Cryopreservation of shoot tips and meristems.

Abstract

Shoot-tip meristem cryopreservation methodologies are reported for the complementary cryoprotective strategies of vitrification and equilibrium freezing using traditional controlled-rate freezing and chemical additive cryoprotection. Pregrowth, pretreatment, and cold acclimation approaches for the improvement of tolerance to liquid nitrogen are also presented. The chapter concludes by reporting an analytical protocol that profiles volatile hydrocarbon stress markers (for ethylene, hydroxyl radicals, and lipid peroxidation products) during cryopreservation. This method uses noninvasive headspace sampling and gas chromatography, and it is widely applicable across cryogenic systems.