Knockout of theSfVipR1gene confers high-level resistance toBacillus thuringiensisVip3Aa toxin inSpodoptera frugiperda

Background Bacillus thuringiensis (Bt) insecticidal proteins, including Cry proteins and vegetative insecticidal proteins (Vips), are extensively utilized in transgenic crops due to their efficacy and safety. The fall armyworm, Spodoptera frugiperda , has evolved practical resistance to Cry1Fa, yet no practical resistance to Vip3Aa has been documented. However, both laboratory selection and field screen studies indicate a high potential for this pest to evolve resistance to Vip3Aa, making it crucial to evaluate potential resistance genes. HaVipR1 has recently been identified as a key determinant of Vip3Aa resistance in the cotton bollworm, Helicoverpa armigera . This study investigated whether the HaVipR1 -homologous gene in S. frugiperda ( SfVipR1 ) is similarly involved in Vip3Aa resistance. Results We employed CRISPR/Cas9 technology to generate a homozygous knockout strain of SfVipR1 . In comparison with the parent susceptible YJ-19 strain, the knockout strain (Sfru-KO) exhibited high-level resistance to Vip3Aa (>1875-fold) but showed no resistance to Cry1Fa. This acquired resistance to Vip3Aa is autosomal, recessive, and genetically linked to the deletion mutation in SfVipR1 within the Sfru-KO strain of S. frugiperda . Conclusion Disruption of SfVipR1 results in high-level resistance to Vip3Aa, highlighting SfVipR1 has a critical role in Vip3Aa toxicity in S. frugiperda , despite the exact mechanism remaining unclear. Early detection of SfVipR1 mutant alleles in the field is essential for developing adaptive resistance management strategies against S. frugiperda .