Cry1 resistance in a CRISPR/Cas9-mediated HaCad1 gene knockout strain of the Australian cotton bollworm Helicoverpa armigera conferta (Lepidoptera: Noctuidae)

BACKGROUND: Helicoverpa armigera is a highly polyphagous species that causes huge losses to agricultural and horticultural crops worldwide. In the cotton industry, H. armigera, including the Australian subspecies Helicoverpa armigera conferta, is largely managed using genetically modified crops that express insecticidal toxins, such as Cry1Ac. Resistance to Cry1 proteins occurs and, in some cases, is mediated by changes to HaCad1, a gene that encodes the midgut protein cadherin. Around the world, numerous resistance-associated polymorphisms have been identified in the HaCad1 gene of H. armigera, but Cry1Ac resistance is rare in the Australian subspecies. We used CRISPR/Cas9 to disrupt the cadherin gene in H. armigera conferta and characterised the resulting phenotype with bioassays and transcriptomics. 

RESULTS: Compared to the parental strain, the newly generated HaCad1 knockout strain is 44-fold and 16-fold more resistant to Cry1Ac and Cry1A.105, respectively, while wild-type and knockout insects were equally insensitive to Cry1F. 

CONCLUSION: The disruption of the HaCad1 gene causes Cry1Ac resistance in Australian H. armigera conferta. However, Cry1Ac resistance remains rare in Australian field populations suggesting that Australia's approach to pest management in cotton has prevented widespread Cry1Ac resistance.