Vitrification in plants as a natural form of cryoprotection

A small group of woody plants from the far northern hemisphere can, while in the dormant state, tolerate freezing and thawing to and from any subzero temperature at rates less than 30 degrees C/hr. In addition, the hardiest of them can tolerate cooling and warming between -20 degrees C and any colder temperature at virtually any combination of rates subsequent to cooling to -20 degrees C at rates less than 5 degrees C/hr. We term this latter capability "quench hardiness." I and my colleagues have shown that the limits of this quench hardiness can be closely correlated to the stability of intracellular glasses formed during the slow cooling of hardy tissues in the presence of extracellular ice. In this paper, I briefly review the evidence for intracellular glass formation and present data indicating that major components of the glass forming solutions are raffinose and stachyose. Evidence from differential scanning calorimetry that sugar-binding soluble proteins are also important is presented. Finally, I correlate data from our work with that of other workers to make the case that, even when most of a cytoplasmic solution is vitrified, fluid microdomains remain which can lead to long-term biodegradation during storage at high subzero temperatures.