Identification of developmentally-specific kinotypes and mechanisms of Varroa mite resistance through whole-organism, kinome analysis of honeybee

Abstract

Recent investigations associate Varroa destructor (Mesostigmata: Varroidae) parasitism and its associated pathogens and agricultural pesticides with negative effects on colony health, resulting in sporadic global declines in domestic honeybee (Apis mellifera) populations. These events have motivated efforts to develop research tools that can offer insight into the causes of declining bee health as well as identify biomarkers to guide breeding programs. Here we report the development of a bee-specific peptide array for characterizing global cellular kinase activity in whole bee extracts. The arrays reveal distinct, developmentally-specific signaling profiles between bees with differential susceptibility to infestation by Varroa mites. Gene ontology analysis of the differentially phosphorylated peptides indicates that the differential susceptibility to Varroa mite infestation does not reflect compromised immunity; rather, there is evidence for mite-mediated immune suppression within the susceptible phenotype that may reduce the ability of these bees to counter secondary viral infections. This hypothesis is supported by the demonstration of more diverse viral infections in mite-infested, susceptible adult bees. The bee-specific peptide arrays are an effective tool for understanding the molecular basis of this complex phenotype as well as for the discovery and utilization of phosphorylation biomarkers for breeding programs.

Differential gene expression of two extreme honey bee (Apis mellifera) colonies showing varroa tolerance and susceptibility

“…A detailed description of the selection of the two contrasting colonies, the tolerant (S88) and the susceptible (G4), selected from a large diverse gene pool (approximately 50 000 colonies), is given in Robertson et al . (). The varroa infestation rates in white‐eyed and dark‐eyed pupae in sealed broods (n = 500) of G4 were 88 and 70%, respectively, and the rate in the adults was 67 mites per hundred bees.…”

“…A complementary study on the same colony phenotypes using kinome arrays showed that varroa mite resistance and susceptibility are also reflected at the signal transduction level (Robertson et al ., ). Distinct kinome profiles were observed between G4 and S88 bees at three developmental stages (pink‐eyed, dark‐eyed and adult).…”

Systems medicine approaches for peptide array-based protein kinase profiling: progress and prospects

“…As opposed to clinical medicine, custommade array designs are much more common in this field, for instance peptide arrays containing bovine peptides have been successfully employed to identify novel drug targets to combat Jone's disease, a granulomatous affliction of the intestine that affects cattle [40][41][42]. Likewise chicken-specific [43], Syrian hamster-specific [13], and even honey bee-specific [44] platforms have been employed to generate specific insight into questions related to veterinary medicine. The results obtained in this field suggest that remarkably species-specific kinotypes appear to exist, supporting the use of custom species-specific peptide arrays [45].…”

Improving the future of honey bee breeding programs by employing recent scientific advances

“…Novel tools to identify specific molecular signatures of parasite and pathogen resistance, such as bee-specific peptide array which characterizes and correlates phosphorylated protein profiles with Varroa resistance [67 ], should be developed and incorporated into breeding efforts. Varroa resistance may be conferred not only by increasing hygienic behavior but also by increased resistance to the specific viruses it vectors [68] thus highlighting the need to identify resistance-associated QTLs.…”