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Biomarkers and gene copy number variation for terpenoid traits associated with insect resistance in Sitka… Bohlmann, Joerg; Hall, Dawn; Robert, Jeanne; Keeling, Christopher; Domanski, Dominik; Quesada, Alfonso L; Jancsik, Sharon; Kuzyk, Michael; Hamberger, Britta; Borchers, Christoph Sep 13, 2011

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ORAL PRESENTATION Open AccessBiomarkers and gene copy number variation forterpenoid traits associated with insect resistancein Sitka spruce: An integrated genomic,proteomic, and biochemical analysis of (+)-3-carene biosynthesisJoerg Bohlmann1*, Dawn Hall1, Jeanne Robert1, Christopher Keeling1, Dominik Domanski2, Alfonso Lara Quesada1,Sharon Jancsik1, Michael Kuzyk2, Britta Hamberger1, Christoph Borchers3From IUFRO Tree Biotechnology Conference 2011: From Genomes to Integration and DeliveryArraial d Ajuda, Bahia, Brazil. 26 June - 2 July 2011Conifers have evolved complex chemical defenses in theform of oleoresin terpenoids to resist attack from patho-gens and herbivores. The large diversity of terpenoidmetabolites is determined by the size and compositionof the terpene synthase (TPS) gene family, and the sin-gle- and multi-product profiles of these enzymes. Themonoterpene (+)-3-carene is associated with resistanceof Sitka spruce (Picea sitchensis) to white pine weevil(Pissodes strobi). We used a combined genomic, proteo-mic and biochemical approach to analyze the (+)-3-car-ene phenotype in two contrasting Sitka sprucegenotypes. Resistant trees produced significantly higherlevels of (+)-3-carene than susceptible trees, in whichonly trace amounts were detected. Biosynthesis of (+)-3-carene is controlled, at the genome level, by a smallfamily of closely related (82-95% amino acid sequenceidentity) (+)-3-carene synthase (PsTPS-3car) genes.Transcript profiling identified one PsTPS-3car gene(PsTPS-3car1) which is expressed in both genotypes,one gene (PsTPS-3car2) expressed only in resistanttrees, and one gene (PsTPS-3car3) expressed only insusceptible trees. The PsTPS-3car2 gene was notdetected in genomic DNA of susceptible trees. Target-specific selected reaction monitoring substantiated thispattern of differential expression of members of thePsTPS-3car family on the proteome level. Kineticcharacterization of the recombinant PsTPS-3carenzymes identified differences in the activities of PsTPS-3car2 and PsTPS-3car3as a factor for the different(+)-3-carene profiles of resistant and susceptible trees.In conclusion, variation of the (+)-3-carene phenotype iscontrolled by PsTPS-3car gene copy number variation,variation of gene and protein expression, and variationof catalytic efficiencies.Author details1Michael Smith Laboratories, University of British Columbia, Canada.2University of Victoria – Genome BC Proteomics Centre, Canada. 3Universityof Victoria – Genome BC Proteomics Centre and Department ofBiochemistry, University of Victoria, Canada.Published: 13 September 2011Reference1. Hall DE, Robert JA, Keeling CI, Domanski D, Qesada AL, Jancsik S, Kuzyk M,Hamberger Br, Borchers CH, J Bohlmann: An integrated genomic,proteomic, and biochemical analysis of (+)-3-carene biosynthesis in Sitkaspruce (Picea sitchensis) genotypes which are resistant or susceptible towhite pine weevil. The Plant Journal 2011, 65:936-948, doi: 10.1111/j.1365-313X.2010.04478.x.doi:10.1186/1753-6561-5-S7-O27Cite this article as: Bohlmann et al.: Biomarkers and gene copy numbervariation for terpenoid traits associated with insect resistance in Sitkaspruce: An integrated genomic, proteomic, and biochemical analysis of(+)-3-carene biosynthesis. BMC Proceedings 2011 5(Suppl 7):O27.* Correspondence: bohlmann@msl.ubc.ca1Michael Smith Laboratories, University of British Columbia, CanadaFull list of author information is available at the end of the articleBohlmann et al. BMC Proceedings 2011, 5(Suppl 7):O27http://www.biomedcentral.com/1753-6561/5/S7/O27© 2011 Bohlmann et al; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the CreativeCommons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, andreproduction in any medium, provided the original work is properly cited.

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