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Functional genomics of mountain pine beetle (Dendroctonus ponderosae) midguts and fat bodies

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 differ...

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Autores principales: Aw, Tidiane, Schlauch, Karen, Keeling, Christopher I, Young, Sharon, Bearfield, Jeremy C, Blomquist, Gary J, Tittiger, Claus
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2858752/
https://www.ncbi.nlm.nih.gov/pubmed/20353591
http://dx.doi.org/10.1186/1471-2164-11-215
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author Aw, Tidiane
Schlauch, Karen
Keeling, Christopher I
Young, Sharon
Bearfield, Jeremy C
Blomquist, Gary J
Tittiger, Claus
author_facet Aw, Tidiane
Schlauch, Karen
Keeling, Christopher I
Young, Sharon
Bearfield, Jeremy C
Blomquist, Gary J
Tittiger, Claus
author_sort Aw, Tidiane
collection PubMed
description 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.
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spelling pubmed-28587522010-04-23 Functional genomics of mountain pine beetle (Dendroctonus ponderosae) midguts and fat bodies Aw, Tidiane Schlauch, Karen Keeling, Christopher I Young, Sharon Bearfield, Jeremy C Blomquist, Gary J Tittiger, Claus BMC Genomics Research Article 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. BioMed Central 2010-03-30 /pmc/articles/PMC2858752/ /pubmed/20353591 http://dx.doi.org/10.1186/1471-2164-11-215 Text en Copyright ©2010 Aw et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Aw, Tidiane
Schlauch, Karen
Keeling, Christopher I
Young, Sharon
Bearfield, Jeremy C
Blomquist, Gary J
Tittiger, Claus
Functional genomics of mountain pine beetle (Dendroctonus ponderosae) midguts and fat bodies
title Functional genomics of mountain pine beetle (Dendroctonus ponderosae) midguts and fat bodies
title_full Functional genomics of mountain pine beetle (Dendroctonus ponderosae) midguts and fat bodies
title_fullStr Functional genomics of mountain pine beetle (Dendroctonus ponderosae) midguts and fat bodies
title_full_unstemmed Functional genomics of mountain pine beetle (Dendroctonus ponderosae) midguts and fat bodies
title_short Functional genomics of mountain pine beetle (Dendroctonus ponderosae) midguts and fat bodies
title_sort functional genomics of mountain pine beetle (dendroctonus ponderosae) midguts and fat bodies
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2858752/
https://www.ncbi.nlm.nih.gov/pubmed/20353591
http://dx.doi.org/10.1186/1471-2164-11-215
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