At eight European field sites, the impact of loss of plant diversity on primary productivity was simulated by synthesizing grassland communities with different numbers of plant species. Results differed in detail at each location, but there was an overall log-linear reduction of average aboveground biomass with loss of species. For a given number of species, communities with fewer functional groups were less productive. These diversity effects occurred along with differences associated with species composition and geographic location. Niche complementarity and positive species interactions appear to play a role in generating diversity-productivity relationships within sites in addition to sampling from the species pool.

Ecosystem effects of biodiversity manipulations in European grasslands AbstractWe present a multisite analysis of the relationship between plant diversity and ecosystem functioning within the European BIODEPTH network of plant-diversity manipulation experiments. We report results of the analysis of 11 variables addressing several aspects of key ecosystem processes like biomass production, resource use (space, light, and nitrogen), and decomposition, measured across three years in plots of varying plant species richness at eight different European grassland field sites. Differences among sites explained substantial and significant amounts of the variation of most of the ecosystem processes examined. However, against this background of geographic variation, all the aspects of plant diversity and composition we examined (i.e., both numbers and types of species and functional groups) produced significant, mostly positive impacts on ecosystem processes.Analyses using the additive partitioning method revealed that complementarity effects (greater net yields than predicted from monocultures due to resource partitioning, positive interactions, etc.) were stronger and more consistent than selection effects (the covariance between monoculture yield and change in yield in mixtures) caused by dominance of species with particular traits. In general, communities with a higher diversity of species and functional groups were more productive and utilized resources more completely by intercepting more light, taking up more nitrogen, and occupying more of the available space. Diversity had significant effects through both increased vegetation cover and greater nitrogen retention by plants when this resource was more abundant through N2 fixation by legumes. However, additional positive diversity effects remained even after controlling for differences in vegetation cover and for the presence of legumes in communities. Diversity effects were stronger on above-than belowground processes. In particular, clear diversity effects on decomposition were only observed at one of the eight sites.The ecosystem effects of plant diversity also varied between sites and years. In general, diversity effects were lowest in the first year and stronger later in the experiment, indicating that they were not transitional due to community establishment. These analyses of our complete ecosystem process data set largely reinforce our previous results, and those from comparable biodiversity experiments, and extend the generality of diversity-ecosystem functioning relationships to multiple sites, years, and processes. Abstract. We present a multisite analysis of the relationship between plant diversity and ecosystem functioning within the European BIODEPTH network of plant-diversity manipulation experiments. We report results of the analysis of 11 variables addressing several aspects of key ecosystem processes like biomass production, resource use (space, light, and nitrogen), and decomposition, measured across three years in plots of varying plant species ri...

Geographic variation can lead to the evolution of different local varieties, even in widespread forage plants. We investigated the performance of common forage plants in relation to their genetic diversity and local adaptation at a continental scale using reciprocal transplants at eight field sites across Europe over a 2‐year period. The overall performance of the three test species, Trifolium pratense, Dactylis glomerata, Plantago lanceolata, was generally highest for plants replanted at their home site and declined with increasing transplanting distance. The three species differed in the fitness components responsible for the increased overall performance and selection advantage at home sites. In addition to the effects of local adaptation, the majority of measured traits in all three species also showed ecotypic variation. However, no single ecotype of any species was able to outperform the locally adapted strains and do best at all sites, highlighting the importance of maintaining these plant genetic resources.

The mechanism of flavin reduction in morphinone reductase (MR) and pentaerythritol tetranitrate (PETN) reductase, and flavin oxidation in MR, has been studied by stopped-flow and steady-state kinetic methods. The temperature dependence of the primary kinetic isotope effect for flavin reduction in MR and PETN reductase by nicotinamide coenzyme indicates that quantum mechanical tunneling plays a major role in hydride transfer. In PETN reductase, the kinetic isotope effect (KIE) is essentially independent of temperature in the experimentally accessible range, contrasting with strongly temperaturedependent reaction rates, consistent with a tunneling mechanism from the vibrational ground state of the reactive C-H/D bond. In MR, both the reaction rates and the KIE are dependent on temperature, and analysis using the Eyring equation suggests that hydride transfer has a major tunneling component, which, unlike PETN reductase, is gated by thermally induced vibrations in the protein. The oxidative half-reaction of MR is fully rate-limiting in steady-state turnover with the substrate 2-cyclohexenone and NADH at saturating concentrations. The KIE for hydride transfer from reduced flavin to the ␣/␤ unsaturated bond of 2-cyclohexenone is independent of temperature, contrasting with strongly temperaturedependent reaction rates, again consistent with groundstate tunneling. A large solvent isotope effect (SIE) accompanies the oxidative half-reaction, which is also independent of temperature in the experimentally accessible range. Double isotope effects indicate that hydride transfer from the flavin N5 atom to 2-cyclohexenone, and the protonation of 2-cyclohexenone, are concerted and both the temperature-independent KIE and SIE suggest that this reaction also proceeds by ground-state quantum tunneling. Our results demonstrate the importance of quantum tunneling in the reduction of flavins by nicotinamide coenzymes. This is the first observation of (i) three H-nuclei in an enzymic reaction being transferred by tunneling and (ii) the utilization of both passive and active dynamics within the same native enzyme.Enzymes are phenomenal catalysts that can achieve rate enhancements of up to ϳ21 orders of magnitude over the uncatalyzed reaction rate (1). Our quest to understand the physical basis of this catalytic power is challenging and has involved sustained and intensive research efforts by many workers in the physical and life sciences (for recent reviews see Refs. 2-5). Recent years have witnessed new activity in this area and extended our theoretical understanding beyond the shortcomings of transition state theory (6) to include roles for protein "motion" (7, 8), low barrier hydrogen bonds (e.g. Refs. 9 -11), active site preorganization (e.g. reviewed in Refs. 4 and 12), and quantum mechanical tunneling (for recent reviews see Refs. 13-15). New theoretical frameworks incorporating quantum mechanical tunneling and protein motion are emerging to address the catalytic potency of enzymes. These invoke motion in the protein and/or subs...

The reaction of pentaerythritol tetranitrate reductase with reducing and oxidizing substrates has been studied by stopped-flow spectrophotometry, redox potentiometry, and X-ray crystallography. We show in the reductive half-reaction of pentaerythritol tetranitrate (PETN) reductase that NADPH binds to form an enzyme-NADPH charge transfer intermediate prior to hydride transfer from the nicotinamide coenzyme to FMN. In the oxidative half-reaction, the two-electron-reduced enzyme reacts with several substrates including nitroester explosives (glycerol trinitrate and PETN), nitroaromatic explosives (trinitrotoluene (TNT) and picric acid), and ␣,␤-unsaturated carbonyl compounds (2-cyclohexenone). Oxidation of the flavin by the nitroaromatic substrate TNT is kinetically indistinguishable from formation of its hydride-Meisenheimer complex, consistent with a mechanism involving direct nucleophilic attack by hydride from the flavin N5 atom at the electrondeficient aromatic nucleus of the substrate. The crystal structures of complexes of the oxidized enzyme bound to picric acid and TNT are consistent with direct hydride transfer from the reduced flavin to nitroaromatic substrates. The mode of binding the inhibitor 2,4-dinitrophenol (2,4-DNP) is similar to that observed with picric acid and TNT. In this position, however, the aromatic nucleus is not activated for hydride transfer from the flavin N5 atom, thus accounting for the lack of reactivity with 2,4-DNP. Our work with PETN reductase establishes further a close relationship to the Old Yellow Enzyme family of proteins but at the same time highlights important differences compared with the reactivity of Old Yellow Enzyme. Our studies provide a structural and mechanistic rationale for the ability of PETN reductase to react with the nitroaromatic explosive compounds TNT and picric acid and for the inhibition of enzyme activity with 2,4-DNP.

Maps of a species' potential range make an important contribution to conservation and invasive species risk analysis. Spatial predictions, however, should be accompanied by an assessment of their uncertainty. Here, we use multimodel inference to generate confidence intervals that incorporate both the uncertainty involved in model selection as well as the error associated with model fitting. In the case of the invasive Argentine ant, we found that it was most likely to occur where the mean daily temperature in mid-winter was 7-14 degrees C and maximum daily temperatures during the hottest month averaged 19-30 degrees C. Uninvaded regions vulnerable to future establishment include: southern China, Taiwan, Zimbabwe, central Madagascar, Morocco, high-elevation Ethiopia, Yemen and a number of oceanic islands. Greatest uncertainty exists over predictions for China, north-east India, Angola, Bolivia, Lord Howe Island and New Caledonia. Quantifying the costs of different errors (false negatives vs. false positives) was considered central for connecting modelling to decision-making and management processes.

The reaction of heterotetrameric sarcosine oxidase (TSOX) of Arthrobactor sp. 1-IN has been studied by stopped-flow spectroscopy, with particular emphasis on the reduction of the enzyme by sarcosine. Expression of the cloned gene encoding TSOX in Escherichia coli enables the production of TSOX on a scale suitable for stopped-flow studies. Treatment of the enzyme with sulfite provides the means for selective formation of a flavin-sulfite adduct with the covalent 8alpha-(N(3)-histidyl)-FMN. Formation of the sulfite-flavin adduct suppresses internal electron transfer between the noncovalent FAD (site of sarcosine oxidation) and the covalent FMN (site of enzyme oxidation) and thus enables detailed characterization of the kinetics of FAD reduction by sarcosine using stopped-flow methods. The rate of FAD reduction displays a simple hyperbolic dependence on sarcosine concentration. Studies in the pH range 6.5-10 indicate there are no kinetically influential ionizations in the enzyme-substrate complex. A plot of the limiting rate of flavin reduction/the enzyme-substrate dissociation constant (k(lim)/K(d)) versus pH is bell-shaped and characterized by two macroscopic pK(a) values of 7.4 +/- 0.1 and 10.4 +/- 0.2: potential candidates for the two ionizable groups are discussed with reference to the structure of monomeric sarcosine oxidase (MSOX). The kinetic data are discussed with reference to potential mechanisms for the oxidation of amine molecules by flavoenzymes. Additionally, kinetic isotope effect studies of the rate of C-H bond breakage suggest that a ground-state quantum tunneling mechanism for H-transfer, facilitated by the low-frequency thermal motions of the protein molecule, accounts for C-H bond cleavage by TSOX. TSOX thus provides another example of C-H bond breakage by ground-state quantum tunneling, driven by protein dynamics [vibrationally enhanced ground-state quantum tunneling (VEGST)], for the oxidation of amines by enzymes.

The number of exotic ant species being dispersed to new regions by human transportation and the trade pathways responsible for this are poorly understood. In this study, the taxonomic diversity, trade pathways, and origin of exotic ants intercepted at the New Zealand border were examined for the period 1955 -2005. Overall, there were a total 4355 interception records, with 115 species from 52 genera. The 10 most frequently intercepted genera, and the 20 most frequently intercepted species contributed > 90% of all records. Many of the species frequently intercepted are regarded as invasive species, and several are established in New Zealand. The most intercepted species was Pheidole megacephala . Despite a relatively low trade relationship, a high proportion (> 64%) of the exotic ants which were intercepted originated from the Pacific region. However, the majority of species intercepted from the Pacific was exotic to the region (71%), or to a lesser extent, wide-ranging Pacific native species. No endemic species from the Pacific were intercepted. The effectiveness of detecting exotic ant species at the New Zealand border ranged from 48 -78% for different trade pathways, indicating a number of species remain undetected. Trade routes associated with specific geographical regions represent a major filter for the arrival of exotic ant species. Despite the importance of the Pacific as a frequent pathway, we suggest that the future establishment of exotic ant species in New Zealand is likely to be mitigated by a renewed focus on trade routes with cool temperate regions, particularly Australia.