A new land cover database of Greater Mesoamerica has been prepared using moderate resolution imaging spectroradiometer (MODIS, 500 m resolution) satellite data. Daily surface reflectance MODIS data and a suite of ancillary data were used in preparing the database by employing a decision tree classification approach. The new land cover data are an improvement over traditional advanced very high resolution radiometer (AVHRR) based land cover data in terms of both spatial and thematic details. The dominant land cover type in Greater Mesoamerica is forest (39%), followed by shrubland (30%) and cropland (22%). Country analysis shows forest as the dominant land cover type in Belize (62%), Cost Rica (52%), Guatemala (53%), Honduras (56%), Nicaragua (53%), and Panama (48%), cropland as the dominant land cover type in El Salvador (60.5%), and shrubland as the dominant land cover type in Mexico (37%). A three-step approach was used to assess the quality of the classified land cover data: (i) qualitative assessment provided good insight in identifying and correcting gross errors; (ii) correlation analysis of MODIS- and Landsat-derived land cover data revealed strong positive association for forest (r² = 0.88), shrubland (r² = 0.75), and cropland (r² = 0.97) but weak positive association for grassland (r² = 0.26); and (iii) an error matrix generated using unseen training data provided an overall accuracy of 77.3% with a Kappa coefficient of 0.73608. Overall, MODIS 500 m data and the methodology used were found to be quite useful for broad-scale land cover mapping of Greater Mesoamerica.