Cloning and characterization of three chemosensory proteins from Spodoptera exigua and effects of gene silencing on female survival and reproduction

Abstract

Insect chemosensory proteins (CSPs) are supposed to transport hydrophobic chemicals to receptors on sensory neurons. However, CSPs are broadly expressed in various insect tissues, suggesting their involvement in the physiological processes beyond chemoreception. So, the exact physiological roles of CSPs in insects still need to be unraveled. In this study, three full-length of CSP genes from Spodoptera exigua have been cloned and characterized. The deduced amino acid sequences of SexiCSP1, SexiCSP2 and SexiCSP3 revealed open reading frames of 128, 128 and 126 amino acids, respectively, with four conserved cysteine residues. The expression patterns of the three SexiCSPs were further investigated by real-time PCR. Three SexiCSPs were expressed in antennae, heads, legs, wings, thoraxes, abdomens, testes and ovaries, with the highest expression level in female and male antennae. Furthermore, all three SexiCSPs mRNA were distributed extensively in the tested development stages with the highest expression level in pupae. RNAi-based gene silencing study resulted in a dramatic reduction of corresponding mRNA in female S. exigua after injection with dsRNA of all three SexiCSPs. Consequentially, 42.5% of mortalities, 68.3% (compare to DEPC water injected control) and 71.4% (compare to uninjected control) oviposition inhibition, and significantly effected egg hatching were observed in the female S. exigua injected with dsSexiCSP3 as compared to control treatments. On the other hand, dsSexiCSP1 and dsSexiCSP2 injected female adults did not show effects on survival and reproduction. Our study confirms the utility of RNAi approach to functional characterization of CSP genes in S. exigua and provides a starting point for further studies on female survival and reproduction in this insect. It also reveals the potential pest controlling method, as insect behavior regulation agent that disrupts the expression of chemosensory proteins.