Genetic knockout and general odorant-binding/chemosensory protein interactions: Revealing the function and importance of GOBP2 in the yellow peach moth's olfactory system

Abstract

The GOBP2 protein has a unique function in the yellow peach moth (Conogethes punctiferalis (Guenée)). Several general odorant-binding proteins (GOBPs) have been identified in various lepidopteran species, but the functional difference between GOBP1 and GOBP2 in recognition of host plant odorants is still unknown. The functions of GOBP1 and GOBP2 in the yellow peach moth were evaluated in this study by using the CRISPR-Cas9 system. The results revealed the importance of GOBP2 in the olfaction mechanism in the yellow peach moth. The perception of the GOBP1-knockout larvae toward feeding decreased but did not reach a significant level while knocking out the GOBP2 and GOBP1/2 genes resulted in huge differences. On the other hand, electroantennograms (EAGs) and wind tunnel tests showed that the sensitivity of GOBP2 knockout adults to odorants decreased more than that of GOBP1 knockout individuals. The results of STRING database text mining grabbed our attention in protein-protein interaction studies. In this research, we first proved the existence of physical interactions between GOBPs and chemosensory proteins (CSPs) through the surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) methods. Interestingly, GOBP1 and GOBP2 could not interact with each other, but they could interact with CSPs. The interaction results indicated that GOBP2 could physically interact with CSP15, CSP5, and OBP17, whereas GOBP1 could bind only with CSP5 and CSP10, and its association constant (ka) was also more substantial than that of GOBP1. These results strongly suggest the importance of the function of GOBP2 in the perception of host plant odorants by the yellow peach moth.