Visualization of vacuoplasts in isolated vacuole preparations from mesophyll protoplasts of periwinkle [Catharanthus roseus (L.) G. Don

Abstract

A procedure was developed for the rapid detection of vacuoplasts in vacuole preparations isolated from mesophyll protoplasts of Catharanthus roseus (L.) G. Don (periwinkle). The procedure relies on the staining of surface carbohydrates on the plasma membrane surrounding vacuoplasts with fluorescein-labeled lectins. When isolated under conditions of constant osmotic strength, approximately 15-20% of the vacuoles isolated showed surface labeling with FITC-agglutinin from Abrus precatorius. Isolation of vacuoles after an initial osmotic shock showed much lower (<5%) surface labeling. This lower level of surface labeling correlated well with a lower level of other non-vacuolar marker enzyme activities. A thin layer of cytoplasm was visible in a small number of these stained structures, indicating that they were vacuoplasts.