Synergistic Interactions Between Cry1Ac and Natural Cotton Defenses Limit Survival of Cry1Ac-resistant Helicoverpa Zea (Lepidoptera: Noctuidae) on Bt Cotton

Larvae of the bollworm Helicoverpa zea (Boddie) show some tolerance to Bacillus thuringiensis (Bt) Cry1Ac, and can survive on Cry1Ac-expressing Bt cotton, which should increase resistance development concerns. However, field-evolved resistance has not yet been observed. In a previous study, a population of H. zea was selected for stable resistance to Cry1Ac toxin. In the present study, we determined in laboratory bioassays if larvae of the Cry1Ac toxin-resistant H. zea population show higher survival rates on field-cultivated Bt cotton squares (= flower buds) collected prebloom-bloom than susceptible H. zea. Our results show that Cry1Ac toxin-resistant H. zea cannot complete larval development on Cry1Ac-expressing Bt cotton, despite being more than 150-fold resistant to Cry1Ac toxin and able to survive until pupation on Cry1Ac toxin concentrations greater than present in Bt cotton squares. Since mortality observed for Cry1Ac-resistant H. zea on Bt cotton was higher than expected, we investigated whether Cry1Ac interacts with gossypol and or other compounds offered with cotton powder in artificial diet. Diet incorporation bioassays were conducted with Cry1Ac toxin alone, and with gossypol and 4% cotton powder in the presence and absence of Cry1Ac. Cry1Ac toxin was significantly more lethal to susceptible H. zea than to resistant H. zea, but no difference in susceptibility to gossypol was observed between strains. However, combinations of Cry1Ac with gossypol or cotton powder were synergistic against resistant, but not against susceptible H. zea. Gossypol concentrations in individual larvae showed no significant differences between insect strains, or between larvae fed gossypol alone vs. those fed gossypol plus Cry1Ac. These results may help explain the inability of Cry1Ac-resistant H. zea to complete development on Bt cotton, and the absence of field-evolved resistance to Bt cotton by this pest.