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Eighteen native species of Nymphaea (waterlilies) inhabit a range of freshwater wetlands in northern Australia, which are threatened by increased development and the potential impacts of climate change. To investigate conservation seed banking of these vulnerable species, we aimed to characterize their seed storage physiology by determining (i) seed desiccation tolerance and (ii) the effects of moisture content and storage temperature on seed germination and viability. Seeds of N. immutabilis, N. lukei, N. macrosperma and N. violacea (including multiple collections of three species) were placed in experimental storage at a range of temperatures (25 degrees C, 5 degrees C, -20 degrees C and -190 degrees C) following pre-equilibration at different RHs (15%, 30%, 50%, 70% or 95%). Seeds were also experimentally aged at 60% RH and 45 degrees C to assess comparative longevity. We found seeds of all species to be desiccation tolerant. However, the responses of seeds to experimental storage conditions were complex and variable between species and collections of the same species, and seeds of many species/collections were short-lived across many of the storage treatments. In many cases decreasing storage temperature did not increase longevity. Additional protocol development is necessary before we can have confidence that ex situ seed banking is a viable long-term germplasm conservation strategy for Nymphaea.