We examined the potential effects of environmental variables, and their interaction, on phytoplankton community succession in spring using long-term data from 1992 to 2012 in Lake Taihu, China. Laboratory experiments were additionally performed to test the sensitivity of the phytoplankton community to nutrient concentrations and temperature. A phytoplankton community structure analysis from 1992 to 2012 showed that Cryptomonas (Cryptophyta) was the dominant genus in spring during the early 1990s. Dominance then shifted to Ulothrix (Chlorophyta) in 1996 and 1997. However, Cryptomonas again dominated in 1999, 2000, and 2002, with Ulothrix regaining dominance from 2003 to 2006. The bloom-forming cyanobacterial genus Microcystis dominated in 1995, 2001 and 2007-2012. The results of ordinations indicated that the nutrient concentration (as indicated by the trophic state index) was the most important factor affecting phytoplankton community succession during the past two decades. In the laboratory experiments, shifts in dominance among phytoplankton taxa occurred in all nutrient addition treatments. Results of both long term monitoring and experiment indicated that nutrients exert a stronger control than water temperature on phytoplankton communities during spring. Interactive effect of nutrients and water temperature was the next principal factor. Overall, phytoplankton community composition was mediated by nutrients concentrations, but this effect was strongly enhanced by elevated water temperatures.