ProPhenolOxidase in Daphnia magna: cDNA sequencing and expression in relation to resistance to pathogens

Invertebrates utilise the innate immune system when defending against pathogenic attack. However, except for some effectors as proPhenolOxidase (proPO), the innate immune response is less well understood outside model insect species, and its role in natural host-pathogen systems is generally not well documented. We have therefore initiated studies on the immune response of the crustacean Daphnia when exposed to the specialist endobacterial pathogen, Pasteuria ramosa. This study was focused on the proPO gene of Daphnia magna. D. magna possesses a single copy of proPO (as does its congener, D. pulex), but there was some evidence of alternative splicing. Analyses of sequence similarity in a range of arthropod taxa suggested that the proPO gene in Daphnia was as dissimilar to other crustaceans as it was to insects, while analysis on intraspecific variation indicated that the gene is highly conserved. ProPO was found to be significantly up-regulated within 1-4h following exposure to the bacteria. This is the first evidence of a Daphnia immune response, and our observations raise the possibility that the PhenolOxidase (PO) cascade is involved in the defence against pathogenic gram-positive bacteria.