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Anaerobic treatability of waste water from pulp and paper industries.

A black liquor evaporator condensate from a Kraft mill and a waste water from production of corrugating medium were anaerobically treated on a laboratory scale. The composition of the waste waters was determined before and after treatment in fixed bed reactors. Toxicity studies by the Microtox-method showed that both waste waters were highly toxic and a slight decrease in toxicity was achieved by anaerobic treatment. Despite the toxicity efficient anaerobic treatment was obtained. Major components of the condensate were methanol, ethanol, acetone, guaiacol, hydrogen sulfide and dimethyl disulfide. Anaerobic treatment reduced the concentration of the major components considerably with one exception. The concentration of hydrogen sulfide was unchanged. Organic overloading of the fixed bed reactor or a temperature drop resulted in an accumulation of acetone, although methanol and ethanol were degraded. Major components of the waste water from the production of corrugating medium were: Klason-lignin, acid-soluble lignin, carbohydrates, extractives and ash. When the fixed bed reactor was operated at a volumetric load of 1.6 kg COD/m(3).d the following reductions were obtained: Klason - lignin (solids fraction 84%; soluble and colloidal fraction 76%), acid-soluble lignin (solids fraction 56%; soluble and colloidal fraction 7%), carbohydrates (100%), extractives (71%), total-S (80%), COD (73%) and BOD(7) (78%). Kinetic studies showed that condensate was more easily degraded anaerobically than corrugating medium waste water.

Journal Article↗

In vitro binding of estrogens by dietary fiber and the in vivo apparent digestibility tested in pigs.

Within the framework of experiments related to the association between dietary fiber and breast cancer an in vitro test system was used to study the binding of estrogens to various fibers (e.g. cholestyramin, lignin and cellulose) and fiber sources (e.g. wheat bran, cereals, seeds and legumes). Furthermore, the in vivo apparent digestibility of the different fiber sources was tested using a mobile nylon bag technique in intestine-cannulated pigs. Estradiol-17 beta (E2) bound more strongly to the various fibers than did estrone (E1), estriol or estrone-3-glucuronide. At increasing pH (greater than 7) binding of both E1 and E2 to wheat bran decreased significantly. Cholestyramine and lignin bound almost all estrogens present in the medium. Linseed (91%), oats (83%), barley chaff (88%) and wheat bran (82%) are other excellent binders of E2. Corn, rye and white wheat flour showed lower binding capacity with a relatively low affinity. Cereals with the highest percentage of lignin in the fiber (greater than 3%) were also the fiber sources with the lowest apparent digestibility. Estrogens bound with the highest affinity (relative to bovine serum albumin) to these fiber sources. Together with wheat bran and lignin, oats, linseed and soybean seem to be products with good perspectives for in vivo evaluation of the lowering effect of dietary fiber on estrogen exposure of estrogen-sensitive tissues.

Animals↗

Regression models of ultimate methane yields of fruits and vegetable solid wastes, sorghum and napiergrass on chemical composition.

Several fractions of fruits and vegetable solid wastes (FVSW), sorghum and napiergrass were analyzed for total solids (TS), volatile solids (VS), total organic carbon, total kjeldahl nitrogen, total soluble carbohydrate, extractable protein, acid-detergent fiber (ADF), lignin, cellulose and ash contents. Their ultimate methane yields (B(o)) were determined using the biochemical methane potential (BMP) assay. A series of simple and multiple regression models relating the B(o) to the various substrate constituents were generated and evaluated using computer statistical software, Statistical Package for Social Sciences (SPSS). The results of simple regression analyses revealed that, only weak relationship existed between the individual components such as carbohydrate, protein, ADF, lignin and cellulose versus B(o). A regression of B(o) versus combination of two variables as a single independent variable such as carbohydrate/ADF and carbohydrate+protein/ADF also showed that the relationship is not strong. Thus it does not appear possible to relate the B(o) of FVSW, sorghum and napiergrass with single compositional characteristics. The results of multiple regression analyses showed promise and the relationship appeared to be good. When ADF and lignin/ADF were used as independent variables, the percentage of variation accounted for by the model is low for FVSW (r(2)=0.665) and sorghum and napiergrass (r(2)=0.746). Addition of nitrogen, ash and total soluble carbohydrate data to the model had a significantly higher effect on prediction of B(o) of these wastes with the r(2) values ranging from 0.9 to 0.99. More than 90% of variation in B(o) of FVSW could be accounted for by the models when the variables carbohydrate, lignin, lignin/ADF, nitrogen and ash (r(2)=0.904), carbohydrate, ADF, lignin/ADF, nitrogen and ash (r(2)=0.90) and carbohydrate/ADF, lignin/ADF, lignin and ash (r(2)=0.901) were used. All the models have low standard error values, which indicate the amount of spread is less. Thus, considering only the higher r(2) values, six models are proposed for predicting the B(o) based on FVSW data and sorghum and napiergrass data. It would be more convenient if B(o) could be predicted by analyzing the chemical composition of the substrate rather than performing the long-term batch fermentation. To test the validity of the regression models, chemical constituents of FVSW that were not included in the regression analyses were determined and their experimental B(o) were determined by BMP assay. All the six models were used to predict the B(o) from the chemical constituents of these FVSW. It was found that most of the predicted values were within 20% of the experimental B(o) in models 1, 3 and 6. Since models 3 and 6 used the same variables namely, total soluble carbohydrate, ADF, lignin/ADF, nitrogen and ash, B(o) can be predicted from these five chemical constituents which accounts for more than 90% of the variation in B(o) (r(2)>90).

Fruit↗

A comparison of chemical pretreatment methods for improving saccharification of cotton stalks.

The effectiveness of sulfuric acid (H(2)SO(4)), sodium hydroxide (NaOH), hydrogen peroxide (H(2)O(2)), and ozone pretreatments for conversion of cotton stalks to ethanol was investigated. Ground cotton stalks at a solid loading of 10% (w/v) were pretreated with H(2)SO(4), NaOH, and H(2)O(2) at concentrations of 0.5%, 1%, and 2% (w/v). Treatment temperatures of 90 degrees C and 121 degrees C at 15 psi were investigated for residence times of 30, 60, and 90 min. Ozone pretreatment was performed at 4 degrees C with constant sparging of stalks in water. Solids from H(2)SO(4), NaOH, and H(2)O(2) pretreatments (at 2%, 60 min, 121 degrees C/15 psi) showed significant lignin degradation and/or high sugar availability and hence were hydrolyzed by Celluclast 1.5L and Novozym 188 at 50 degrees C. Sulfuric acid pretreatment resulted in the highest xylan reduction (95.23% for 2% acid, 90 min, 121 degrees C/15 psi) but the lowest cellulose to glucose conversion during hydrolysis (23.85%). Sodium hydroxide pretreatment resulted in the highest level of delignification (65.63% for 2% NaOH, 90 min, 121 degrees C/15 psi) and cellulose conversion (60.8%). Hydrogen peroxide pretreatment resulted in significantly lower (p<or=0.05) delignification (maximum of 29.51% for 2%, 30 min, 121 degrees C/15 psi) and cellulose conversion (49.8%) than sodium hydroxide pretreatment, but had a higher (p<or=0.05) cellulose conversion than sulfuric acid pretreatment. Ozone did not cause any significant changes in lignin, xylan, or glucan contents over time. Quadratic models using time, temperature, and concentration as continuous variables were developed to predict xylan and lignin reduction, respectively for sulfuric acid and sodium hydroxide pretreatments. In addition, a modified severity parameter (log M(0)) was constructed and explained most of the variation in xylan or lignin reduction through simple linear regressions.

Biotechnology↗

Storage stability of Anagrapha falcifera nucleopolyhedrovirus in spray-dried formulations.

A multiply embedded nucleopolyhedrovirus isolated from Anagrapha falcifera (Kirby) (AfMNPV) can lose insecticidal activity during months of dry storage in ambient room conditions. We tested the spray-dried AfMNPV formulations after storage for up to 1 year at room temperatures for insecticidal activity against neonate Trichoplusia ni (Hübner). Experimental formulations were made using combinations of corn flours, lignin, and sucrose, and were selected based on previous work which demonstrated that these formulations resisted solar degradation in field experiments. Twelve experimental formulations (organized in three groups of four formulations) compared the effect of (1) the ratio of formulation ingredients (lignin and corn flour) to virus concentration, (2) different sources of lignin, or (3) different corn flours and sugar. Based on a single-dose plant assay with these 12 formulations, none of the formulations lost significant activity due to the drying process, when compared with the unformulated wet AfMNPV. Samples of the 12 dried formulations were stored at room (22+/-3 degrees C) and refrigerated (4 degrees C) temperatures. Insecticidal activity (LC(50)) was determined with a dosage-response assay for neonates fed on treated cotton-leaf disks. After 6 (or 9) and 12 months storage, refrigerated samples maintained insecticidal activity better than corresponding samples stored at room temperatures with LC(50)s that averaged 2.0 x 10(6) polyhedral inclusion bodies per milliliter (pibs/ml) for refrigerated samples and 5.4 x 10(6) pibs/ml for samples stored at room temperatures. Compared with unformulated stock virus stored frozen, six formulations stored at room temperature and 10 formulations stored in the refrigerator did not lose significant insecticidal activity after 1 year based on overlapping 90% confidence intervals. Changing the ratio of virus to formulation ingredients did not provide a clear trend over the range of concentrations tested, and may be less important for shelf-life of virus activity compared with formulations made with different ingredients. Two of the four formulations made with different lignins were about 15 times less active after 1 year at room temperature compared with refrigerated samples, indicating that specific formulation ingredients can affect storage stability. Formulations that contained sugar generally maintained activity during storage better than formulations without sugar. Unformulated virus stock maintained insecticidal activity (ranged from 0.20 to 2.5 x 10(6) pibs/ml) better during storage than dried formulations with LC(50)s that ranged from 0.39 to 27 x 10(6) pibs/ml. Unformulated virus stock, which is essentially a suspension of virus occlusion bodies in homogenized insect cadavers, did not lose activity when stored at refrigerated or room temperature. We believe that stability of AfMNPV insecticidal activity during storage as dry formulations is related to the general composition of the formulation and that sugar may play a critical role in maintaining insecticidal activity.

Animals↗

Enzyme production activity of Phanerochaete chrysosporium and degradation of pentachlorophenol in a bioreactor.

Lignin peroxidase production by a white rot fungus, Phanerochaete chrysosporium, was experimentally investigated using a batch system and a reactor system with various carriers. Immobilization of mycelia cell culture was more effective in promoting cell growth and lignin peroxidase production compared to conventional stationary liquid culture. Biostage carrier, commonly used for biochemical treatment in a fluidized bed disposal system, greatly improved production of lignin peroxidase up to 8.1 U/mL in the batch system. The packed bed reactor system was operated using a repeated batch technique, consisting of alternating growth and production phases, to sustain lignin peroxidase growth and production during the entire experiment period. Steady-state continuous PCP degradation over an extended period was accomplished with a mineralization ratio exceeding 80%. These systems and operation methods are promising techniques for the treatment of hazardous waste.

Bioreactors↗

Removal of phenolics in olive mill wastewaters using the white-rot fungus Pleurotus ostreatus.

Olive mill wastewaters (OMW) present a major environmental problem. The large amounts generated, combined with the high phenols and chemical oxygen demand concentrations, are the main difficulties in finding a solution for the management of these wastewaters, which are dangerous for the environment. The phenols, which are contained in the OMW have a structure similar to lignin, which makes them difficult to biodegrade. Lignin can be degraded only by a few microorganisms, such as "white-rot" basidiomycete, which produce manganese (MnPs) and lignin peroxidases (LiPs) and laccases that are responsible for the oxidisation of lignin compounds. The capability of Pleurotus ostreatus to degrade phenols of OMW in different conditions such as in sterilized and thermally processed (at 100 degrees C) wastewater, with and without dilution, is investigated in this work. According to the experimental results P. ostreatus removed phenols from the culture medium, under all different conditions that were examined. The degradation of phenols reached up to 78.3% for the sterilized and 50% diluted OMW, 66.7% and 64.7% for the thermally processed OMW, with and without dilution, respectively. The effect of pre-treatment of OMW on the performance of anaerobic digestion is also assessed, as methanogenic bacteria are seriously affected by the presence of phenol compounds. The pre-treated wastewater was shown to be more amenable to a subsequent anaerobic digestion.

Biodegradation, Environmental↗

Hydroxyl radical generation by an extracellular low-molecular-weight substance and phenol oxidase activity during wood degradation by the white-rot basidiomycete Trametes versicolor.

One-electron oxidation activity, as measured by ethylene generation from 2-keto-4-thiomethylbutyric acid, phenol oxidase activity, and the generation of hydroxyl radical were examined in cultures of the lignin-degrading white-rot basidiomycete fungus, Trametes (Coriolus) versicolor. The activity levels of specific lignin-degrading enzymes and cellulases, as well as the rate of wood degradation, also were examined. The fungus secreted a low-molecular-weight substance (M(r) 1000-5000) that catalyzed a redox reaction between molecular oxygen and an electron donor, to produce the hydroxyl radical via hydrogen peroxide. During wood decay, T. versicolor also produced significant amounts of laccase and lignin peroxidase, carboxymethyl cellulase, and Avicelase. The roles of the hydroxyl radical, phenol oxidases, and cellulases in wood degradation by white-rot fungi are discussed. That the hydroxyl radical produced by the low-molecular-weight substance secreted by T. versicolor results in new phenolic substructures on the lignin polymer, making it susceptible to attack by laccase or manganese peroxidase is suggested.

Journal Article↗

Chemometrics: an important tool for the modern chemist, an example from wood-processing chemistry

This study briefly outlines the idea of principal component analysis and cross-correlation calculations (applied chemometrics) and presents an illustrative example from wood-processing chemistry. The applicability of chemometric data analysis was demonstrated by investigating the various structural changes that take place in dissolved and degraded lignin ("kraft lignin") during laboratory-scale kraft pulping of Scots pine (Pinus sylvestris) and silver birch (Betula pendula). The structural data (31P NMR and size exclusion chromatographic data) on kraft lignin were further processed by chemometric multivariate techniques (PCA and 2DCC), confirming, for example, that the cleavage of beta-aryl ether structures, the most prominent linkages between monomeric units, is directly related to the decrease in the average molecular mass of lignin.

Journal Article↗

Molecular characterization of biodegradable dissolved organic matter using bioreactors and [12C/13C] tetramethylammonium hydroxide thermochemolysis GC-MS.

Little is known about the molecular composition of the biodegradable fraction of dissolved organic matter (BDOM) in stream ecosystems. We combined plug-flow biofilm reactors, tetramethylammonium hydroxide (TMAH) thermochemolysis GC-MS, and 13C-labeled TMAH thermochemolysis GC-MS to study the molecular composition of BDOM from two stream ecosystems. TMAH products derived from fatty acids, lignin, and other aromatic molecules were quantified using an internal standard approach. We applied the 13C-TMAH thermochemolysis procedure to differentiate between compounds in dissolved organic matter (DOM) that had natural methoxyl groups from those that acquired methoxyl groups during the TMAH reaction. In Rio Tempisquito, a stream draining a tropical evergreen forest, and White Clay Creek, a stream draining a temperate deciduous woodlands, carbohydrates, fatty acids, and lignin contributed to the DOM and BDOM molecular composition. We observed 97 different peaks in the chromatograms of streamwater, with 57% of the peaks common to both streams. The DOM and BDOM pools from each site also contained a unique suite of compounds. Our combined use of TMAH and 13C-TMAH thermochemolysis revealed that heterotrophic bacteria can selectively degrade and demethylate different types of compounds in the lignin residues of DOM. This demonstration of bacterial demethylation of lignin, an abundant and refractory plant molecule, has potential implications for global carbon cycling.

Biodegradation, Environmental↗

Oxidation of spruce wood sawdust by MnO(2) plus oxalate: a biochemical investigation.

This paper reports the modification/degradation of lignin within spruce sawdust by manganese complexes formed by the association of MnO(2) and oxalate. The Mn oxidants formed are shown to modify both the chemical and physical properties of the wood cell wall. Scanning electron microscopy analysis of oxidized tracheids revealed a smoothing of the cell wall surface from the lumen side due to the removal of some material. Thioacidolysis analysis of the oxidized lignin showed reductions of up to 30% in the recovery of ether-linked guaiacyl monomers and up to 45% for the some dimers composing the polymer. The MnO(2)/oxalate system also slightly modified polysaccharides, corresponding to a 10% loss in weight of arabinose and glucose in the oxidized sample. However, no delignification occurred, according to the acid insoluble lignin content of spruce. Oxalic acid at pH 2.5 did not induce detectable changes in the chemical structures of the lignin or of the polysaccharides.

Manganese Compounds↗

A novel aromatic alcohol dehydrogenase in higher plants: molecular cloning and expression.

Cinnamyl alcohol dehydrogenase (CAD; EC 1.1.195) catalyses the conversion of p-hydroxy-cinnamaldehydes to the corresponding alcohols and is considered a key enzyme in lignin biosynthesis. In a previous study, an atypical form of CAD (CAD 1) was identified in Eucalyptus gunnii [12]. We report here the molecular cloning and characterization of the corresponding cDNA, CAD 1-5, which encodes this novel aromatic alcohol dehydrogenase. The identity of CAD 1-5 was unambiguously confirmed by sequence comparison of the cDNA with peptide sequences derived from purified CAD 1 protein and by functional expression of CAD 1 recombinant protein in Escherichia coli. Both native and recombinant CAD 1 exhibit high affinity towards lignin precursors including 4-coumaraldehyde and coniferaldehyde, but they do not accept sinapaldehyde. Moreover, recombinant CAD 1 can also utilize a wide range of aromatic substrates including unsubstituted and substituted benzaldehydes. The open reading frame of CAD 1-5 encodes a protein with a calculated molecular mass of 35,790 Da and an isoelectric point of 8.1. Although sequence comparisons with proteins in databases revealed significant similarities with dihydroflavonol-4-reductases (DFR; EC 1.1.1.219) from a wide range of plant species, the most striking similarity was found with cinnamoyl-CoA reductase (CCR; EC 1.2.1.44), the enzyme which directly precedes CAD in the lignin biosynthetic pathway. RNA blot analysis and immunolocalization experiments indicated that CAD 1 is expressed in both lignified and unlignified tissues/cells. Based on the catalytic activity of CAD 1 in vitro and its localization in planta, CAD 1 may function as an 'alternative' enzyme in the lignin biosynthetic pathway. However, additional roles in phenolic metabolism are not excluded.

3-Hydroxysteroid Dehydrogenases↗

Wood digestion in Pselactus spadix Herbst--a weevil attacking marine timber structures.

Pselactus spadix tunnels timber structures in the marine environment. Recent studies reported a cosmopolitan distribution for this weevil, which is frequently found in harbour and port areas. P. spadix feeds on timber (hardwood and softwood) in immature and adult life stages, but its digestion of wood components had not been investigated. Using dry weight analyses of tunnel walls and frass produced, P. spadix adults consumed Scots pine with soft rot decay at a rate of 1.59 +/- 0.37 mg d-1 and the digestibility of this substrate was 57.96 +/- 5.89 (i.e. for 100 mg consumed SR-pine, 58 mg was digested). Using gravimetric analysis to quantify structural wood components in tunnel walls and frass, P. spadix adults were found to digest cellulose, lignin and hemicellulose with digestibility coefficients of 82.2, 41.2 and 14.5 respectively. Fourier Transform Infrared (FTIR) spectroscopy analyses of tunnel walls and frass of adults and larvae from soft rotted pine also indicated digestion of all structural components, with larvae digesting cellulose and lignin more efficiently than adults. When FTIR was employed to analyse adult tunnel walls and frass from undecayed pine, cellulose and hemicellulose were digested, but no evidence of lignin digestion was found. This study shows that adults digest lignin when soft rot is present and suggests a symbiotic function of wood degrading microorganisms.

Animals↗

Stoichiometry of foliar carbon constituents varies along light gradients in temperate woody canopies: implications for foliage morphological plasticity.

Foliar morphology and chemical composition were examined along a light gradient in the canopies of five deciduous temperate woody species, ranked according to shade-tolerance as Populus tremula L. < Fraxinus excelsior L. < Tilia cordata Mill. = Corylus avellana L. < Fagus sylvatica L. Foliar carbon was divided between structural (cell-wall polysaccharides, lignin) and nonstructural (proteins, ethanol-soluble carbohydrates, starch) fractions. Foliar morphology of all species was strongly affected by irradiance. Both leaf dry mass per area (M(A)), a product of leaf density and thickness, and leaf dry to fresh mass ratio (D(w)), characterizing the apoplastic leaf fraction, increased with increasing relative irradiance (I(sum), calculated as the weighted mean of fractional penetration of diffuse and direct irradiance). Though the relationships were qualitatively identical among the taxa, more shade-tolerant species generally had lower values of M(A) than shade-intolerant species, and their morphological relationships with irradiance were curvilinear; however, there were no signs of saturation even at the highest irradiances in shade-intolerant species. In all species, lignin concentrations increased and cell-wall polysaccharide concentrations decreased with increasing irradiance. Consequently, biomass investment in structural leaf components appeared to be relatively constant along light gradients. The relationship between irradiance and structural compounds tended to be asymptotic in the more shade-tolerant species, whereas M(A) was linearly correlated with concentrations of structural leaf components, suggesting that similar factors were responsible for the curvature in the morphological and chemical relationships with irradiance. Because lignin increases tissue elastic modulus thereby rendering leaves more resistant to low leaf water potentials, we conclude that changes in stoichiometry of cell wall components were related to foliage acclimation to the gradients of water deficit that develop in the canopy and inherently accompany light gradients. We also conclude that increased lignification decreased leaf expansion growth, and that species differences in lignification were partly responsible for the observed interspecific variability in morphological plasticity. Analysis of structural leaf compounds provided no indication of how shade-intolerant species with low investments in lignin acclimated to gradients of water availability in the canopy. Because shade-intolerant species generally had higher capacities for photosynthesis than shade-tolerant species, we postulated that they should also have a greater ability for osmotic adjustment of leaf water potential with photosynthates. The concentrations of soluble carbohydrates increased with increasing irradiance in all species; however, the osmotic adjustment achieved in this way was similar in all species, except for shade-intolerant F. excelsior, which had a lower potential for osmotic adjustment with carbohydrates than the other taxa. Although we did not determine whether the gradients of stem water potential and leaf water deficits were similar in canopies of different species, we demonstrated that water relations play a central role in determining foliar structure and composition along light gradients in the canopy.

Journal Article↗

Differences in chemical composition relative to functional differentiation between petioles and laminas of Fraxinus excelsior.

Differences in structural and nonstructural carbohydrates, lignin and chlorophyll, and Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase) activity between petioles and leaflets were studied along a canopy light gradient in Fraxinus excelsior L., which has pinnate compound leaves and up to 20% of foliar biomass invested in petioles. Long-term light conditions at the sampling locations were characterized by values of seasonal mean integrated quantum flux density (Q(int), mol m(-2) day(-1)) estimated by combining data from hemispherical photographs at the sampling locations with measurements of global solar radiation above the canopy during the growing season. The contribution of petioles to leaf carbon assimilation was disproportionally lower than that of leaf laminas. Though the light relationships of assimilative compounds-foliar chlorophyll concentration increasing with decreasing Q(int) to improve leaf absorptance, foliar N concentration and Rubisco activity being relatively constant along the light gradient-were similar for both petioles and leaflets, petiole nitrogen and chlorophyll concentrations were only 30% and 10%, respectively, of those of leaflets. Nonstructural carbohydrate concentration was about 20% higher in petioles than in leaf laminas, indicating that petioles also serve as storage tissues for photosynthates. Relationships between foliar structural carbon components and irradiance-increasing lignin (L) and decreasing structural polysaccharide (SP) concentrations with increasing Q(int)-were qualitatively similar for petioles and leaflets. However, petioles had lower L, but higher SP and total investment in structural compounds (L + SP) than leaflets. Greater lignification at high irradiances in leaflets than in petioles was attributed to greater water stresses at high light, and to more variable water contents of actively transpiring leaflets. Low lignin concentration in combination with high osmotically active carbohydrate concentrations in petioles suggest that turgor plays an important role in the mechanical properties of petioles. As a result of lower lignin and protein concentrations, the glucose cost of petiole construction (g glucose per g dry mass) was about 5% lower than that of leaf laminas.

Journal Article↗

Effects of elevated concentrations of atmospheric CO2 and tropospheric O3 on leaf litter production and chemistry in trembling aspen and paper birch communities.

Human activities are increasing the concentrations of atmospheric carbon dioxide ([CO2]) and tropospheric ozone ([O3]), potentially leading to changes in the quantity and chemical quality of leaf litter inputs to forest soils. Because the quality and quantity of labile and recalcitrant carbon (C) compounds influence forest productivity through changes in soil organic matter content, characterizing changes in leaf litter in response to environmental change is critical to understanding the effects of global change on forests. We assessed the independent and combined effects of elevated [CO2] and elevated [O3] on foliar litter production and chemistry in aspen (Populus tremuloides Michx.) and birch-(Betula papyrifera Marsh.) aspen communities at the Aspen free-air CO2 enrichment (FACE) experiment in Rhinelander, WI. Litter was analyzed for concentrations of C, nitrogen (N), soluble sugars, lipids, lignin, cellulose, hemicellulose and C-based defensive compounds (soluble phenolics and condensed tannins). Concentrations of these chemical compounds in naturally senesced litter were similar in aspen and birch-aspen communities among treatments, except for N, the C:N ratio and lipids. Elevated [CO2] significantly increased C:N (+8.7%), lowered mean litter N concentration (-10.7%) but had no effect on the concentrations of soluble sugars, soluble phenolics and condensed tannins. Elevated [CO2] significantly increased litter biomass production (+33.3%), resulting in significant increases in fluxes of N, soluble sugars, soluble phenolics and condensed tannins to the soil. Elevated [O3] significantly increased litter concentrations of soluble sugars (+78.1%), soluble phenolics (+53.1%) and condensed tannins (+77.2%). There were no significant effects of elevated [CO2] or elevated [O3] on the concentrations of individual C structural carbohydrates (cellulose, hemicellulose and lignin). Elevated [CO2] significantly increased cellulose (+37.4%) input to soil, whereas elevated [O3] significantly reduced hemicellulose and lignin inputs to soil (-22.3 and -31.5%, respectively). The small changes in litter chemistry in response to elevated [CO2] and tropospheric [O3] that we observed, combined with changes in litter biomass production, could significantly alter the inputs of N, soluble sugars, condensed tannins, soluble phenolics, cellulose and lignin to forest soils in the future.

Betula↗

Cytoplasmic Acidification and Secondary Metabolite Production in Different Plant Cell Suspensions (A Comparative Study).

In this study, a correlation is described between low cytoplasmic pH, measured with the fluorescent probes 2[prime],7[prime]-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (acetoxymethyl ester) and bis- [3-propyl-5-oxoisoxazol-4-yl]pentamethine oxonol, and the production of secondary metabolites for several plant cell-suspension systems. Anthraquinone production in Morinda citrifolia suspensions is negligible in the presence of 2,4-dichlorophenoxyacetic acid (2,4-D), whereas with naphthalene acetic acid (NAA) a significant accumulation is realized. NAA-grown cells showed a lower cytoplasmic pH than did 2,4-D-grown cells. Addition of 2,4-D or parachlorophenoxy acetic acid to NAA-grown cells resulted in an inhibition of anthraquinone production and an increase of the cytoplasmic pH, whereas addition of parachlorophenyl acetic acid had no effect on either parameter. Lignin production in Petunia hybrida cells could be induced by subculturing them in a medium without iron. These cells showed a lower cytoplasmic pH than control cells. Addition of Fe3+ led to a decreased lignin content and an increased cytoplasmic pH. Two cell lines of Linum flavum showed a different level of coniferin and lignin concentration in their cells. Cells that accumulated coniferin and lignin had a lower cytoplasmic pH than cells that did not accumulate these secondary metabolites. Apparently, in different species and after different kinds of treatment there is a correlation between acidification of the cytoplasm and the production of different secondary metabolites. The possible role of this acidification in secondary metabolite production is discussed.

Journal Article↗

Wound-induced deposition of polyphenols in transgenic plants overexpressing peroxidase.

Tobacco (Nicotiana tabacum) plants transformed with a chimeric tobacco anionic peroxidase gene have previously been shown to synthesize high levels of peroxidase in all tissues throughout the plant. One of several distinguishable phenotypes of transformed plants is the rapid browning of pith tissue upon wounding. Pith tissue from plants expressing high levels of peroxidase browned within 24 hours of wounding, while tissue from control plants did not brown as late as 7 days after wounding. A correlation between peroxidase activity and wound-induced browning was observed, whereas no relationship between polyphenol oxidase activity and browning was found. The purified tobacco anionic peroxidase was subjected to kinetic analysis with substrates which resemble the precursors of lignin or polyphenolic acid. The purified enzyme was found to readily polymerize phenolic acids in the presence of H(2)O(2) via a modified ping-pong mechanism. The percentage of lignin and lignin-related polymers in cell walls was nearly twofold greater in pith tissue isolated from peroxidase-overproducer plants compared to control plants. Lignin deposition in wounded pith tissue from control plants closely followed the induction of peroxidase activity. However, wound-induced lignification occurred 24 to 48 hours sooner in plants overexpressing the anionic peroxidase. This suggests that the availability of peroxidase rather than substrate may delay polyphenol deposition in wounded tissue.

Journal Article↗