Abstract
Publisher Summary This chapter discusses characterization of tonoplast enzyme activities and their transport. One artifact of particular importance in relation to the separation of authentic tonoplasts is vacuoplasts, such as vacuoles that are surrounded by the plasmalemma in addition to the tonoplast and contains very few cytoplasmic remnants among the membranes. They appear in some preparations of vacuoles that are obtained by lysis of protoplasts and can be detected by fluorescein diacetate (FDA) staining. Authentic tonoplast has been obtained from these vacuoles and their properties are compared to those of microsomal fractions separated on density gradients. Such comparison resulted in the identification of a tonoplast-enriched light microsomal fraction from microsomes. This fraction is characterized by the presence of an NO 3 - inhibited and vanadate-insensitive Mg 2+ -ATPase, which serves as a marker enzyme for the tonoplast.
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