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TheCaMV 35S promoter is highly active on floral organs and pollen of transgenic strawberry plants

  • Genetic Transformation and Hybridization
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Abstract

We have evaluated the expression of the reporter β-glucuronidase (GUS) gene driven by the cauliflower mosaic virus 35S (CaMV 35S) promoter in flowers and pollen from 14 independent transgenic strawberry lines. Of the 14 lines evaluated, 13 (92.8%) showed GUS activity—as estimated by the histochemical GUS assay—in some floral organs, with expression being most common in the flower stem, sepals, petals, ovary and stigma. Ten of these thirteen transgenic lines (77%) showed GUS activity in pollen, although the percentages of positive pollen per flower varied greatly among the different lines. A study of the GUS expression during pollen maturation showed that the (CaMV 35S) promoter showed low expression in pollen from flower buds before anthesis but was activated in mature pollen following anther dehiscence. The percentages of pollen grains that showed GUS activity ranged from 2.1% to 46.3%. These percentages were similar or even higher when mature pollen was stored dry at room temperature for 2 weeks. After 5 weeks of storage, the percentages of GUS-positive pollen decreased in two of the six lines analysed but remained at similar values in the other four lines. GUS activity was also measured in protein extracts of mature pollen by means of the fluorometric GUS assay, with the values obtained ranging from 3.8 μmol MU mg protein−1 h−1 to 0.26 μmol MU mg protein−1 h−1. Contrary to the generally held view that the CaMV 35S promoter is virtually silent in pollen, we conclude that it is highly expressed in transgenic strawberry pollen.

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Abbreviations

CaMV 35S :

Cauliflower mosaic virus promoter

GUS :

β-Glucuronidase (EC 3.2.1.31)

MU:

4-Methyl umbelliferone

nos :

Nopaline synthase promoter

nptII :

Neomycin phosphotransferase

X-Gluc:

5-Bromo-4-chloro-3-indolyl-β-d-glucuronic acid

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Acknowledgements

This work was supported by the Ministerio de Ciencia y Tecnología, Spain and EU FEDER funds (grant nos. IFD97/0843C0504 and AGL 2001-1897).

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Authors and Affiliations

  1. Departamento de Biología Vegetal, Universidad de Málaga, 29071, Málaga, Spain

    M. Cordero de Mesa, N. Santiago-Doménech, F. Pliego-Alfaro, M. A. Quesada & J. A. Mercado

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  1. M. Cordero de Mesa
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  2. N. Santiago-Doménech
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  3. F. Pliego-Alfaro
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  4. M. A. Quesada
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  5. J. A. Mercado
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Correspondence to J. A. Mercado.

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Communicated by L. Peña

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de Mesa, M.C., Santiago-Doménech, N., Pliego-Alfaro, F. et al. TheCaMV 35S promoter is highly active on floral organs and pollen of transgenic strawberry plants. Plant Cell Rep 23, 32–38 (2004). https://doi.org/10.1007/s00299-004-0776-0

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  • DOI: https://doi.org/10.1007/s00299-004-0776-0

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