ATP Synthase Subunit α from Helicoverpa armigera Acts as a Receptor of Bacillus thuringiensis Cry1Ac and Synergizes Cry1Ac Toxicity

Insect resistance to Bacillus thuringiensis (Bt) toxins has led to an urgent need to explore the insecticidal mechanisms of Bt. Previous studies indicated that Helicoverpa armigera ATP synthase subunit α (HaATPs-α) is involved in Cry1Ac resistance. In this study, a real-time quantitative polymerase chain reaction (RT-PCR) confirmed that HaATPs-α expression was significantly reduced in the Cry1Ac-resistant strain (BtR). Cry1Ac feeding induced the downregulated expression of HaATPs-α in the susceptible strain, but not in the BtR strain. Furthermore, the interaction between HaATPs-α and Cry1Ac was verified by ligand blotting and homologous competition experiments. The in vitro gain and loss of function analyses showed HaATPs-α involved in Cry1Ac toxicity by expressing endogenous HaATPs-α and HaATPs-α double-stranded RNAs in Sf9 and midgut cells, respectively. Importantly, purified HaATPs-α synergized Cry1Ac toxicity to H. armigera larvae. These findings provide the first evidence that HaATPs-α is a potential receptor of Cry1Ac, it shows downregulated participation in Cry1Ac resistance, and it exhibits higher enhancement of Cry1Ac toxicity to H. armigera larvae.