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Optimisation of protoplast production in white lupin

Sinha, A., Wetten, A. C. and Caligari, P. D. S. (2003) Optimisation of protoplast production in white lupin. Biologia Plantarum, 47 (1). pp. 21-25. ISSN 0006-3134

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To link to this item DOI: 10.1023/A:1027316511865

Abstract/Summary

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.

Item Type:Article
Refereed:Yes
Divisions:Faculty of Life Sciences > School of Agriculture, Policy and Development
ID Code:9177
Uncontrolled Keywords:Lupinus albus, protoplast isolation, protoplast viability, protoplast, yield, ANGUSTIFOLIUS CV KUBESA, TRIFOLIUM-PRATENSE L, PLANT-REGENERATION, MESOPHYLL PROTOPLASTS, LEAF PROTOPLASTS, CULTURE, LEAVES, CLOVER, CELLS, MEDICAGO

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