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A comparison of methods for direct gene transfer into maize (Zea mays L.)
Authors:E. M. Southgate  M. R. Davey  J. B. Power  R. J. Westcott
Affiliation:(1) Plant Research Group, Department of Life Science, University of Nottingham, University Park, NG7 2RD Nottingham, UK;(2) Unilever Research, Colworth House, MK44 1LQ Sharnbrook, Bedford, UK
Abstract:Summary Techniques for transforming intact tissues of cereals were evaluated for their efficacy in transforming immature embryos and Type II callus of maize (Zea mays L.). The techniques used were particle bombardment, tissue electroporation, tissue electrophoresis, and silicon carbide fibers. Each method was assessed in terms of transient β-glucuronidase (GUS) expression. High levels of GUS expression were observed in A188 Type II callus using both tissue electroporation and particle bombardment, with means of 417.8 and 954.5 blue expression units (beu) per g fresh weight (FW) callus, respectively. Only particle bombardment resulted in high transient gene expression in immature embryos, with a mean transformation frequency of 34.8 b.e.u. per embryo. Very low levels of GUS expression were achieved with silicon carbide-mediated gene transfer, even when employing tissues used in the original publication (Black Mexican Sweet suspension cells). GUS expression was not obtained following tissue electrophoretic gene delivery.
Keywords:particle bombardment  tissue electroporation  tissue electrophoresis  silicon carbide transformation
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