Greenhouse-cultured, container-grown ponderosa pine (Pinus ponderosa var. scopulorum Engelm.), interior Douglas-fir (Pseudotsuga menziesii var. glauca (Beissn.) Franco), and Engelmann spruce (Picea engelmannii (Parry) Engelm.) were cold acclimated and deacclimated in growth chambers over 19 weeks. Cold hardiness was measured weekly by a whole-plant freeze test and by two quick tissue tests: freeze-induced electrolyte leakage of needles, and differential thermal analysis of buds. The whole-plant freeze test provided results in 7 days, and indicated differences in cold hardiness among stems, buds, and needles. Although the whole-plant freeze test could accurately measure cold hardiness, it was not precise, and it required destructive sampling. Results from freeze-induced electrolyte leakage and differential thermal analysis were available in 2 days and 1 hour, respectively. The freeze-induced electrolyte leakage test was a precise, sensitive and objective predictor of changes or differences in tissue cold hardiness. To determine actual cold hardiness, results could be calibrated to the response of the same tissue in the whole-plant freeze test. The speed and objectivity of differential thermal analysis made this test useful for rapid, general assessment of cold hardiness status, but calibration was difficult, and precision varied.

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Greenhouse-cultured, container-grown seedlings of Aleppo pine (Pinus halepensis Mill.), radiata pine (Pinus radiata D. Don), and interior Douglas-fir (Pseudotsuga menziesii var. glauca (Beissn.) Franco) were cold acclimated and deacclimated in growth chambers over 24 weeks. Needle and root cold hardiness and root growth potential (RGP) were measured weekly. Root, needle and stem analyses for soluble sugars and starch were performed biweekly. In all tissues, there was a close correspondence between cold hardiness and the absolute concentration of soluble sugars, as well as between the increase and decrease in concentration of soluble sugars during cold hardening and dehardening, respectively, supporting the theory that soluble sugars function as cryoprotectants in plant tissues. The magnitude of starch concentration did not parallel the magnitude of the cold hardiness attained, and changes in starch concentration were related to production and consumption factors, rather than timing of changes in cold hardiness. The rise and fall of RGP paralleled the rise and fall of total carbohydrate concentration in roots. The behavior of the three species was surprisingly similar, considering the different climates to which they are adapted.

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