Carbohydrate requirements for the development of black spruce (Picea mariana (Mill.) B.S.P.) and red spruce (P. rubens Sarg.) somatic embryos

Different carbohydrates were investigated for somatic embryo development of black spruce and red spruce. They were tested in a basal maturation medium consisting of Litvay's salts at half-strength containing 1 g l^-1 glutamine, 1 g l^-1 casein hydrolysate, 7.5 M abscisic acid, and 0.9% Difco Bacto-agar. A comparison of different sucrose concentrations showed that 6% was optimal for embryo development. Among the nine carbohydrates tested, sucrose, fructose, glucose, maltose, and cellobiose supported embryo development while arabinose, mannitol, myo-inositol, and sorbitol did not. A comparison of sucrose, glucose, and fructose at three concentrations showed that the general pattern of response for both species followed concentration expressed as a percentage, independent of the molarity of carbohydrate in the medium. Interspecific differences were observed concerning carbohydrate requirements. For red spruce, 6% fructose was found best for embryo development, while no such preference was observed for black spruce. No significant difference was observed in the number of embryos produced with 6% sucrose or 3% sucrose plus an equimolar concentration of either mannitol, sorbitol, or myo-inositol in the maturation medium, suggesting that the effect of the carbohydrate on the maturation was partly osmotic.     

Nitrogen mineralization in high elevation forests of the Appalachians. I. Regional patterns in southern spruce-fir forests

Annual and seasonal rates of net nitrogen mineralization were determined for 19 sites in the spruce-fir forests of the Southern Appalachian Mountains. These sites included high and low elevation stands of red spruce (Picea rubens Sarg.) and Fraser fir (Abies fraseri (Pursh.) Poir.) on east and west exposures on Whitetop Mountain, Virginia; Mt. Mitchell, North Carolina; and Clingman's Dome in the Great Smoky Mountains National Park. Mineralization rates were determined using in situ soil incubations in PVC tubes with ion exchange resin bags placed in the bottom of the tubes to collect leachate. Throughfall was collected in resin bags placed in the top of the tubes. Average initial NH₄-N + NO₃-N ranged from 0.6 to 4.8 kg N/ha across all plots, and average mineralization rates ranged from 26 to 180 kg-N ha⁻¹ yr⁻¹. Throughfall ranged from 18 to 32 kg-N ha⁻¹ yr⁻¹ with NH₄-N accounting for about two-thirds of the throughfall N across all sites. Throughfall and mineralization rates were not related to elevation or exposure. The high rates of N mineralization and relatively high nitrate concentrations indicate that leaching losses of nitrogen and associated cations could be substantial. Requests for offprints     

Biochemical symptoms of stress in the mycorrhizal roots of Norway spruce (Picea abies)

Summary The mycorrhizal activity of spruce in a mixed-wood forest was monitored over 1 year by measuring biochemical characters in fine roots of six canopy trees and of a regrowth stand. The concentration of adenosine 5-triphosphate (ATP), a measure of living biomass, showed two peaks per year, one at bud break and one after main shoot growth. The concentration of storage polysaccharides in mycorrhizae showed the same cycles even more pronouncedly. It is proposed that these changes reflect growth and senescence of mycorrhizae and that the timing of the cycles is controlled by translocation of assimilates from the shoot. Differences between mycorrhizae collected from canopy trees and the regrowth stand were small and not significant. Characters known to be related to fungal activity of the mycorrhizal symbiosis (concentration of trehalose, glucose uptake, respiration) also varied little among the six canopy trees. Large differences among fine-root samples from different canopy trees, however, were detected in the concentrations of ATP and storage polysaccharides, measures which seemed to be physiologically integrated within trees. If low concentrations in roots precede losses of foliage from trees, these two symptoms could be used as early indicators of growth decline in individual spruce trees.     

Food consumption and utilization by 6th-instar larvae of spruce budworm, Choristoneura fumiferana: a comparison on three Picea (spruce) species

The effects of tree species, shoot age, and sex of larvae on food consumption and utilization were investigated in a factorial experiment. Diets prepared from lyophilized and powdered current-year shoots, from two age classes (sample dates) of Picea glauca (Moench) Voss, Picea rubra, Sarg., and Picea mariana (Mill.) B.S.P., were evaluated for eight performance criteria. All factors had significant effects on all criteria except for tree species on assimilation efficiency, shoot age on development time, and sex on net growth efficiency. Eight of the possible 24 two-factor interactions and one of the possible eight three-factor interactions were significant. Food utilization efficiencies and relative growth rate were in the low and moderate characterization limits for immature arthropods; relative consumption rates were high. The higher performance values on Picea glauca were related to this species greater vulnerability to spruce budworm. Performance criteria of similar larvae on an artificial diet were compared with those on the foliage-diets.

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Résumé L'auteur a déterminé la quantité d'aliments utilisées, à 22±0,5 °C, par des larves de Tordeuse des bourgeons de l'épinette parvenues à leur sixième stade de développement qui s'étaient nourries d'un régime artificiel et d'aiguilles de l'année en cours lyophilisées et réduites en poudre, prélevées à deux dates différentes, à 10 jours d'intervalle, et provenant d'épinettes blanchés (Picea glauca [Moench] Voss), d'épinettes rouges (Picea rubra Sarg.) et d'épinettes noires (Picea mariana [Mill.] B.S.P.). Les femelles ont invariablement ingéré une quantité significativement plus importante de tous les régimes que les mâles, consommant 24% de plus d'aiguilles d'épinette noire et 73% de plus d'aiguilles d'épinette blanche.Les femelles prenaient en général plus de temps pour parvenir au stade de chrysalide et avaient un taux d'utilisation des aliments significativement plus élevé. L'efficacité nette de la croissance chez les deux sexes ne présentait toutefois aucune différence significative. La performance des larves ayant ingéré des aiguilles provenant de deux classes d'âge différait de façon significative mais non pas uniforme d'un régime à l'autre: les aiguilles d'épinette blanche plus récentes donnaient une performance généralement plus élevée contrairement aux aiguilles plus jeunes d'épinette rouge et d'épinette noire. Les larves ingéraient une quantité significativement plus élevée, avaient un taux de consommation et de croissance supérieur et augmentaient davantage leur biomasse avec des aiguilles d'épinette blanche. L'efficacité d'utilisation des aliments et le taux de croissance relatif étaient de faibles à modérément faibles pour un lépidoptère arboricole phyllophage; la vitesse d'ingestion relative était élevée. La qualité des aiguilles des trois essences d'épinettes était faible. Les aiguilles d'épinette blanche permettaient aux larves d'avoir une performance significativement plus élevée, performance mesurée à partir de plusieurs critères, alors que les aiguilles d'épinette rouge et d'épinette noire donnaient une performance égale. La supériorité des performances obtenues avec l'épinette blanche explique la plus grande vulnérabilité de cette essence à la Tordeuse des bourgeons de l'épinette.

     

Fine-root vitality in a Norway spruce stand subjected to various nutrient supplies

The vitality of fine roots in a Norway spruce stand subjected to application of ammonium sulphate (NS), wood ash (A) and nitrogen-free fertilizer (V) respectively, was investigated using i) vitality classification of fine roots based on morphological characteristics and ii) the triphenyl tetrazolium chloride (TTC) method of estimating dehydrogenase activity.Although the NS-treated areas showed a 30% increase in above-ground production in response to the NS-application, the vitality of the fine-root system in the humus layer appeared to be in a state of deterioration, as indicated by a decrease in fine-root biomass, an increased amount of dead fine (0–1 mm) and small (1–2 mm) roots, a decreased specific root length (SRL = fine root length/fine root dry weight) and an increased dehydrogenase activity. The impact of the the A and V treatments was reflected in a decrease in fine-root biomass and an increase in SRL. The results make it clear that in order to study the vitality of forest trees, both fine-root studies and studies of above-ground tree parts are necessary.     

A measuring device to quantify deposition of fogwater and ionic input by fog on small spruce trees

Summary A measuring device is described that allows the monitoring of fogwater deposition on to small spruce trees. The mass of fogwater deposition ranged from 41.9 to 60.5 ml/m² of needle surface area. Fogwater was collected, and ion concentrations in fogwater and calculated masses of deposited ions are presented.     

Performance of two Picea abies (L.) Karst. stands at different stages of decline

Summary A declining Picea abies (L.) Karst. stand produced as much foliage and branches as a healthy stand but less stemwood at a similar leaf area index and climate. Nutrient analyses revealed that most biomass components at the declining site had lower concentrations of calcium and magnesium, but similar nitrogen and potassium (except for lower potassium in younger needles) and higher phosphorus, manganese and aluminum than the respective components at the healthy site. Comparison of these data with the results from studies on the nutrition and growth of P. abies seedlings (Ingestad 1959) led to the conclusion that the healthy stand is in a balanced nutritional state, while trees at the declining stand have only 56% of the foliar magnesium concentration required to permit growth at a rate which could be achieved at their nitrogen status. It appears that acidic deposition, which involves an input of nitrogen and a leaching of cations from the soil, causes an imbalance in the availability of nitrogen and magnesium. Growth is eventually reduced as magnesium becomes limiting.     

Seasonal changes in the pigments, carbohydrates and growth of red spruce as affected by ozone

Two-year-old red spruce seedlings were exposed to various levels ot ozone, from 0.4 to 3 times ambient levels, in open-top chambers in Ithaca, NY, USA. Exposures, which varied with changes in day length, commenced on May 30, 1987 and continued until December 14, 1987. Seedling biomass, carbohydrate contents, pigment contents, and rate of electron transport were assessed twice monthly during the fumigation period. Orthogonal quadratic or cubic polynomials were used to model the response through time each variable measured. A one-way analysis of variance model was fitted to every regression coefficient in each polynomial model to test for ozone effects on seasonal physiological patterns. Ozone did not influence growth, foliar pigment content, foliar starch content, root carbohydrate content, or rate of electron transport. The seasonal change of needle raffinose content differed between exposed to low (0.4 ×, 1×) and high (2×, 3×) ozone levels. There was also a trend towards reduced total soluble sugar content foliage during late autumn in higher ozone treatments.     

Changes in root growth patterns of (Picea abies) spruce roots by inoculation with an ectomycorrhizal fungusPisolithus tinctorius and jasmonic acid treatment

Symbiosis between fungi and plant roots forming a mycorrhiza involves extensive interactions at the molecular level between both partners. The role of plant hormones in the regulation of mycorrhizal infection is not known to involve jasmonates. Their endogenous levels increase during pathogen attack; however, little has been done on their involvement in mycorrhizae. In our recent work, root growth patterns of 2-month-old spruce seedlings after inoculation withPisolithus tinctorius and/or jasmonic acid (JA) treatment were studied using a paper-sandwich technique. Changes in root length, the degree of branching, presence and length of root hairs, and infection parameters were followed using a stereomicroscope. The first mycorrhizal contact of hyphae with roots was significantly accelerated upon treatment with 0.5 M JA. Interactions between root hairs and fungal hyphae were seen by scanning electron microscopy. The multiplication of root hairs of non-mycorrhized seedlings treated with 5.0 M JA and changes of the root surface were observed by the same technique.     

Activity and activation state of ribulose-1,5-bisphosphate carboxylase of spruce trees with varying degrees of damage relative to the occurrence of novel forest decline

The aim was to determine whether a reduced carboxylation efficiency in needles of damaged spruce trees (Picea abies), is derived from a direct impairment of the ribulose-1,5-bisphosphate carboxylase (RuBP carboxylase) or there is an indirect inhibition of the RuBP carboxylase. In 1985, 1986 and 1987 measurements of RuBP carboxylase activity were carried out at three locations. Trees of different ages and degrees of damage were examined. RuBP carboxylase was assayed using both a rapid extraction method to determine the initial activity and an in vitro test after total activation to determine the total activity. The activation state was calculated as the ratio of initial activity to total activity.Within three vegetation periods the total activity in needles of damaged and apparently healthy or slightly damaged spruce trees indicated no definite difference in the annual average. On the other hand, in damaged needles a continued decline of the actual activation of RuBP carboxylase was established. The observation of continued depression of the activation state of the enzyme in needles of damaged spruce trees can possibly be due to a reduced photosynthetic electron transport rate.The measurements of the soluble protein content indicate a tendency to increased amounts in the needles of damaged trees. In accordance, a considerable increase of the activity of some enzymes like glutamine synthethase, phosphoenol-pyruvate carboxylase, and catalase could be noticed. However, there is no clear connection between the RuBP carboxylase and the content of soluble proteins.Abbreviations chl chlorophyll a+b, dw-dry weight, i.a-initial activity - P-700 reaction center of photosystem I - PVP polyvinylpyrrolidone 25 - RuBP ribulose-1,5-bisphosphate - RuBPCase ribulose-1,5-bisphosphate carboxylase - t.a. total activity     

Life expectancy of spruce needles under extremely high air pollution stress: performance of trees in the Ore Mountains

Summary Photosynthetic performance of Norway spruce needles [Picea abies (L.) Karst.] was measured over a 1-year period. The trees grew in an area of heavy air pollution and forest decline on a mountain ridge in the eastern Ore Mountains (Czech Republic). Photosynthetic capacity, as well as light use efficiency, decreased dramatically with time, starting in July (2 months after bud-break) to finally reach zero (respiration only) by February of the following year. Two months later all needles from upper crown parts were shed. Needles from lower crown parts, on the other hand, were undamaged. The chlorophyll and Mg content decreased transiently during the cold season, with Mg reaching deficiency thresholds during winter. However, total sulfur, as well as organic and sulfate S increased with time. The increase was higher in needles from the upper parts of the crown, which were exposed to windy air throughout the year, than in the lower parts of the crown, which were covered by grass during summer and by snow during most of the winter.     

Genotype dependent blue and red light inhibition of the proliferation of the embryogenic tissue of Norway spruce

Summary The influence of light quality on the proliferation of embryogenic tissue of three genotypes of Norway spruce (Picea abies [L.] Karst), with different capacities for mature somatic embryo production, was studied. The proliferating tissues were subjected to light from commercially available light sources: Philips TLD Warm White 36W/29, Philips TLD Blue 18W/18, Philips TLD Red 36W/15, Osram L Fluora 36W/77 and Sylvania Far Red 7080, for 18 h a day with the photon flux (PAR) at 30 μmol m⁻²s⁻¹. The effect of light quality on the growth of embryogenic tissue was strongly genotype dependent. In genotype 164-4 tissue proliferation was strongly inhibited by blue and red light. Genotype 86∶52 reacted in a similar way, but not as strongly as 164-4, whereas the tissue of genotype 186-3 was almost insensitive to light quality and grew fast in all light conditions.     

Species-specific RAPD fingerprints for the closely related Picea mariana and P. rubens

Species-specific molecular markers were designed to assist in the identification of closely related black spruce (Picea mariana [B.S.P.] Mill.) and red spruce (P. rubens Sarg.) in northeastern North America. Trees from six provenances of black spruce and three provenances of red spruce were sampled from outside the sympatric zone. They were first classified using a composite index of five qualitative morphological traits. The species-specific genetic markers were developed using random amplified polymorphic DNAs (RAPD) and a combination of bulk sample and individual tree analyses. Each species bulk sample was constructed from DNAs obtained from 12 trees that were from outside the sympatric zone and showed a morphological composite index specific of each species. A total of 161 primers were screened with the bulk samples. From these, 52 primers showing segregating fingerprints were further screened with the individual trees. Most of the markers observed were shared by the two species, and there was less diversity in P. rubens. A small number of markers were found to be monomorphic or nearly monomorphic and specific to either P. mariana or P. rubens. These markers remained species-specific when F₁ progenies derived from independent intraspecific crosses were screened, and they were subsequently found to co-segregate in hybrids derived from independent interspecific crosses here used as controls.     

ABA consumption in norway spruce (Picea abies) and white spruce (Picea glauca) somatic embryo cultures

Summary We investigated abscisic acid (ABA) metabolism among Norway and white spruce somatic embryo cultures which exhibited differences in maturation response when placed on racemic abscisic acid [(±)-ABA]. Differences in metabolic rate among the spruce genotypes could affect the ABA pool available for the maturation process, and might therefore be responsible for the differences in maturation response. The production of cotyledonary (stage 3) somatic embryos in cultures (genotypes) of Norway spruce (PA86:26A and PA88:25B) and of white spruce (WS1F cryoD and WS46) was compared. In each species pair one of the two genotypes failed to show stage 3 embryo development (respectively, PA88:25B and WS46). The investigation of ABA metabolism of each species pair showed that no substantial differences in ABA consumption or in the production of metabolites occurred. In each case ABA was metabolized to phaseic acid and dihydrophaseic acid over the 42-day culture period, metabolites were recoverable from the agar-solidified medium, and the sum of residual ABA and metabolites were equivalent to the ABA initially supplied. The results indicate that the process of ABA metabolism occurs essentially independently of somatic embryo maturation. NRCC no. 37345.     

Contents of ATP and ADP in needles of Norway spruce in relation to their development,age, and to symptoms of forest decline

Summary Pool sizes of ATP and ADP were analysed in freeze-stopped, lyophilised homogenates of needles from Norway spruce [Picea abies (L.) Karst.]. Control experiments in which possible changes in adenylate pools during sample acquisition were investigated did not reveal significant differences between needles taken from branches in situ or within a 30-min period after cutting off a branch. In addition, pool sizes of ADP and ATP were not affected by changes in light intensity (between 60 and 1500 E^*-m^-2*s^-1), which inevitably occur when samples have to be taken from the upper region of older trees. Levels of ATP and ADP showed considerable seasonal changes (May through October) with the highest ratios of ATP/ADP in developing needles. In general, there was a tendency towards increased ratios of ATP/ADP with increasing needle age. This observation was corroborated by analyses of needles from spruce trees of different age and growing under different conditions. Needles from declining trees or from trees specifically fumigated with low concentrations of ozone and sulphur dioxide had significantly increased ratios of ATP/ADP compared to controls. The results are discussed with respect to physiological responses connected with natural senescence and induced ageing.