Evaluation of two approaches to the quantitative histochemical localization of sucrose-P synthase in leaves.

Two methods for determining the quantitative localization of sucrose-P synthase in plant tissues were evaluated. The single-cell method (rapid freezing, freeze-drying, microdissection, micro-analysis) was validated in several ways, including comparative biochemistry, comparative histochemistry, and kinetics. In contrast, bulk isolation of cells by protoplast-forming methods resulted in loss of sucrose-P synthase activity. This latter approach is widely used and, as far as we are aware, can be successfully used for measurement of other enzymes. Thus, our observations form the basis for a specific caution against the use of protoplast-forming methods in an assay protocol for sucrose-P synthase.