Seeds from ten terrestrial orchid species, nine from the south-west Australian biodiversity hotspot (Caladenia arenicola, Caladenia flava, Caladenia huegelii, Diuris laxiflora, Microtis media ssp. media, Pterostylis recurva, Pterostylis sanguinea, Thelymitra crinita and Thelymitra macrophylla) and one from south-east Australia (Diuris fragrantissima), were placed into experimental storage to assess their relative longevity and likely optimal conditions for long-term conservation seed banking. Seeds from all species were desiccation tolerant, germinating after drying at 23% relative humidity (C. arenicola, C. huegelii, P. sanguinea and T. macrophylla) or 5% relative humidity (C. flava, D. laxiflora, M. media ssp. media, P. recurva and T. crinita) at 23 °C. From automatedly determined moisture adsorption and desorption isotherms at 23 °C, these equate to tolerance of drying to 0.03–0.06 g water g−1 dry weight or 0.013–0.028 g water g−1 dry weight, respectively. Results of storage experiments at a range of moisture contents and temperatures suggest conventional seed bank storage at −18 °C after equilibration at c. 23% relative humidity (at 23 °C) may be suitable for most of the species, although there was higher germination of P. recurva seeds stored at −80 °C and of M. media ssp. media seeds equilibrated at 75% relative humidity. However, there was considerable variation in germination of seeds sampled after different storage periods, making it difficult to identify optimal storage conditions definitively. Results of comparative longevity storage experiments at 60% relative humidity and 40 °C suggest seeds from these orchid species are short-lived compared with non-orchid species, and with Australian species in particular. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 164, 26–41.