In-situ fluorescence is a widely used method to estimate the chlorophyll-a (Chla) concentration, a proxy of the phytoplankton biomass. With the emergence of autonomous platforms such as BioGeoChemical-Argo (BGC-Argo) profiling floats, its use has expanded to global scale observations. However, the relationship between in-situ fluorescence and Chla may vary significantly, leading to major discrepancies between oceanic regions. This study aims to investigate the main sources of the natural variability in the in-situ fluorescence signal in the global open ocean, specifically the influence of the phytoplankton community composition. In this view, we analyzed a combination of three datasets comprising concomitant measurements of in-situ fluorescence, pigment concentrations and phytoplankton absorption spectra. Two datasets cover several contrasted bioregions of the global ocean whereas the third one consists of a regional time series in the northwestern Mediterranean Sea, which allows to examine the effect of phytoplankton community composition on the fluorescence signal on the global, seasonal and vertical scales. We studied the variability of the two major drivers of the natural variability of the fluorescence process, i.e. the light absorption and the fluorescence quantum yield of phytoplankton, in regards of the variability of the pigment composition of the communities. The community composition correlates substantially with the Chla-to-fluorescence ratio, with high fluorescence values associated with phytoplankton communities dominated by large cells. This trend may be explained by the combined effects of the community composition on the phytoplankton absorption coefficient and the fluorescence quantum yield, and is consistently observed globally, seasonally and vertically. Non-photosynthetic pigments also appear to play a critical role in oligotrophic surface waters, leading to a reduction of the quantum yield of fluorescence. The results indicate that the phytoplankton community composition plays a key role in the relationship between the in-situ fluorescence signal and Chla concentration. Therefore, we suggest that taking into account the composition of phytoplankton communities in the retrieval of the Chla concentration from current in-situ fluorometers, those mounted on BGC-Argo floats in particular, would lead to a better estimation of the phytoplankton biomass on a wide range of spatial and temporal scales.