The spatiotemporal distribution of phytoplankton biomass drives the marine ecosystem. Chlorophyll-a concentration (Chla) is a biomass index for microalgae in seawater that is commonly used to study phytoplankton by means of satellite remote sensing. The St. Lawrence Estuary and Gulf (SLEG), in Eastern Canada, is a highly dynamic subpolar environment characterized by complex marine optical properties that make it difficult to distinguish Chla from the background signal caused by a strong freshwater discharge. In this study, we implement an inverse model based on a set of in situ Chla processed through a principal component analysis, making it specifically designed for the local marine optical conditions. We used this model to convert a multi-mission remote sensing reflectance dataset to daily Chla between 1998 and 2019 at a 4 km spatial resolution. From the resulting Chla time series, we computed the climatology, phenology and trend over the SLEG. The Chla climatology reveals the presence of relatively high Chla in the Gaspé Current, along the Gulf's North Shore and in areas of strong tidal mixing. Important differences in the phytoplankton phenology between the various subregions are found, with a prevailing shift towards earlier spring blooms of larger intensity. Finally, we found a positive mean Chla increase of 1.1% yr −1 over the SLEG, with strong positive trends in the Magdalen Shallows and west of the Anticosti Island. This description of the surface Chla in the SLEG provides important baseline information for the marine ecosystem. • Our regional algorithm produces realistic chlorophyll-a in the St. Lawrence Estuary and Gulf. • Chlorophyll-a is higher in coastal areas with upwellings or freshwater input. • Spring blooms develop from south to north, with the estuarine portion last. • Between 1998 and 2019, chlorophyll-a increased by 1.1% yr −1 .