The “Measurements of Pollution in the Troposphere” (MOPITT) satellite record is applied to study the geographical and temporal variability of carbon monoxide (CO) from biomass burning in the Amazon Basin. The presented analysis demonstrates the use of satellite observations for interpreting the effects of deforestation and climate on past and future emissions of CO. The study exploits the MOPITT “multispectral” retrieval product which effectively resolves tropospheric CO into two independently measured layers. New validation results based on in situ measurements during the ACRIDICON‐CHUVA aircraft campaign in 2014 are used for bias correction. Contrasting CO monthly climatologies are presented for the Amazon Basin for the lower and upper troposphere (“LT” and “UT”) with an emphasis on the Amazonian dry season. Climatologically, spatial patterns of UT CO over the Amazon Basin appear to be related to both deep convection and anticyclonic flow. Strongly enhanced LT basin‐mean CO concentrations are observed for the dry season months in 2005, 2007, 2010, and 2015, while the record also indicates a decreasing long‐term trend. These observations are consistent with the expected effects of falling deforestation rates since 2004, punctuated by CO spikes in drought years due to large‐scale wildfires.