Negative air ions are natural components of the air we breathe. Forests are the main continuous natural source of negative air ions (NAI). The spatio-temporal patterns of negative air ions were explored in Shanghai, based on monthly monitoring in 15 parks from March 2009 to February 2010. In each park, sampling sites were selected in forests and open spaces. The annual variation in negative air ion concentrations (NAIC) showed peak values from June to October and minimum values from December to January. NAIC were highest in summer and autumn, intermediate in spring, and lowest in winter. During spring and summer, NAIC in open spaces were significantly higher in rural areas than those in suburban areas. However, there were no significant differences in NAIC at forest sites among seasons. For open spaces, total suspended particles (TSP) were the dominant determining factor of NAIC in sum-mer, and air temperature and air humidity were the dominant determining factors of NAIC in spring, which were tightly correlated with Shanghai’s ongoing urbanization and its impacts on the environment. It is suggested that urbanization could induce variation in NAIC along the urban-rural gradient, but that may not change the temporal variation pattern. Fur-thermore, the effects of urbanization on NAIC were limited in non-vegetated or less-vegetated sites, such as open spaces, but not in well-vegetated areas, such as urban forests. Therefore, we suggest that urban greening, especially urban forest, has significant resistance to theeffect of urbanization on NAIC.