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The alpine grassland ecosystem accounts for 53% of the Qinghai-Tibet Plateau (QTP) area, which is an important ecological protection barrier, but fragile and highly vulnerable to climate change. Therefore, continuous monitoring of the aboveground biomass (AGB) of grassland is necessary. Although many studies have mapped the spatial distribution of AGB over the QTP, the results vary widely due to the limited ground samples and mismatches with satellite pixel scales. This paper proposed a new algorithm using unmanned aerial vehicles (UAVs) as a bridge to re-estimate the grassland AGB over 20 the QTP from 2000 to 2019. The innovations were as follows: 1) In the aspect of ground data collection, the spatial scale matching among the traditional ground quadrat sampling, UAV photos, and MODIS pixels was fully considered. From 2015 to 2019, 906 pairs of ground-UAV sample data at the quadrat scale and 2,602 sets of UAV data matching the MODIS pixel scale were collected. A total of more than 37,000 UAV photos were captured at the height of 20 meters. Therefore, the ground validation samples was sufficient and scale matched. 2) In terms of model construction, the traditional quadrat scale 25 (0.25m2) was successfully upscaled to the MODIS pixel scale (6,2500 m2) based on the random forest method and stepwise upscaling scheme. Compared with previous studies, the scale matching of independent and dependent variables was realized, effectively reducing the impact of scale mismatch. At the pixel scale, the AGB value estimated by UAV had a more linear correlation with the MODIS vegetation indices than the traditional sampling method. The multi-year independent crossvalidation results showed that the constructed pixel scale AGB estimation had good robustness, with an average R2 of 0.83 30 and RMSE of 34.13 g/m2. Our dataset provides an important input parameter for a comprehensive understanding of the QTP in the process of global climate change. [ABSTRACT FROM AUTHOR]