The influence of cold acclimation on the behavior of the plasma membrane following osmotic contraction of isolated protoplasts

Abstract

Osmotic contraction of protoplasts isolated from cold acclimated leaves ofSecale cereale L. cv. Puma results in the formation of exocytotic extrusions of the plasma membrane. Numerous knobs or polyps were observed on the surface of the protoplasts with scanning electron microscopy. In thin sections, the extrusions were bounded by the plasma membrane with a densely osmiophilic interior. Cross-fracturing of the extrusions revealed aparticulate bodies within, a further indication that the interior of the extrusions was predominantly lipid material. Freeze-fracture of the plasma membrane suggests a possible source of this lipid material. Following osmotic contraction, the particle density on the plasma membrane protoplasmic face (PFp) increased, being reflected in both a substantial increase in paracrystalline arrays and an increase in the particle density in non-crystalline regions. This increase in particle density indicates that lipid material is preferentially lost from the plasma membrane during contraction. The density on the exoplasmic face (EFp) did not change. Together, these findings suggest that during hypertonic contraction of acclimated protoplasts, lipid material is preferentially subducted from the plasma membrane and sequestered into lipid bodies (the osmiophilic regions). The formation of lipid bodies and extrusions was readily reversible. Following osmotic expansion of acclimated protoplasts, the extrusions were retracted back into the plane of the plasma membrane.