An unusual phenomenon of anaerobic phosphate uptake under alternating anaerobic/aerobic condition was observed in a granular sludge sequencing batch reactor, fed with acetate as sole organic substrate. Anaerobic phosphate uptake efficiencies remained at 50-70% as the influent P/COD was increased from 2/100 to 4/100, and results showed that anaerobic uptake of phosphate was correlated with anaerobic absorption of acetate. Excluding the main possibility of chemical phosphate removal, it appeared that phosphate uptake during the anaerobic phase was associated with organisms enriched in the reactor. Moreover, results indicated that intracellular glycogen was used as the main energy source of organics anaerobic absorption and intracellular polymers storage. Measuring and analysing the variation of phosphate, organic substrate, intracellular glycogen and pH in the anaerobic phase, a preliminary explanation was developed that anaerobic uptake of phosphate was the demand of intracellular glycogen degradation, and extracellular phosphate was transported to intracellular by pH gradient-sensitive phosphate carrier protein.