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This article is part of the supplement: IUFRO Tree Biotechnology Conference 2011: From Genomes to Integration and Delivery

Open Access Oral presentation

Biomarkers and gene copy number variation for terpenoid traits associated with insect resistance in Sitka spruce: An integrated genomic, proteomic, and biochemical analysis of (+)-3-carene biosynthesis

Joerg Bohlmann1*, Dawn Hall1, Jeanne Robert1, Christopher Keeling1, Dominik Domanski2, Alfonso Lara Quesada1, Sharon Jancsik1, Michael Kuzyk2, Britta Hamberger1 and Christoph Borchers3

Author Affiliations

1 Michael Smith Laboratories, University of British Columbia, Canada

2 University of Victoria – Genome BC Proteomics Centre, Canada

3 University of Victoria – Genome BC Proteomics Centre and Department of Biochemistry, University of Victoria, Canada

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BMC Proceedings 2011, 5(Suppl 7):O27  doi:10.1186/1753-6561-5-S7-O27


The electronic version of this article is the complete one and can be found online at: http://www.biomedcentral.com/1753-6561/5/S7/O27


Published:13 September 2011

© 2011 Bohlmann et al; licensee BioMed Central Ltd.

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.

Oral presentation

Conifers have evolved complex chemical defenses in the form of oleoresin terpenoids to resist attack from pathogens and herbivores. The large diversity of terpenoid metabolites is determined by the size and composition of the terpene synthase (TPS) gene family, and the single- and multi-product profiles of these enzymes. The monoterpene (+)-3-carene is associated with resistance of Sitka spruce (Picea sitchensis) to white pine weevil (Pissodes strobi). We used a combined genomic, proteomic and biochemical approach to analyze the (+)-3-carene phenotype in two contrasting Sitka spruce genotypes. Resistant trees produced significantly higher levels of (+)-3-carene than susceptible trees, in which only trace amounts were detected. Biosynthesis of (+)-3-carene is controlled, at the genome level, by a small family of closely related (82-95% amino acid sequence identity) (+)-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 resistant trees, and one gene (PsTPS-3car3) expressed only in susceptible trees. The PsTPS-3car2 gene was not detected in genomic DNA of susceptible trees. Target-specific selected reaction monitoring substantiated this pattern of differential expression of members of the PsTPS-3car family on the proteome level. Kinetic characterization of the recombinant PsTPS-3car enzymes 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 is controlled by PsTPS-3car gene copy number variation, variation of gene and protein expression, and variation of catalytic efficiencies.

References

  1. 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 Sitka spruce (Picea sitchensis) genotypes which are resistant or susceptible to white pine weevil.

    The Plant Journal 2011, 65:936-948.

    doi: 10.1111/j.1365-313X.2010.04478.x

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