Perfluorochemicals and plant biotechnology: an improved protocol for protoplast culture and plant regeneration in rice ( Oryza sativa L.)

• Published in: Journal of biotechnology Vol. 50; no. 1; pp. 47 - 54
• Main Authors: Wardrop, J., Lowe, K.C., Power, J.B., Davey, M.R.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 01.01.1996; Amsterdam Elsevier; New York, NY
• Subjects: Biological and medical sciences; biotechnologie vegetale; Biotechnology; biotecnologia vegetal; cell culture; chemicals; cultivo de celulas; culture de cellule; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; in vitro regeneration; Methods. Procedures. Technologies; Oryza sativa; Orzya sativa L; Perfluorochemical; plant biotechnology; Plant cells and fungal cells; Plant regeneration; productos quimicos; produit chimique; Protoplast preparation, cell fusion and regeneration; protoplaste; protoplastos; Protoplasts; regeneracion in vitro; regeneration in vitro; BAP; Kin; Perfluorochemical; 2,4-D; MS; Protoplasts; Orzya sativa L; Plant regeneration; PDA; PFC; NAA; Culture medium; Cell culture; Monocotyledones; Vegetals; Organic perhalocompound; Optimization; Cereal crop; Oryza sativa; Regeneration; Gramineae; Protoplast; Angiospermae; Spermatophyta; Oxygenation
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/0168-1656(96)01548-9

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.