Polyamines in embryogenic cultures of Norway spruce (Picea abies) and red spruce (Picea rubens)

Embryogenic cultures of red spruce (Picea rubens Sarg.) and Norway spruce (Picea abies (L.) Karst.) were initiated from dissected mature zygotic embryos. The tissues were grown on either proliferation medium or maturation medium. On proliferation medium, the embryogenic tissue continued to produce early stage somatic embryos (organized meristems attached to elongated, suspensor-like cells), whereas on maturation medium fully mature embryos developed from the embryonic tissue. Analysis of polyamines in tissues grown on these two media showed that: (1) both putrescine and spermidine concentrations were always higher in cultures grown on proliferation medium than in cultures grown on maturation medium; (2) in both species, spermidine concentrations declined with time in the tissues grown on maturation medium; and (3) spermine was present in only minute quantities and showed only a small change with time. The presence of difluoromethylornithine in the culture medium had little effect on polyamine concentration, whereas the presence of difluoromethylarginine caused a decrease in putrescine concentrations in both red spruce and Norway spruce tissues grown on proliferation medium or maturation medium.     

Frost hardiness of 16 European provenances of sessile oak growing in Scotland

The frost hardiness of 16 European provenances of sessile oakQuercus petraea (Matt.)Liebl. originating from six Europeancountries was examined from autumn until budburst in springusing the method of relative conductivity. There were significant differences of frost hardiness betweenprovenances and a strong relationship between phenology andfrost hardiness. In spring, provenances that burst bud earlydehardened earlier than provenances that burst bud later. Inautumn, provenances that stopped growing early were more frosthardy than provenances that continued to grow. The interprovenance ranges of frost hardiness were greatestin spring and autumn and least in midwinter. Consequently, itwas not possible to demonstrate significant differences of frosthardiness between provenances in December and January. Overall, German, Polish and Danish provenances were more frosthardy than French, Austrian and British provenances. Frenchprovenances were least hardy at all times.     

Seasonal variation in photochemical activity and hardiness in clones of Norway spruce (Picea abies)

We investigated changes in photochemical activity and cold hardiness of detached needles of three clones of Picea abies (L.) Karst. by measuring variable chlorophyll fluorescence (F(v)/F(m)), before and after artificial freezing, from September to June. Photochemical activity varied considerably during the study, but only minor differences in photochemical activity among the clones were observed before freezing. Photochemical activity was high during early fall and then declined from November until April. Photochemical activity was at a minimum in April and then increased quickly to high values in May. During the period from late September to October, and also during the winter, differences in F(v)/F(m) ratios after artificial freezing to below -10 degrees C were observed among clones, indicating clonal differences in cold hardiness and hardiness development. The clone having an average height of 2.3 m after 11 years showed consistently lower cold hardiness than clones that had reached average heights of 4.0 and 5.0 m. There were also differences in the temperature requirement for bud flushing among clones.     

Seasonal air and soil temperature effects on photosynthesis in red spruce (Picea rubens) saplings

Net photosynthesis and stomatal conductance were measured in ten red spruce (Picea rubens Sarg.) saplings, growing near Ithaca, New York, throughout the early spring and late-fall growing periods. Gas exchange and daily minimum and maximum soil and air temperatures were also measured. Linear regression analysis showed that rates of net photosynthesis were positively correlated with both minimum daily soil and air temperatures but that minimum soil temperature was a better predictor of net photosynthesis. Moreover, net photosynthesis was more sensitive to changes in soil temperature than to changes in air temperature, and photosynthesis was approximately twice as sensitive to temperature changes during the fall than during the spring.     

Effects of elevated nitrate and aluminum on the growth and nutrition of red spruce (Picea rubens) seedlings

Acidic deposition in high-elevation forests in the Appalachian Mountains of the eastern United States has been implicated in the decline of red spruce (Picea rubens Sarg.). Elevated soil acidity may increase soil Al availability and toxicity to roots. Enhanced soil solution NO(3) (-) concentrations, resulting from precipitation inputs and enhanced soil organic matter mineralization, may exacerbate Al toxicity by increasing root Al uptake. We exposed red spruce seedlings to 350, 500, 800 or 1400 micro M NO(3) (-) and 0 or 200 micro M Al in a factorial design in sand-nutrient solution culture to test if increased NO(3) (-) concentrations enhance Al uptake and toxicity. In addition to significant reductions in seedling growth parameters resulting from Al exposure, we found significant interactions between NO(3) (-) and Al for seedling height growth rate, needle weight, shoot weight and root weight. Differences in these parameters between Al treatments became more pronounced as solution NO(3) (-) concentration increased and reflected an Al-mediated inhibition of seedling response to increasing NO(3) (-) concentration. Solution NO(3) (-) concentrations above 500 micro M induced root nitrate reductase (NR) activity, whereas shoot NR activity increased in response to NO(3) (-) up to 500 micro M and declined above that concentration. In contrast, exposure to Al depressed NR activity of roots but tended to stimulate needle NR activity. Foliar N concentrations increased in seedlings grown in cultures containing between 350 and 500 micro M NO(3) (-), with no change above 500 micro M. Increasing concentrations of NO(3) (-) depressed foliar P concentrations, with reductions being greatest in seedlings exposed to 1400 micro M NO(3) (-). Exposure to Al increased foliar Ca, K and Al concentrations, decreased foliar P concentrations, and inhibited increases in foliar Mg concentration in response to increasing NO(3) (-). The consistent interactions between NO(3) (-) and Al for growth, root NR activity and foliar Mg concentration were the result of an inhibition of seedling response to NO(3) (-) mediated by Al in solution, rather than enhanced Al toxicity resulting from growth in the presence of elevated NO(3) (-) concentrations.     

Freezing cycles enhance winter injury in Picea rubens

We examined changes in chlorophyll absorbency in red spruce (Picea rubens Sarg.) foliage in response to simulated freezing cycles. Current-year branch tips were collected from 16 trees on January 8, January 20, February 8 and February 26, 1996. Tissue was subjected to freezing cycle treatments with a minimum of -35 degrees C and a maximum of 3 degrees C for a one-cycle treatment, and -9, -6, -3, 0 or 3 degrees C for four-cycle treatments. Samples were frozen at a rate of 5 degrees C h(-1), and warmed at 12 to 15 degrees C h(-1). Controls were held at -9 degrees C. Temperatures during the three-day periods preceding each sample date averaged -18, 4.7, -9.6 and 3.7 degrees C, respectively. On January 8, treated trees showed no significant (P > 0.1) increase in the breakdown of chlorophyll, as measured by the ratio of chlorophyll a absorbency (435 nm) to phaeophytin a absorbency (415 nm), compared with control branch tips. On later sampling dates, seven trees consistently exhibited needle reddening and nine exhibited few symptoms (< 10% of total needle surface reddened) after four-cycle treatments. On February 26, chlorophyll degradation in trees with needle reddening differed (P < 0.05) from the control by 26, 26, 16, 14 and 15% for the 3, 0, -3, -6 and -9 degrees C maxima, respectively. No detectable chlorophyll degradation occurred after a one-cycle treatment in any trees on any sampling date. Freezing cycles with sub-zero maxima and a -35 degrees C minimum enhanced winter injury in red spruce after a midwinter thaw had rendered the trees susceptible to freezing damage.     

Differential adsorption of Al, Ca, and Mg by roots of red spruce (Picea rubens Sarg.)

The surface adsorption characteristics of red spruce (Picea rubens) roots were examined as a function of changes in external acidity and cation concentrations. Root cation exchange capacity varied significantly with changes in pH, increasing from 110 micromol(c) g(-1) at pH 3.5 to 155 micromol(c) g(-1) at pH 4.5, and reaching 250 micromol(c) g(-1) at pH 7.0. In general, Al adsorption by spruce roots was much greater than either Ca or Mg adsorption under the same initial conditions. However, root affinity for the divalent cations was proportionately much more sensitive to pH changes than was root affinity for Al. The fractions of adsorbed Ca and Mg increased by 50 to 100% as pH increased from 3.5 to 4.5, whereas the fraction of adsorbed Al remained relatively constant at both initial pH conditions. Competition experiments at pH 3.5 and 4.5 indicated that Al adsorption was strongly favored over Ca adsorption, except at low Al concentrations (~10 micromol l(-1)), high solution Ca(2+)/Al(3+) ion activity ratios (> 1.5 to 5.0), and at the higher pH. These results suggest that cell wall exchange sites in red spruce roots will tend to become progressively saturated with Al under the prevailing conditions of many acidic forest soils. To the extent that root adsorbed Al interferes with the active uptake of Ca and Mg, this process of competitive cation adsorption can contribute to impaired mineral nutrition in the spruce forest community.     

Impact of eastern dwarf mistletoe (Arceuthobium pusillum) infection on the needles of red spruce (Picea rubens) and white spruce (Picea glauca): oxygen exchange, morphology and composition

Eastern dwarf mistletoe (Arceuthobium pusillum Peck) is a hemiparasitic angiosperm that infects white spruce (Picea glauca (Moench) Voss) and red spruce (P. rubens Sarg.) in northeastern North America. The effects of mistletoe infection differ substantially between white and red spruce, with white spruce suffering greater infection-induced mortality. In the present study, we sought to determine the role that species-specific differences in needle-scale responses to parasitism may play in the observed differences in the effect of infection on host tree health. Based on the measurements made, the most apparent effect of parasitism was a reduction in needle size distal to infections. The magnitude of this reduction was greater in white spruce than in red spruce. Eastern dwarf mistletoe was a sink for host photosynthate in red spruce and white spruce; however, there were no adjustments in needle photosynthetic capacities in either host to accommodate the added sink demands of the parasite. Needle total nonstructural carbohydrate concentrations (TNC) were also unaltered by infection. Red spruce needles had higher TNC concentrations despite having lower overall photosynthetic capacities, suggesting that red spruce may be more sink limited and therefore better able to satisfy the added sink demands of parasitic infection. However, if carbon availability limits the growth of the mistletoe, one may expect that the extent of the parasitic infection would be greater in red spruce. Yet in the field, the extent of infection is generally greater in white spruce. Taken together, these results suggest that dwarf mistletoe may not substantially perturb the carbon balance of either host spruce species and that species-specific differences in needle-scale responses to the parasite cannot explain the contrasting effects of infection on white spruce and red spruce.     

Influence of temperature and leaf-to-air vapor pressure deficit on net photosynthesis and stomatal conductance in red spruce (Picea rubens)

The roles of temperature (T) and leaf-to-air vapor pressure deficit (VPD) in regulating net photosynthesis (A(net)) and stomatal conductance (G(s)) of red spruce (Picea rubens Sarg.) were investigated in a field study and in a controlled environment experiment. Both A(net) and G(s) exhibited a relatively flat response to temperatures between 16 and 32 degrees C. Temperatures between 32 and 36 degrees C markedly decreased both A(net) and G(s). Vapor pressure deficits above 2 kPa had significant effects on both A(net) and G(s). The influence of VPD on A(net) and G(s) fit a linear response model and did not interact significantly with T effects.     

Modeling shoot water contents in high-elevation Picea rubens during winter

During the winter of 1990-1991, a meteorological tower was established at an 880-m elevation site within the spruce-fir zone on Mt. Moosilauke, New Hampshire, USA. Hourly means of air, needle and trunk temperatures, wind velocity, relative humidity and solar radiation were recorded. On a weekly basis, shoots that had elongated during the preceding growing season were collected from four red spruce (Picea rubens Sarg.) trees and their relative water contents (RWC) determined. Cuticular resistances of needles from these shoots were measured four times during the winter.Measured meteorological parameters were used in a previously developed model to simulate changes in red spruce shoot RWC during the winter. The modeled results were compared to measured shoot RWCs. The predictive power of the model was improved when it was modified to include measured values of cuticular resistance and needle and trunk temperatures. The new version of the model accurately predicted RWC from late December 1990 to the beginning of April 1991, after which spring recharge appeared to occur. We conclude that water lost from foliage was easily replaced by stored reserves and that uptake of water by the roots was not required to maintain an adequate foliar water content during the winter.     

Effect of elevated temperatures on carbon dioxide exchange in Picea rubens

We examined some of the physiological reasons that may underlie past and expected future migrations of red spruce (Picea rubens Sarg.) by evaluating the effects of high temperatures on photosynthesis and respiration of trees growing on Whiteface Mountain, NY. At temperatures of 35-40 degrees C, the trees exhibited a zero or negative carbon balance. Higher temperatures resulted in cellular disorganization and death. Temperatures around 30 degrees C resulted in reduced CO(2) uptake, a condition that could decrease future reproductive output and competitive stature. We conclude that thermal intolerance explains, at least in part, the absence of red spruce at low elevations and latitudes where temperatures of >/= 30 degrees C occur. We suggest that the thermosensitivity of this species is important with respect to global climate trends and migration patterns.     

Performance and Frost Hardiness of Picea sitchensis x Picea glauca Hybrids in Scotland

The growth of seven Picea sitchensis x Picea glauca hybridswas compared with the growth of two P. sitchensis provenancesand Picea glauca var.albertiana at two sites in northern Scotland.The sites were at Aultmore (an exposed, dry site with a mineralsoil) and Shin (a frosty, wet site with deep peat). They wereof the type considered more suited to Pinus contorta than P.sitchensis. At age 10, in 1984, most of the hybrids, at both sites, wereabout 10 and 20 per cent taller than P. sitchensis of Masset(Q.C.I.) and Ketchikan (Alaska) provenance, respectively. P.glauca var. albertiana grew very poorly, especially at Aultmore. At Aultmore, the frost hardiness of three of the tallest hybrids,the two P. sitchensis provenances, and P. glauca var.albertiana,was tested at about 3-weekly intervals throughout 1982 and 1983.Detached shoots were subjected to artificial frosts in a programmablechamber. P. glauca var. albertiana was frost susceptible atbudburst, but at all other times it was relatively very frosthardy (eg. to^–10°C in mid-August). Also, the hybridswere consistently more hardy than P. sitchensis of even Ketchikan(Alaska) provenance from July onwards. However, the hybridswere less frost hardy than P. sitchensis of Masset (Q.C.I.)provenance in early spring (they dehardened a week earlier inMarch-April) and their buds were equally as frost susceptibleat the time of budburst. In 1983, trees of P. glauca var. albertianaburst their buds about a week sooner than P. sitchensis. It was concluded that P. sitchensis x P. glauca hybrids canperform better than P. sitchensis at sites considered ‘marginal’for P. sitchensis, and that their good performance may be partlyattributed to, or associated with, their greater summer andautumn frost hardiness. A programme of inter-specific hybridizationis being pursued.     

Frost hardiness, bud phenology and growth of containerized Picea mariana seedlings grown at three nitrogen levels and three temperature regimes

We studied the influence of temperature and near- and sub- optimal mineral nutrition of black spruce seedlings (Picea mariana [Mill.] B.S.P.) during their second growing period on bud set, bud development, growth, mineral content and cold tolerance. Bud break and growth after bud break were also studied. Seedlings were grown for 106 d in growth chambers under three temperature regimes in combination with three concentrations of a fertilizer. They were then cold hardened for 56 d and dehardened for 66 d.Under these near- and sub-optimal N levels, bud formation occurred during the growing season. Bud formation was accelerated with decreasing fertilization, but was not affected by temperature treatments. Needles from seedlings with 0.64% N (dry mass basis) before hardening did not harden. Those with 0.87% N showed a lesser degree of hardiness than those with 1.28% N. Stem diameter increased at the beginning of the hardening period. During this acclimation period, shoot dry mass decreased with time at a constant rate and at the same rate over time for all treatments whereas root dry mass was more variable. Total number of needle primordia was low and no difference was observed among growing conditions. Bud break was similar in all treatments. Following bud break, shoot height and stem diameter increases were small but their magnitude varied with the nutritional regimes applied during the previous growing period. During hardening, nitrogen concentration of shoot tissues first increased and then decreased; phosphorus concentration first increased and then remained stable; potassium concentration remained stable. Concentration of these three elements generally decreased in the roots during this hardening.     

Frost hardiness gradients in shoots and roots of picea mariana seedlings

Frost hardiness of tissues along the length of the stem and the root was investigated in first‐year black spruce (Picea mariana (Mill.) B.S.P.) seedlings. Frost hardiness of 1 cm long stem and root segments was evaluated based on Index of Injury, calculated from post‐freezing electrolyte leakage. Frost hardiness was tested approximately weekly beginning seven weeks after seedlings were transferred from an 18 to a 10 h photoperiod, both at day/night temperatures of 26°C/16°C. Trees were transferred to temperatures of 10°C day and 5°C night at a 10 h photoperiod after a further 18 days. Frost hardiness was greater at the terminal bud and least at the root tips. Although shoots were generally more frost hardy than roots, differences in hardiness along the stem and root axes were gradual, rather than abruptly differing at the shoot‐root interface. All tissues, including root tips, increased in frost hardiness after conditioning for 18 days under short photoperiods (10 h) and warm temperatures (26?C/16°C, day/night). Under cold temperatures (10°C/5°C, day/night) all tissues, excepting the root tips, tolerated — 16°C with little subsequent electrolyte leakage.     

Solution aluminum and transpiration in Picea rubens and Gleditsia triacanthos seedlings

Soil solution Al may contribute to red spruce (Picea rubens Sarg.) decline by inhibiting transpiration (Klein 1985). This study examines how Al affects transpiration in red spruce and honeylocust (Gleditsia triacanthos L.) seedlings and explores mechanisms for the observed responses. Red spruce seedlings were grown in dilute nutrient solutions containing 0 to 925 microM Al at pH 4.5 or 3.8. Solution Al had no effect on the transpiration, root hydraulic conductivity (K(r)) or shoot water potential (Psi) of red spruce. Transpiration of honeylocust was higher in the presence of 100 to 350 microM Al than 0, 500, or 600 microM Al. The Al-induced increases in transpiration of honeylocust were unrelated to shoot Psi or root K(r), but were closely correlated with the greater root lengths produced by intermediate levels of Al. In neither species was transpiration correlated with tissue concentrations of Al, P, K, Ca or Mg.     

Calcium addition at the Hubbard Brook Experimental Forest increases sugar storage, antioxidant activity and cold tolerance in native red spruce (Picea rubens)

In fall (November 2005) and winter (February 2006), we collected current-year foliage of native red spruce (Picea rubens Sarg.) growing in a reference watershed and in a watershed treated in 1999 with wollastonite (CaSiO(3), a slow-release calcium source) to simulate preindustrial soil calcium concentrations (Ca-addition watershed) at the Hubbard Brook Experimental Forest (Thornton, NH). We analyzed nutrition, soluble sugar concentrations, ascorbate peroxidase (APX) activity and cold tolerance, to evaluate the basis of recent (2003) differences between watersheds in red spruce foliar winter injury. Foliar Ca and total sugar concentrations were significantly higher in trees in the Ca-addition watershed than in trees in the reference watershed during both fall (P=0.037 and 0.035, respectively) and winter (P=0.055 and 0.036, respectively). The Ca-addition treatment significantly increased foliar fructose and glucose concentrations in November (P=0.013 and 0.007, respectively) and foliar sucrose concentrations in winter (P=0.040). Foliar APX activity was similar in trees in both watersheds during fall (P=0.28), but higher in trees in the Ca-addition watershed during winter (P=0.063). Cold tolerance of foliage was significantly greater in trees in the Ca-addition watershed than in trees in the reference watershed (P<0.001). Our results suggest that low foliar sugar concentrations and APX activity, and reduced cold tolerance in trees in the reference watershed contributed to their high vulnerability to winter injury in 2003. Because the reference watershed reflects forest conditions in the region, the consequences of impaired physiological function caused by soil Ca depletion may have widespread implications for forest health.     

Autumn Frost Damage on Young Picea sitchensis 2. Shoot Frost Hardening, and the Probability of Frost Damage in Scotland

The natural increase in frost hardiness of detached shoots ofPicea sltchensis during August to November was measured usinga programmable freezing chamber. Oregon, Queen Charlotte Islandsand Alaskan provenances were compared, and the effects on hardeningof long days, warm temperatures and frosts were determined.A computer model was constructed to mimic the observed patternsof autumn frost hardening, as functions of air minimum temperatures,daylengths and the occurrence of frosts. The model was used(a) to describe the pattern of autumn frost hardening at differentsites in northern Britain, using past meteorological records,and hence (b) to determine when frosts occurred that might havedamaged young trees. The model accurately predicted known instancesof autumn frost damage at Kirroughtree and Carnwath. The predicted probability of autumn frost damage on young treesof P. sitchensis in upland areas of Scotland was much lowerthan that previously predicted for spring frost damage. Theestimated return time for autumn frost damage to an Oregon provenanceat Eskdalemuir was 8.3 years, and the return time for a Q.C.I.provenance was longer than 10 years. Most damaging frosts occurredin October, but frosts like those on 13–15 October 1971,which followed warm weather and caused wide spread damage inScotland, have been quite rare. Alaskan provenances would rarelybe damaged by autumn frosts, nor would trees of Q.C.I. provenancegrowing in lowland areas of Scotland, or at Masset on the QueenCharlotte Islands.     

The importance of seedling competition in the segregation of spruce and fir in the southern Appalachians

Fraser fir (Abies fraseri [Pursh] Poir.) and red spruce (Picea rubens Sarg.) are codominants of southern Appalachian spruce-fir forests. Fraser fir generally dominates above 1740 m, while red spruce usually dominates below this elevation. This study was designed to determine whether the present segregation of the two species along elevational gradients is associated with seedling competition or contrasting physiological responses to environmental factors. Seedlings were grown for two years in a replacement series experiment along two elevational transects extending from 1300 m to 1900 m, and harvested for growth analysis.Competition increased with decreasing elevation and Fraser fir was apparently the stronger competitor. Mortality was inversely correlated with elevation and was most evident in red spruce in May, and in fir in late summer. Fir growth increased with elevation and red spruce grew most at the middle elevation (1600 m). Neither competitive interactions nor growth responses completely explained the elevational segregation of red spruce and Fraser fir. We hypothesize that the early-successional Fraser fir quickly dominates at higher elevations following disturbance and that continued disturbance will favor its dominance at higher elevations. However, the late-successional noncompetitive red spruce slowly establishes itself and, while consistently present, may only dominate in the absence of disturbance and where other species are at the margins of their distribution.     

Frost hardening spruce container stock for overwintering in Ontario

Difficulties overwintering container stock in northern Ontario led to the development of the extended greenhouse culture hardening regime for spruce seedlings. Laboratories to measure shoot frost hardiness and evaluate terminal bud development were established to monitor nursery crops being hardened using this regime. Information on frost hardiness and bud development provided by these laboratories has been used by nursery managers to determine readiness of container seedlings for overwintering. Since 1982, over 200 stock lots have been monitored by these operational laboratories. This database can be used to determine the importance of nursery cultural factors and seed source on frost hardening. The database shows large differences between nurseries in approach to hardening seedlings which were reflected in levels of freezing damage, winter desiccation and overwintering success. Rates of frost hardening (i.e., the interval between terminal bud initiation and attainment of a --15°C level of shoot frost hardiness) of crops produced in north central Ontario failed to show significant seed source effects. The rate of frost hardening was faster in crops producing fewer needle primordia in terminal buds.     

Influence of nutrient supply on spring frost hardiness and time of bud break in Norway spruce (Picea abies (L.) Karst.) seedlings

When spring frosts occur on recently planted forest sites, severe damage may occur to the seedlings. The aim of the present study was to test how different low levels of nutrient concentrations in Norway spruce (Picea abies (L.) Karst.) seedlings affected spring frost hardiness and time of bud break. Seedlings were grown in a greenhouse for one season and supplied with fertiliser containing 22, 43 and 72 mg N l^–1, respectively. The treatments resulted in needle nitrogen concentrations ranging from 0.9 to 1.8% in autumn. After winter storage at 0 °C, bud break was recorded on seedlings growing in the greenhouse, outdoors and in growth chambers at 12 °C and at 17 °C. Freezing tests were performed on seedlings directly removed from winter storage and following one week growth in the greenhouse. Seedlings receiving fertiliser with 43 mg N l^–1 had less freezing injury than the two other fertilisation treatments in the present study. The earliest bud break occurred in seedlings receiving 72 mg N l^–1.