Stable transformation of barley via PEG-induced direct DNA uptake into protoplasts.
Protoplasts isolated from a barley cell suspension (cv Dissa) were transformed with plasmid DNA containing the neomycinphosphotransferase II (NPT) and β-glucuronidase (GUS) genes, using polyethyleneglycol (PEG) to induce DNA uptake. Transformed microcalli were selected in media containing G418 sulphate. NPT activity was detected in all antibiotic-resistant cell lines, but not all NPT-positive cell lines had GUS activity. Southern analysis confirmed the presence of sequences homologous to the APT and GUS genes in DNA of G418-resistant callus.
Institute for General Botany (AMP II), University of Hamburg, Ohnhorststrasse 18, 52, Hamburg, FRG.
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Lazzeri
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Brettschneider
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Lührs
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Lörz