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Heterogeneous genetic structure in a Fagus crenata population in an old-growth beech forest revealed by microsatellite markers.

The within-population genetic structure of Fagus crenata in a 4-ha plot (200 x 200 m) of an old-growth beech forest was analysed using microsatellite markers. To assess the genetic structure, Moran's I spatial autocorrelation coefficient was calculated. Correlograms of Moran's I showed significant positive values less than 0.100 for short-distance classes, indicating weak genetic structure. The genetic structure within the population is created by limited seed dispersal, and is probably weakened by overlapping seed shadow, secondary seed dispersal, extensive pollen flow and the thinning process. Genetic structure was detected in a western subplot of 50 x 200 m with immature soils and almost no dwarf bamboos (Sasa spp.), where small and intermediate-sized individuals were distributed in aggregations with high density because of successful regeneration. By contrast, genetic structure was not found in an eastern subplot of the same size with mature soils and Sasa cover, where successful regeneration was prevented, and the density of the small and intermediate-sized individuals was low. Moreover, genetic structure of individuals in a small-size class (diameter at breast height < 12 cm) was more obvious than in a large-size class (diameter at breast height >/= 12 cm). The apparent genetic structure detected in the 4-ha plot was therefore probably the result of the structure in the western portion of the plot and in small and intermediate-sized individuals that successfully regenerated under the favourable environment. The heterogeneity in genetic structure presumably reflects variation in the density that should be affected by differences in regeneration dynamics associated with heterogeneity in environmental conditions.

Demography↗

Half-sib family structure of Fagus crenata saplings in an old-growth beech-dwarf bamboo forest.

Half-sib family structure of Fagus crenata saplings was examined in an old-growth beech-dwarf bamboo (Sasa spp.) forest using microsatellite genotypes in parentage analysis to identify the half-sib families in two 50 x 50 m plots: one with 36 adults, 641 saplings and no Sasa cover, the other with 21 adults, 61 saplings and Sasa cover. For large proportions of the saplings (44.6% and 75.4%, respectively) both of their parents were found within the same plot, indicating that pollination events frequently involved short-range pollen dispersal, probably because of the high density of adults in the study population. Although almost all of the adults had half-sib families, the number of offspring in the families was highly variable. In the plot with no Sasa cover, the variation in the number of offspring was significantly explained by the size of parents, i.e. the reproductive success is higher for large adults than for small adults. The half-sibs were aggregately distributed around their parents and the distribution overlapped among different half-sib families, which may be due to the limited seed dispersal and overlapping seed shadows of this species. As expected, there was weak genetic structure in the plot. By contrast, in the plot with Sasa cover, the half-sibs were distributed sparsely with a low density, and the degree of genetic structure was very weak. The difference in the half-sib family structure and genetic structure among saplings presumably reflects the difference in density that should be affected by regeneration dynamics associated with environmental conditions.

Demography↗

Natural selection and climate change: temperature-linked spatial and temporal trends in gene frequency in Fagus sylvatica.

Rapid increases in global temperature are likely to impose strong directional selection on many plant populations, which must therefore adapt if they are to survive. Within populations, microgeographic genetic differentiation of individuals with respect to climate suggests that some populations may adapt to changing temperatures in the short-term through rapid changes in gene frequency. We used a genome scan to identify temperature-related adaptive differentiation of individuals of the tree species Fagus sylvatica. By combining molecular marker and dendrochronological data we assessed spatial and temporal variation in gene frequency at the locus identified as being under selection. We show that gene frequency at this locus varies predictably with temperature. The probability of the presence of the dominant marker allele shows a declining trend over the latter half of the 20th century, in parallel with rising temperatures in the region. Our results show that F. sylvatica populations may show some capacity for an in situ adaptive response to climate change. However as reported ongoing distributional changes demonstrate, this response is not enough to allow all populations of this species to persist in all of their current locations.

Alleles↗

Plant-mediated nitrous oxide emissions from beech (Fagus sylvatica) leaves.

Nitrous oxide (N2O) emission estimates from forest ecosystems are based currently on emission measurements using soil enclosures. Such enclosures exclude emissions via tall plants and trees and may therefore underestimate the whole-ecosystem N2O emissions. Here, we measured plant-mediated N2O emissions from the leaves of potted beech (Fagus sylvatica) seedlings after fertilizing the soil with 15N-labelled ammonium nitrate (15NH4(15)NO3), and after exposing the roots to elevated concentrations of N2O. Ammonium nitrate fertilization induced N2O + 15N2O emissions from beech leaves. Likewise, the foliage emitted N2O after beech roots were exposed to elevated concentrations of N2O. The average N2O emissions from the fertilization and the root exposure experiments were 0.4 and 2.0 microg N m(-2) leaf area h(-1), respectively. Higher than ambient atmospheric concentrations of N2O in the leaves of the forest trees indicate a potential for canopy N2O emissions in the forest. Our experiments demonstrate the existence of a previously overlooked pathway of N2O to the atmosphere in forest ecosystems, and bring about a need to investigate the magnitude of this phenomenon at larger scales.

Fagus↗

Early detection of Biscogniauxia nummularia in symptomless European beech (Fagus sylvatica L.) by TaqMan quantitative real-time PCR.

AIMS: To develop a quantitative real-time PCR (Rt PCR) assay for the early detection of Biscogniauxia nummularia, a xylariaceous fungus that causes strip-canker and wood decay on European beech (Fagus sylvatica L.). METHODS AND RESULTS: The molecular assay was based on TaqMan chemistry using species-specific primers and a fluorogenic probe designed on the ITS1 sequence of rRNA gene clusters. The specificity of the oligonucleotides and the probe were tested using the DNA of B. nummularia isolates from different geographic areas, of phylogenetically related species, and of some fungi commonly colonizing European beech bark and wood. A total of 31 symptomless and symptomatic shoots of European beech were collected from three forest sites in the Apennine Mountains of Italy. The percentage of positive detections of B. nummularia with the TaqMan assay was 78.6%, compared with only 14.3% of positive isolations on growth media for two sites. CONCLUSIONS: In shoots, the quantitative Rt PCR assay detected down to 8.0-fg fungal DNA per microgram of total DNA extracted. SIGNIFICANCE AND IMPACT OF THE STUDY: The assay developed in quantitative Rt PCR, by using TaqMan chemistry, revealed a rapid and sensitive method useful for the early detection of B. nummularia in symptomless European beech twigs.

DNA Primers↗

Protective effect of lignophenol derivative from beech (Fagus crenata Blume) on copper- and zinc-mediated cell death in PC12 cells.

Lignophenol, prepared using a phase-separation system, is a derivative of lignin, which is one of the components in the plant cell wall, and possesses high phenolic function, high stability and antioxidant properties. However, little is known about the beneficial effect of lignophenol. In this study, we investigated the protective effect of lignophenol from the beech tree (Fagus crenata Blume) on copper- and zinc-mediated apoptosis in PC12 cells by using DNA fragmentation and TUNEL assays. In DNA fragmentation assays, the DNA ladder patterns in the PC12 cells treated with 200 microM Cu and 200 microM Zn were enhanced, whereas the DNA ladder pattern was hardly observed in these cells treated with 20 mM lignophenol. In the TUNEL assay, TUNEL signals increased significantly in the untreated PC12 cells exposed to 200 microM Cu compared with the control. In contrast, the degree of apoptosis in the 20 mM lignophenol-treated cells was significantly lower than in the untreated cells, indicating that lignophenol inhibited Cu-induced apoptotic cell death in PC 12 cells. In the 200 microM Zn-exposed group, the degree of apoptosis in the 20 mM lignophenol-treated cells was also low compared with the untreated cells. In conclusion, these results suggest that lignophenol plays a role in protecting against Cu- and Zn-mediated PC12 apoptotic cell death.

Animals↗

Changes in the activity of amylase, peroxidase and catalase in beech (Fagus orientalis Lipsky) during dormancy and growth.

Activities of peroxidase, amylase and catalase were analyzed on beech (Fagus orientalis Lipsky) during one year to determine relationship between these enzymes with the period of dormancy and growing season. Results showed that amylase activity was high but catalase and peroxidase activities were low during the growing season. Peroxidase and catalase activities increased from July to November whereas amylase activity started to decrease at the same time, which means that the plants were prepared themselves to be dormant and the hardening was happened. The peroxidase maximum activity was in November, which is an important sign of dormancy and completing hardening in plant. The dormancy released from February along with decrease of catalase activity. At the beginning of growing season, correlation between amylase and catalase was increased. Amylase activity also increased gradually. No significant correlation between amylase-catalase and amylase-peroxidase activities was found at the period of dormancy. In our study, we found that the seasonal activity of enzymes such as peroxidase, catalase, and amylase, also correlation between these enzymes could be an important factor for the detection the period of dormancy and growing season.

Amylases↗

[Seedling establishment of Fagus engleriana, a dominant in mountain deciduous forests].

The survival and growth of Fagus engleriana seedlings in three light levels, and with and without supplying additional fertilizer (F1 and F0, respectively) for each light level were studied. The three light levels were to simulate the light regimes in the understorey, small gaps and clearings (L1, L2 and L3 about 1%, 18% and 100% of full sunlight, respectively). The seedling development in L1 was severely inhibited by low light intensity. Seedling mortality was remarkably higher in L1 than in L2 and L3, and in F1 than in F0. The death of the seedlings seemed to result from attack of fungal pathogens. Although seedling survival and growth were significantly improved as light intensities increased from L1 to L2, seedlings in L3 developed similarly as those in L2. It is suggested that the young seedlings could not tolerate the shade of closed forest canopy, and fertile patches on the forest floor might not improve seedling establishment. Successful regeneration of the species in the forests needs better light conditions such as those in gaps than those under the closed forest canopy, at least during the phase of seedling establishment.

Biomass↗

Genetic variability among beech (Fagus sylvatica L.) populations from the Sudety Mountains, in respect of peroxidase and malate dehydrogenase loci.

Individual trees growing in five populations of European beech (Fagus sylvatica L.) in the Sudety Mountains were investigated in respect of variability of peroxidases (2 loci) and malate dehydrogenase (1 locus). Differences between populations were illustrated by a dendrogram constructed on the basis of Hedrick's (1974) genetic distances. The mean GST coefficient (=0.0333) value demonstrated the higher level of intra-population variability, as compared to the inter-population (DST = 0.0149) variability.

Alleles↗

Induction kinetics of delayed fluorescence of sun and shade leaves of Fagus sylvatica in the ms-range.

Induction kinetics of luminescence (= delayed chlorophyll fluorescence or delayed light emission) were measured with sun and shade leaves of a tall beech tree (Fagus sylvatica 'pendula', weeping beech). The kinetics detected in the ms-range are contrasted for the upper and the lower leaf side. The influence of the following parameters is demonstrated: time of dark-adaptation prior to the measurement, intensity of the excitation light and photoinhibitory treatment. The effects are discussed with respect to chlorophyll concentration, absorption of the excitation light, reabsorption of the luminescence and photosynthetic activity of the leaf tissue. It is shown that the luminescence signal and its kinetic are determined mainly by the properties of the mesophyll parenchyma facing the detector. Thus the more densely packed palisade parenchyma at the upper leaf side exhibits a lower luminescence and a slower kinetic than the spongy parenchyma at the lower leaf side, which is characterized by many aerial interspaces. Our study shows that luminescence kinetics can be applied to interpret the physiological state of a specific leaf tissue. They may serve as an indicator of disorders in the photosynthetic function.

Kinetics↗

Exclusive accumulation of Z-isomers of monolignols and their glucosides in bark of Fagus grandifolia.

In addition to Z-coniferyl and Z-sinapyl alcohols, bark extracts of Fagus grandifolia also contain significant amounts of the glucosides, Z-coniferin, Z-isoconiferin (previously called faguside) and Z-syringin. The corresponding E-isomers of these glucosides do not accumulate to a detectable level. The accumulation of the Z-isomers suggests that either they are not lignin precursors or that they are reservoirs of monolignols for subsequent lignin biosynthesis; it is not possible to distinguish between these alternatives. The co-occurrence of Z-coniferin and Z-isoconiferin demonstrate that glucosylation of monolignols can occur at either the phenolic or the allylic hydroxyl groups.

Acetylation↗

Ultrastructure in leaves of Fagus silvatica and Carpinus betulus individuals tolerant and susceptible to SO2 and heavy metal pollutants.

The ultrastructure characteristics of leaves during the development of cell organelles, in the "greening process" of spring, have been studied in pollutant-tolerant and susceptible individuals of beech (Fagus silvatica) and hornbeam (Carpinus betulus) in an area polluted with SO2 (the main pollutant may reach 0.06-2.72 mg/m3), heavy metals, and other gases discharged by non-iron metal processing factories. Leaves from tolerant beech and hornbeam individuals did not undergo ultrastructural changes compared to control leaves in an unpolluted area. The susceptible and very susceptible individuals of both species gradually undergo modification in organelle ultrastructure not only during the first steps of greening and momentary exposure to air pollution, but also as a result of long-term exposure to chronic pollution in the forest.

Air Pollutants↗

Ozone-induced nitrate formation in needles and leaves of Picea abies, Fagus sylvatica and Quercus robur.

Much attention has been paid to ozone as a major cause of novel forest decline in Europe. In combination with acidic mist, O(3) has been observed to increase ion leaching. Besides cations lake Mg(2+), Ca(2+), K(+), NH(4)(+), considerable amounts of nitrate were found to be leached by acidic mist from needles of Norway spruce. Controlled fumigation experiments, with 100, 300, and 600 microg O(3)m(-3) over 22 days continuously, have led to a nitrate accumulation of 94.1 +/- 14.8, 119.4 +/- 28.7 and 198.9 +/- 14.9 microg NO(3)(-1) g(-1) FW, respectively, in leaves of Quercus robur. Similar values were found in leaves of Fagus sylvatica and current and previous year needles of Picea abies. Nitrate levels of controls receiving charcoal filtered air were well below 40 microg NO(3)(-) g (-1) FW. Statistically significant elevated nitrate levels were observed after only 48 h of continuous fumigation with 600 microg O(3)m(-3), in all tree species tested, and after 144 h in the 100 microg O(3)m(-3) treatment. In another experiment, trees of Picea abies were kept in two charcoal (C) and two Purafil plus charcoal (P/C) ventilated chambers, and fumigated with O and 500 microg O(3)m(-3) in cabinets of each filter-type in order to eliminate NO(x) from chamber air. After 29 days of continuous ozone fumigation, NO(3)(-) accumulation in needles amounted to 102.0 +/- 37.7 and 137.4 +/- 40.5 microg g(-1) FW in P/C and C-filtered chambers, respectively. Nitrate contents of controls were below 30 microg NO(3)(-)g(-1) FW at the end of the experiment. No significant differences in NO(3)(-) accumulation between filter treatments were observed. Since NO(x) was reduced by more than 95% in the Purafil/charcoal versus the charcoal treatment, NO(3)(-) accumulation in needles can be attributed predominantly to the influence of ozone and not to direct NO(2) uptake of needles by the possible oxidation of NO to NO(2) in the presence of ozone.

Journal Article↗

Tree species (Picea abies and Fagus sylvatica) effects on soil water acidification and aluminium chemistry at sites subjected to long-term acidification in the Ore Mts., Czech Republic.

The effect of European beech (Fagus sylvatica) and Norway spruce (Picea abies) on acid deposition and soil water chemistry was studied at a site in the Ore Mts., Czech Republic, that has been subjected to decades of elevated acidic deposition. Dry deposition onto the spruce canopy significantly increased acid input to the soil in comparison to the beech canopy. As a result soil waters were more acidic; Al, SO4(2-), and NO3- concentrations were significantly higher; and Ca and K concentrations were lower in the spruce stand than in the beech stand. The concentrations of potentially toxic inorganic aluminium (Al(in)) were, on average, three times higher in the spruce stand than in the beech stand. Thus, Al played a major role in neutralizing acid inputs to mineral soils in the spruce stand. Despite the higher dissolved organic carbon (DOC) concentrations in spruce organic soil solutions, organic Al (Al(org)) accounted for only 30% of total Al (Al(tot)), whereas in beech organic soil solutions Al(org) was 60% of Al(tot). Soil waters in the beech stand exhibited Al(in) concentrations close to solubility with jurbanite (Al(SO4)OH.5H2O). The more acidic soil waters in the spruce stand were oversaturated with respect to jurbanite. The Bc/Al(in) ratio (Bc = Ca + Mg + K) in O horizon leachate was 4.6 and 70 in spruce and beech stands, respectively. In beech mineral soil solutions, the Bc/Al(in) ratio declined significantly to about 2. In the spruce stand, mineral soil solutions had Bc/Al(in) values below the critical value of 1. The observed Bc/Al(in) value of 0.4 at 30 cm depth in the spruce stand suggests significant stress for spruce rooting systems. A more favourable value of 31 was observed for the same depth in the beech stand. The efficiency of the spruce canopy in capturing acidic aerosols, particulates, and cloud water has resulted in the long-term degradation of underlying soils as a medium for sustainable forest growth.

Journal Article↗

Variations of wood delta(13)C for the past 50 years in declining Siebold's beech (Fagus crenata) forests.

Markedly damaged stands have been observed in Siebold's beech (Fagus crenata) forests on the Tanzawa Mountains, Japan, which are located close to densely populated and heavily industrial areas. We measured the delta(13)C records for the past 50 years (1944-1997) in wood cellulose sampled from relatively healthy trees in declining forests on Mt. Hinokiboramaru (1600 m). This may provide information on the history of stresses related to environmental changes, which have caused the decline symptoms. The results showed that, for all of the trees studied, wood delta(13)C has decreased with time. Also, the difference in delta(13)C among trees grew abruptly after the mid-1960s, which almost coincides with the time when the decline symptoms were markedly observed. Some trees with large reduction of wood delta(13)C exhibited the strong decreases in radial growth. This suggests that the reduction of tree growth may have been more greatly influenced by decreasing carboxylation rate than by stomatal limitation. It is unlikely that water stress and SO(2) and O(3) stresses have induced the growth reduction, because those stresses cause increasing wood delta(13)C. This is supported by the facts that wet conditions and relatively low SO(2) and O(3) levels have been observed near Mt. Hinokiboramaru. In addition, analyses of wood Ca showed no evidence that acid fog and soil acidification have affected the wood delta(13)C and growth through effects on the nutrient uptake of trees. However, what type of stresses have induced the large reduction of wood delta(13)C and growth for some of the trees studied remains unknown because of the lack of sufficient data for evaluation. In contrast, lesser reduction of wood delta(13)C from the other trees may be related to an increase in the plant water-use efficiency with increasing atmospheric CO(2) concentration.

Journal Article↗

Seasonal ozone response of mature beech trees (Fagus sylvatica) at high altitude in the Bavarian forest (Germany) in comparison with young beech grown in the field and in phytotrons.

Mature beech trees (Fagus sylvatica) grown at two different altitudes in the Bavarian forest were compared with young beech trees grown at nearby field sites or in phytotrons for their macroscopic and physiological responses to different ozone (O(3)) exposures. Cumulative O(3) exposure expressed as the sum of hourly mean concentrations above the canopy ranged between 100 and 150 microl l(-1) h, with the vertical O(3) profiles at the higher altitude site being enhanced by 30%. O(3) profiles at all sites were reduced by up to 20% with increasing depth within and beneath the canopy. The leaf discoloration that developed in the absence of premature leaf loss was similar in the sun foliage of mature and young trees (including plant grown in the phytotron). Injury became apparent at low O(3) exposures, expressed as accumulated hourly means over a threshold of 40 nl l(-1) (AOT40 <3.5 microl l(-1) h) at the lower site in both the mature trees and the young beech at the field site, but only occurred when AOT40 values reached 7 microl l(-1) h at the upper site, and 6 microl l(-1) h in the phytotrons. However, the association between injury and O(3) exposure was improved when cumulative ozone uptake to sun leaves was the ozone index, used with values of about 3 mmol m(-2) resulting in visible injury in both mature and young beech growing in phytotrons. Under high ozone exposure levels of inositol were lowered, whilst concentrations of lignin-like materials were enhanced in mature beech. Similar responses were observed in young beech grown in phytotrons. As the sun foliage was affected by only a small and variable extent each year, the seasonal O(3) impact at high altitude did not appear to pose an acute risk to mature beech trees.

Journal Article↗

Beech (Fagus sylvatica) response to ozone exposure assessed with a chlorophyll a fluorescence performance index.

This paper describes a relationship between ozone exposure, biomass, visual symptoms and a chlorophyll a fluorescence performance index for young beech trees (Fagus sylvatica). The plants were exposed to four levels of ozone in open-top fumigation chambers (50, 85, 100% of ambient, and 50% of ambient+30 nl l(-1) ozone) that fluctuated in parallel with ambient ozone during a single growing season. The trees were fumigated in the four treatments with ozone levels corresponding to an AOT40 (accumulated exposure above a threshold of 40 nl l(-1)) of 0.01, 3.35, 7.06 and 19.70 microl l(-1) h, respectively. Highly significant differences were found between the 50% of ambient+30 nl l(-1) ozone treatment and all other treatments, with a 70.5% reduction in primary photosynthetic performance, as measured with the PI index. The reduction of the PI values demonstrated a high correlation with visual symptom development (r(2)=0.98), and by the end of September with biomass loss (r(2)=0.99). A significant ozone exposure-response relationship was found between AOT40 and primary photochemistry (r(2)=0.97). Thus, analysis of PI provides an alternative method for regional monitoring of tree health within the context of the currently employed AOT40.

Journal Article↗

Effects of ozone exposure in open-top chambers on poplar (Populus nigra) and beech (Fagus sylvatica): a comparison.

Rooted cuttings of poplar (Populus nigra) and seedlings of beech (Fagus sylvatica) were exposed to ozone in open-top chambers for one growing season. Three treatments were applied: charcoal-filtered (CF), non-filtered (NF) and non-filtered air plus 30 ppb (nl l(-1)) ozone (NF+). Extra ozone was only added on clear days, from 09:00 until 17:00-20:00. The AOT40s (accumulated exposure over a threshold of 40 ppb), calculated from April to September were 4055 ppb.h for the NF and 8880 ppb.h for the NF+ treatments. For poplar ozone exposure caused highly significant reductions in growth rate, light-saturated net CO(2) assimilation rate, stomatal conductance, F(v)/F(m) and chlorophyll content. The largest effects were observed in August at which time ozone concentrations were elevated. A reduction was noticed in new leaf production, while accelerated ageing and visible damage to leaves caused high leaf losses. For beech the responses were similar but less pronounced: ozone exposure resulted in non-significant growth reductions, slight changes in light-saturated photosynthesis and accelerated leaf abscission. The chlorophyll content of beech leaves was not affected by the ozone treatments. The results confirmed previous observations that fast-growing tree species, such as most poplar species and hybrids, are more sensitive and responsive to tropospheric ozone than slower-growing species, such as beech. The growth reductions observed and reported here for beech were within the range of those reported in relationship to the AOT40 (accumulated exposure over a threshold of 40 ppb) critical level for ozone.

Journal Article↗