The changes in grain-filling, starch accumulation and activity of enzymes associated with starch synthesis in two different hybrid rice varieties were analyzed at different N supplying dates (earlier-date-emphasized, mean-date-emphasized and later-date- emphasized). The results showed that the N application of later-date-emphasized could promote grain-filling rate, increase grain weight and amylopectin content. The peak of activity in three enzymes of ADPglusoce pyriphosphorylase (ADPG), starch synthesis enzyme (SSS) and starch branching enzyme (SBE) in grains of two different rice varieties was not changed obviously, but the mean and maximum activity of these three enzymes changed, and the changes of SSS and ADPG were bigger than that of SBE as N supplying date changed. The N application of earlier-date-emphasized increased SSS activity and the N application of later-date-emphasized increased ADPG and SBE activities. The mean SSS activity during whole grain-filling period, and ADPG and SBE activities at middle and late period of grain-filling were significantly or very significantly correlated with grain-filling rate and accumulating rate of amylose and amylopectin. Both of ADPG and SBE played an equal important role in the changes of amylose and amylopectin content. The N application of later-date-emphasized increased amylose and amylopectin accumulating rate. The changes in grain-filling, starch accumulation and activity of enzymes associated with starch synthesis in two different hybrid rice varieties were analyzed at different N supplying dates (earlier-date-emphasized, mean-date-emphasized and later-date-emphasized). The results of that N application of later-date-emphasized could promote grain-filling rate, increase grain weight and amylopectin content. The peak of activity in three enzymes of ADPglusoce pyriphosphorylase (ADPG), starch synthesis enzyme (SSS) and starch branching enzyme (SBE) in grains of two different rice varieties was not changed obviously, but the mean and maximum activity of these three enzymes changed, and the changes of SSS and ADPG were bigger than that of SBE as N supplying date changed. The N application of earlier-date-emphasized increased SSS activity and the N application of later-date-emphasized increased ADPG and SBE activities. The mean SSS activity during whole grain-filling period, and ADPG and SBE activities at middle and late period of grain-filling were significantly or very significantly with grain-filling rate and accumulating rate of amylose and amylopectin. Both of ADPG and SBE played an equal important role in the changes of amylose and amylopectin content. The N application of later-date-emphasized increased amylose and amylopectin accumulating rate.