Understanding temporal niche separation between C-3 and C-4 species (e. g. C-3 species. ourishing in a cool spring and autumn while C-4 species being more active in a hot summer) is essential for exploring the mechanism for their co-existence. Two parallel pot experiments were conducted, with one focusing on water and the other on nitrogen (N), to examine growth responses to water or nitrogen (N) seasonality and competition of two co-existing species Leymus chinensis (C-3 grass) and Chloris virgata (C-4 grass) in a grassland. The two species were planted in either monoculture (two individuals of one species per pot) or a mixture (two individuals including one L. chinensis and one C. virgata per pot) under three different water or N seasonality regimes, i.e. the average model (AM) with water or N evenly distributed over the growing season, the one-peak model (OPM) with more water or N in the summer than in the spring and autumn, and the two-peak model (TPM) with more water or N in the spring and autumn than in the summer. Seasonal water regimes significantly affected biomass in L. chinensis but not in C. virgata, while N seasonality impacted biomass and relative growth rate of both species over the growing season. L. chinensis accumulated more biomass under the AM and TPM than OPM water or N treatments. Final biomass of C. virgata was less impacted by water and N seasonality than that of L. chinensis. Interspecific competition significantly decreased final biomass in L. chinensis but not in C. virgata, suggesting an asymmetric competition between the two species. The magnitude of interspecific competition varied with water and N seasonality. Changes in productivity and competition balance of L. chinensis and C. virgata under shifting seasonal water and N availabilities suggest a contribution of seasonal variability in precipitation and N to the temporal niche separation between C-3 and C-4 species.