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Sequence analysis and functional characterization of the promoter of the Picea glauca Cinnamyl Alcohol Dehydrogenase gene in transgenic white spruce plants

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Abstract

The enzyme Cinnamyl Alcohol Dehydrogenase (CAD) catalyses the last step of lignin monomer synthesis, and is considered as a molecular marker of cell wall lignification in different plants species. Here, we report the isolation and analysis of 5′ flanking genomic DNA regions upstream to the CAD gene, from two conifers, i.e. white spruce (Picea glauca (Moench) Voss) and loblolly pine (Pinus taeda L.). Sequence comparisons with available CAD gene promoters from angiosperms highlighted the conservation of cis-elements matching MYB, WRKY and bHLH binding sites. Functional characterization of the P. glauca CAD promoter used P. glauca seedlings stably transformed with a DNA fragment of 1,163 base pairs (PgCAD) fused to the β-glucuronidase (GUS) gene. Histochemical observations of different vegetative organs of the transgenic trees showed that this sequence was sufficient to drive GUS expression in lignifying tissues, and more specifically in differentiating xylem cells. Quantitative RT-PCR experiments also indicated that the native CAD gene was preferentially expressed in differentiating xylem both in stems and roots. In addition, GUS expression driven by the PgCAD promoter was wound-inducible which was consistent with the accumulation of CAD mRNA in response to jasmonate application and mechanical wounding. The spruce CAD promoter represents a valuable tool for research and biotechnology applications related to xylem and wood.

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Acknowledgments

We are grateful to Françoise Pelletier and Laurence Tremblay for excellent assistance in tissue culture and growth of the plants in greenhouse. We acknowledge Denis Lachance for helpful advice for the jasmonate application methods. This research was supported by funding from Genome Canada and Génome Québec to JM and AS for the ARBOREA project.

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  1. Frank Bedon

    Present address: UMR Biodiversité Gènes Communauté 1202, INRA, Equipe de Génétique, 69 route d’Arcachon, 33612, Cestas cedex, France

Authors and Affiliations

  1. Centre d’Étude de la Forêt, Université Laval, Quebec, QC, G1V 0A6, Canada

    Frank Bedon & John MacKay

  2. UMR UPS/CNRS 5546, Pôle de Biotechnologies Végétales, 24 chemin de Borde Rouge, BP42617, Auzeville Tolosane, 31326, Castanet Tolosan, France

    Frank Bedon & Jacqueline Grima-Pettenati

  3. Natural Resources Canada, Canadian Forest Service, Laurentian Forestry Centre, Quebec, QC, G1V 4C7, Canada

    Caroline Levasseur & Armand Séguin

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  1. Frank Bedon
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  2. Caroline Levasseur
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  3. Jacqueline Grima-Pettenati
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  4. Armand Séguin
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Correspondence to John MacKay.

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Communicated by L. Jouanin.

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Bedon, F., Levasseur, C., Grima-Pettenati, J. et al. Sequence analysis and functional characterization of the promoter of the Picea glauca Cinnamyl Alcohol Dehydrogenase gene in transgenic white spruce plants. Plant Cell Rep 28, 787–800 (2009). https://doi.org/10.1007/s00299-009-0688-0

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