Optimization of protoplast production in white lupin

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