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Effect of Heat Shock Treatment on Hordeum Vulgare Protoplast Transformation Mediated by Polyethylene Glycol

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Biologia Plantarum

Abstract

The stable transformation of barley protoplasts has been observed in cv. Dissa by PEG-mediated pGL2 plasmid DNA uptake containing hygromycin phosphotransferase (hph) gene under the control of 35S transcript of CaMV. Hygromycin resistant colonies were obtained, when protoplasts were incubated with hygromycin B (25 μg cm−3) at 3 – 4 cell stage. Heat shock treatment, prior to polyethylene glycol (PEG) treatment, markedly increased the protoplast transformation frequency. The presence of active hph gene product, and integration of hph gene was confirmed by hygromycin assay and Southern blotting, respectively. Analysis indicated that rearrangement had possibly occurred during the integration of plasmid DNA into genomic DNA of barley callus. These transformed calli showed no morphogenic response onto the regeneration medium.

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Authors and Affiliations

  1. Plant Genetic Manipulation Group, Department of Life Science, University of Nottingham, University Park, Nottingham, NG7 2RD, UK

    V.K. Tiwari, J. Zhang, T.J. Golds, E.C. Cocking, M.R. Davey & J.B. Power

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  1. V.K. Tiwari
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  2. J. Zhang
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  3. T.J. Golds
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  6. J.B. Power
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Tiwari, V., Zhang, J., Golds, T. et al. Effect of Heat Shock Treatment on Hordeum Vulgare Protoplast Transformation Mediated by Polyethylene Glycol. Biologia Plantarum 44, 25–31 (2001). https://doi.org/10.1023/A:1017957902042

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