title: Transcriptional response of honey bee larvae infected with the bacterial pathogen Paenibacillus larvae.
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 inoculated with P. larvae at 12 hours post-emergence and incubated for 72 hours, 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 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 post infection allowed further discrimination of genes 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 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.
authors.affiliation: Bee Research Laboratory, Agricultural Research Service of the United States Department of Agriculture, Beltsville, Maryland, United States of America. scott.cornman@gmail.com
authors.forename: Robert Scott
authors.initials: RS
authors.lastname: Cornman
authors.forename: Dawn
authors.initials: D
authors.lastname: Lopez
authors.forename: Jay D
authors.initials: JD
authors.lastname: Evans
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000276:immunology; D001516:Bees / Q000235:genetics* / Q000276:immunology / Q000382:microbiology; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005786:Gene Expression Regulation; D019476:Insect Proteins / Q000235:genetics / Q000276:immunology; D007814:Larva / Q000235:genetics* / Q000276:immunology / Q000382:microbiology; D008969:Molecular Sequence Data; D056507:Paenibacillus / Q000254:growth & development* / Q000472:pathogenicity; D017434:Protein Structure, Tertiary; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics* / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D015533:Transcriptional Activation
chemical_list: D023181:Antimicrobial Cationic Peptides; D029721:Drosophila Proteins; D019476:Insect Proteins; D012333:RNA, Messenger; C080586:hymenoptaecin protein, Apis mellifera; C061361:apidaecin; D011505:Protein-Tyrosine Kinases