Abstract
The influence, was investigated, of abiotic parameters on the isolation of protoplasts from in vitro seedling cotyledons of white lupin. The protoplasts were found to be competent in withstanding a wide range of osmotic potentials of the enzyme medium, however, −2.25 MPa (0.5 M mannitol), resulted in the highest yield of protoplasts. The pH of the isolation medium also had a profound effect on protoplast production. Vacuum infiltration of the enzyme solution into the cotyledon tissue resulted in a progressive drop in the yield of protoplasts. The speed and duration of orbital agitation of the cotyledon tissue played a significant role in the release of protoplasts and a two step (stationary-gyratory) regime was found to be better than the gyratory-only system.
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Sinha, A., Wetten, A. & Caligari, P. Optimisation of Protoplast Production in White Lupin. Biologia Plantarum 47, 21–25 (2003). https://doi.org/10.1023/A:1027316511865
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DOI: https://doi.org/10.1023/A:1027316511865