Changes in climate and land cover are potential drivers of land surface phenology changes. Here, we investigate whether subpixel percent tree cover (PTC) change is an important driver of trends in satellite derived vegetation spring greenup date (GUD) across the Hulunbuir temperate forest-grassland ecotone in northeastern China. GUD was estimated using the MODIS-derived enhanced vegetation index time series during 2001–2020 with a spatial resolution of 500 m. To understand the influential mechanisms of PTC on GUD, we examined relationships between the spatial variations in GUD and PTC at multiple spatio-temporal extents. Forested pixels with greater PTC were found to have generally earlier GUDs for all forest types. The GUD of forests was also generally earlier than that of grassland. On the other hand, we observed approximately 23.7% and 1.2% significantly earlier and later trends in GUD across the region, respectively. Meanwhile, widespread increases in preseason land surface temperature (LST) and PTC were detected. Both increases in LST and PTC contributed to the earlier GUD in the forested region. Specifically, we found negative correlations (Spearman correlation coefficient -0.17 to -0.55) between the change slopes of GUD and PTC in every forest and grassland type. The results highlight the important impacts of subpixel PTC on GUD variations, and improve the understanding of ecosystem changes under the effects of climate and human activities (e.g., afforestation) over the Hulunbuir temperate forest-grassland ecotone.