Isoëtes are iconic but understudied wetland plants, despite having suffered severe losses globally mainly because of alterations in their habitats. We therefore provide the first global ecological assessment of aquatic Isoëtes to identify their environmental requirements and to evaluate if taxonomically related species differ in their ecology. The assessment resulted in an extensive new database on aquatic Isoëtes, ecological niche analyses, and descriptive species accounts. We compiled a global database that includes all known environmental data collected from 1935 to 2023 regarding aquatic Isoëtes. We then evaluated the environmental drivers of 16 species using 2,179 global records. Additionally, we used hypervolume analysis to quantify the ecological niches of the two species with the greatest number of records, finding significant differences and evidence that Isoëtes echinospora occupies a wider ecological niche than Isoëtes lacustris. Fifty-nine species (30% of the c. 200 Isoëtes species known today) were categorised as aquatic and were mainly reported in the Americas and northern Europe. About 38% of the aquatic species are threatened with extinction or are endemic to a small region, according to the International Union for Conservation of Nature's Red List in 2023. Many species were determined to be sensitive to certain water physical and chemical factors, generally preferring oligotrophic conditions such as low total phosphorus, moderate total nitrogen, moderate to low pH, and low conductivity. This analysis includes ecological data in the assessment of rare/threatened aquatic plants globally. This new database and the ecological analyses completed defined the ecological requirements of several species and identified knowledge gaps, which can aid management actions and future research. This paper highlights ecological significance and environmental sensitivities of aquatic Isoëtes. The current level of knowledge is inadequate for a large proportion of known taxa. We affirm the extreme need to support global, collaborative initiatives on which to build future conservation strategies.