Cloning And Experession Of Csaqp1Genes In Rice Stem Borer, Chilo Suppressalis Walker(Lepidoptera:Pyralidae)

The rice stem borer, Chilo suppressalis (Walker), is one of the key pests of rice, which distributes widely in China, and has strong adaption to environment. Previous studies have indicated that physical activity of insects was associated with water channel proteins (AQPs), which involved in transportation of water and regulation of osmotic pressure.. In present study, two CDNAs are cloned from C. suppressalis by RACE, then their expression patterns in different life stage and distribution in tissues from5th larvae were monitored by real-time PCR. In addition, the potential role of AOPs in C.suppressalis development was disccused. The main results are as following:Two full length of CsAQP1CDNAs (CsAQP1transcript variant A and CsAQP1transcript variant B) were cloned from C. suppressalis larvae by RT-PCR and RACE. The full length of CsAQP1transcript variant A consists of1416bp with a777bp of opening reading fragment (ORF), which encodes258amino acids residues. The5’UTR and3’UTR are342bp and300bp, respectively. The full length of CsAQPl transcript variant B is1825bp encoding777amino acids residues, with a777bp of ORF. The5’UTR and3’UTR are327bp and724bp, respectively. Homology analysis suggests that the deduced amino acids of AOP1from C.suppressalis has80%identity with those of silkworm and75%identity with oriental fruit moth.Compared with amino acids sequence from other insects by DNAMAN software, we find that CsAQP1have two NPA motifs and a signature sequence of SYDF at the C-terminus and a Ar/R constriction regions, which is composed of four conserved regions that adjacent to each other in space(Phe, His, Ser and Arg). Hydropathy plot of C. suppressalis aquaporin indicates that CsAQP1consists of six Hydropathy transmembrane spanning and five loops regions. Analysis from CBS prediction system proves that CsAQPl has one N-linked glycosylation site and two phosphorylation sites.The expression patterns of CsAQPl in eggs,1-6larvae,female and male pupa, female and male adults and foregut, midgut, handgut, Malpighian tubule, head, epidermis, fat body and hemolymph of5th larvae was investigated by real-time PCR. Results suggested that the highest expression levels of CsAQPl was found in Malpighian tubule, the second is handgut, but with low expression levels in head, hemolymph, epidermis, fat body, hemolymph. In all life stages, the highest expression of CsAQP1was detected in eggs and the expression levels declined in later stages, the lowest expression was found in fifth larvae. In pupa and adults stages, the male individuals have higher expression levels than female...