Diarrhetic shellfish poisoning (DSP) toxins and pectenotoxins (PTXs) produced by endemic species of Dinophysis, mainly D. acuta and D. acuminata, threaten public health and negatively impact the shellfish industry worldwide. Despite their socioeconomic impact, research on the environmental drivers of DSP outbreaks in the Chilean fjords is scanty. From 22 to 24 March 2017, high spatial–temporal resolution measurements taken in Puyuhuapi Fjord (Northern Patagonia) illustrated the short-term (hours, days) response of the main phytoplankton functional groups (diatoms and dinoflagellates including toxic Dinophysis species) to changes in water column structure. Results presented here highlight the almost instantaneous coupling between time–depth variation in density gradients, vertical shifts of the subsurface chlorophyll maximum, and its evolution to a buoyancy-driven thin layer (TL) of diatoms just below the pycnocline the first day. A second shallower TL of dinoflagellates, including Dinophysis acuta, was formed on the second day in a low-turbulence lens in the upper part of the pycnocline, co-occurring with the TL of diatoms. Estimates of in situ division rates of Dinophysis showed a moderate growth maximum, which did not coincide with the cell density max. This suggests that increased cell numbers resulted from cell entrainment of off-fjord populations combined with in situ growth. Toxin profiles of the net tow analyses mirrored the dominance of D. acuminata/D. acuta at the beginning/end of the sampling period. This paper provides information about biophysical interactions of phytoplankton, with a focus on Dinophysis species in a strongly stratified Patagonian fjord. Understanding these interactions is crucial to improv predictive models and early warning systems for toxic HABs in stratified systems.