。本研究通过反向遗传学手段证实SfABCC2是Cry1F受体。

Abstract:

ATP-binding cassette transporter C2 (ABCC2) is known to be a receptor for Bacillus thuringiensis (Bt) toxins in several lepidopteran insects. Mutations in the ABCC2 gene have been genetically linked to field-evolved resistance to the Cry1F toxin from Bt in Spodoptera frugiperda. Here we generated a SfABCC2 knockout strain of S. frugiperda using the CRISPR/Cas9 system to provide further functional evidence of the role of this gene in susceptibility and resistance to Cry1F. Results from bioassays showed that the SfABCC2 knockout S. frugiperda strain displayed 118-fold resistance to Cry1F compared with the parental DH19 strain, but no resistance to Vip3A toxin from Bt. These results provide the first reverse genetic evidence for SfABCC2 as a functional receptor for Cry1F.

Key words: Spodoptera frugiperda , ABCC2 , CRISPR/Cas9 , Bt receptor , Cry1F

. [J]. Journal of Integrative Agriculture, 2021, 20(3): 815-820.

JIN Ming-hui, TAO Jia-hui, LI Qi, CHENG Ying, SUN Xiao-xu, WU Kong-ming, XIAO Yu-tao . Genome editing of the SfABCC2 gene confers resistance to Cry1F toxin from Bacillus thuringiensis in Spodoptera frugiperda[J]. Journal of Integrative Agriculture, 2021, 20(3): 815-820.

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Genome editing of the SfABCC2 gene confers resistance to Cry1F toxin from Bacillus thuringiensis in Spodoptera frugiperda

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