Understanding the desiccation and freezing tolerance of bryophyte spores is vital to explain how plants conquered land and current species distribution patterns and help to develop efficient ex situ conservation methods. However, knowledge of these traits is scarce. We investigated tolerance to drying (at 15% relative humidity [RH] for two weeks) and freezing (1 h exposure to liquid nitrogen) on the spores of 12 bryophyte species (23 accessions) from the UK. The presence of storage lipids and their thermal fingerprint, and the levels of unfrozen water content, were determined by differential scanning calorimetry (DSC). The presence of chlorophyll in dry spores was detected by fluorescence microscopy. All species and accessions tested tolerated the drying and freezing levels studied. DSC suggested that 4.1−29.3% of the dry mass is storage lipids, with crystallization and melting temperatures peaking at around −30 °C. Unfrozen water content was determined <0.147 g H2O g−1 dry weight (DW). Most of the spores investigated showed the presence of chlorophyll in the cytoplasm by red autofluorescence. Bryophyte spores can be stored dry at low temperatures, such as orthodox seeds, supporting the creation of bryophyte spore banks. However, the presence of storage lipids and chlorophyll in the cytoplasm may reduce spore longevity during conventional storage at −20 °C. Alternatively, cryogenic spore storage is possible.
Keywords: chlorophyll; cryopreservation; desiccation tolerance; differential scanning calorimetry; ex situ conservation; in vitro germination; lipid crystallization; liquid nitrogen; unfrozen water content.