title: Functional genomics of mountain pine beetle (Dendroctonus ponderosae) midguts and fat bodies.
abstract: BACKGROUND
  The mountain pine beetle (Dendroctonus ponderosae) is a significant coniferous forest pest in western North America. It relies on aggregation pheromones to colonize hosts. Its three major pheromone components, trans-verbenol, exo-brevicomin, and frontalin, are thought to arise via different metabolic pathways, but the enzymes involved have not been identified or characterized. We produced ESTs from male and female midguts and associated fat bodies and used custom oligonucleotide microarrays to study gene expression patterns and thereby made preliminary identification of pheromone-biosynthetic genes.
  
  
  RESULTS
  Clones from two un-normalized cDNA libraries were directionally sequenced from the 5' end to yield 11,775 ESTs following sequence cleansing. The average read length was 550 nt. The ESTs clustered into 1,201 contigs and 2,833 singlets (4,034 tentative unique genes). The ESTs are broadly distributed among GO functional groups, suggesting they reflect a broad spectrum of the transcriptome. Among the most represented genes are representatives of sugar-digesting enzymes and members of an apparently Scolytid-specific gene family of unknown function. Custom NimbleGen 4-plex arrays representing the 4,034 tentative unique genes were queried with RNA from eleven different biological states representing larvae, pupae, and midguts and associated fat bodies of unfed or fed adults. Quantitative (Real-Time) RT-PCR (qRT-PCR) experiments confirmed that the microarray data accurately reflect expression levels in the different samples. Candidate genes encoding enzymes involved in terminal steps of biosynthetic pathways for exo-brevicomin and frontalin were tentatively identified.
  
  
  CONCLUSIONS
  These EST and microarray data are the first publicly-available functional genomics resources for this devastating forestry pest.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Nevada, Reno, Reno, NV 89557, USA.
authors.forename: Tidiane
authors.initials: T
authors.lastname: Aw
authors.forename: Karen
authors.initials: K
authors.lastname: Schlauch
authors.forename: Christopher I
authors.initials: CI
authors.lastname: Keeling
authors.forename: Sharon
authors.initials: S
authors.lastname: Young
authors.forename: Jeremy C
authors.initials: JC
authors.lastname: Bearfield
authors.forename: Gary J
authors.initials: GJ
authors.lastname: Blomquist
authors.forename: Claus
authors.initials: C
authors.lastname: Tittiger
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001517:Coleoptera / Q000737:chemistry / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D020224:Expressed Sequence Tags; D005216:Fat Body / Q000378:metabolism; D005260:Female; D041981:Gastrointestinal Tract / Q000378:metabolism; D049750:Genome, Insect; D023281:Genomics; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D005810:Multigene Family; D010675:Pheromones / Q000096:biosynthesis; D016415:Sequence Alignment
chemical_list: D019476:Insect Proteins; D010675:Pheromones