Construction Of Normalized Full-length CDNA Library Of Apolygus Lucorum And Exploration Of AlucCSPs

Chemical communication is a crucial way to contact with their partner as well as thesurrounding environment for insects who can’t communicate with each other by usinglanguage and words. Olfactory system is one of the most important physiological systems forthe sense of chemical communication. Clarifying the mechanism of insect olfactionrecognition, will help usdevelop more effective and friendly means for pests control. The green plant bugs havebecome major pests instead of cotton bollworm after the commercial planting of BT cotton inChina cotton ecosystem. Olfactory related proteins make the indispensable material basis inthe process of recognition mechanism, including chemosensory proteins (CSPs)、odorantbinding proteins (OBPs）、olfactory receptors (ORs)、sensory neuron membrane proteins(SNMPs)、odorant degrading enzymes (ODEs), they can be used as new potential target genesin the prevention of green plant bugs. Among them, CSPs exercise the function of combiningwith odors as well as some other functions in the terms of other physiological. In this article,we contructed a high-quality cDNA library from the antennae of A. lucorum,8novel putativeCSP genes were identified from the antennal cDNA library, and we named them AlucCSP1-AlucCSP8. And then we analysed the library by particular bioinformatics, and the expressionprofiles of8CSPs in different tissues (antennae, heads, thoraxes, abdomens, legs and wings)were analyzed by real-time quantitative PCR.1. We constructed a cDNA library from the antennae of A. lucorum using SMARTmethod, and then we analysed them by bioinformatics using cluster analysis, and we obtained3881unigenes, And the average length of the assembled unigenes was1143.87bp. Then theseunigenes were assigned into three different GO-termes, including cellular location, molecularfunction and biological process. In particular,560unigenes (53.7%of the1042unigenes)encoding proteins were assigned to have putative binding function in the molecular functioncategory. And the analysis of KEGG told us they produced1590pathways, which involvedfive categories: metabolism, genetic information processing, environmental informationprocessing, cellular processes, human diseases. These results reflect it was a the high qualitylibrary that involved comprehensive genes.2. Eight new CSP genes were discovered from the ESTs sequence in the library, and we builded the Neighbor-joining tree to compare the homology of these sequence, and the resultrevealed that AlucCSP3had a long genetic distance with other genes. And we then analyzesthe CSPs amino acid structure according to their amino acid sequence.3. The expression profiles of the8CSPs in different tissues (antennae, heads, thoraxes,abdomens, legs and wings) were analyzed by real-time quantitative PCR. The results revealedthatAlmost all of the CSPs were rarely expressed in the thoraxes snd abdomens, while theywere highly expressed in antennae and legs, then they also had much expression in wings, andthen they were merely espressed in heads. AlucCSP3were specifically expressed in theantennae, and AlucCSP1and AlucCSP8were mainly expressed in legs, and AlucCSP2,AlucCSP4, AlucCSP5, AlucCSP6, AlucCSP7were highly expressed in the female antennaand legs.These8CSPs were divided into three classes based on this result together with theamino acid structure. ClassI included AlucCSP1and AlucCSP8, and Class II includedAlucCSP3, and Class III included AlucCSP2, AlucCSP4, AlucCSP5, AlucCSP6, AlucCSP7,maybe they all had their particular functions in olfactory.