Molecular characterization and functional analysis of sucrose-cleaving enzymes in carrot (Daucus carota L.).

The amount of carbon transported into storage organs of crop plants to a large degree determines crop yield. The role of sucrose-cleaving enzymes in this process is not clear and it is the main goal of our work to tackle this question. Sucrose cleavage is catalysed either by invertase or sucrose synthase both of which exist in several isoforms with different subcellular locations. Carrot (Daucus carota L.) contains three major isoenzymes of acid invertase, which either accumulate as soluble polypeptides in the vacuole (isoenzymes I and II) or are ionically bound to the cell wall. Carrot sucrose synthase is thought to be a cytoplasmic enzyme encoded by two genes. cDNA clones have been isolated and characterized for cell wall invertase, for isoenzymes I and II of vacuolar invertase, and for sucrose synthase. Gene-specific fragments of these clones were used to determine the steady-state levels of transcripts in the prominent sink and source organs of developing carrot plants. The expression patterns of each gene were different and were organ- and development-specific. Developing tap roots contained only transcripts for isoenzyme II of vacuolar invertase and sucrose synthase. The source/sink balance of these plants was manipulated and only the expression of these two genes was markedly altered, indicating their importance in sucrose partitioning. Based on these results, a model is proposed for sucrose partitioning in carrot plants with developing tap roots in which sucrose synthase regulates sucrose utilization, whereas isoenzyme II of vacuolar invertase controls sucrose storage and sugar composition.