Targeted proteomics using selected reaction monitoring reveals the induction of specific terpene synthases in a multi-level study of methyl jasmonate-treated Norway spruce (Picea abies)

Authors

  • Katherine G. Zulak,

    1. Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada
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    • These authors contributed equally to this work.

  • Dustin N. Lippert,

    1. Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada
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    • These authors contributed equally to this work.

    • Present address: Manitoba Centre for Proteomics and Systems Biology, University of Manitoba, Winnipeg, MB, Canada.

  • Michael A. Kuzyk,

    1. University of Victoria – Genome BC Proteomics Centre, Victoria, BC, Canada
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  • Dominik Domanski,

    1. University of Victoria – Genome BC Proteomics Centre, Victoria, BC, Canada
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  • Tina Chou,

    1. Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada
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  • Christoph H. Borchers,

    1. University of Victoria – Genome BC Proteomics Centre, Victoria, BC, Canada
    2. Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada
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  • Jörg Bohlmann

    Corresponding author
    1. Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada
      *For correspondence (fax +01 604 822 2114; e-mail bohlmann@msl.ubc.ca).
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*For correspondence (fax +01 604 822 2114; e-mail bohlmann@msl.ubc.ca).

Summary

Induction of terpene synthase (TPS) gene expression and enzyme activity is known to occur in response to various chemical and biological stimuli in several species of spruce (genus Picea). However, high sequence identity between TPS family members has made it difficult to determine the induction patterns of individual TPS at the protein and transcript levels and whether specific TPS enzymes respond differentially to treatment. In the present study we used a multi-level approach to measure the induction and activity of TPS enzymes in protein extracts of Norway spruce (Picea abies) bark tissue following treatment with methyl jasmonate (MeJA). Measurements were made on the transcript, protein, enzyme activity and metabolite levels. Using a relatively new proteomics application, selective reaction monitoring (SRM), it was possible to differentiate and quantitatively measure the abundance of several known TPS proteins and three 1-deoxy-d-xylulose 5-phosphate synthase (DXS) isoforms in Norway spruce. Protein levels of individual TPS and DXS enzymes were differentially induced upon MeJA treatment and good correlation was generally observed between induction of transcripts, proteins, and enzyme activities. Most of the mono- and diterpenoid metabolites accumulated with similar temporal patterns of induction as part of the coordinated multi-compound chemical defense response. Protein and enzyme activity levels of the monoTPS (+)-3-carene synthase and the corresponding accumulation of (+)-3-carene was induced to a higher fold change than any other TPS or metabolite measured, indicating an important role in the induced terpenoid defense response in Norway spruce.

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