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Isolation, Purification, Scanning Electron Microscopy And Bacterial Dna Uptake Of Plant Protoplasts, Bronwyn G. Hughes
Isolation, Purification, Scanning Electron Microscopy And Bacterial Dna Uptake Of Plant Protoplasts, Bronwyn G. Hughes
Theses and Dissertations
Protoplasts were isolated from tobacco and barley leaves in sucrose or mannitol using commercially available cellulases and macerozymes. Barley growth and protoplast isolation and purification conditions were optimized so that protoplasts were obtained in high yields free of unwanted debris and organelles. A technique for processing barley and tobacco protoplasts for examination by scanning electron microscopy was developed in which protoplasts seem to have maintained their structural integrity. Barley and tobacco protoplasts took up 3H-B. subtilis DNA, 125I-B. subtilis DNA or 125I-M. luteus DNA as a linear function of time (0-6 hr) and DNA concentration (0-200 μg/ml). Up to 16 …