Chemosense-related Genes Identification And Expression Analysis In Phenacoccus Solenopsis Tinsley

In the long-term evolution, insects have gradually formed a complete set of chemosense system, by which insects can systematically sense the chemical information in the surrounding environment and complete the behavior of host orientation, mate, food location, avoidance of predators and toxic substances. In the chemosense system, the function odorant binding proteins(OBPs), chemosensory proteins(CSPs) and olfactory receptors(ORs) are pretty important. Phenacoccus solenopsis Tinsley, is one of the important invasive pests in China. In recent years, many researches have been done about the ecology and behavior of the P.solenopsis, but molecular mechanism research of chemosense system is less. Thus, we have identified the CSPs, OBPs and ORs genes,investigated the expression patterns of CSPs and ORs, cloned and analysis the expression of Ps CSP10. The main results are as follows:1 Chemosense-related gene identificationThe transcriptome profile databases revealed 12 chemosensory proteins(CSPs)( Psol CSP1-Psol CSP12), 15 odorant binding proteins(OBPs)( Psol OBP1-Psol OBP15)and 1 olfactory receptor(ORs)( Psol OR1).2 Expression patterns of the chemosensense-related genes in P.solenopsisUsing comparative quantitative real-time PCR to analyze the expression of the Psol CSP1-Psol CSP12 genes at different developmental stages(egg, 1st instar larvae, 2nd instar larvae, 3rd instar larvae, male adult and female adult) of P.solenopsis were detected.The data indicated Psol CSP2 got a high level in the 2nd instar larvae. The expression level of Psol CSP3, Psol CSP4, Psol CSP6 and Psol CSP9 are higher than in other developmental stages. Expression of Psol CSP5 and Psol CSP7 have no significant difference in different developmental stages, while Psol CSP11 distributed with a higher level in the 3rd insar larvae. We also found that the expression of Psol CSP1, Psol CSP8, Psol CSP10, Psol CSP12 in every developmental stage, and they got a highest level in the male adult. Furthermore,we found Psol OR1 got a high level in the egg.3 Molecular Cloning and Expression Analysis of Chemosensory Protein Gene Ps CSP10In this study, one CSP gene of P.solenopsis was cloned and characterized, which was named Ps CSP10(Gen Bank accession no.: KT958555). The Ps CSP10 c DNA was 640 bp in length, encoding 128 amino acid residues with the predicated molecular mass of approximately 14.99 k D and p I of 6.61, of which four conservative cysteines are included.The alignment of Ps CSP10 showed moderate similarity(50%- 56%) with the CSP genes of other insects analyzed with the deduced amino acid sequences. Real-time PCR analysis revealed that Ps CSP10 was expressed across various development stages, the expressions were more apparent in the male adult. The expression levels of Ps CSP10 had not significant difference(p > 0.05) in male adult feed on Chinese rose(Hibiscus rosa-sinensis), cotton(Gossypium spp) and Lantana camara respectively. These results suggest that the Ps CSP10 gene is most likely involved in mate location and other chemosensory functions of the insect.By analyzing the transcriptome of P.solenopsis, a number of chemosense-related genes were identified. Meanwhile the differentially expression of the chemosense-related genes between different developmental stages were investigated. By rapid-amplification of c DNA ends(RACE), Ps CSP10 full length sequence was cloned. These results are important to reveal the chemosense mechanism.