Co-Authors:
Shneyour, Y., Department of Plant Genetics and Breeding, Agricultural Research Organization, The Volcani Center, Bet Dagan 50-250, Israel
Zelcer, A., Department of Plant Genetics and Breeding, Agricultural Research Organization, The Volcani Center, Bet Dagan 50-250, Israel
Izhar, S., Department of Plant Genetics and Breeding, Agricultural Research Organization, The Volcani Center, Bet Dagan 50-250, Israel
Beckmann, J.S., Department of Plant Genetics and Breeding, Agricultural Research Organization, The Volcani Center, Bet Dagan 50-250, Israel
Abstract:
A convenient procedure for the efficient plating of cells and protoplasts at low densities (as low as 200/ml) was developed. Feeder cells from exponentially growing Petunia suspension cell cultures are embedded in solid medium and overlaid with a cellophane membrane on which the nursed cells are plated. This method allows for the rapid transfer of cells from one set of experimental conditions to another, without the need to wash the cells. Being transparent, the membrane allows for an easy macro- as well as microscopic follow-up of both feeder and nursed cells. The method was successfully utilized for the isolation and rescue of Petunia hybrida cells resistant to 6-aza-uridine (6-aza-U), methotrexate (MTX) or the aminoglycoside G-418. Finally, the possibility of utilizing feeders from distant species has been demonstrated for tobacco mesophyll protoplasts using petunia feeder cells, suggesting that this technique could also be useful in other plant cell culture systems. © 1984.
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