Towards disrupting Varroa -honey bee chemosensing: A focus on a Niemann-Pick type C2 transcript.

We focused our study on the 12 recently identified putative odorant carrier proteins in the ectoparasitic mite, Varroa destructor. Here we show, via an exclusion of the chemosensory appendages (forelegs and gnathosoma) that transcripts of five of the 12 genes were significantly lower, suggesting that they are likely involved in carrying host volatiles. Specifically, three transcripts were found to be foreleg-specific while the other two transcripts were expressed in both the forelegs and gnathosoma. We focused on one of the highly expressed and foreleg-specific transcript Vd40090, which encodes a Niemann-Pick disease protein type C2 (NPC2) protein. Effects of dsRNA-mediated silencing of Vd40090 were first measured by quantifying the transcript levels of genes that encode other putative odorant carrier proteins as well as reproduction related proteins. In addition, the impact of silencing on mites behaviour and survival was tested. Silencing of Vd40090 effectively disrupted Varroa host selection, acceptance and feeding and significantly impaired the expression of genes that regulate its reproduction in brood cells, resulting in reduced reproduction and survival.

Varroa chemosensory proteins: some are conserved across Arthropoda but others are arachnid specific.

The tight synchronization between the life cycle of the obligatory parasitic mite Varroa destructor (Varroa) and its host, the honeybee, is mediated by honeybee chemical stimuli. These stimuli are mainly perceived by a pit organ located on the distal part of the mite's foreleg. In the present study, we searched for Varroa chemosensory molecular components by comparing transcriptomic and proteomic profiles between forelegs from different physiological stages, and rear legs. In general, a comparative transcriptomic analysis showed a clear separation of the expression profiles between the rear legs and the three groups of forelegs (phoretic, reproductive and tray-collected mites). Most of the differentially expressed transcripts and proteins in the mite's foreleg were previously uncharacterized. Using a conserved domain approach, we identified 45 transcripts with known chemosensory domains belonging to seven chemosensory protein families, of which 14 were significantly upregulated in the mite's forelegs when compared to rear legs. These are soluble and membrane bound proteins, including the somewhat ignored receptors of degenerin/epithelial Na+ channels and transient receptor potentials. Phylogenetic clustering and expression profiles of the putative chemosensory proteins suggest their role in chemosensation and shed light on the evolution of these proteins in Chelicerata.

Identification and gene-silencing of a putative odorant receptor transcription factor in Varroa destructor: possible role in olfaction.

The ectoparasitic mite Varroa destructor is one of the major threats to apiculture. Using a behavioural choice bioassay, we determined that phoretic mites were more successful in reaching a bee than reproductive mites, suggesting an energy trade-off between reproduction and host selection. We used both chemo-ecological and molecular strategies to identify the regulation of the olfactory machinery of Varroa and its association with reproduction. We focused on transcription regulation. Using primers designed to the conserved DNA binding region of transcription factors, we identified a gene transcript in V. destructor homologous to the pheromone receptor transcription factor (PRTF) gene of Pediculus humanus corporis. Quantitative PCR (qPCR) revealed that this PRTF-like gene transcript is expressed in the forelegs at higher levels than in the body devoid of forelegs. Subsequent comparative qPCR analysis showed that transcript expression was significantly higher in the phoretic as compared to the reproductive stage. Electrophysiological and behavioural studies revealed a reduction in the sensitivity of PRTF RNA interference-silenced mites to bee headspace, consistent with a reduction in the mites' ability to reach a host. In addition, vitellogenin expression was stimulated in PRTF-silenced mites to similar levels as found in reproductive mites. These data shed light upon the regulatory mechanism of host chemosensing in V. destructor.