Abstract
A novel approach is described for the culture of protoplasts isolated from cell suspensions of the Japonica rice ( Oryza sativa L.) cultivar Taipei 309 involving supplementation with an oxygenated perfluorochemical (PFC) liquid, perfluorodecalin. Protoplasts cultured in the presence of oxygenated perfluorodecalin exhibited a 5-fold increase in mean plating efficiency ( P < 0.01) compared to controls. This beneficial effect was sustained throughout culture with a 12% increase ( P < 0.01) in shoot regeneration efficiency, leading to intact plants with morphometric characteristics similar to those of seed-derived individuals. Further refinement of the system, involving 24-well culture plates, similarly revealed a 7.5-fold increase in plating efficiency in response to PFC supplementation ( P < 0.01). The enhanced growth of protoplast-derived colonies was confirmed by image analysis assessments of the cultures. Overall, these results demonstrate a beneficial effect of supplementing rice protoplast culture systems with oxygenated perfluorochemical liquid which will be of broader value in plant biotechnology.
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