Vegetation serves as a crucial indicator of ecological environment and plays a vital role in preserving ecosystem stability. However, as urbanization escalates rapidly, natural vegetation landscapes are undergoing continuous transformation. Paradoxically, vegetation is pivotal in mitigating the ecological and environmental challenges posed by urban sprawl. The middle and lower Yangtze River Basin (MLYRB) in China, particularly its economically thriving lower reaches, has witnessed a surge in urbanization. Consequently, this study explored the spatiotemporal variations of normalized difference vegetation index (NDVI) in the MLYRB, with an emphasis on elucidating the impact of climate change and urbanization on vegetation dynamics. The results indicate that a significant increasing trend in NDVI across the MLYRB from 2000 to 2020, a pattern that is expected to persist. An improvement in vegetation was observed in 94.12% of the prefecture-level cities in the study area, predominantly in the western and southern regions. Temperature and wind speed stand out as dominant contributors to this improvement. Nevertheless, significant vegetation degradation was detected in some highly urbanized cities in the central and eastern parts of the study area, mainly attributed to the negative effects of escalating urbanization. Interestingly, a positive correlation between NDVI and the urbanization rate was observed, which may be largely related to proactive ecological preservation policies. Additionally, global climatic oscillations were identified as a key force driving periodic NDVI variations. These findings hold significant importance in promoting harmonious urbanization and ecological preservation, thereby providing invaluable insights for future urban ecological planning efforts.