To assess whether inadequate cold hardiness could be a contributor to yellow-cedar (Chamaecyparis nootkatensis (D. Don) Spach) decline, we measured the freezing tolerance of foliage from yellow-cedar trees in closedcanopy (nondeclining) and open-canopy (declining at elevations below 130 m) stands at three sites along an elevational gradient in the heart of the decline in southeastern Alaska. Foliar freezing tolerance was also assessed for sympatric nondeclining western hemlock (Tsuga heterophylla (Raf.) Sarg.). Measurements were made in the fall, winter, and spring to evaluate if seasonal differences in cold hardiness help explain species-specific injury. Significant differences in freezing tolerance attributable to site, canopy closure, species, and the interaction of canopy closure and species were each detected for at least one sample period. However, only two results were consistent with field reports of yellow-cedar decline: (1) between winter and spring measurements, yellow-cedar trees dehardened almost 13°C more than western hemlock trees, so that yellow-cedar trees were more vulnerable to foliar freezing injury in spring than western hemlock; and (2) stands below 130 m appeared more vulnerable to freezing injury than stands above 130 m.Résumé : Afin d'évaluer si une résistance inadéquate au froid pouvait contribuer au dépérissement du faux-cyprès de Nootka (Chamaecyparis nootkatensis (D. Don) Spach), les auteurs ont mesuré la tolérance au gel du feuillage de fauxcyprès dans des peuplements fermés (non dépérissant) et ouverts (dépérissant à une altitude inférieure à 130 m), dans trois stations situées le long d'un gradient altitudinal au coeur de la zone de dépérissement dans le sud-est de l'Alaska. La tolérance au gel du feuillage a également été évaluée chez la pruche de l'Ouest (Tsuga heterophylla (Raf.) Sarg.), une espèce sympatrique qui ne dépérit pas. Des mesures ont été prises à l'automne, à l'hiver et au printemps pour dé-terminer si des différences saisonnières de résistance au froid pouvaient contribuer à expliquer pourquoi les dommages sont limités à une espèce. Des différences significatives dans la tolérance au gel attribuables au site, à la fermeture du couvert, à l'espèce et à l'interaction entre la fermeture du couvert et l'espèce ont été détectées pendant au moins une période d'échantillonnage. Cependant, seulement deux résultats sont consistants avec les rapports de terrain sur le dépé-rissement du faux-cyprès : (1) entre les mesures prises en hiver et au printemps, le faux-cyprès s'est désendurci de presque 13°C de plus que la pruche de l'Ouest, de telle sorte que le faux-cyprès était plus vulnérable que la pruche de l'Ouest aux dommages causés par le gel du feuillage au printemps et (2) les peuplements situés à moins de 130 m d'altitude semblaient plus vulnérables aux dommages causés par le gel que les peuplement situés à plus de 130 m d'altitude.[Traduit par la Rédaction] Schaberg et al. 2070

Laboratory experiments have verified that acid-deposition-induced calcium (Ca) leaching reduces the foliar cold tolerance of red spruce (Picea rubens Sarg.) current-year foliage, increasing the risk of winter injury and crown deterioration. However, to date no studies have shown that ambient losses in soil Ca have resulted in increased winter injury in the field. In 2003, a year of severe region-wide winter injury to red spruce, we measured the nutrition and winter injury of current-year foliage and bud mortality for red spruce on two watersheds at the Hubbard Brook Experimental Forest in Thornton, New Hampshire: (1) a reference watershed that has undergone considerable Ca loss attributed to acid-deposition-induced leaching and (2) a watershed that was fertilized with CaSiO3 in 1999 to replace lost Ca. For all crown classes combined, winter injury was significantly greater (P = 0.05) for red spruce on the reference watershed than for spruce on the Ca-addition watershed. Differences in foliar injury were particularly evident for dominant and codominant trees. For these crown classes, red spruce on the reference watershed lost about 75% of their current-year foliage to winter injury, about three times more than foliar losses for the Ca-addition watershed (P = 0.01). Patterns of bud mortality followed that of foliar injury. The only difference in foliar cation nutrition detected was a significantly greater concentration of Ca in red spruce foliage from the Ca-addition watershed relative to spruce from the reference watershed (P = 0.001). Differences in Ca concentration, foliar winter injury, and bud mortality that occurred coincident with watershed Ca treatment provide the first evidence that ambient Ca depletion is associated with elevated winter injury of red spruce trees.

We examined the effects and potential interactions of acid mist and soil solution Ca and Al treatments on foliar cation concentrations, membrane-associated Ca (mCa), ion leaching, growth, carbon exchange, and cold tolerance of red spruce (Picea rubens Sarg.) saplings. Soil solution Ca additions increased foliar Ca and Zn concentrations, and increased rates of respiration early in the growing season (July). Soil Al treatment had a broad impact, reducing foliar concentrations of Ca, Mg, Mn, P and Zn, and resulting in smaller stem diameters, sapling heights and shoot lengths compared with soil treatments with no added Al. Aluminum treatment also reduced respiration when shoots were elongating in July and decreased net photosynthesis at the end of the growing season (September). Three lines of evidence suggest that Al-induced alterations in growth and physiology were independent of foliar Ca status: (1) Ca concentrations in foliage of Al-treated saplings were within the range of sufficiency established for red spruce; (2) mCa concentrations were unaffected by Al treatment; and (3) no Al x Ca interactions were detected. Acid mist treatment increased foliar Fe and K concentrations and increased leaching of Ca, Mg, Mn, Zn, Fe, and Al from foliage. Leaching losses of Ca were more than twice those of the element with the next highest amount of leaching (Zn), and probably led to the reductions in mCa concentration and membrane stability of acid-treated saplings. Acidic mist resulted in enhanced shoot growth, and consistent reductions in foliar cold tolerance in the fall and winter. Of the few significant interactions among treatments, most involved the influence of mist pH and Al treatment on foliar nutrition. In general, reductions in cation concentration associated with Al addition were greater for pH 5.0-treated saplings than for pH 3.0-treated saplings. We propose that H(+)-induced leaching of mCa from mesophyll cells is the mechanism underlying acid-induced reductions in foliar cold tolerance of red spruce.

We examined the pattern of seasonal variation in total foliar calcium (Ca) pools and plasma membrane-associated Ca (mCa) in mesophyll cells of current-year and 1-year-old needles of red spruce (Picea rubens Sarg.) and the relationship between mCa and total foliar Ca on an individual plant and seasonal basis. Foliar samples were collected from seedlings and analyzed on 16 dated at 2- to 3-week intervals between June 1994 and March 1995. Concentrations of mCa in current-year needles were more seasonally dynamic and responsive to temporal environmental changes than either mCa concentrations of 1-year-old needles, which were largely stable, or total foliar Ca concentrations in both tissues. In current-year needles, mCa was barely evident in early summer, increased steadily through summer, and then increased dramatically in early fall and surpassed the concentration in 1-year-old needles. Coincident with the first severe frost, mCa concentrations in current-year needles declined significantly and subsequently maintained concentrations comparable to those of 1-year-old needles. Following an extended January thaw, which included 5 days of minimum temperatures > 5 degrees C, mCa concentrations of current-year needles temporarily, but significantly, declined. However, there was no change in mCa concentrations of 1-year-old needles or total Ca concentrations of either tissue. Total Ca concentrations were stable through midsummer in both tissues, doubled in late summer, and then were stable in both tissues throughout fall and winter. Total Ca concentrations were consistently higher in 1-year-old than in current-year needles. Correlations between concentrations of mCa and total foliar Ca were consistently low and mostly nonsignificant. Thus, the dominant, but insoluble, extracellular Ca pool reflected in commonly measured total foliar Ca concentrations is not a meaningful surrogate for the physiologically important and labile pool associated with the plasma membrane-cell wall compartment of red spruce mesophyll cells. It is likely that shifts in the critical mCa compartment would not be detected by analysis of total foliar Ca pools. Seasonal changes in mCa concentration seemed to parallel seasonal changes in membrane structure, and possibly the important role of extracellular Ca in transducing messages associated with environmental signals.

Acidic deposition can leach essential pools of calcium (Ca) directly from plant foliage. Because of the central role of Ca in environmental signal transduction, disruptions of labile foliar Ca pools could impair physiological responses to a variety of environmental stimuli and stressors. We investigated the possibility that acidic mist-induced depletion of membrane-associated Ca (mCa), which is one form of labile Ca, may alter stomatal responsiveness to water stress, a process known to include Ca in signal transduction cascades. Red spruce (Picea rubens Sarg.) seedlings were exposed to either pH 3.0 or pH 5.0 mist treatments for one growing season. Foliar nutrition was assessed following treatments, and declines in stomatal conductance and net photosynthesis were measured on current-year shoots following stem excision. Seedlings exposed to pH 3.0 acidic mist treatments had reduced mCa relative to the pH 5.0 treated seedlings. Seedlings subjected to the pH 3.0 acidic mist treatment exhibited impaired stomatal functions, including a smaller maximum aperture, slower closure and an increased lag time between stomatal closure and photosynthetic decline following experimental water stress. Delayed stomatal closure could undermine desiccation avoidance mechanisms. Previous work has demonstrated that acidic mist treatments deplete mCa in red spruce and impair cold tolerance, with similar effects in other species. The results we present provide further evidence that acidic mist-induced mCa depletion may cause disruption of a broad range of plant stress responses.