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PubMed2002

Impact of four years exposure to different levels of ozone, phosphorus and drought on chlorophyll, mineral nutrients, and stem volume of Norway spruce, Picea abies.

Saplings of one clone of Norway spruce, Picea abies (L.) Karst, were planted in 120 l pots in 1991 and exposed to three levels of ozone, two levels of phosphorus and two levels of water supply in 42 open-top chambers (OTCs), during 1992-1996. The effects of pots and OTCs were also tested. Nutrient concentrations of the needles were not affected by ozone, while the low phosphorus supply (LP) and drought stress (D) treatments had significant effects on several mineral nutrients, e.g. phosphorus, calcium, magnesium, manganese, sulphur and boron. Ozone reduced the chlorophyll concentration in the 2- and 3-year-old needles in 1994 and 1995. The highest ozone concentration reduced the stem volumes (- 8%), as well as the stem lengths (- 5%), of the saplings in 1993 and 1994, after two and three years of exposure. After the fourth growing season this ozone-induced reduction in stem volume disappeared which might be caused by pot limitation. LP supply and D both caused large decreases in the stem volume and length. The needles from LP treatment had as high P concentration as 1.2-1.5 mg g-1, implying a need for increasing the critical value for phosphorus. The OTC enclosure stimulated the stem volume growth significantly compared to saplings growing in ambient plots. This was suggested to be attributed to the slightly higher temperature in the OTCs. The overall result is that ozone in southern Sweden is likely to have negative effects on Norway spruce trees, although much less than other environmental factors, e.g. water and phosphorus.

Journal Article↗
PubMed2002

Lignification related enzymes in Picea abies suspension cultures.

Activity of a number of enzymes related to lignin formation was measured in a Picea abies (L) Karsten suspension culture that is able to produce native-like lignin into the nutrient medium. This cell culture is an attractive model for studying lignin formation, as the process takes place independently of the complex macromolecular matrix of the native apoplast. Suspension culture proteins were fractionated into soluble cellular proteins, ionically and covalently bound cell wall proteins and nutrient medium proteins. The nutrient medium contained up to 5.3% of total coniferyl alcohol peroxidase (EC 1.11.1.7) activity and a significant NADH oxidase activity that is suggested to be responsible for hydrogen peroxide (H2O2) production. There also existed some malate dehydrogenase (EC 1.1.1.37) activity in the apoplast of suspension culture cells (in ionically and covalently bound cell wall protein fractions), possibly for the regeneration of NADH that is needed for peroxidase-catalysed H2O2 production. However, there is no proof of the existence of NADH in the apoplast. Nutrient medium peroxidases could be classified into acidic, slightly basic and highly basic isoenzyme groups by isoelectric focusing. Only acidic peroxidases were found in the covalently bound cell wall protein fraction. Several peroxidase isoenzymes across the whole pI range were detected in the protein fraction ionically bound to cell walls and in the soluble cellular protein fraction. One laccase-like isoenzyme with pI of approximately 8.5 was found in the nutrient medium that was able to form dehydrogenation polymer from coniferyl alcohol in the absence of H2O2. The total activity of this oxidase towards coniferyl alcohol was, however, several orders of magnitude smaller than that of peroxidases in vitro. According to 2D 1H-13C correlation NMR spectra, most of the abundant structural units of native lignin and released suspension culture lignin are present in the oxidase produced dehydrogenation polymer but in somewhat different amounts compared to peroxidase derived synthetic lignin preparations. A coniferin beta-glucosidase (EC 3.2.1.21) was observed to be secreted into the culture medium.

Journal Article↗
PubMed2002

Winter-drought induced embolism in Norway spruce (Picea abies) at the Alpine timberline.

At the timberline in the Central Alps, climatic conditions during winter frequently cause excessive drought stress (frost drought, 'Frosttrocknis'), which we hypothesized to induce cavitation in trees. We investigated the extent of winter-embolism in Norway spruce (Picea abies (L.) Karst.) growing near the timberline and analysed adaptations in vulnerability and anatomy. We found conductivity losses of up to 100% at the highest elevation (2020 m) correlated with low water potentials down to - 4.0 MPa. Vulnerability thresholds (50% loss in conductivity) decreased from - 3.39 MPa at 800 m to - 3.88 MPa at 1600 m corresponding to a decrease in tracheid cross-sectional area as well as pit and pit pore diameters. These thresholds were lower than potentials measured in embolized twigs near the timberline at the sampling dates probably due to lower potentials and/or a role of freeze-thaw events earlier in winter. Data indicated refilling processes, which may be of particular relevance for trees at the timberline, since adaptations in drought-induced vulnerability failed to prevent winter-embolism.

Journal Article↗
PubMed2002

Consequences of elevated CO2, augmented nitrogen-deposition and soil type on the soluble nitrogen and sulphur in the phloem of beech (Fagus sylvatica) and spruce (Picea abies) in a competitive situation.

Mixed spruce-beech plantations grown in large open-top chambers (OTC) were used to study consequences of elevated CO2, nitrogen-deposition and soil type on plant internal nitrogen and sulphur cycling of juvenile beech (Fagus sylvatica L.) and spruce (Picea abies Karst.) in a competitive situation. Processes of re-cycling as a consequence of protein turnover during leaf senescence in autumn were of further interest. For this purpose, phloem sap was collected in September 1998 and analysed for the composition and concentrations of organic and inorganic nitrogen and sulphur compounds. The phloem exudate of spruce showed higher total soluble non-protein nitrogen (TSNN) concentration on calcareous soil than on acidic soil, independent of the treatment. N-fertilization increased the N-concentration of phloem exudate significantly on both soil types, mainly by an increase of Arg and Gln concentrations. Elevated CO2 slightly increased TSNN on calcareous, but not on acidic soil. The combination of elevated CO2 and augmented N-deposition induced a further increase of TSNN on calcareous soil, but caused a lower N-effect on TSNN on acidic soil. Arg, the main TSNN component in phloem exudate, mediated this effect. Since Arg is considered to be a major nitrogen storage compound, it is concluded that in autumn elevated CO2 and augmented N-deposition, influence storage of N rather than N-supply of spruce. An effect of elevated CO2 and augmented N-deposition on GSH and sulphate concentrations in phloem exudate of spruce was not observed on acidic soil. On calcareous soil augmented N-deposition enhanced, elevated CO2 decreased phloem exudate GSH contents. In combination, elevated CO2 compensated the positive effect of N-deposition. The effects of elevated CO2 and augmented N-deposition on phloem sap N- and S-contents described above were not observed for beech trees. Apparently, elevated CO2 and augmented N-deposition did not affect plants internal S and N cycling of beech grown in spruce-beech plantations.

Journal Article↗
PubMed2002

Induction of tolerance to fast desiccation in black spruce (Picea mariana) somatic embryos: relationship between partial water loss, sugars, and dehydrins.

Events associated with the induction of tolerance to fast desiccation in black spruce (Picea mariana) somatic embryos were investigated. An experimental approach using an initial period of partial water loss was developed to induce either no, partial, or complete tolerance to fast desiccation. Tolerance to subsequent fast desiccation was not promoted by decreasing embryo water content from 1.5 to 1.1 g H2O g-1 DW (g g-1) throughout the first 24 h of slow desiccation. However, tolerance increased from 10 to 95% germination during the second 24-h period of slow desiccation after partial water loss from 1 to 0.55 g g-1. Emphasis was also placed on the relationship between observed tolerance, and sugar and dehydrin contents. Compared to controls, sucrose content in embryos doubled after 24 h of slow desiccation and more than tripled after 48 h. Conversely, starch content was decreased by one half after 24 h and by three quarters after 48 h. Sucrose abundance and raffinose occurrence after 48 h of slow desiccation were congruent with complete tolerance to fast desiccation. The period of slow desiccation between 24 and 48 h also increased the content of a 24-kDa dehydrin and the appearance of a 42-kDa dehydrin. The relationship between partial water loss, sugars and dehydrins is discussed with respect to tolerance to fast desiccation in black spruce somatic embryos.

Journal Article↗
PubMed2005

Analytical approaches to the determination of simple biophenols in forest trees such as Acer (maple), Betula (birch), Coniferus, Eucalyptus, Juniperus (cedar), Picea (spruce) and Quercus (oak).

Analytical methods are reviewed for the determination of simple biophenols in forest trees such as Acer (maple), Betula (birch), Coniferus, Eucalyptus, Juniperus (cedar), Picea (spruce) and Quercus (oak). Data are limited but nevertheless clearly establish the critical importance of sample preparation and pre-treatment in the analysis. For example, drying methods invariably reduce the recovery of biophenols and this is illustrated by data for birch leaves where flavonoid glycosides were determined as 12.3 +/- 0.44 mg g(-1) in fresh leaves but 9.7 +/- 0.35 mg g(-1) in air-dried samples (data expressed as dry weight). Diverse sample handling procedures have been employed for recovery of biophenols. The range of biophenols and diversity of sample types precludes general procedural recommendations. Caution is necessary in selecting appropriate procedures as the high reactivity of these compounds complicates their analysis. Moreover, our experience suggests that their reactivity is very dependent on the matrix. The actual measurement is less contentious and high performance separation methods particularly liquid chromatography dominate analyses whilst coupled techniques involving electrospray ionization are becoming routine particularly for qualitative applications. Quantitative data are still the exception and are summarized for representative species that dominate the forest canopy of various habitats. Reported concentrations for simple phenols range from trace level (<0.1 microg g(-1)) to in excess of 500 microg g(-1) depending on a range of factors. Plant tissue is one of these variables but various biotic and abiotic processes such as stress are also important considerations.

Chromatography, High Pressure Liquid↗
PubMed1998

The postfire discomycete Geopyxis carbonaria (Ascomycota) is a biotrophic root associate with Norway spruce (Picea abies) in nature.

The hypothesis that the postfire discomycete Geopyxis carbonaria (Ascomycota, Pezizales, Pyronemataceae) has a biotrophic association with roots of Norway spruce (Picea abies) in nature was tested by isolation of fungal strains from fresh, brown, smooth mycorrhiza-like root tips of Norway spruce collected from below the depth of detrimental heat penetration in a postfire site. The morphology of seven culture isolates originating from the smooth mycorrhiza-like root tips of two different spruce trees was congruent with the morphology of axenic culture isolates obtained from ascospores of G. carbonaria. DNA sequences of the nuclear ribosomal internal transcribed spacers ITS1 and ITS2 from these root-derived cultures and the ascosporic G. carbonaria culture isolates were found to be identical, further supporting the conclusion that the isolates were conspecific. The extensive ascocarp and ascospore formation of G. carbonaria which succeeds a forest fire may be explained in terms of a fungal escape from a moribund tree associate. Possible ecological adaptations of G. carbonaria to the pre- and postfire community are discussed.

Ascomycota↗
PubMed2001

Haplotype variation in a mitochondrial tandem repeat of Norway spruce (Picea abies) populations suggests a serious founder effect during postglacial re-colonization of the western Alps.

Populations from 13 elevational transects of Norway spruce [Picea abies (L.) Karst] across the Alpine range were sampled to elucidate the geographical pattern of genetic variation in relation to postglacial re-colonization and to study elevational effects on haplotypic diversity. We assessed fragment length variation in a tandem repeat region of the mitochondrial (mt) nad1 intron 2. This maternally inherited genetic marker is suited to infer migration as it is dispersed by seed only. A total of 10 haplotypes was found, most of which were due to repeat copy number variation. An analysis of molecular variance (amova) showed that overall population differentiation was high (F(ST)=0.41), and it revealed a significant differentiation between monomorphic western and moderately to highly variable eastern Alpine populations. This phylogeographic pattern may be explained by a founder effect during postglacial re-colonization. An early arriving haplotype, assumed to originate from a western Carpathian refugium, could expand into suitable habitats, reducing the chances for establishment of subsequently arriving haplotypes. On the other hand, the high variation in populations within an Italian transect of the south-eastern Alps may be the consequence of merging migration pathways from and close distance to putative glacial refugia, most likely those assumed in the Carpathian mountains and on the Balkan peninsula or possibly in the central plains of Italy. An effect of elevation on haplotypic diversity was not evident, though a low, but significant, partition of total genetic variation was attributed to among-population variation in one Italian transect. Various factors, such as vertical seed dispersal and forest management, may account for blurring an otherwise established pattern of genetic variation on a small geographical scale.

Analysis of Variance↗
PubMed1998

Characterization of an AGAMOUS homologue from the conifer black spruce (Picea mariana) that produces floral homeotic conversions when expressed in Arabidopsis.

Advances in elucidating the molecular processes controlling flower initiation and development have provided unique opportunities to investigate the developmental genetics of non-flowering plants. In addition to providing insights into the evolutionary aspects of seed plants, identification of genes regulating reproductive organ development in gymnosperms could help determine the level of homology with current models of flower induction and floral organ identity. Based upon this, we have searched for putative developmental regulators in conifers with amino acid sequence homology to MADS-box genes. PCR cloning using degenerate primers targeted to the MADS-box domain revealed the presence of over 27 MADS-box genes within black spruce (Picea mariana), including several with extensive homology to either AP1 or AGAMOUS, both known to regulate flower development in Arabidopsis. This indicates that like angiosperms, conifers contain a large and diverse MADS-box gene family that probably includes regulators of reproductive organ development. Confirmation of this was provided by the characterization of an AGAMOUS-like cDNA clone called SAG1, whose conservation of intron position and tissue-specific expression within reproductive organs indicate that it is a homologue of AGAMOUS. Functional homology with AGAMOUS was demonstrated by the ability of SAG1 to produce homeotic conversions of sepals to carpels and petals to stamens when ectopically expressed in transgenic Arabidopsis. This suggests that some of the genetic pathways controlling flower and cone development are homologous, and antedate the 300-million-year-old divergence of angiosperms and gymnosperms.

AGAMOUS Protein, Arabidopsis↗
PubMed2000

Bioactive triterpenoids from the stem bark of Picea glehni.

Two new triterpenoids, 3 alpha-methoxyserrat-14-en-21 beta-yl formate (1), and 24-methylcycloartenone (2), were isolated from the stem bark of Picea glehni (Fr. Schm.) Masters together with three known triterpenoids, 3 alpha-methoxyserrat-14-en-21 beta-ol, 3 beta-methoxyserrat-14-en-21 beta-ol, and piceanonol A. Compounds 1, 2, and a synthetic sample, 3 alpha-methoxyserrat-13-en-21 beta-yl formate showed potent inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA).

Antigens, Viral↗
PubMed1998

A homeobox gene with potential developmental control function in the meristem of the conifer Picea abies.

Many homeobox genes control essential developmental processes in animals and plants. In this report, we describe the first cDNA corresponding to a homeobox gene isolated from a gymnosperm, the HBK1 gene from the conifer Picea abies (L.) Karst (Norway spruce). The sequence shows distinct similarities specifically to the KNOX (knotted-like homeobox) class of homeobox genes known from different angiosperm plants. The deduced amino acid sequence of HBK1 is strikingly similar within the homeodomain (84% identical) to the maize gene Knotted1 (Kn1), which acts to regulate cell differentiation in the shoot meristem. This similarity suggested that the phylogenetic association of HBK1 with the KNOX genes might be coupled to a conservation of gene function. In support of this suggestion, we have found HBK1 to be expressed in the apical meristem in the central population of nondifferentiated stem cells, but not in organ primordia developing at the flanks of the meristem. This pattern of expression is similar to that of Kn1 in the maize meristem. We show further that HBK1, when expressed ectopically in transgenic Arabidopsis plants, causes aberrations in leaf development that are similar to the effects of ectopic expression of angiosperm KNOX genes on Arabidopsis development. Taken together, these data suggest that HBK1 has a role, similar to the KNOX genes in angiosperms, in the control of cellular differentiation in the apical meristem of spruce. The data also indicate that KNOX-gene regulation of vegetative development is an ancient feature of seed plants that was present in the last common ancestor of conifers and angiosperms.

Amino Acid Sequence↗
PubMed2001

Two APETALA2-like genes of Picea abies are differentially expressed during development.

The EREBP/AP2 gene family codes for plant specific transcription factors. The first two gymnosperm genes of this family were isolated from Picea abies and shown to be structurally related, but not orthologous, to the angiosperm AP2-like genes. The two P. abies genes are differentially expressed in different organs and may be important developmental regulators.

Amino Acid Sequence↗
PubMed1986

Effects of gibberellin A(4 + 7) and bark-ringing on cone initiation in mature Picea sitchensis grafts.

Large numbers of cones (strobili) were induced in a 10-year-old plot of mature grafts of Picea sitchensis (Bong.) Carr. All trees injected with 20 mg GA(4 + 7) in June initiated female and male cones in the same year. This treatment increased the number of female cones per plant 12-fold above the controls, more than doubling the percentage of cones that were female. Complete bark-ringing (done in May of the previous year) showed an additive effect with GA on the number of female cones formed, but a negative interaction on the number of male cones induced. Ringing promoted male cone production most when used alone. Treatments, singly or combined, also increased the proportion of cones that were lateral, compared with the preponderance of terminal male cones in the controls. The effects were apparently not directly associated with alterations in vegetative vigor, although these occurred causing a reduction in the proportion of buds containing vegetative shoots the following year. The clones differed in most characteristics, but both sparse and prolific clones were induced to reproductive activity. Viable seed yields per tree, and notional production from seed-orchards were enhanced almost 10-fold by GA injection, and about 4-fold by bark-ringing and GA + bark-ringing.

Journal Article↗
PubMed1986

Genetic and environmental control of crown development in Picea sitchensis and its relation to stem wood production.

Differences in stem wood production were found among four clones of Picea sitchensis (Bong.) Carr. at both an agricultural and a forest site. Clonal rankings were not consistent between the sites. Four variables were identified that may influence stem wood production, foliage amount, total aboveground production per unit foliage, proportional allocation of production to new foliage and to branch wood thickening. Comparison of clonal performance between sites showed that stem wood production could be influenced by differences in each of these variables. The four variables were themselves determined by components of crown structure, these were (i) production of new branches from the main stem, (ii) the numbers and lengths of branchlets supported on branches from the main stem, (iii) foliage production per unit branchlet length and foliage longevity, and (iv) branch wood thickening per unit branch length. These components varied both among clones and between sites. Branch production from the mainstem varied among clones, and for some clones it varied substantially between sites. In some clones branch production was positively correlated with mainstem height increment. There were differences among clones in the way that branchlet production varied between the agricultural and forest sites. In the absence of needle fall, foliage weight/branch length varied threefold among clones, but for each clone, varied little between sites. Branch wood weight/branch length was generally greater when foliage weight/branch length was large, but total branch wood increment/tree was strongly influenced by total branch length. The dynamics of crown development and its relation to stem wood production are discussed with reference to (i) the importance of needle longevity, (ii) the importance of tallness and narrowness of crowns and (iii) the importance of branch thickening, a process that competes with stem growth in the utilization of photosynthate. It is concluded that although each of these factors may be important, none singly accounts for the observed differences in stem wood production among genotypes and the way these change in response to the environment.

Journal Article↗
PubMed1992

Laser microprobe mass analysis (LAMMA) of aluminum and lead in fine roots and their ectomycorrhizal mantles of Norway spruce (Picea abies (L.) Karst.).

Fine roots and ectomycorrhizal root tips were sampled in a Norway spruce (Picea abies (L.) Karst.) stand in the eastern part of the Belgian Ardennes. The cellular and partly subcellular localizations of aluminum and lead were identified by the micro-analytical laser microprobe mass analysis (LAMMA) technique. In fine roots with secondary structure, localization of aluminum was limited to the peripheral cell layers. Lead was found in the outer layers, and also in the primary phloem. Aluminum penetrated the mycorrhizal mantle, but lead was seldom detected in ectomycorrhizae.

Journal Article↗
PubMed1992

Influence of a high Mn supply on Norway spruce (Picea abies (L.) Karst.) seedlings in relation to the nitrogen source.

Effects of 3, 25, 100, 200 and 800 microM Mn on biomass and pigment, starch and nitrate concentrations were studied in Norway spruce (Picea abies (L.) Karst.) seedlings grown with either NO(3) (-) or NH(4) (+) as the sole nitrogen source. After 77 days of exposure to 800 microM Mn, shoot growth had ceased in about 50% of the seedlings independently of the N source. Despite high Mn concentrations in roots and shoots of the Mn-treated seedlings, no visible symptoms of Mn toxicity were evident. The rate of root elongation was decreased by treatment with >/= 200 microM Mn when N was supplied as NO(3) (-), but not when it was supplied as NH(4) (+). This difference could be attributed to the higher Mn concentrations in root tips of the NO(3) (-)-grown seedlings compared with the NH(4) (+)-grown seedlings. In Mn-treated seedlings, the concentration of Mg, and to a lesser extent that of Ca, decreased. Depletion of these elements might account for the observed growth depression. Potassium concentrations were similar in the control and Mn-treated seedlings. Treatment of seedlings with 800 microM Mn for 50 days led to several physiological changes: starch accumulated, the concentrations of nitrate and phenolic compounds increased, pigment concentrations decreased, and in vivo nitrate reductase activity in roots was reduced.

Journal Article↗
PubMed1992

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.

Journal Article↗
PubMed1993

Ecophysiology of shade needles of Picea glauca saplings in relation to removal of competing hardwoods and degree of prior shading.

We studied shade needles of Picea glauca (Moench) Voss saplings that were growing in 15, 45 or 90% shade from competing vegetation at northern boreal forest sites in Alberta and Saskatchewan. At each site, in late May or early June 1990, all hardwoods were removed within a 2-m radius of each of eight saplings in each shade treatment (released saplings), and eight saplings in each shade treatment were left as controls. Light-saturated net assimilation (NA), stomatal and mesophyll conductance and water use efficiency of one-year-old needles were measured four times during the 1990 growing season and in the spring of 1991. There was a trend of increased photosynthetic capacity within one week following release. By August 1990 and in the following spring, NA was higher in released trees than in controls. The increase in NA appeared to be related to increased stomatal conductance to water vapor and to increased foliar nitrogen and resulting increases in mesophyll conductance to CO(2). There was no measurable effect of degree of shading prior to release on NA following release. Foliage of the released saplings appeared capable of rapid acclimation to the open conditions.

Journal Article↗