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Freeze induced phase transitions in the plasma membrane of isolated protoplasts


, : Freeze induced phase transitions in the plasma membrane of isolated protoplasts. Physiologia Plantarum 69(4): 666-674

Protoplasts were enzymically isolated from 2-week-old non-acclimated rye (Secale cereale L. cv. Puma) seedlings. They were resuspended in isotonic sorbitol with different concentrations (0-10%) of dimethyl sulfoxide (DMSO). The survival of the protoplasts frozen in isotonic sorbitol solutions declined at temperatures below the freezing point with the LT50 being -8.degree. C. Addition of DMSO to the osmoticum increased survival at freezing temperatures. The optimum concentration of DMSO was 4% and lowered the LT50 to 19.degree. C. Freeze-fracture studies of the plasma membrane revealed a particulate lipid lamellae at -4.degree. C, but the first appearance of lateral phase separations, striations, and inverted cylindrical micelles (hexagonalII-type structures) occurred at -6.degree. C. At lower temperatures, -8 and -10.degree. C, the occurrence of nonbilayer structures became more common. The addition of DMSO decreased the incidence of the ultrastructural changes. With 2 or 4% DMSO, non-bilayer structures were not observed at temperatures above -10.degree. C. Instead, striations and HII-type structures were observed at -15 and 20.degree. C.

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