biologia plantarum

International journal on Plant Life established by Bohumil Nìmec in 1959

Biologia plantarum 46:21-25, 2003 | DOI: 10.1023/A:1027316511865

Optimisation of Protoplast Production in White Lupin

A. Sinha1, A.C. Wetten1, P.D.S. Caligari1
1 Department of Agricultural Botany, School of Plant Sciences, The University of Reading, Whiteknights, Reading, United Kingdom

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.

Keywords: Lupinus albus; protoplast isolation; protoplast viability; protoplast yield
Subjects: in vitro culture, protoplast isolation; Lupinus albus; protoplast isolation, viability, yield; white lupin, protoplast production

Published: July 1, 2003  Show citation

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Sinha, A., Wetten, A.C., & Caligari, P.D.S. (2003). Optimisation of Protoplast Production in White Lupin. Biologia plantarum, 46(1), 21-25. doi: 10.1023/A:1027316511865
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