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Production, standing biomass and natural abundance of 15N and 13C in ectomycorrhizal mycelia collected at different soil depths in two forest types.

Nutrient uptake by forest trees is dependent on ectomycorrhizal (EM) mycelia that grow out into the soil from the mycorrhizal root tips. We estimated the production of EM mycelia in root free samples of pure spruce and mixed spruce-oak stands in southern Sweden as mycelia grown into sand-filled mesh bags placed at three different soil depths (0-10, 10-20 and 20-30 cm). The mesh bags were collected after 12 months and we found that 590+/-70 kg ha(-1) year(-1) of pure mycelia was produced in spruce stands and 420+/-160 kg ha(-1) year(-1) in mixed stands. The production of EM mycelia in the mesh bags decreased with soil depth in both stand types but tended to be more concentrated in the top soil in the mixed stands compared to the spruce stands. The fungal biomass was also determined in soil samples taken from different depths by using phospholipid fatty acids as markers for fungal biomass. Subsamples were incubated at 20 degrees C for 5 months and the amount of fungal biomass that degraded during the incubation period was used as an estimate of EM fungal biomass. The EM biomass in the soil profile decreased with soil depth and did not differ significantly between the two stand types. The total EM biomass in the pure spruce stands was estimated to be 4.8+/-0.9 x 10(3) kg ha(-1) and in the mixed stands 5.8+/-1.1 x 10(3) kg ha(-1) down to 70 cm depth. The biomass and production estimates of EM mycelia suggest a very long turnover time or that necromass has been included in the biomass estimates. The amount of N present in EM mycelia was estimated to be 121 kg N ha(-1) in spruce stands and 187 kg N ha(-1) in mixed stands. The delta13C value for mycelia in mesh bags was not influenced by soil depth, indicating that the fungi obtained all their carbon from the tree roots. The delta13C values in mycelia collected from mixed stands were intermediate to values from pure spruce and pure oak stands suggesting that the EM mycelia received carbon from both spruce and oak trees in the mixed stands. The delta15N value for the EM mycelia and the surrounding soil increased with soil depth suggesting that they obtained their entire N from the surrounding soil.

Biomass↗

Examining the strength and possible causes of the relationship between fire history and Sudden Oak Death.

Fire can be a dominant process in the ecology of forest vegetation and can also affect forest disease dynamics. Little is known about the relationship between fire and an emerging disease epidemic called Sudden Oak Death, which is caused by a new pathogen, Phytophthora ramorum. This disease has spread across a large, fire-prone portion of California, killing great numbers of oaks and tanoaks and infecting most associated woody plants. Suitable hosts cover a much broader geographic range, raising concern over where the disease may spread. To understand the strength and potential sensitivities of a fire-disease relationship, we examined geographic patterns of confirmed P. ramorum infections in relation to past fire history. We found these infections to be extremely rare within the perimeter of any area burned since 1950. This finding is not caused by spatial bias in sampling for the disease, and is robust to variation in host abundance scenarios and to aggregation of closely spaced sampling locations. We therefore investigated known fire-related factors that could result in significantly lower incidence of the disease in relatively recently burned landscapes. Chemical trends in post-fire environments can influence the success of pathogens like P. ramorum, either by increasing plant nutrient stress or by reducing the occurrence of chemicals antagonistic to Phytophthoras. Succession in the absence of fire leads to greater abundance of host species, which will provide increased habitat for P. ramorum; this will also increase intraspecific competition where these trees are abundant, and other density-dependent effects (e.g. shading) can reduce resource allocation to defenses. Despite these findings about a fire-disease relationship, a much deeper understanding is necessary before fire can be actively used as a tool in slowing the epidemic.

California↗

Whole-stream nitrate addition affects litter decomposition and associated fungi but not invertebrates.

We assessed the effect of whole-stream nitrate enrichment on decomposition of three substrates differing in nutrient quality (alder and oak leaves and balsa veneers) and associated fungi and invertebrates. During the 3-month nitrate enrichment of a headwater stream in central Portugal, litter was incubated in the reference site (mean NO3-N 82 microg l-1) and four enriched sites along the nitrate gradient (214-983 microg NO3-N l-1). A similar decomposition experiment was also carried out in the same sites at ambient nutrient conditions the following year (33-104 microg NO3-N l-1). Decomposition rates and sporulation of aquatic hyphomycetes associated with litter were determined in both experiments, whereas N and P content of litter, associated fungal biomass and invertebrates were followed only during the nitrate addition experiment. Nitrate enrichment stimulated decomposition of oak leaves and balsa veneers, fungal biomass accrual on alder leaves and balsa veneers and sporulation of aquatic hyphomycetes on all substrates. Nitrate concentration in stream water showed a strong asymptotic relationship (Michaelis-Menten-type saturation model) with temperature-adjusted decomposition rates and percentage initial litter mass converted into aquatic hyphomycete conidia for all substrates. Fungal communities did not differ significantly among sites but some species showed substrate preferences. Nevertheless, certain species were sensitive to nitrogen concentration in water by increasing or decreasing their sporulation rate accordingly. N and P content of litter and abundances or richness of litter-associated invertebrates were not affected by nitrate addition. It appears that microbial nitrogen demands can be met at relatively low levels of dissolved nitrate, suggesting that even minor increases in nitrogen in streams due to, e.g., anthropogenic eutrophication may lead to significant shifts in microbial dynamics and ecosystem functioning.

Alnus↗

A preliminary reconstruction (A.D. 1635-2000) of spring precipitation using oak tree rings in the western Black Sea region of Turkey.

Tree-ring data for Turkey are crucial to the understanding of the climatological effect of drought and rainfall from one era to the next. To this end, the present study reconstructed precipitation patterns in the western Black Sea region of Turkey. Tree-ring widths of oak trees were used to reconstruct March-June precipitation patterns for the years A.D. 1635-2000. According to the findings, during the past four centuries drought events in this region persisted for no more than 2 years, and extreme dry and wet events occurred generally in 1-year intervals. Historical records of droughts in Anatolia and neighboring countries corroborate the data furnished by tree-ring widths to indicate that major droughts and famine events occurred in 1725, 1757, 1887, 1890-1891, 1893-1894 and 1927-1928.

Climate↗

Phenophases alter the soil respiration-temperature relationship in an oak-dominated forest.

Soil respiration (SR) represents a major component of forest ecosystem respiration and is influenced seasonally by environmental factors such as temperature, soil moisture, root respiration, and litter fall. Changes in these environmental factors correspond with shifts in plant phenology. In this study, we examined the relationship between canopy phenophases (pre-growth, growth, pre-dormancy, and dormancy) and SR sensitivity to changes in soil temperature (T(S)). SR was measured 53 times over 550 days within an oak forest in northwest Ohio, USA. Annual estimates of SR were calculated with a Q(10) model based on T(S) on a phenological (PT), or annual timescale (AT), or T(S) and soil volumetric water content (VWC) on a phenological (PTM) or annual (ATM) timescale. We found significant (p<0.01) difference in apparent Q(10) from year 2004 (1.23) and year 2005 (2.76) during the growth phenophase. Accounting for moisture-sensitivity increased model performance compared to temperature-only models: the error was -17% for the ATM model and -6% for the PTM model. The annual models consistently underestimated SR in summer and overestimated it in winter. These biases were reduced by delineating SR by tree phenophases and accounting for variation in soil moisture. Even though the bias of annual models in winter SR was small in absolute scale, the relative error was about 91%, and may thus have significant implications for regional and continental C balance estimates.

Models, Theoretical↗

Soil modification by different tree species influences the extent of seedling ectomycorrhizal infection.

Established vegetation can facilitate the ectomycorrhizal infection of seedlings, but it is not known whether this interaction is limited by the phylogenetic relatedness of trees and seedlings. We use a series of bioassay experiments to test whether soil modification by different ectomycorrhizal tree species causes different levels of seedling infection, whether the extent of seedling infection is a function of the relatedness of tree and seedling, and whether the effect of trees on seedlings is mediated by biotic or abiotic soil factors. We found that soils from under different tree species do vary in their mycorrhizal infectiveness. However, this variation is not related to the genetic relatedness of trees and seedlings but instead, appears to be an attribute of the overstory species, irrespective of seedling species, mediated through a suite of humus- and base-cation-related abiotic effects on soils. Modification of abiotic soil properties by overstory trees should be considered as an important factor in the effect of different overstory trees on the extent of seedling mycorrhizal infection.

Ecosystem↗

Intra- and inter-specific variations in the copy number of two types of retrotransposons from the ectomycorrhizal basidiomycete Tricholoma matsutake.

To explore intra- and inter-specific variations of the ectomycorrhizal basidiomycete Tricholoma matsutake that produces the fruit body "matsutake", we carried out real-time PCR analysis based on two types of retrotransposons, one designated marY1, which resembles a retrovirus carrying the long terminal repeat (LTR) and the other marY2N, which resembles mRNA carrying the polyadenylated tail. Calculation based on the average genome size of homobasidiomycetes (34 Mbp) shows that ca. 5.5% of the total genome of T. matsutake isolated from Asia is made up of these retrotransposons, whereas they occupy ca. 1.4% in the isolates from Morocco, ca. 0.8% in isolates from Mexico, and ca. 0.5% in Tricholoma magnivelare, the species which produces "American matsutake". Other Tricholoma spp. that produce fruit bodies similar to those of T. matsutake, such as T. bakamatsutake, T. fulvocastaneum, and T. robustum, carry them in the region less than 0.05% of their total genome. Copy number of LTR of marY1 is consistently and markedly higher than that of the coding regions of marY1 and marY2N. Data suggest that retrotransposons are deeply involved in evolution of the ectomycorrhizal symbiont.

Agaricales↗

Survival analysis for a large-scale forest health issue: Missouri oak decline.

Survival analysis methodologies provide novel approaches for forest mortality analysis that may aid in detecting, monitoring, and mitigating of large-scale forest health issues. This study examined survivor analysis for evaluating a regional forest health issue - Missouri oak decline. With a statewide Missouri forest inventory, log-rank tests of the effects of covariates on the survivor function and equality of the survivor function among classes were conducted for selected oak species. Additionally, hazard functions were determined for diameter classes for damaged and undamaged oaks. Results indicate that mortality appears to vary significantly among some inventory classes such as oak species, but not among other classes such as ownership class. Indicators of individual tree vigor (i.e., crown class and ratio) were more significant predictors of oak tree mortality than site/stand attributes (i.e., density and aspect). Finally, results indicate that even fast-growing oak trees are at high risk of mortality if damaged by disease. Survival analyses, such as those applied in this study, may enable testing of forest health hypotheses using large-scale inventories. In the context of Missouri's oak forest decline, study results suggest management efforts should focus on limiting the spread of disease damage, increasing the vigor of residual trees, and emphasizing small trees when developing stand prescriptions.

Conservation of Natural Resources↗

An assessment of tree health and trace element accumulation near a coal-fired generating station, Manitoba, Canada.

A forest health assessment was performed in stands dominated by bur oak and trembling aspen to study the potential effects of airborne emissions from a 132 MW coal-fired station. Forty-two stands were sampled within a 16-km radius of the station for both foliar stress symptoms and trace element toxicology. The concentrations of tracer elements (As, Ba, Sr, and V) in the leaf litter were not spatially congruent with airborne emission deposition models (except Ba, which showed elevated levels immediately SE of the station), nor were they at phytotoxic levels. Elemental concentrations were significantly related to soil parameters including organic matter and texture. No patterns were found in forest health along directional or distance gradients from the generating station. Trembling aspen stands demonstrated little decline in general, but three of the 19 bur oak plots, all located on thin sandy soils developed on calcareous till, demonstrated branch dieback. In addition to poor soil conditions, two of these sites also had high water tables, and exhibited tree mortality. The bur oak decline did not appear to be related to emissions from the station, but is suspected to be a result of poor site quality, with urban development as a confounding factor.

Air Pollutants↗

Measuring photosynthetic parameters at a distance: laser induced fluorescence transient (LIFT) method for remote measurements of photosynthesis in terrestrial vegetation.

We have developed a laser induced fluorescence transient (LIFT) technique and instrumentation to remotely measure photosynthetic properties in terrestrial vegetation at a distance of up to 50 m. The LIFT method uses a 665 nm laser to project a collimated, 100 mm diameter excitation beam onto leaves of the targeted plant. Fluorescence emission at 690 nm is collected by a 250 mm reflective telescope and processed in real time to calculate the efficiency of photosynthetic light utilization, quantum efficiency of PS II, and the kinetics of photosynthetic electron transport. Operating with peak excitation power of 125 W m-2, and duty cycle of 10-50%, the instrument conforms to laser safety regulations. The LIFT instrument is controlled via an Internet connection, allowing it to operate from remote locations or platforms. Here we describe the theoretical basis of the LIFT methodology, and demonstrate its applications in remote measurements of photosynthetic properties in the canopy of cottonwood and oak trees, and in the rosette of Arabidopsis mutants.

Fluorescence↗

Hinnuliquinone, a C2-symmetric dimeric non-peptide fungal metabolite inhibitor of HIV-1 protease.

HIV-1 protease is one of several key enzymes required for the replication and maturation of HIV-1 virus. An almost two-decade research effort by academic and pharmaceutical institutions resulted in the successful commercialization of seven drugs that are potent inhibitors of HIV-1 protease activity and which, if used correctly, are highly effective in managing viral load. However, identification of clinical viral isolates that are resistant to these drugs indicates that this is a significant problem and that new classes of inhibitors are continually needed. Screening of microbial extracts followed by bioassay-guided isolation led to the discovery of a natural hinnuliquinone, a C(2)-symmetric bis-indolyl quinone natural product that inhibited the wild-type and a clinically resistant (A44) strain of HIV-1 protease with K(i) values of 0.97 and 1.25microM, respectively. Crystallographic analysis of the inhibitor-bound HIV-1 protease helped explain the importance of the C(2)-symmetry of hinnuliquinone for activity. Details of the isolation, biological activity, and crystallographic analysis of the inhibitor-bound protease are herein described.

Benzoquinones↗

Yield, size and bacterial blotch resistance of Pleurotus eryngii grown on cottonseed hulls/oak sawdust supplemented with manganese, copper and whole ground soybean.

Experiments were performed to determine effects of supplementation of cottonseed hull/sawdust substrate with Mn, Cu, and ground soybean on yield, mushroom size, and bacterial blotch resistance of two commercial strains of Pleurotus eryngii. A basal formulation (d.w.) of cottonseed hulls (62%), aged red oak sawdust (27%), whole ground soybean (6%), corn distiller's waste (4%) and calcium sulfate (1%) was supplemented to 50, 150 or 250 microg/g Mn or Cu and to 4%, 8% and 12% whole ground soybean. The cottonseed hulls content in the basal substrate was adjusted to compensate for the addition of ground soybean. Formulated substrates were mixed, placed in 1050ml bottles, and sterilized at 121 degrees C for 90min. Mushroom yields were significantly higher from substrates containing Mn at 50 microg/g and soybean at 8% and 12% supplementation compared to the basal substrate. As the level of soybean addition to substrate increased, yield also increased. The addition of Mn at levels of 150 and 250 microg/g significantly enhanced yield as well, although less than did the 50 microg/g treatment. To assess the influence of mushroom strain and substrate composition on blotch disease severity, pilei of P. eryngii were inoculated with Pseudomonas tolaasii. Strain WC888 was more resistant to disease than WC846. Disease severity was greater when substrates were amended with Cu to 150 or 250 microg/g. There was a significant difference in inherent levels of Cu in the basidiomata of different strains, but P. eryngii did not accumulate Cu and disease severity was not correlated with Cu content of the basidiomata.

Analysis of Variance↗

Ground wheat straw as a substitute for portions of oak wood chips used in shiitake (Lentinula edodes) substrate formulae.

Oak woodchips, used for production of shiitake Lentinula edodes (Berk) Pegler, are increasingly difficult to obtain due to dwindling supplies. We investigated the effect of adding ground wheat straw as a substitute for portions of oak woodchips in substrate formulae on mushroom yield and size. We also determined the effect of mushroom cropping on relative feed value (RFV) by chemical analysis of the substrate at spawning (AS) and after cropping (AC). Three formulae containing 0%, 8% and 16% ground wheat straw and 52%, 44% and 36% oak sawdust, respectively, were bulk pasteurized (111 degrees C for 20 min) in an autoclaving mixer, subjected to spawn run (21 d), browning (28 d) and a production cycle of three breaks (38 d). Mean (4 crops) mushroom yields were 11% higher when 8% wheat straw was used in the medium and 19% higher when 16% wheat straw was substituted for portions of oak sawdust. There were no significant differences in mushroom sizes between any of the treatments. Relative feed values of shiitake substrates AC increased more dramatically as more wheat straw was added to the formulae. Using mature alfalfa (full bloom) as a base value of 100%, RFVs for substrate AS were 98%, 92%, and 92% for 0%, 8% and 16% straw, respectively; RFVs AC were 118%, 120% and 133%, respectively. Substrate AC containing 16% straw had a RFV comparable to corn silage (well-eared). Fat contents of the substrates decreased by 50-62% AC, whereas potassium contents decreased by 40%. Use of ground wheat straw in synthetic medium would not only increase mushroom yield by up to 19%, but may help alleviate periodic shortages of oak sawdust. In addition, growers would avoid the added expense of aging the wheat straw (for 8-12 week) as is typically done for oak sawdust in the industry. This is the first report of RFVs for spent shiitake substrate (SSS) predicting its excellent potential for use as animal feed.

Agriculture↗

Aleppo tannin: structural analysis and salivary amylase inhibition.

The effectiveness and specificity of a tannin inhibition on human salivary amylase (HSA) catalyzed hydrolysis was studied using 2-chloro-4-nitrophenyl 4-O-beta-D-galactopyranosyl-alpha-maltoside (GalG(2)-CNP) and amylose substrates. Aleppo tannin was isolated from the gall nut of Aleppo oak. This tannin is a gallotannin, in which glucose is esterified with gallic acids. This is the first kinetic report, which details the inhibitory effects of this compound on HSA. A mixed non-competitive type inhibition has been observed on both substrates. The extent of inhibition is markedly dependent on the substrate-type. Kinetic constants were calculated from Lineweaver-Burk secondary plots for GalG(2)-CNP (K(EI) 0.82 microg mL(-1), K(ESI) 3.3 microg mL(-1)). This indicates a 1:1 binding ratio of inhibitor-enzyme and/or inhibitor-enzyme-substrate complex. When amylose was the substrate the binding ratio of inhibitor to enzyme-substrate complex was found to be 2:1, with the binding constants of K(EI) 17.4 microg mL(-1), K(ESI) 14.9 microg mL(-1), K(ESI(2)) 9.6 microg mL(-1). Presumably, the tannin inhibitor can bind not only to HSA, but to the amylose substrate, as well. Kinetic data suggest that Aleppo tannin is a more efficient amylase inhibitor than the recently studied other tannin with quinic acid core (GalG(2)-CNP: K(EI) 9.0 microg mL(-1), K(ESI) 47.9 microg mL(-1)).

Amylases↗

Treatment of cork boiling wastewater using chemical oxidation and biodegradation.

Three cultures were enriched from cork boiling wastewater using tannic acid as the selective carbon substrate, at 25 degrees C and pH 7.2, 25 degrees C and pH 4.7 and 50 degrees C and pH 4.7. The enrichment culture obtained at neutral pH was composed of five culturable isolates, whereas from each acidic enrichment two bacterial strains were isolated. Mesophilic isolates were Gram negative bacteria belonging to the genera Klebsiella, Pseudomonas, Stenotrophomonas and Burkholderia. Thermophilic isolates were members of the genus Bacillus. Despite the capability of the enrichment cultures to use tannic acid as single carbon and energy source, those cultures were unable to reduce the total polyphenols or the total organic carbon content of cork boiling wastewater. In order to increase the bioavailability of the organic carbon in cork boiling wastewater, biodegradation was preceded by Fenton oxidation. It was demonstrated that the combined process, using small amounts of Fenton reagents and biodegradative inoculum added almost simultaneously to cork boiling wastewater, leads to TOC reductions of more than 90%.

Biodegradation, Environmental↗

Multiple solid-phase microextraction in a non-equilibrium situation. Application in quantitative analysis of chlorophenols and chloroanisoles related to cork taint in wine.

Multiple HS-solid-phase microextraction (MHS-SPME) is a modification of SPME developed for quantitative analysis that avoids possible matrix effects based on an exhaustive analyte extraction from the sample. In this paper, the theory of this process associated with a non-equilibrium situation has been presented. The application of an optimised HS-SPME-based method in the analysis of chloroanisoles and chlorophenols, previously acetylated, associated with the occurrence of cork taint in different red, white and rosé wine samples, has revealed the existence of matrix effects. This fact determines the choice of standard addition as the adequate technique for the quantification of these compounds in real samples. MHS-SPME is proposed as a good alternative technique with respect to HS-SPME because it avoids matrix effects, simplifies the quantification of these compounds in real samples and reduces analysis time, providing sensitivity below chloroanisole sensory threshold with acceptable precision.

Anisoles↗

Stir bar sorptive extraction for the determination of volatile compounds in oak-aged wines.

Stir bar sorptive extraction (SBSE) followed by a thermal desorption-gas chromatography-mass spectrometry analysis has been optimized for the determination of 13 of the most important oak volatiles in wine samples, all in a single run. The stir bar sorptive extraction method was optimized in terms of temperature, time, pH and NaCl addition, and the best results were obtained after stirring the wine sample with the polydimethylsiloxane stir bar during 90min at room temperature. The addition of sodium chloride did not enhance the volatile extraction. The method proposed showed good linearity over the concentration range tested, with correlation coefficients ranging from 0.92 to 0.99 for all the analytes. The reproducibility and repeatability of the method was estimated between 0.11 and 5.45%. The detection and quantification limits of all analytes were lower than their respective olfactory threshold values, and, most importantly, no artifacts have been observed during the analysis as described in most extractions using other current methodologies.

Flavoring Agents↗

Multiple response optimisation based on desirability functions of a microwave-assisted extraction method for the simultaneous determination of chloroanisoles and chlorophenols in oak barrel sawdust.

A microwave-assisted extraction (MAE) method was optimised for extracting 2,4,6-trichloroanisole (TCA), 2,3,4,6-tetrachloroanisole (TeCA), pentachloroanisole (PCA), 2,4,6-trichlorophenol (TCP), 2,3,4,6-tetrachlorophenol (TeCP) and pentachlorophenol (PCP) from oak barrels. The method was optimised by using a central composite experimental design with extraction time, temperature and solvent volume as influential parameters. A desirability function was then employed in addition to the simultaneous optimisation of the compounds. The optimal conditions identified were temperature 130 degrees C, solvent volume 35 mL and extraction time 50 min. The compounds were determined by gas chromatography with electron-capture detection. MAE was compared with conventional Soxhlet extraction; the results obtained with MAE were in good agreement with those obtained by Soxhlet extraction.

Anisoles↗