Nitrogen application at the beginning flowering stage (R1 stage) increased the soybean grain yield, however, the rapid effect of enriched nitrogen at R1 growth stage on soybean dry matter accumulation and sugar metabolism is still unclear. Continuous high nitrogen (CHN), Continuous low nitrogen (CLN), Enriched nitrogen supply at R1 stage (ENS) treatments were applied on two soybean cultivars (Liaodou11, Liaodou14), to investigate the effect of enriched nitrogen on plant biomass accumulation and sucrose metabolism. After 12 h of ENS treatment, the root/shoot rate of both cultivars were lower than that of CLN, but at 24 h it was no significant difference between ENS and CLN. Enriched N at R1 stage, soybean kept a balance of sucrose synthesis and decomposition in leaf by affecting sucrose synthetase (SS) and sucrose phosphate synthase (SPS) activities. Under N limitation condition the plant dry matter accumulation supported root growth priority. Enriched N at R1 stage resulted in the rapid shoot biomass accumulation. In high yield cultivar, the shoot growth was priority to root growth, the common yield cultivar was on the contrary. Our result suggest that enrich N at R1 stage resulted in the accumulation of biomass in shoot rapidly.