Quantitative estimation of wetland aboveground biomass (AGB) is an essential aspect in evaluating the health and conservation of this valuable ecosystem. We combine AGB field measurements and remote sensing data to establish a suitable model for estimating wetland AGB in the Poyang Lake National Nature Reserve (PLNNR), which is included in the Ramsar Convention's List of Wetlands of International Importance. All field sampling points cover four dominant vegetation communities (Carex cinerascen, Phalaris arundinacea, Artemisia selengensis, and Miscanthus sacchariflorus) in the PLNNR. Wetland AGB is retrieved from the Landsat-8 OLI image. To improve the accuracy of wetland AGB estimation, we compare the performances of three machine learning algorithms, namely, random forest (RF), back-propagation neural network (BPNN), and support vector regression (SVR), with linear regression (LR) in estimating the AGB in the PLNNR. Results are as follows: (1) the RF model with a root-mean-square error of 0.25 kg m −2 performs better than BPNN (0.29 kg m −2), SVR (0.27 kg m −2), and LR (0.31 kg m −2) in our testing dataset, and AGB density in the PLNNR is between 0 and 1.973 kg m −2. (2) The four most important features for AGB modeling are near-infrared, shortwave infrared 1 band, enhanced vegetation index, and red band. Our study presents an effective and operational RF model that estimates wetland AGB from Landsat data, providing a scientific basis for floodplain wetland carbon accounting and possible future studies, such as the linkage between wetland AGB and the great water level fluctuations. © The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.