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Characterisation of PtMYB1, an R2R3-MYB from pine xylem

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Abstract

A cDNA encoding a member of the R2R3-MYB family of transcription factors was cloned from a library constructed from differentiating Pinus taeda (loblolly pine) xylem RNA. This MYB family member, Pinus taeda MYB1 (PtMYB1), was most abundantly expressed in differentiating xylem, as assessed by both ribonuclease protection assays, and by northern blot analysis with poly(A)-enriched RNA. Similar to other plant R2R3-MYB family members, recombinant PtMYB1 protein was able to bind to AC elements in electrophoretic mobility shift assays (EMSAs). AC elements are DNA motifs rich in adenosine and cytosine that have been implicated in the xylem-localised regulation of genes encoding lignin biosynthetic enzymes. PtMYB1 not only bound to AC elements, but was also able to induce AC-element-dependent shifts in the electrophoretic mobility of a plant promoter that contains three AC elements, the minimal PHENYLALANINE AMMONIA-LYASE 2 (PAL2) promoter from Phaseolus vulgaris. Transcriptional activation assays conducted using yeast showed that PtMYB1 also activated transcription, and that it did so in an AC-element-dependent fashion. PtMYB1 also activated transcription from the minimal PAL2 promoter in plant cells in an AC-element-dependent fashion, as revealed by transient transcriptional activation assays with microprojectile-bombarded tobacco NT-1 cells. Taken together, these finding are consistent with the hypothesis that PtMYB1 may regulate transcription from cis-acting AC elements in pine xylem.

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Authors and Affiliations

  1. Department of Plant Sciences, University of Oxford, South Parks Road, Oxford, OX1 3RB, UK

    Astrid Patzlaff, Lisa J. Newman, Christian Dubos, Stephanie McInnis & Malcolm M. Campbell

  2. Pioneer Hi-Bred International, Inc., 7250 NW 62nd Avenue, PO Box 552, Johnston, IA, 50131-0552, USA

    Lisa J. Newman

  3. Forest Biotechnology Laboratory, Department of Forestry, North Carolina State University, 2500 Partners II, 840 Main Campus Drive, Centennial Campus, Box 7247, Raleigh, NC, 27695, USA

    Ross W. Whetten & Ronald R. Sederoff

  4. Department of Cell & Developmental Biology, John Innes Centre, Norwich Research Park, Colney, Norwich, Norfolk, NR4 7UH, UK

    Caroline Smith & Michael W. Bevan

  5. School of Biological Sciences, University of Bristol, Woodland Road, Bristol, BS8 1UG, UK

    Stephanie McInnis

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  1. Astrid Patzlaff
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  2. Lisa J. Newman
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Correspondence to Malcolm M. Campbell.

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these authors contributed equally to the work

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Patzlaff, A., Newman, L.J., Dubos, C. et al. Characterisation of PtMYB1, an R2R3-MYB from pine xylem. Plant Mol Biol 53, 597–608 (2003). https://doi.org/10.1023/B:PLAN.0000019066.07933.d6

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