Assessment of holocellulose for the production of bioethanol by conserving Pinus radiata cones as renewable feedstock.

Renewable and green energy sources are much sought. Bioethanol is an environmentally friendly transportation fuel. Pine cones from Pinus radiata were shown to be a potential feedstock for the production of bioethanol. Alkaline (NaOH) pretreatment was carried out to delignify the lignocellulosic material and generate holocellulose (72 wt. % yield). The pretreated biomass was hydrolysed using HCl as catalyst under microwave irradiation and hydrothermal conditions. Microwave irradiation was found to be better than the hydrothermal process. Microwave irradiation accelerated the hydrolysis of biomass (42 wt. % conversion) with the reaction conditions being 3 M HCl and 5 min of irradiation time. Interestingly, even the xylose, which is the major component of the hydrolyzate was found to be metabolized to ethanol using Baker's yeast (Saccharomyces cerevisiae) under the experimental conditions. 5.7 g of ethanol could be produced from 100 g of raw pine cones.

Alterations of chemical composition, construction cost and payback time in needles of Masson pine (Pinus massoniana L.) trees grown under pollution.

Previous studies show that Masson pine (Pinus massoniana L.) stands grown at the industrially-polluted site have experienced unprecedented growth decline, but the causal mechanisms are poorly understood. In this study, to understand the mechanisms of growth decline of Mason pine strands under pollution stresses, we determined the reactive oxygen species levels and chemical composition of the current-year (C) and one-year-old (C + 1) needles, and calculated the needle construction costs (CCmass) of Masson pine trees grown at an industrially-polluted site and an unpolluted remote site. Pine trees grown at the polluted site had significantly higher levels of hydroxyl radical and superoxide anion in their needles than those grown at the unpolluted site, and the former trees eventually exhibited needle early senescence. The contents of lipids, soluble phenolics and lignins in C and C + 1 needles were significantly higher at the polluted site than at the unpolluted site, but the total amounts of non-construction carbohydrates were lower in non-polluted needles than in polluted needles. Elevated levels of the reactive oxygen species and early senescence in polluted needles together led to significant increases in CCmass and a longer payback time. We infer that the lengthened payback time and needle early senescence under pollution stress may reduce the Masson pine tree growth and consequently accelerate tree decline.

In situ assessment of the velocity of carbon transfer by tracing 13 C in trunk CO2 efflux after pulse labelling: variations among tree species and seasons.

Phloem is the main pathway for transferring photosynthates belowground. In situ(13) C pulse labelling of trees 8-10 m tall was conducted in the field on 10 beech (Fagus sylvatica) trees, six sessile oak (Quercus petraea) trees and 10 maritime pine (Pinus pinaster) trees throughout the growing season. Respired (13) CO2 from trunks was tracked at different heights using tunable diode laser absorption spectrometry to determine time lags and the velocity of carbon transfer (V). The isotope composition of phloem extracts was measured on several occasions after labelling and used to estimate the rate constant of phloem sap outflux (kP ). Pulse labelling together with high-frequency measurement of the isotope composition of trunk CO2 efflux is a promising tool for studying phloem transport in the field. Seasonal variability in V was predicted in pine and oak by bivariate linear regressions with air temperature and soil water content. V differed among the three species consistently with known differences in phloem anatomy between broadleaf and coniferous trees. V increased with tree diameter in oak and beech, reflecting a nonlinear increase in volumetric flow with increasing bark cross-sectional area, which suggests changes in allocation pattern with tree diameter in broadleaf species. Discrepancies between V and kP indicate vertical changes in functional phloem properties.

[Effects of CO2 storage flux on carbon budget of forest ecosystem].

Carbon dioxide (CO2) storage flux in the air space below measurement height of eddy covariance is very important to correctly evaluate net ecosystem exchange of CO2 (NEE) between forest ecosystem and atmosphere. This study analyzed the dynamic variation of CO2 storage flux and its effects on the carbon budget of a temperate broad-leaved Korean pine mixed forest at Changbai Mountains, based on the eddy covariance flux data and the vertical profile of CO2 concentration data. The CO2 storage flux in this forest ecosystem had typical diurnal variation, with the maximum variation appeared during the transition from stable atmospheric layer to unstable atmospheric layer. The CO2 storage flux calculated by the change in CO2 concentration throughout a vertical profile was not significantly different from that calculated by the change in CO2 concentration at the measurement height of eddy covariance. The NEE of this forest ecosystem was underestimated by 25% and 19% at night and at daytime, respectively, without calculating the CO2 storage flux at half-hour scale, and was underestimated by 10% and 25% at daily scale and annual scale, respectively. Without calculating the CO2 storage flux in this forest ecosystem, the parameters of Michaelis-Menten equation and Lloyd-Taylor equation were underestimated, and the ecosystem apparent quantum yield (alpha) and the ecosystem respiration rate (Rref) at the reference temperature were mostly affected. The gross primary productivity (GPP) and ecosystem respiration (Re) of this forest ecosystem were underestimated about 20% without calculating the CO2 storage flux at half-hour, daily scale, and annual scale.

Assessment of saccharification efficacy in the cellulase system of the brown rot fungus Gloeophyllum trabeum.

Brown rot fungi uniquely degrade wood by creating modifications thought to aid in the selective removal of polysaccharides by an incomplete cellulase suite. This naturally successful mechanism offers potential for current bioprocessing applications. To test the efficacy of brown rot cellulases, southern yellow pine wood blocks were first degraded by the brown rot fungus Gloeophyllum trabeum for 0, 2, 4, and 6 weeks. Characterization of the pine constituents revealed brown rot decay patterns, with selective polysaccharide removal as lignin compositions increased. G. trabeum liquid and solid state cellulase extracts, as well as a commercial Trichoderma reesei extract (Celluclast 1.5 L), were used to saccharify this pretreated material, using beta-glucosidase amendment to remove limitation of cellobiose-to-glucose conversion. Conditions varied according to source and concentration of cellulase extract and to pH (3.0 vs. 4.8). Hydrolysis yields were maximized using solid state G. trabeum extracts at a pH of 4.8. However, the extent of glucose release was low and was not significantly altered when cellulase loading levels were increased threefold. Furthermore, Celluclast 1.5 L continually outperformed G. trabeum cellulase extracts, although extent of glucose release never exceeded 22.0%. Results suggest methodological advances for utilizing crude G. trabeum cellulases and imply that the suboptimal hydrolysis levels obtained with G. trabeum and Celluclast 1.5 L cellulases, even at high loading levels, may be due to brown rot modifications insufficiently distributed throughout the pretreated material.

Potential enzyme cost reduction with the addition of surfactant during the hydrolysis of pretreated softwood.

The potential economic benefits of surfactants addition on enzymatic hydrolysis of steam-exploded lodgepole pine (SELP) and ethanol-pretreated lodgepole pine (EPLP) were investigated in this study. Free cellulase readsorption on fresh substrate was used to recover and recycle cellulase enzymes during the hydrolysis of SELP and EPLP substrate. Supplementing Tween 80 during the hydrolysis could facilitate enzyme recycling for EPLP substrate. A logarithmic correlation was established between surfactant concentration and free cellulase content after lignocellulosic hydrolysis, which was used to compute enzyme cost savings over various Tween 80 concentrations. A simple economic analysis of enzyme cost savings versus the cost of surfactant was undertaken. The results indicated that the addition of Tween 80 (priced at US $0.25/kg) during the hydrolysis of the EPLP substrate could save 60% of the total enzyme cost at concentrations in the 0.025% to 0.2% range. The addition of Tween for the hydrolysis of the SELP substrate significantly reduced the material cost by 24% per 1 gal of ethanol produced, and the ethanol production cost could be reduced by 8.6% with the addition of Tween and enzymes recycle for the hydrolysis of SELP substrate. A schematic concept of recycling enzyme and surfactant was also presented with a recirculation of process streams during hydrolysis. Further analysis indicated a 66% reduction in total enzyme cost could potentially be achieved under the concept.

The carbon budget of Pinus radiata plantations in south-western Australia under four climate change scenarios.

We conducted a comprehensive modelling study to estimate future stem wood production and net ecosystem production (NEP) of Pinus radiata D. Don plantations in south-western Australia, a region that is predicted to undergo severe rainfall reduction in future decades. The process-based model CenW was applied to four locations where it had previously been tested. Climate change scenarios under four emission scenarios for the period from 2005 to 2066 were considered, in addition to simulations under the current climate. Results showed that stem wood production and NEP were little affected by moderate climate change. However, under the most pessimistic climate change scenario (Special Report on Emission Scenarios A2), stem wood production and NEP decreased strongly. These results could be explained by the trade-off between the positive effect of rising atmospheric CO(2) on plant water use efficiency and the negative effects of decreasing rainfall and increasing temperatures. Because changes in heterotrophic respiration (R(H)) lagged behind changes in plant growth, and because R(H) rates were increased by higher temperatures, NEP was more negatively affected than stem wood production. Stem wood production and NEP also strongly interacted with location, with the site currently having the wettest climate being least affected by climatic change. These results suggest that realistic predictions of forest production and carbon sequestration potential in the context of climate change require (1) the use of modelling tools that describe the important feedbacks between environmental variables, plant physiology and soil organic matter decomposition, (2) consideration of a range of climate change scenarios and (3) simulations that account for a gradual climate change to capture transient effects.

Mountain pine beetle and forest carbon feedback to climate change.

The mountain pine beetle (Dendroctonus ponderosae Hopkins, Coleoptera: Curculionidae, Scolytinae) is a native insect of the pine forests of western North America, and its populations periodically erupt into large-scale outbreaks. During outbreaks, the resulting widespread tree mortality reduces forest carbon uptake and increases future emissions from the decay of killed trees. The impacts of insects on forest carbon dynamics, however, are generally ignored in large-scale modelling analyses. The current outbreak in British Columbia, Canada, is an order of magnitude larger in area and severity than all previous recorded outbreaks. Here we estimate that the cumulative impact of the beetle outbreak in the affected region during 2000-2020 will be 270 megatonnes (Mt) carbon (or 36 g carbon m(-2) yr(-1) on average over 374,000 km2 of forest). This impact converted the forest from a small net carbon sink to a large net carbon source both during and immediately after the outbreak. In the worst year, the impacts resulting from the beetle outbreak in British Columbia were equivalent to approximately 75% of the average annual direct forest fire emissions from all of Canada during 1959-1999. The resulting reduction in net primary production was of similar magnitude to increases observed during the 1980s and 1990s as a result of global change. Climate change has contributed to the unprecedented extent and severity of this outbreak. Insect outbreaks such as this represent an important mechanism by which climate change may undermine the ability of northern forests to take up and store atmospheric carbon, and such impacts should be accounted for in large-scale modelling analyses.

Assessment and implications of intraspecific and phenological variability in monoterpenes of Scots pine (Pinus sylvestris) foliage.

Scots pine populations contain individuals with widely differing amounts and composition of monoterpenes and exist as one of two chemotypes: with or without delta3-carene. We investigated the significance for ecological studies of two types of variation in monoterpenes: (1) the inherent variability in the concentration of monoterpenes or their relative amounts in needles of seedlings, saplings, and mature trees; and (2) phenological variation in developing needles. The relative composition of needle monoterpenes in 5-year-old saplings changed during the needle development period until the final composition was reached upon needle maturity. Changes in composition depended on chemotype. Needles of the "no-delta3-carene" chemotype had higher absolute concentrations of alpha-pinene, beta-pinene, camphene, and total monoterpenes than "delta3-carene" chemotype. For the "delta3-carene" chemotype, the relative concentration of delta3-carene during the needle growing season and immediately after emergence of seedlings was higher compared to that reached at needle maturity. Repeated removal of single needles (at weekly intervals during growth) from 5-year-old saplings did not influence the composition of monoterpenes. Within a natural Scots pine dominated woodland, 18% of mature Scots pines (N=574) belonged to the "no-delta3-carene" chemotype. Chemotypic variation within populations means that the statistical power with which differences in monoterpene concentrations can be detected is lower when sampling from the whole population compared to sampling within chemotypes. Reduction of this background variation and accounting for chiral variation if present, would significantly aid efficiency, interpretation, and understanding of processes in chemical and ecological research. One method for achieving this is the screening of plants for chemotypes before the establishment of experiments or field sampling regimes. We present a summary of suitable analytical methods for needle tissue that facilitates this prior screening.

Root-associated bacteria contribute to mineral weathering and to mineral nutrition in trees: a budgeting analysis.

The principal nutrient source for forest trees derives from the weathering of soil minerals which results from water circulation and from plant and microbial activity. The main objectives of this work were to quantify the respective effects of plant- and root-associated bacteria on mineral weathering and their consequences on tree seedling growth and nutrition. That is why we carried out two column experiments with a quartz-biotite substrate. The columns were planted with or without pine seedlings and inoculated or not with three ectomycorrhizosphere bacterial strains to quantify biotite weathering and pine growth and to determine how bacteria improve pine growth. We showed that the pine roots significantly increased biotite weathering by a factor of 1.3 for magnesium and 1.7 for potassium. We also demonstrated that the inoculation of Burkholderia glathei PML1(12) significantly increased biotite weathering by a factor of 1.4 for magnesium and 1.5 for potassium in comparison with the pine alone. In addition, we observed a significant positive effect of B. glathei PMB1(7) and PML1(12) on pine growth and on root morphology (number of lateral roots and root hairs). We demonstrated that PML1(12) improved pine growth when the seedlings were supplied with a nutrient solution which did not contain the nutrients present in the biotite. No improvement of pine growth was observed when the seedlings were supplied with all the nutrients necessary for pine growth. We therefore propose that the growth-promoting effect of B. glathei PML1(12) mainly resulted from the improved plant nutrition via increased mineral weathering.

DESPOT, a process-based tree growth model that allocates carbon to maximize carbon gain.

We present a new model of tree growth, DESPOT (Deducing Emergent Structure and Physiology Of Trees), in which carbon (C) allocation is adjusted in each time step to maximize whole-tree net C gain in the next time step. Carbon gain, respiration and the acquisition and transport of substitutable photosynthetic resources (nitrogen, water and light) are modeled on a process basis. The current form of DESPOT simulates a uniform, monospecific, self-thinning stand. This paper describes DESPOT and its general behavior in comparison to published data, and presents an evaluation of the sensitivity of its qualitative predictions by Monte Carlo parameter sensitivity analysis. DESPOT predicts determinate height growth and steady stand-level net primary productivity (NPP), but slow declines in aboveground NPP and leaf area index. Monte Carlo analysis, wherein the model was run repeatedly with randomly different parameter sets, revealed that many parameter sets do not lead to sustainable NPP. Of those that do lead to sustainable growth, the ratios at maturity of net to gross primary productivity and of leaf area to sapwood area are highly conserved.

Rapid molecular assessment of the bioturbation extent in sandy soil horizons under pine using ester-bound lipids by on-line thermally assisted hydrolysis and methylation-gas chromatography/mass spectrometry.

Each plant species has a unique chemical composition, and also within a given plant the various tissues differ from one another in their chemistry. These different compositions can be traced back after decay of the plant parts when they are transformed into soil organic matter (SOM). As a result, the composition of SOM reflects not only the plant origin, but also the various tissues, and the composition consequently provides an estimate of the contribution of above-ground vs. below-ground litter. From the latter distribution the extent of bioturbation (mixing of above-ground litter with the mineral soil) can be assessed. Application of thermally assisted hydrolysis and methylation (THM) using tetramethylammonium hydroxide (TMAH) and subsequent analysis by gas chromatography/mass spectrometry (GC/MS) releases all typical cutin- and suberin-derived aliphatic monomers (mono-, di- and trihydroxyalkanoic acids, alpha,omega-alkanedioic acids) as their methyl esters and/or ethers in a rapid manner. Using the distribution of omega-hydroxyalkanoic acids that are present in pine needle cutin (C(12) and C(14)) and not in root suberin, and those that are present in roots but not in needles (C(20) and C(22)), the extent of bioturbation (mixing of above-ground plant litter with the mineral soil) can be assessed. Similarly, the (9,16-dihydroxyhexadecanoic acid+9,10,18-trihydroxyoctadecanoic acid)/(C(20) + C(22) alpha,omega-alkanedioic acids) ratio reflects the degree of bioturbation. Three mineral soil profiles under Corsican pine with an A horizon that exhibited extensive bioturbation phenomena, and underlying C horizons with hardly any or no bioturbation, were investigated in order to examine the applicability of such an approach. It appeared that the A horizons contained all four mentioned omega-hydroxyalkanoic acids, while the C horizons contained virtually only the C(20) and C(22) members. The results not only suggest that bioturbation occurs in the A horizons, but also that possible illuviation or other transport mechanisms of omega-hydroxyalkanoic acids seem hardly ever or never to occur, which is a prerequisite for applying this biomarker approach in assessing degrees of bioturbation.