Identification of Defensin genes and analysis of expression patterns in response against bacteria in Lasioderma serricorne
The purpose of this study was to elucidate the function of Lasioderma serricorne Defensin(LsDef1 and LsDef2)genes in response to bacterial infection.The structure,characteristics and phylogenetic relationship of LsDef1 and LsDef2 were analyzed by bioinformatics.Real-time quantitative PCR(RT-qPCR)was used to detect the tissue-specific expression(Fat body,Integument,Midgut,Hemocyte,Malpighian tubule),developmental stages(egg,1st instar larva,2nd instar larva,3rd larva,4th instar larva,pupa,adult)of LsDef1 and LsDef2.RT-qPCR was also to detect the expression pattern of LsDef1 and LsDef2 response against different pathogen,bacteria induction pattern and bacterial cell walls induction pattern.The sequence comparison showed that induction pattern had 6 typical cysteines and formed 3 pairs of disulfide bonds.RT-qPCR showed that LsDef1 and LsDef2 gene in L.serricornes was mainly expression in fat body and hemocytes,followed by integument and midgut,and the expression level was lowest in the Malpighian tubules tissues,and the expression level was high in the 4th instar larvae.It could be significantly stimulated by foreign pathogens,and LsDef1 and LsDef2 were the most sensitive to Staphylococcus aureus.Bacterial and cell wall component induced expression patterns were further analysed,and the expression peak appears at 6 h after induction.When the S.aureus PGN-SA stimulated L.serricorne,the peak expression peak appeared at 3 h after induction.These results indicated that antimicrobial peptides LsDef1 and LsDef2 genes could be induced by pathogen and laid a foundation for further research on the function of LsDef1 and LsDef2.
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