The temporal relationship between sucrose (Suc) accumulation and carbon partitioning was investigated in developing sugarcane internodes. Radiolabeling studies on tissue slices, which contained Suc concentrations ranging from 14 to 42% of the dry mass, indicated that maturation coincided with a redirection of carbon from water-insoluble matter, respiration, amino acids, organic acids, and phosphorylated intermediates into Suc. It is evident that a cycle of Suc synthesis and degradation exists in all of the internodes. The decreased allocation of carbon to respiration coincides with a decreased flux from the hexose pool. Both the glucose and fructose (Fru) concentrations significantly decrease during maturation. The phosphoenolpyruvate, Fru-6-phosphate (Fru-6-P), and Fru-2,6-bisphosphate (Fru-2, 6-P2) concentrations decrease between the young and older internodal tissue, whereas the inorganic phosphate concentration increases. The calculated mass-action ratios indicate that the ATP-dependent phosphofructokinase, pyruvate kinase, and Fru-1,6-bisphosphatase reactions are tightly regulated in all of the internodes, and no evidence was found that major changes in the regulation of any of these enzymes occur. The pyrophosphate-dependent phosphofructokinase reaction is in apparent equilibrium in all the internodes. Substrate availability might be one of the prime factors contributing to the observed decrease in respiration.