Retrotransposon-mediated variation of a chitin synthase gene confers insect resistance toBacillus thuringiensisVip3Aa toxin

Abstract

Bacillus thuringiensis(Bt) crops expressing Vip3Aa are highly efficacious in controlling major lepidopteran pests and delaying evolution of pest resistance. Although practical resistance to Vip3Aa in the field has not been reported, to proactively manage the pest resistance, there is an urgent need to better understand the genetic basis of resistance to Vip3Aa. This is particularly important for the fall armyworm (Spodoptera frugiperda), one of the most destructive pests around the world, which has evolved practical resistance toBtcrystal (Cry) toxins. Here, a highly Vip3Aa-resistant (resistance ratio: 5,562-fold) strain ofS. frugiperdawas selected in the laboratory. Results from bulked segregant analysis, fine-scale mapping, and genetic linkage analysis indicate that a mutation in the midgut-specific chitin synthase gene,SfCHS2, is strongly associated with high-level resistance to Vip3Aa. The resistance is ascribed to the transcriptional variation caused by retrotransposon insertion. The same variation ofSfCHS2was also detected in a field population. Importantly, knockout ofSfCHS2via CRISPR/Cas9 in susceptibleS. frugiperdaconfers its complete resistance (>10,000-fold) to Vip3Aa. Also, we demonstrate that knockout ofCHS2can result in complete resistance to Vip3Aa in additional lepidopteran species, suggesting a general role of this gene in Vip3Aa resistance among lepidopteran pests. These results reported here would contribute to monitor and management of pest resistance to Vip3Aa.