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PubMed2006

Gene expression profiles of different breast cancer cells compared with their responsiveness to fermented mistletoe (Viscum album L.) extracts Iscador from oak (Quercus), pine (Pinus), white fir (Abies) and apple tree (Malus) in vitro.

Cytotoxicity assays in vitro (MTT test) showed that the different breast cancer cell lines Kpl-1, MCF-7 and Mfm-223 respond differently to the mistletoe (Viscum album L.) preparations Iscador. Quercus (Qu), Abies (A), Malus (M) and Pinus (P). In order to determine the differences in the responsiveness of the cells more exactly, the gene expression profiles were determined by cells, which were treated with Mistletoe extracts, compared with untreated control cells. Such differences can be analysed in more detail by looking at the gene expression using Human Whole Genome microarray chips (41,000 genes). The results of the transcriptome analyses suggested that Iscador preparations influenced the overregulation of genes regarding immune defense, stress response, apoptosis and cell-cell adhesion pathways. Within the Mfm-223-Zellen was the Genexpression in MCF-7 and Kpl-1. The MCF-7 cells were affected on the genes which are involved in cell-cell contacts whereas Kpl-1 responded to the mistletoe extracts by changing the mRNA levels of the immune and stress response pathways. Concerning the effects of the mistletoe extract, we conclude that Iscador Qu and M have a greater influence on the immune defense and stress response genes whereas Iscador A tends to affect the cell-cell adhesion and cytoskeleton pathways. In summary, cDNA microarray analyses give us information on whether a cancer cell is sensitive to mistletoe extracts in relation to how many genes are significantly overrepresented after mistletoe treatment, and whether a particular mistletoe extract is more effective on a specific cancer cell than the other preparation.

Abies↗

PubMed2001

Triterpene constituents from the stem bark of Pinus luchuensis and their DNA topoisomerase II inhibitory effect.

Nine lanostane and serratane-type triterpenes including two unknown compounds were isolated from the stem bark of Pinus luchuensis. These new compounds were characterized as 3-oxolanost-9(11)-ene-24S,25-diol (1) and 29-acetoxy-3beta-methoxyserrat-14-en-21alpha-ol (2) on the basis of spectroscopic evidence. Some of these triterpenes were tested for the inhibitory effect on DNA topoisomerase II activity. Compound 1 showed a slightly less potent inhibitory activity with an IC(50) value of 186 microM.

Acetyl-CoA C-Acyltransferase↗

PubMed2006

Gas exchange and growth responses of ectomycorrhizal Picea mariana, Picea glauca, and Pinus banksiana seedlings to NaCl and Na2SO4.

Black spruce (PICEA MARIANA), white spruce (PICEA GLAUCA), and jack pine (PINUS BANKSIANA) seedlings were inoculated with HEBELOMA CRUSTULINIFORME or LACCARIA BICOLOR and subjected to NaCl and Na (2)SO (4) treatments. The effects of ectomycorrhizas on salt uptake, growth, gas exchange, and needle necrosis varied depending on the tree and fungal species. In jack pine seedlings, ectomycorrhizal (ECM) fungi reduced shoot and root dry weights and in the ECM white spruce, there was a small increase in dry weights. Sodium chloride treatment reduced net photosynthesis and transpiration rates in the three studied tree species. However, NaCl-treated black spruce and jack pine colonized by H. CRUSTULINIFORME maintained relatively high photosynthetic and transpiration rates and needle necrosis of NaCl-treated black spruce seedlings was reduced by the ECM fungi. Higher concentrations of Na (+) were found in shoots compared with roots of the three examined conifer species. ECM fungi reduced the concentrations of Na (+) mainly in the shoots and this reduction was greater in plants treated with NaCl compared with Na (2)SO (4). Shoots contained generally higher concentrations of Cl (-) compared with roots. In the NaCl-treated black spruce and white spruce, both ECM species significantly reduced Cl (-) concentrations. Our results point to overall greater phytotoxicity of NaCl compared with Na (2)SO (4) and support our earlier findings which demonstrated beneficial effects of ECM fungi for woody plants exposed to NaCl stress.

Mycorrhizae↗

PubMed2002

A stilbene synthase from Japanese red pine (Pinus densiflora): implications for phytoalexin accumulation and down-regulation of flavonoid biosynthesis.

Stilbene synthase (STS) and chalcone synthase (CHS) are plant-specific polyketide synthases that play key roles in the stilbenoid and flavonoid biosyntheses, respectively. We have recently isolated from Pinus densiflora three STS cDNAs (PDSTS1, PDSTS2, and PDSTS3) and one CHS cDNA (PDCHSX). We then heterologously expressed these cDNAs in Escherichia coli and characterized their properties. An unusual STS isozyme, PDSTS3, lacks the common C-terminal extension of STS because of a frame-shift mutation and shows the highest pinosylvin-forming activity among the STSs tested. Pinosylvin was shown to be a potent inhibitor of PDCHSX (K(i) = 6 microM) as well as PDSTS2 (K(i) = 13 microM), which presumably maintains the balance between the stilbenoid and flavonoid biosyntheses. PDSTS3 was insensitive to product inhibition. We identified PDSTS3 in the pine seedlings as well as full-length STS. The data provide evidence that PDSTS3 is involved in the potential regulation of the stilbenoid and flavonoid biosynthetic pathways in pine trees.

Acyltransferases↗

PubMed1996

Nitrogen metabolism in Lignifying Pinus taeda cell cultures.

The primary metabolic fate of phyenylalanine, following its deamination in plants, is conscription of its carbon skeleton for lignin, suberin, flavonoid, and related metabolite formation. Since this accounts for approximately 30-40% of all organic carbon, an effective means of recycling the liberated ammonium ion must be operative. In order to establish how this occurs, the uptake and metabolism of various 15N-labeled precursors (15N-Phe, 15NH4Cl, 15N-Gln, and 15N-Glu) in lignifying Pinus taeda cell cultures was investigated, using a combination of high performance liquid chromatography, 15N NMR, and gas chromatograph-mass spectrometry analyses. It was found that the ammonium ion released during active phenylpropanoid metabolism was not made available for general amino acid/protein synthesis. Rather it was rapidly recycled back to regenerate phenylalanine, thereby providing an effective means of maintaining active phenylpropanoid metabolism with no additional nitrogen requirement. These results strongly suggest that, in lignifying cells, ammonium ion reassimilation is tightly compartmentalized.

Amino Acids↗

PubMed2003

Alterations in endogenous levels of cytokinins following grafting of Pinus radiata support ratio of cytokinins as an index of ageing and vigour.

Markers for ageing or maturation in woody plants provides the possibility of identifying genotypes with a prolonged juvenile phase, or to evaluate reinvigoration procedures, such as grafting of adult material. This study focuses specifically on cytokinins (Cks) as markers, due to their role in the transition process from juvenile to adult states. Previous reports from our group disclosed that the ratio of isopentenyladenine-type (iP-type) to zeatin-type (Z-type) Cks decreases during tree maturation, a maturation and ageing index being likely in Pinus radiata. Based on these results, Cks were tested in terminal buds of adult and grafted adult P. radiata material to corroborate the importance of the aforementioned balance as an index in the reinvigoration process of adult trees associated with grafting procedures. Results revealed the reversion of this index pattern parallel to the reactivation of some juvenile traits, namely, increasing values of the ratio throughout the reinvigoration process. According to previous results, however, the opposite was the case during the reverse process. This fact allowed us to validate the iP-type/Z-type Ck ratio as an ageing and vigour index.

Adenine↗

PubMed2005

A new diterpenoid and active stilbenes from Pinus armandii heartwood.

8(17),13-Labdadien-16,14-olid-18-oic acid (1), a new diterpenoid, has been isolated from the heartwood of Pinus armandii Francher, along with seven known diterpenoids (2-8) and four known stilbenes (9-12). Their structures have been elucidated by spectral evidence. (E)-3-Hydroxy-5-methoxystilbene (9) showed significant inhibition against white-rot fungi.

Diterpenes↗

PubMed1999

Antioxidants and protective pigments of Pinus ponderosa needles at gradients of natural stresses and ozone in the San Bernardino Mountains in California.

At the San Bernardino Mountains, California, a well documented gradient of ozone pollution overlays a natural stress gradient from mesic to dry and from lower elevation to higher elevation sites. In contrast to gradient studies in European regions, the highest ambient ozone levels are observed at low elevation and more mesic locations. In the present study, antioxidative and photoprotective systems in Pinus ponderosa needles were investigated at three plots--DW (1725 m, high ozone impact, mesic site), SW (1200 m, clean air, xeric site) and CO (above 2000 m, clean air and xeric site). Needles from the CO site contained significantly more total GSH (500 vs 300 nmol g(-1) dw in c needles), less alpha-carotene (6-10 vs 14-19 microg mg(-1) total chlorophyll) and chlorophyll (1.7-2 vs 2.5-2.6 mg g(-1) dw in c + 1 needles) than those at the DW site. Furthermore, their xanthophyll cycle pool was in a more de-epoxidized state at midday (up to 60% in c needles), and the carotenoid/chlorophylls ratios were generally higher. These patterns correspond to those observed at higher elevation plots in the Alps. On the other hand, needles from the high ozone site (DW) had a higher proportion of GSSG, indicating the onset of biochemical injury to needles. Needles from the SW site had intermediate proportions of GSSG. The results show the potential of environmental stressors to induce antioxidative and photoprotective responses in the absence of elevated ozone concentrations, but support the oxidative effects of ozone injury to ponderosa pine.

Air Pollutants↗

PubMed2006

Analysis of telomere length and telomerase activity in tree species of various lifespans, and with age in the bristlecone pine Pinus longaeva.

Normal somatic cells have a finite replicative capacity, and with each cell division telomeres progressively shorten, unless the telomerase enzyme is present. The bristlecone pine, Pinus longaeva, is the oldest known living eukaryotic organism, with the oldest on record turning 4770 years old in 2005. The results from our study of telomere length and telomerase activity in samples (needle, root, core) from P. longaeva with age, and in other tree species of various lifespans, support the hypothesis that both increased telomere length and telomerase activity may contribute to the increased lifespan and longevity evident in long-lived pine trees (i.e., 2000- to 5000-year lifespan) compared with medium-lived (400- to 500-year lifespan) and short-lived (100- to 200-year lifespan) pine trees, as well as in P. longaeva with age.

Aging↗

PubMed2002

X-ray microanalytical studies of mineral localization in the needles of white pine (Pinus strobus L.).

Eastern white pine (Pinus strobus L.) shoots from mature trees were collected from two sites of contrasting soil pH: the Glendon campus of York University in Toronto, Ontario (pH 6.7 at 40 cm); and Muskoka near Huntsville, Ontario (pH 4.2 at 40 cm). Needles of ages 1-3 years were removed from the shoots, and the percentage of ash and silica was determined for all ages. Other needles were frozen in liquid nitrogen and kept in a cryo-biological storage system before x-ray microanalysis. Percentages of ash and silica were higher in the needles from Muskoka. Ash and silica increased with needle age for trees from the Muskoka site, but less so at the Toronto site. Of the 13 elements (Na, Mg, Al, Si, P, S, Cl, K, Ca, Mn, Fe, Cu and Zn) detected by microanalysis, Mn, Fe, Cu and Zn were detected in small amounts in the epidermis, endodermis and transfusion tissue (the layer of tracheids and parenchyma immediately surrounding the vascular bundles), and K, P, S and Cl were almost ubiquitous in distribution. Sodium was occasionally detected in the transfusion tissue, and magnesium was concentrated in the endodermal cells. The epidermal walls, transfusion tissue and endodermis were major sites of calcium localization. Silicon was concentrated in the extreme tips of the needles in all tissues, but particularly in the transfusion tissue, and more so in the Muskoka samples. Microanalysis revealed a higher Al content in the Muskoka needles, that Al was concentrated in the needle tips and that the transfusion tissues were major sites of accumulation.

Aluminum↗

PubMed2004

Seasonal variation of organogenetic activity and reserves allocation in the shoot apex of Pinus pinaster Ait.

BACKGROUND AND AIMS: To understand better the basic growth characteristics of pines and the fundamental properties of the shoot apical meristem (SAM), variations within the shoot apex of buds were studied. METHODS: A detailed structural comparison of meristem dimensions, organogenetic activity, and the presence of lipids, starch grains and tannins was performed on shoot apices of juvenile, and male and female adult Pinus pinaster at five different times in the annual growth cycle. KEY RESULTS: There were significant correlations among traits and differences in the pattern for juvenile and adult shoots. In juvenile shoots, peaks of organogenesis were present in spring and autumn, but not in summer. In adult shoots, one peak, characterized by an increase in meristem dimensions, was present in summer. The accumulation of starch grains beneath the SAM and of tannin in sub-apical pith parenchyma were at their maximum when organogenetic activity was high in spring and autumn in juvenile plants, and in summer in adult plants. In juvenile and adult plants, lipids were stored within the SAM in autumn, filling a large part of the bud in winter, and were depleted in the cortical parenchyma and then in the pith during shoot elongation. CONCLUSIONS: Depending on the sites of accumulation within the SAM and on the stage of the annual growth cycle, lipids, starch and tannins may be involved in different processes. In spring, energy and structural materials released by lipid hydrolysis may contribute to stem elongation and/or cell-to-cell communication. During organogenesis, energy and structural materials released by starch hydrolysis may influence developmental programmes in the SAM and adjacent cells. Tannins may be involved in cellular detoxification. At the end of the growing season, accumulation of lipid and starch is positively correlated with the onset of dormancy.

Lipid Metabolism↗

PubMed2006

Modelling environmental variation in Young's modulus for Pinus radiata and implications for determination of critical buckling height.

BACKGROUND AND AIMS: Although density-specific stiffness, E/rho, (where E is Young's modulus and rho is wood density) is often assumed constant by the elastic similarity model, and in determination of critical buckling height (H(crit)), few studies have tested this assumption within species. Here this assumption is tested for Pinus radiata growing across an environmental gradient, and theory is combined with data to develop a model of Young's modulus. METHODS: Analyses use an extensive series of environmental plots covering the range of climatic and edaphic conditions over which P. radiata is grown in New Zealand. Reduced major axis regression was used to determine scaling exponents between log-log plots of H(crit) vs. groundline diameter (D), and E/rho vs. D. Path analysis was used to identify significant direct and indirect (through stem slenderness) edaphic and climatic influences on E. KEY RESULTS: Density-specific stiffness exhibited 3-fold variation. As E/rho scaled positively with D, the exponent of 0.95 between H(crit) and D exceeded the assumed value of 0.67 under constant E/rho. The final path analysis model included mean air temperature in early autumn (T(aut)) and slenderness as significant (P < 0.05) positive direct influences on E. Tree leaf area index and T(aut) were indirectly associated with E through their significant (P < 0.05) positive direct relationship with stem slenderness. Young's modulus was most sensitive to T(aut), followed by stem slenderness then leaf area index, and the final model explained 76 % of the variance in E. CONCLUSIONS: The findings suggest that within species E/rho variation may influence H(crit) and the scaling exponent between D and H(crit) so important in assumptions regarding allometric relationships. The model presented may provide a useful means of determining variation in E, E/rho and H(crit) across environmental gradients.

Ecosystem↗

PubMed2006

Short delay in timing of emergence determines establishment success in Pinus sylvestris across microhabitats.

BACKGROUND AND AIMS: The date of emergence may have far-reaching implications for seedling performance. Seedlings emerging early in the growing season often have a greater rate of survival or grow better if early emergence provides advantages with respect to an environmental cue. As a result, the benefits of early emergence may be lost if the environmental stress creating the differences among cohorts disappears. The experimental manipulation under field conditions of the factors that constitute the main sources of stress for seedling establishment is thus a straightforward method to evaluate the impact of date of emergence on seedling establishment under realistic conditions. METHODS: Two field experiments were performed to analyse the effect of emergence date on survival and first-year growth of Scots pine seedlings in natural mountain forests in south-east Spain. Two main environmental factors that determine seedling success in these mountains were considered: (1) microhabitat type (monitoring the effect of date of emergence in the three most common microhabitats where seedlings recruit); (2) summer drought (monitored by an irrigation treatment with control and watered sampling points). KEY RESULTS: Overall, early emergence resulted in a higher probability of survival and better growth in the two experiments and across microhabitats. However, the reduction in summer drought did not diminish the differences observed among cohorts: all cohorts increased their survival and growth, but early cohorts still had a clear advantage. CONCLUSIONS: Date of emergence determines establishment success of Pinus sylvestris seedlings, even if cohorts are separated by only a few days, irrespective of the intensity of summer drought. The experimental design, covering a gradient of light intensity and soil moisture that simulates conditions of the regeneration niche of Scots pine across its geographical range, allows the results to be extrapolated to other areas of the species. Date of emergence is thus likely to have a large impact on the demography of Scots pine across its geographical range.

Ecosystem↗

PubMed1999

A consensus map for loblolly pine (Pinus taeda L.). I. Construction and integration of individual linkage maps from two outbred three-generation pedigrees.

A consensus map for loblolly pine (Pinus taeda L.) was constructed from the integration of linkage data from two unrelated three-generation outbred pedigrees. The progeny segregation data from restriction fragment length polymorphism, random amplified polymorphic DNA, and isozyme genetic markers from each pedigree were recoded to reflect the two independent populations of parental meioses, and genetic maps were constructed to represent each parent. The rate of meiotic recombination was significantly greater for males than females, as was the average estimate of genome length for males (1983.7 cM [Kosambi mapping function (K)]) and females [1339.5 cM(K)]. The integration of individual maps allows for the synthesis of genetic information from independent sources onto a single consensus map and facilitates the consolidation of linkage groups to represent the chromosomes n = 12 of loblolly pine. The resulting consensus map consists of 357 unique molecular markers and covers approximately 1300 cM(K).

Chromosome Mapping↗

PubMed2001

Genetic composition and diploid hybrid speciation of a high mountain pine, Pinus densata, native to the Tibetan plateau.

Pinus densata has been suggested to have originated from hybridization events involving P. tabulaeformis and P. yunnanensis. In this study, allozyme differentiation at 12 loci was studied in 14 populations of P. tabulaeformis, P. densata, and P. yunnanensis from China. The observed genetic composition of P. densata supported the hybrid hypothesis and showed varying degrees of contribution from P. yunnanensis and P. tabulaeformis among its populations. These data, together with previous chloroplast DNA results, indicated different evolutionary histories among P. densata populations. To examine the possibility of ongoing hybridization among the three species, we analyzed patterns of linkage disequilibria between allozyme loci in ovule, pollen, and zygote pools. None of these tests suggested that there is significant ongoing gene exchange, implying that populations of P. densata have a stabilized hybrid nature. The normal fertility and high fecundity of P. densata indicate that this hybrid is maintained through sexual reproduction. P. densata represents an example of diploid hybrid speciation in an extreme ecological habitat that is both spatially and ecologically separated from that of its parents.

Alleles↗

PubMed2000

Effect of different daytime and night-time temperature regimes on the foliar respiration of Pinus taeda: predicting the effect of variable temperature on acclimation.

The objectives of this study were to determine the acclimation of loblolly pine (Pinus taeda L.) foliar respiration to different night-time low temperatures, daytime high temperatures, and daily mean temperatures, and then to use the responses of temperature acclimation to various temperature regimes to predict acclimation under fluctuating temperatures. Experiments were conducted on two-year-old seedlings in growth chambers using different combinations of day and night-time temperatures. The first experiment exposed trees to 22/22, 29/22, 22/15, and 29/15 degrees C day/night (d/n). When measured at a common temperature (15, 22 or 29 degrees C), respiration rates were lower for trees exposed to higher treatment temperatures and acclimation was influenced by both day and night-time temperature. However, the extent of acclimation did not relate to mean temperature, i.e. respiration rates measured at a common temperature ranked as follows for seedlings exposed to different temperature regimes, 22/15>22/22>29/15 congruent with29/22 degrees C d/n. Rather, acclimation of foliar respiration was linearly related to mean daily respiration rate, where mean daily respiration rate is the average of the respiration rates measured at the day and night-time treatment temperatures. The discrepancy between mean daily respiration rate and mean daily temperature occurred because respiration increased exponentially with increasing temperature. In a second experiment, the same seedlings were exposed to 22/22, 15/15, 25.5/18.5, and 25.5/15 degrees C d/n to test the relationship between mean daily respiration rate and acclimation. As in the first experiment, acclimation was linearly related to mean daily respiration rate. The concept of effective acclimation temperature, which is the temperature at which the mean daily respiration rate occurs, was derived from these results as a means to predict the extent that foliar respiration acclimates to treatment temperature.

Adaptation, Physiological↗

PubMed2000

The electrical impedance spectroscopy of Scots pine (Pinus sylvestris L.) shoots in relation to cold acclimation.

Electrical impedance spectroscopy (EIS) was applied to stems of Scots pine (Pinus sylvestris L.) in a provenance field trial during frost hardening to find an EIS parameter for assessing frost hardiness (FH) without a controlled freezing test. The FH of stems and needles assessed by controlled freezing tests was compared with the equivalent circuit EIS parameters of a distributed model of stems (not exposed to controlled freezing treatment) and with dry matter (DM) content of stems. Significant differences in the equivalent circuit parameters, FH and DM content were found between provenances. The relaxation time (tau(1)), describing the peak of the high frequency arc of the impedance spectrum, and the intracellular resistance (r(i)) of stems increased with increasing FH. According to the linear regression, the coefficient of determination (R(2)) between the FH of stems and needles with tau(1) of the stem was 0.87 and 0.89, and with r(i) of the stem 0.74 and 0.85, respectively. The relation between FH and tau(1) changed with the degree of hardiness. The highest coefficient of determination was 0.95 in September when the FH of needles, ranging from -10 degrees C to -25 degrees C, was predicted with an accuracy of +/-2.0 degrees C. The resistance parameter r(2), describing the width of the low frequency arc of the impedance spectrum, decreased prior to and during the initial hardening: significant differences were found between provenances. This indicates that r(2) was not related to frost hardening per se. It is concluded that it is possible to distinguish the hardening patterns of different provenances by tau(1) in the rapid phase of hardening without controlled freezing tests.

Acclimatization↗

PubMedDate not supplied

Inheritance and linkage of allozymes in a Balkan endemic, Pinus peuce Griseb.

This article presents a study of isozyme variation in Pinus peuce Griseb., a Balkan endemic. Among the enzyme systems studied, five were monomorphic and eight were polymorphic in at least one locus. The segregation analysis of the polymorphic loci were consistent with a Mendelian mode of inheritance. No significant deviation from the expected ratio was observed both at the individual and pooled segregation data levels. Segregation patterns were homogeneous across individuals. Two significant linkage groups were found in P. peuce: FEST-2:LAP-2 and 6PG-1:6PG-2, which correspond to the results obtained for other pine species.

Acid Phosphatase↗