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Functional analysis of the DAT gene promoter using transient Catharanthus roseus and stable Nicotiana tabacum transformation systems

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Abstract

The Catharanthus roseus DAT gene encodes the enzyme acetyl-CoA:deacetylvindoline-4-O-acetyltransferase involved in the last step of the indole alkaloid pathway leading to vindoline. This gene is characterized by specific cell type expression in idioblasts and laticifers. To understand the specific transcriptional regulation mechanism(s) of DAT, several DAT promoter GUS constructs were cloned into pCAMBIA1305.1. Agroinfiltration of different explant types of C. roseus resulted in organ-specific accumulation of GUS, albeit at various levels. Heterologous accumulation of GUS in transgenic tobacco revealed both general and non-specific expression with the exception of a stomata-specific expression when 2.3 kb of the DAT promoter was coupled with a portion of the DAT ORF. These results suggest that in addition to the 2.3 kb upstream of the DAT transcriptional start site, additional cis-acting elements may be responsible for the specific spatial expression of DAT in vivo. Furthermore, hairy roots transformed with DAT promoter GUS constructs demonstrated GUS expression in root tissues (visualized through GUS enzyme activity), even though DAT is repressed in non-transformed roots.

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Acknowledgments

The authors gratefully acknowledge financial support from the Department of Plant Physiology, Tours University (BSt-P) and the Natural Sciences and Engineering Research Council (NSERC) of Canada (MAB). Additional financial support to ABM, from the Ministry of Higher Education in Syria is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Biology and The Biotron, The University of Western Ontario, London, ON, N6A 5B7, Canada

    Abdullah Makhzoum & Mark A. Bernards

  2. Biomolécules et Biotechnologies Végétales, Universite Francois-Rabelais, 37200, Tours, France

    Geneviève Petit-Paly & Benoit St. Pierre

Authors
  1. Abdullah Makhzoum
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  2. Geneviève Petit-Paly
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  3. Benoit St. Pierre
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  4. Mark A. Bernards
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Correspondence to Abdullah Makhzoum.

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Communicated by J. Register.

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Makhzoum, A., Petit-Paly, G., St. Pierre, B. et al. Functional analysis of the DAT gene promoter using transient Catharanthus roseus and stable Nicotiana tabacum transformation systems. Plant Cell Rep 30, 1173–1182 (2011). https://doi.org/10.1007/s00299-011-1025-y

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  • DOI: https://doi.org/10.1007/s00299-011-1025-y

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