Insights into the different functions of multiple peptidoglycan recognition proteins in the immune response against bacteria in the mosquito, Armigeres subalbatus

•Bacterial challenges of Ar. subalbatus induced the expression of AsPGRP–S1, but not other AsPGRPs.•RNAi of the four AsPGRPs has different effects on mosquito survival after bacterial challenges.•Four antimicrobial peptide genes were differently regulated after AsPGRP RNAi.•AsPGRP-LC RNAi dramatically suppressed the transcription of the four antimicrobial peptides.•AsPGRP-LC is not a key factor in mediating phagocytosis in Ar. subalbatus.

Peptidoglycan recognition proteins (PGRPs) are a group of proteins that recognize and/or bind to peptidoglycan on the surface of a number of pathogens. To understand the roles of multiple PGRPs in the mosquito Armigeres subalbatus (AsPGRPs), we studied the effects of infection of two bacteria, the gram negative Escherichia coli and the gram positive Micrococcus luteus, on the transcriptional expression of AsPGRPs and RNA interference (RNAi) of AsPGRPs on the immune responses of mosquitoes against the two bacteria. Injection of E. coli or M. luteus into adult mosquitoes both significantly increased the transcription of AsPGRP-S1, but not the other AsPGRPs. A mosquito survival assay using injection of E. coli or M. luteus into AsPGRP double-stranded RNA (dsRNA) injected mosquitoes showed that RNAi of AsPGRPs had different impacts on the survival abilities of mosquitoes, and that AsPGRP-LCs seem to be the most critical ones. Real-time Polymerase Chain Reaction (real-time PCR) analysis indicated that the expression of four antimicrobial peptides (AMPs) was dramatically changed after AsPGRP-LB and AsPGRP-LC RNAi, although AsPGRP-S1 and AsPGRP-LE had slight, but significant, effects, suggesting that the changes in survival abilities were potentially due to the changes in AMP expression after AsPGRP RNAi. In addition, bacterial challenges following AsPGRP-LC RNAi did not induce the expression of AMPs to their normal level as in control experiments. An in vivo assay indicated that AsPGRP-LC RNAi had no significant effects on the phagocytic ability of the hemocytes, suggesting that AsPGRP-LC is not a key factor mediating phagocytosis of bacteria in this mosquito.

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