Submitted to: PLoS One
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: April 15, 2013
Publication Date: June 6, 2013
Citation: Cornman, R.S., Lopez, D.L., Evans, J.D. 2013. Transcriptional response of honey bee larvae infected with the bacterial pathogen Paenibacillus larvae. Developmental and Comparative Immunology. DOI: 10.1371/journal.pone.0065424. Interpretive Summary: American foulbrood disease remains a significant threat to beekeeping worldwide. Honey bees can mount an immune response against the bacterium that causes this disease. This response can be enhanced through selective breeding, and also offers a general tool for understanding the bee immune system. Here we use a genomic approach to show how bees respond to bacterial infection. We found changes in both predicted immune genes and proteins that are involved in development and detoxification. We discuss the possible roles of these proteins in bee health and disease resistance, and suggest future targets for bee breeding and immunity.
Technical Abstract: American foulbrood disease of honey bees is caused by the bacterium Paenibacillus larvae. Infection occurs per os in larvae and systemic infection requires a breaching of the host peritrophic matrix and midgut epithelium. Genetic variation exists for both bacterial virulence and host resistance, and a general immunity is achieved by larvae as they age, the basis of which has not been identified. To quickly identify a pool of candidate genes responsive to P. larvae infection, we sequenced transcripts from larvae at 72 hours post emergence that had been inoculated with P. larvae and compared expression levels to a control cohort. We identified 75 genes with significantly higher expression and six genes with significantly lower expression. In addition to several antimicrobial peptides, two genes encoding peritrophic-matrix domains (Pfam 01607) were also up-regulated. Extracellular matrix proteins, proteases/protease inhibitors, and members of the Osiris gene family were prevalent among differentially regulated genes. However, analysis of Drosophila homologs of differentially expressed genes revealed spatial and temporal patterns consistent with developmental asynchrony as a likely confounder of our results. We therefore used qPCR to measure the consistency of gene expression changes for a subset of differentially expressed genes. A replicate experiment sampled at both 48 and 72 hours allowed further discrimination of genes responsive likely to be involved in host response. The consistently responsive genes in our test set included a hymenopteran-specific protein tyrosine kinase, a hymenopteran specific serine endopeptidase, a cytochrome P450 (CYP9Q1), and a homolog of trynity, a zona pellucida (ZP) domain protein. Of the known honey bee antimicrobial peptides, apidaecin was responsive at both time-points studied whereas hymenoptaecin was more consistent in its level of change between biological replicates and had the greatest increase in expression by RNA-seq analysis.