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Dimethylsulfoniopropionate (DMSP) is produced by many species of marine phytoplankton and has been reported to provide a variety of beneficial functions including osmoregulation. Dinoflagellates are recognized as majorDMSPproducers; however, accumulation has been shown to be highly variable in this group. We explored the effect of hyposaline transfer inGambierdiscus belizeanusbetween ecologically relevant salinities (36 and 31) onDMSPaccumulation, Chla, cell growth, and cell volume, over 12 d. Our results showed thatG. belizeanusmaintained an intracellularDMSPcontent of 16.3 pmol cell⁻¹and concentration of 139 mMin both salinities. Although this intracellular concentration was near the median reported for other dinoflagellates, the cellular content achieved byG. belizeanuswas the highest reported of any dinoflagellate thus far, owing mainly to its large size.DMSPlevels were not significantly affected by salinity treatment but did change over time during the experiment. Salinity, however, did have a significant effect on the ratio ofDMSP:Chla, suggesting that salinity transfer ofG. belizeanusinduced a physiological response other thanDMSPadjustment. A survey ofDMSPcontent in a variety ofGambierdiscusspecies and strains revealed relatively highDMSPconcentrations (1.0–16.4 pmol cell⁻¹) as well as high intrageneric and intraspecific variation. We conclude that, althoughDMSPmay not be involved in long‐term (3–12 d) osmoregulation in this species,G. belizeanusand otherGambierdiscusspecies may be important contributors toDMSPproduction in tropical benthic microalgal communities due to their large size and high cellular content.