Transgene Expression in the Vegetative Tissues of Apple Driven by the Vascular-specific rolC and CoYMV Promoters |
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Authors: | Gittins John R. Pellny Till K. Biricolti Stefano Hiles Elizabeth R. Passey Andrew J. James David J. |
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Affiliation: | (1) Horticulture Research International, East Malling, West Malling, Kent, ME19 6BJ, UK;(2) Present address: Laboratory of Plant Molecular Biology, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawiskiego 5A, 02-106 Warsaw, Poland;(3) Present address: Crop Performance and Improvement Division, Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, UK;(4) Present address: Dipartimento di Ortoflorofrutticoltura, Università degli Studi di Firenze, 50019 Firenze, Italy |
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Abstract: | The ability of the heterologous promoters, rolCP and CoYMVP, to drive expression of the gusA reporter gene in the vegetative tissues of apple (Malus pumila Mill.) has been studied using transgenic plants produced by Agrobacterium-mediated transformation. Replicate plants of each transgenic clone were propagated in soil to a uniform size and samples of leaf, petiole, stem, and root were taken for the measurement of -glucuronidase (GUS) activity by fluorometric assay. The levels of expression were compared with those in tissues of a representative clone containing the CaMV 35S promoter. These quantitative GUS data were related to the copy number of transgene loci assessed by Southern blotting. The CoYMV promoter was slightly more active than the rolC promoter, although both expressed gusA at a lower level than the CaMV 35S promoter. In clones containing the rolC promoter with multiple transgene loci, expression values were generally among the highest or lowest in the range. The precise location of GUS activity in each tissue was identified by staining of whole leaves and tissue sections with a chromogenic substrate. This analysis demonstrated that with both the rolC and CoYMV promoters the reporter gene activity was primarily localised to vascular tissues, particularly the phloem. Our results indicate that both promoters would be suitable to drive the expression of transgenes to combat pests and diseases of apple that are dependent on interaction with the phloem. |
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Keywords: | apple CoYMV gusA expression promoters rolC vascular-specific |
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