Phytoplankton are key players in the aquatic environment and they can be effectively used to understand and predict the functioning and production of aquatic ecosystems. Given that phytoplankton is affected by natural and human-induced perturbations, such as eutrophication and global climate change, it is pressing to understand which factors regulate phytoplankton communities. The main goal of this work was, therefore, to understand how phytoplankton growth and production in the turbid Guadiana estuary, particularly in the freshwater tidal zone, is regulated by bottom-up factors, namely nutrients and light. Enrichment bioassays were carried out to evaluate nutrient and light limitation of phytoplankton growth. Nutrient limitation, specifically by nitrogen, was observed during the productive period. Nitrogen, as nitrate, became limiting to phytoplankton growth at concentrations lower than 20 WM. Although nitrate was the main nitrogen source in the Guadiana estuary, an overall preference for ammonium was observed, mainly by cyanobacteria and green algae. Diatoms were the most nutrient-limited group, and they clearly preferred nitrate as their N-source. Regarding light availability, phytoplankton was not acclimated to the low light conditions that prevail in the Guadiana estuary and light limitation occurred throughout the year. Diatoms were the most light-limited group, whilst cyanobacteria seemed to be more acclimated to low light. Primary production was higher in the more turbid regions, where light availability was the lowest, but nutrient concentrations, although occasionally limiting, were the highest. Therefore, phytoplankton in such turbid regions were the most efficient in using limiting resources. River flow was a major regulator of nutrient and suspended matter inputs to the estuarine zone. Tidallyinduced variability of phytoplankton and environmental drivers in the freshwater tidal estuarine zone was low and resulted from seasonal and fortnightly variability in river flow and tidal currents.