A Mechanistic Model of Cry2Ab12 Toxicity Against Myzus persicae via HSP60-Mediated OLA1 Inhibition

Bacillus thuringiensis Cry toxins are well known for their high insecticidal activity against Lepidoptera, Diptera, and Coleoptera and have been widely used in Bt transgenic crops. However, their activity against Hemipteran aphids remains relatively low. Identifying novel Cry proteins and elucidating their action mechanisms can facilitate the development of effective aphid control strategies. In this study, we found that ingestion of Cry2Ab12 did not kill Myzus persicae adults but significantly reduced their offspring number and exerted a lethal effect on M. persicae nymphs. After identifying Cry2Ab12 toxin-binding proteins in M. persicae, we further characterized the interaction with Obg-like ATPase 1 (OLA1), a conserved protein involved in growth regulation. Bio-layer interferometry (BLI), ELISA, and enzyme activity assays revealed that Cry2Ab12 and OLA1 do not interact directly. Interestingly, heat shock protein 60 (HSP60) was shown to mediate the interaction among Cry2Ab12, HSP60, and OLA1, leading to inhibition of OLA1 enzymatic activity. Based on these findings and bioinformatics simulations, we proposed a mechanistic model for Cry2Ab12 toxicity against M. persicae: upon ingestion of a sufficient amount of Cry2Ab12, the formation of the Cry2Ab12–HSP60–OLA1 complex impairs the cellular stress response, disrupts normal OLA1 expression, and ultimately restricts larval growth and development, resulting in lethality. This study provides new insights into the action of Cry toxins in aphids and offers a basis for developing enhanced aphid biocontrol strategies.