Longevity of Preserved Germplasm: The Temperature Dependency of Aging Reactions in Glassy Matrices of Dried Fern Spores

Abstract

This study explores the temperature dependency of the aging rate in dry cells over a broad temperature range encompassing the fluid to solid transition (Tg) and well below. Spores from diverse species of eight families of ferns were stored at temperatures ranging from +45�C to approximately -176�C (vapor phase above liquid nitrogen), and viability was monitored periodically for up to 4,300 d (∼12 years). Accompanying measurements using differential scanning calorimetry (DSC) provide insights into structural changes that occur, such as Tg between +45 and -20�C (depending on moisture), and triacylglycerol (TAG) crystallization between -5 and -35�C (depending on species). We detected aging even at cryogenic temperatures, which we consider analogous to unscheduled degradation of pharmaceuticals stored well below Tg caused by a shift in the nature of molecular motions that dominate chemical reactivity. We occasionally observed faster aging of spores stored at -18�C (conventional freezer) compared with 5�C (refrigerator), and linked this with mobility and crystallization within TAGs, which probably influences molecular motion of dried cytoplasm in a narrow temperature range. Temperature dependency of longevity was remarkably similar among diverse fern spores, despite widely disparate aging rates; this provides a powerful tool to predict deterioration of germplasm preserved in the solid state. Future work will increase our understanding of molecular organization and composition contributing to differences in longevity.

Keywords: Cryopreservation; Differential scanning calorimetry; Ex situ conservation; Moisture; Molecular mobility; Triacylglycerol.