Abstract
Plants' mechanisms for surviving freezing stresses are essential adaptations that allow their existence in environments with extreme winter temperatures. Although it is known that Vaccinium macrocarpon Ait. buds can acclimate in fall and survive very cold temperatures during the winter, the mechanism for survival of these buds is not known. The main objective of this study was to determine which of the two major mechanisms of freezing stress survival, namely, deep supercooling or freeze-induced dehydration, are employed by V. macrocarpon terminal buds. In the present study, no low-temperature exotherms (LTEs) were detected by differential thermal analysis. Furthermore, a gradual reduction of relative liquid water content in the inner portions of buds during magnetic resonance imaging (MRI) scans performed between 0 and -20 °C (where no damage was detected in controlled freezing tests (CFT)) indicates these buds may not deep supercool. The higher ice nucleation activity of outer bud scales and the appearance of large voids in this structure in early winter, in conjunction with the MRI observations, are evidence supportive of a freeze-induced dehydration process. In addition, the presence of tissue browning in acclimated buds as a result of freezing stress was only observed in CFT at temperatures below -20 °C, and this damage gradually increased as test temperatures decreased and at different rates depending on the bud structure. Ours is the first study to collect multiple lines of evidence to suggest that V. macrocarpon terminal buds survive long periods of freezing stress by freeze-induced dehydration. Our results provide a framework for future studies of cold hardiness dynamics for V. macrocarpon and other woody perennial species and for the screening of breeding populations for freezing stress tolerance traits.