Two ABC transporters are differentially involved in the toxicity of two Bacillus thuringiensis Cry1 toxins to the invasive crop-pest Spodoptera frugiperda (J. E. Smith)
Two ABC transporters are differentially involved in the toxicity of two Bacillus thuringiensis Cry1 toxins to the invasive crop-pest Spodoptera frugiperda (J. E. Smith)
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DOI：
10.1002/ps.6170
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发表时间：
2020-11-19
影响因子：
4.1
通讯作者：
Xiao, Yutao
中科院分区：
文献类型：
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作者：
Jin, Minghui;Yang, Yanchao;Xiao, Yutao
秋粘虫是一种主要的农业害虫，自2016年以来入侵东半球，对包括非洲和亚洲在内的全球粮食安全构成严重威胁。苏云金芽孢杆菌（Bacillus thuringiensis, Bt）产生的Cry毒素已被证明对这种害虫有效。在不同的昆虫中，ABC转运体（ABCC2或ABCC3）已被证明是一些Cry1毒素的受体。我们分析了SfABCC2和SfABCC3在Cry1Fa和Cry1Ab毒素对该害虫的毒性中的作用。结果利用CRISPR/Cas9基因组编辑系统，构建了2株编码潜在功能Bt受体的S. frugiperda SfABCC2和SfABCC3基因敲除菌株。与敏感菌株相比，敲除菌株对Cry1Fa和Cry1Ab毒素均有抗性。SfABCC2敲除菌株对这两种Cry毒素的抗性均高于SfABCC3敲除菌株，表明SfABCC2在这两种Cry毒素的作用模式中起主要作用。此外，SfABCC2和SfABCC3基因在毛癣菌Hi5细胞中的表达也增加了对Cry1Ab和Cry1Fa毒素的易感性，与基因组编辑结果一致。与其他鳞翅目相比，双敲除SfABCC2和SfABCC3菌株是不可行的。此外，我们在这里报道SfABCC2或SfABCC3敲除菌株增加了对阿维菌素和spinosad杀虫剂的敏感性。结论这两种ABCC转运蛋白在frugiperda中作为Bt Cry1Fa和Cry1Ab毒素的受体。
BACKGROUND The fall armyworm Spodoptera frugiperda is a major agricultural pest that has invaded the East Hemisphere since 2016, generating a serious threat to food security worldwide including Africa and Asia. The Cry toxins produced by Bacillus thuringiensis (Bt) have been shown to be effective against this insect pest. In different insect ABC transporters (ABCC2 or ABCC3) have been shown to be involved as receptors of some Cry1 toxins. Here we analyzed the role of SfABCC2 and SfABCC3 in the toxicity of Cry1Fa and Cry1Ab toxins in this insect pest.RESULTS Two S. frugiperda SfABCC2 and SfABCC3 knockout strains, coding for potential functional Bt receptors, were created using CRISPR/Cas9 genome editing system. Both knockout strains showed resistance to both Cry1Fa and Cry1Ab toxins compared with the susceptible strain. SfABCC2 knockout strain showed higher resistance to both Cry toxins than SfABCC3 knockout strain, suggesting a major role of SfABCC2 in the mode of action of these Cry toxins. In addition, expression of SfABCC2 and SfABCC3 genes in Trichoplusia ni Hi5 cells also increased the susceptibility to Cry1Ab and Cry1Fa toxins, in agreement with the genome editing results. The double knockout of SfABCC2 and SfABCC3 strain was not viable in contrast to other lepidopteran species. Furthermore, we report here that SfABCC2 or SfABCC3 knockout strains increased their susceptibility to abamectin and spinosad insecticides.CONCLUSION We provide functional evidence that in S. frugiperda these two ABCC transporters serve as receptors of Bt Cry1Fa and Cry1Ab toxins.