In phytoplankton primary production studies, samples for the determination of photosynthesis versus irradiance relationship ( P -I) are usually incubated at several irradiance levels during a fixed time period, commonly 2 -4 h. However, it is not clear if the use of this fixed incubation time is appropriate to study the P -I relationship in any given ecosystem. The aim of this work was to study the influence of incubation time on the P -I relationship in natural phytoplankton populations from three different coastal ecosystems: an open coastal area, an estuary, and a coastal lagoon. Physical and chemical variables, phytoplankton biomass, species composition, and P -I curves were analysed. The results showed that, in the coastal area and in the estuary, P -I relationships were time dependent, whereas in the coastal lagoon different incubation periods produced the same P -I curve. An underestimation of daily primary production, ranging from 13% to 42.5%, was calculated when data from standard incubation times (2 -4 h) were used in ecosystems where P -I curves present a dynamic time-dependent behaviour. This work suggests that the response of the P -I curves to the incubation time varies with the characteristics of the ecosystem and is related to the light regime to which phytoplankton cells are adapted. D