Transgenic plants as a source for the bioscavenging enzyme,human butyrylcholinesterase

Organophosphorous pesticides and nerve agents inhibit the enzyme acetylcholinesterase at neuronal synapses and in neuromuscular junctions. The resulting accumulation of acetylcholine overwhelms regulatory mechanisms, potentially leading to seizures and death from respiratory collapse. While current therapies are only capable of reducing mortality, elevation of the serum levels of the related enzyme butyrylcholinesterase (BChE) by application of the purified protein as a bioscavenger of organophosphorous compounds is effective in preventing all symptoms associated with poisoning by these toxins. However, BChE therapy requires large quantities of enzyme that can easily overwhelm current sources. Here, we report genetic optimization, cloning and high‐level expression of human BChE in plants. Plant‐derived BChE is shown to be biochemically similar to human plasma‐derived BChE in terms of catalytic activity and inhibitor binding. We further demonstrate the ability of the plant‐derived bioscavenger to protect animals against an organophosphorous pesticide challenge.     

Modern analogues from the Southern Urals provide insights into biodiversity change in the early Holocene forests of Central Europe

Aim The diversity changes that occurred in Central European forests during the early Holocene can be better understood using ecological knowledge of modern analogues of these forests, which occur in far Eastern Europe. Here we compare the diversity of vascular plants, bryophytes and snails among different forest types of the Southern Urals to provide insights into the palaeoecology of the vanished Central European forests. Location Southern Ural Mountains, Bashkortostan, Russia. Methods We recorded all species of vascular plants, ground-dwelling bryophytes and land snails, and measured biotic and abiotic variables, in 100-m² plots located in four forest types dominated by different trees: Pinus sylvestris–Larix sibirica, Betula pendula–Populus tremula, Quercus robur and Acer platanoides–Tilia cordata–Ulmus glabra. These types correspond to the chronosequence of forest communities that occurred in Central Europe in the early Holocene. Results The species richness of herb-layer plants was most affected by the canopy-transmitted light. The former three forest types had open canopy and were rich in species, whereas the Acer–Tilia–Ulmus forests were darker and poor in species. The species richness of ground-dwelling bryophytes decreased from coniferous to broad-leaved deciduous forests. In contrast, the highest species richness of snails was in the Acer–Tilia–Ulmus forests owing to the higher calcium content in the litter of these trees. Main conclusions Based on the modern analogue of the Southern Ural forests, we propose the hypothesis that the late-glacial open-canopy pine and larch forests of Central European lowlands were rich in light-demanding species of vascular plants, many of which were also typical of tall-grass steppes or mesic grasslands. They also contained several species of ground-dwelling bryophytes. The spread of birch, aspen and oak in the early Holocene reduced the local species richness of ground-dwelling bryophytes but not of vascular plants. The subsequent spread of elm, lime, maple and ash caused canopy closure, a retreat of the light-demanding herbs and a decline in the local species richness of vascular plants. Besides the increased shading by these tree species, their litter enriched soils in calcium, and enhanced decomposition and nutrient cycling. This supported an increase in the species richness of land snails.     

Herbarium records identify the role of long‐distance spread in the spatial distribution of alien plants in New Zealand

Aim To use herbarium records to characterize important correlates of spatial spread, areal occupancy and clustering of 100 alien plant species of conservation concern in the North and South Islands of New Zealand. Location New Zealand. Methods Using herbarium data of 6294 records representing 100 alien plant species, we assessed spatial heterogeneity in the distribution of alien species by examining the role of major urban areas as sources of sampling bias. A novel method to account for spatial biases in sampling effort was developed and applied to two simple distance metrics: nearest‐ and furthest‐neighbour spread rate. The relative importance of these two distance metrics in determining the range, areal extent and dispersion of alien species across both the North and South Islands of New Zealand was also assessed. Results The spatial distribution of herbarium records was highly clustered with a significant bias towards the more populated regions. Once sampling bias was taken into account, there was no indication that species were found closer to these urban centres than might be expected based on sampling effort. The nearest‐neighbour spread rates were usually 1–5 km yr^?1 and correlated positively with the furthest‐neighbour spread rates that were an order of magnitude higher. Range and area increased and clustering decreased with higher spread rates and longer recording time span. The spread rates divided species into five groups that were clearly distinguishable in terms of the extent of their distribution and the degree of clustering. Species occurring on both islands did not exhibit similar spread rates or spatial patterns. Main conclusions The nearest‐ and furthest‐neighbour spread rates from herbarium records can explain the area and pattern of alien plant distributions and improve the understanding of the dynamics of their spread. Five groups emerge from the spread rates in relation to a null model. Fast‐spreading plants had the widest, least clustered distribution, which suggests widespread chronic problems; slow‐spreading plants had localized, but dense, clustered distributions, indicating acutely problematic weeds. These patterns appear robust and may be useful in predicting the future patterns of spread and in planning long‐term management strategies in New Zealand.     

Harmonia axyridis in Great Britain: analysis of the spread and distribution of a non-native coccinellid

Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) is native to Asia, and was widely introduced as a biocontrol agent of aphids and coccids in Europe and North America. In Europe H. axyridis is considered to be an invasive alien species. Although not known to have been deliberately introduced to Great Britain, it was first recorded there in 2004, in south-east England. Harmonia axyridis arrived in Great Britain by various means, primarily from mainland Europe, but also from Canada. Extensive national and local media coverage, and a dedicated website (), facilitated public involvement in recording H. axyridis in Great Britain; in excess of 4,000 verified records of the species were received between 2004 and 2006. Through detailed mapping, the objective of our study was to quantify and analyse the spread of H. axyridis in its early stages of invasion in Great Britain. Our data shows that between 2004 and 2006, the species spread north through Great Britain at the rate of 58 km year^-1 and west at the rate of 144.5 km year^-1. In England H. axyridis spread north to Yorkshire and west to Cornwall by the end of 2006, and also reached Wales. Recorded occurrence (of one or more H. axyridis individuals at larval, pupal and/or adult stage) in 10 km squares in Great Britain was: 2004—51; 2005—149; 2006—447. Records of juvenile H. axyridis extend through most of the recorded British range of the species, and we present evidence of bi-voltinism in the population in 2006.     

Effectiveness of methods to control the invasive non-native pitcherplant Sarracenia purpurea L. on a European mire

Pitcherplant Sarracenia purpurea is a North American carnivorous plant that has become invasive on mires throughout Europe. It appears to have limited dispersal ability and so eradication may be feasible given early detection and effective control. We used a field experiment to assess the effectiveness of three control methods – hand-pulling, spot-spraying with glyphosate, and turf-stripping – on the largest established population of S. purpurea in Great Britain. After one year all three treatments had significantly reduced the abundance and flowering performance of S. purpurea. Turf-stripping was the most effective technique but had the greatest impact on non-target species. Spot-spraying and hand-pulling caused significant reductions in adults with only a small reduction in associated species diversity. However, hand-pulling significantly increased the numbers of seedlings and juveniles due to increased recruitment and competitive release following the removal of adults. Spot-spraying caused some damage to associated species but this was not significant. In conclusion, spot-spraying with glyphosate provides the most cost-effective method for eradicating S. purpurea but needs to be carried out with great care to avoid damaging sensitive mire ecosystems. Hand-pulling offers a more labour intensive alternative but its success will rely on monitoring for at least four years to ensure that the recruitment of juveniles is exhausted. Turf-stripping is unlikely to be an acceptable technique given the global importance of mire ecosystems invaded by S. purpurea in Europe.     

Historical relationships of the Mexican cloud forests: a preliminary vicariance model applying Parsimony Analysis of Endemicity to vascular plant taxa

Mexican cloud forests, situated between 600 and 3000 m of elevation, exhibit a remarkable high biotic diversity. They follow a fragmented pattern, similar to that of an archipelago, that makes them suitable to vicariance modelling. A Parsimony Analysis of Endemicity (PAE) was applied to the presence/absence of 1267 species of vascular plants (gymnosperms, angiosperms, and pteridophytes) from twenty-four patches of Mexican cloud forests, in order to postulate a preliminary hypothesis of relationships. The single cladogram obtained grouped the twenty-four cloud forests into five clades. These results indicate that the Sierra Madre Oriental, Sierra Madre del Sur, and Serranías Meridionales floristic provinces do not represent natural units. A preliminary vicariance model is presented to explain the sequence of fragmentation of the Mexican cloud forests.     

Exceptional carbon uptake in European forests during the warm spring of 2007: a data–model analysis

Temperate and boreal forests undergo drastic functional changes in the springtime, shifting within a few weeks from net carbon (C) sources to net C sinks. Most of these changes are mediated by temperature. The autumn 2006–winter 2007 record warm period was followed by an exceptionally warm spring in Europe, making spring 2007 a good candidate for advances in the onset of the photosynthetically active period. An analysis of a decade of eddy covariance data from six European forests stands, which encompass a wide range of functional types (broadleaf evergreen, broadleaf deciduous, needleleaf evergreen) and a wide latitudinal band (from 44° to 62°N), revealed exceptional fluxes during spring 2007. Gross primary productivity (GPP) of spring 2007 was the maximum recorded in the decade examined for all sites but a Mediterranean evergreen forest (with a +40 to +130 gC m^?2 anomaly compared with the decadal mean over the January–May period). Total ecosystem respiration (TER) was also promoted during spring 2007, though less anomalous than GPP (with a +17 to +93 gC m^?2 anomaly over 5 months), leading to higher net uptake than the long‐term mean at all sites (+12 to +79 gC m^?2 anomaly over 5 months). A correlative analysis relating springtime C fluxes to simple phenological indices suggested spring C uptake and temperatures to be related. The CASTANEA process‐based model was used to disentangle the seasonality of climatic drivers (incoming radiation, air and soil temperatures) and biological drivers (canopy dynamics, thermal acclimation of photosynthesis to low temperatures) on spring C fluxes along the latitudinal gradient. A sensitivity analysis of model simulations evidenced the roles of (i) an exceptional early budburst combined with elevated air temperature in deciduous sites, and (ii) an early relief of winter thermal acclimation in coniferous sites for the promotion of 2007 spring assimilation.     

Soil seed banks in church forests of northern Ethiopia: Implications for the conservation of woody plants

Church forests are sanctuaries for different organisms, ranging from microbes to large animals, which have almost disappeared in most parts of northern Ethiopia. Despite the actual and potential significance of these forests, studies and documented information on their bio-physical features and socio-economic setting are either scanty or totally lacking. A study was, therefore, carried out in seven of these church forests, namely Hiruy, Zahara, Gibtsawit, Gelawdiwos, Dengolt, Debresena and Ascha with the objective of assessing the composition, densities and spatial heterogeneity of soil seed banks. The number of viable seeds in the soil samples, from both germination and sieving, corresponded to a seed bank density down to 9 cm in the soil of 7594 seeds m⁻² at Ascha, 2064 seeds m⁻² at Debresena, 4208 seeds m⁻² at Dengolt, 3158 seeds m⁻² at Gelawdiwos, 2754 seeds m⁻² at Gibtsawit, 2759 seeds m⁻² at Hiruy, and 1909 seeds m⁻² at Zahra. The total number of species recorded was 50, representing at least 22 plant families, in the seven church forests, of which 13 were recorded from Ascha, 29 from Debresena, 26 from Dengolt, 19 from Gibtsawit, 22 from Gelawdiwos, 20 from Hiruy and 21 from Zahra. There was a significant difference in seed densities and number of species recovered from the soil seed banks of the seven church forests. Ascha had a significantly higher seed bank density than all other church forests. Dengolt exhibited a significantly higher number of species compared with other church forests. Of the identified species recorded from the seven church forests, 83% were herbs, 11% trees and 6% shrubs. Our results revealed that church forests accumulate large quantities of persistent seeds of herbaceous species in the soil while only five (6%) of the 91 woody species recorded in the standing vegetation of the seven church forests were represented in the soil seed banks. The fact that most of the dominant tree species do not accumulate seeds in the soil suggests that their regeneration from seeds would be prevented by removal of mature individuals in the standing vegetation. The results also provided further evidences that consolidate the conclusions of previous studies, i.e. the future existence of the woody flora characteristic of dry Afromontane areas in Ethiopia depends on the conservation and sustainable utilization of the few remnant natural forests.     

Robinia pseudoacacia as a surrogate for native tree species for saproxylic beetles inhabiting the riparian mixed forests of northern Italy

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Importance of semi-natural habitats for the conservation of butterfly communities in landscapes dominated by pine plantations

While the area of plantation forests continues to increase worldwide, their contribution to the conservation of biodiversity is still controversial. There is a particular concern on the central role played by natural habitat remnants embedded within the plantation matrix in conserving species-rich insect communities. We surveyed butterflies in maritime pine plantation landscapes in south-western France in 83 plots belonging to seven habitat types (five successional stages of pine stands, native deciduous woodlands and herbaceous firebreaks). The effect of plot, habitat and landscape attributes on butterfly species richness, community composition and individual species were analysed with a General Linear Model (GLM), partial Canonical Correspondence Analysis (CCA) and the IndVal method. The most important factors determining butterfly diversity and community composition were the presence of semi-natural habitats (deciduous woodlands and firebreaks) at the landscape scale and the composition of understorey vegetation at the plot scale. Pure effects of plot variables explained the largest part of community variation (12.8%), but landscape factors explained an additional, independent part (6.7%). Firebreaks were characterized by a higher species richness and both firebreaks and deciduous woodlands harboured species not or rarely found in pine stands. Despite the forest-dominated landscape, typical forest butterflies were rare and mainly found in the deciduous woodlands. Threatened species, such as Coenonympha oedippus and Euphydryas aurinia, were found in pine stands and in firebreaks, but were more abundant in the latter. In the studied plantation forest, the conservation of butterflies depends mainly on the preservation of semi-natural habitats, an adequate understorey management and the maintenance of soil moisture levels.     

Predicting invasions by woody species in a temperate zone: a test of three risk assessment schemes in the Czech Republic (Central Europe)

To assess the validity of previously developed risk assessment schemes in the conditions of Central Europe, we tested (1) Australian weed risk assessment scheme (WRA; Pheloung et al . 1999); (2) WRA with additional analysis by Daehler et al . (2004); and (3) decision tree scheme of Reichard and Hamilton (1997) developed in North America, on a data set of 180 alien woody species commonly planted in the Czech Republic. This list included 17 invasive species, 9 naturalized but non-invasive, 31 casual aliens, and 123 species not reported to escape from cultivation. The WRA model with additional analysis provided best results, rejecting 100% of invasive species, accepting 83.8% of non-invasive, and recommending further 13.0% for additional analysis. Overall accuracy of the WRA model with additional analysis was 85.5%, higher than that of the basic WRA scheme (67.9%) and the Reichard–Hamilton model (61.6%). Only the Reichard–Hamilton scheme accepted some invaders. The probability that an accepted species will become an invader was zero for both WRA models and 3.2% for the Reichard–Hamilton model. The probability that a rejected species would have been an invader was 77.3% for both WRA models and 24.0% for the Reichard–Hamilton model. It is concluded that the WRA model, especially with additional analysis, appears to be a promising template for building a widely applicable system for screening out invasive plant introductions.     

Facilitation by pioneer shrubs for the ecological restoration of riparian forests in the Central Andes of Colombia

Riparian vegetation has great functional importance in agroscapes because it establishes physical connections between natural ecosystems embedded in an agricultural matrix. Throughout the tropics, the historical demand for cropland and pastures in the lands adjacent to streams has led to the replacement of forests by exotic grasses. In order to find new methods for initiating the restoration of riparian forests, we evaluated the use of the pioneer shrubs Tithonia diversifolia and Piper auritum as nurse species for their effects on (1) competition with dominant grasses; (2) natural regeneration and the ecological facilitation of six native tree species of different successional stages; and (3) influence on microclimate. Over a period of 15 months, 4.4% coverage of P. auritum was insufficient to inhibit grasses. In contrast, 81% coverage of T. diversifolia limited the growth of dominant grasses such as Cenchrus purpureus, Paspalum paniculatum, and Cynodon plectostachyus likely by intercepting more than 90% of photosynthetically active radiation (PAR), even though other factors cannot be ruled out. T. diversifolia showed simultaneous effects of facilitation and competition by promoting higher survival of the planted native trees while slowing the growth of pioneer species and inhibiting the regeneration of native woody plants probably as a result of high light interception. This study suggests that planting T. diversifolia as a nurse shrub may facilitate early stages of restoration by inhibiting the growth of pastures, but requires pruning, both to increase light availability and promote the growth of planted trees and the recruitment of woody plants.     

The role of vegetative spread and seed dispersal for optimal life histories of clonal plants: a simulation study

Sexual and vegetative reproduction of clonal plants phenotypically differ in dispersal distance, in the phenology of offspring production and establishment, and in the success of establishment. We applied a combination of analytical and of spatially explicit individual-based simulation modelling to calculate long-term fitness. We predicted optimal clonal plant life histories in a parameter space spanned by stolon number, stolon internode length, and relative allocation to sexual and vegetative reproduction. For a given allocation to sexual reproduction and number of stolons, fitness was optimised for rather short internode lengths under small disturbances, and for the longest possible internodes under larger disturbances. A trade-off between length and number of vegetative spacers drew parameters away from their unrestricted optima. Now, intermediate length and number of spacers led to maximum fitness under large disturbances. Simultaneous trade-offs between sexual and vegetative reproduction and between the length and number of spacers could also lead to fitness optima at intermediate parameter values, depending on the success of seedling establishment. We demonstrated that spatial habitat structure (1) selects for an efficient use of available space either by optimum internode length or by investment into seeds, which disperse farther than vegetative spacers, and (2) leads to an interaction between trade-offs. We conclude, that dispersal distance, i.e. a spatial life-history component, and trade-offs must be included in considerations on adaptive evolution of clonal life histories.     

The variation of REE (rare earth elements) patterns in soil-grown plants: a new proxy for the source of rare earth elements and silicon in plants

Rare earth elements (REEs) in five species of soil-grown plants (Taxodium japonicum, Populus sieboldii, Sasa nipponica, Thea sinensis and Vicia villosa) and in the soil on which each plant grew were determined with an inductively coupled plasma mass spectrometer (ICP-MS) in order to observe the variation in the distribution of REEs and to elucidate their source in soil-grown plants. The plant samples were divided into root (secondary root and main root), trunk (stem) and leaf; the soils into water soluble (soil_{soluble fraction}), HCl and HNO₃ soluble (soil_{non-silicate fraction}) and HF soluble (soil_{silicate fraction}). The REE abundances of samples were compared using REE patterns where the abundances were normalized to those of a chondrite and plotted on a logarithmic scale against the atomic number. All the plants showed similar REE patterns independent of species and location, and a W-shape variation (W-type tetrad effect) and abundance depletion of cerium (negative Ce anomaly) were found in each REE patterns of plants, more conspicuous tetrad effect being observed in HREE (heavier rare earth elements) region than in LREE (lighter rare earth elements) region. The overall variation of REE patterns of each secondary root was not similar to that of soil_{soluble fraction}, but similar to that of soil_{silicate fraction} except for the tetrad effect and Ce anomaly. The REE patterns can be interpreted by the idea that plants of different species take in REEs and Si from different parts in the soil. The results of this study seem to imply that Sasa nipponica and Vicia villosa take in free REEs and Si rather directly from silicate in the soil, and that a majority of REEs and Si in Taxodium japonicum and Thea sinensis are originated from the soluble fraction in the soil.     

Development and validation of a phenological model for the univoltine European corn borer

Biological control of a univoltine race of European corn borer, Ostrinia nubilalis Hübner (Lepidoptera: Crambidae), by Trichogramma in Switzerland is currently timed according to repeated, semi‐field observations of the post‐diapause development of the pest at one site. We developed a phenology model with the aims of replacing this costly labour practice and by considering the Swiss landscape. Observations collected over 29 years were used for model calibration and validation. We parameterized a time‐varying distributed delay model based on published laboratory observations of development durations at constant temperatures. The model was driven with hourly temperature recordings beginning on January 1. The calibration of the mean development rates and their variations was based on semi‐field data of larval and pupal development. The model, with its calibrated parameters and their variations, allowed the simulation of mean predictions and prediction intervals. We validated the model predictions of emergence with flight data (obtained via light traps) from several sites in western and northern Switzerland. The simulated mean emergence was 6.9 degree‐days earlier than the observed flight at the calibration sites and only 0.5 degree‐days earlier than the observed flight at other sites. Our simulation of pupation explained half of the variation in emergence time, whereas semi‐field observations of pupation explained three‐quarters of this variation. Our model simulations are not subjected to the local potential biases. Simulation errors from a year with an extremely dry spring were explained by the lack of consideration of humidity by the model. Our simulations provide a valid and less labour‐intensive alternative to observations for timing biological control in the maize‐growing areas of Switzerland and likely other areas of Europe.     

An efficient and reliable method for determining the number of transgenes inserted into transgenic plants

A simple and reliable method is described for determining the number of transgenes inserted into transgenic plants. Analysis of tomato plants transformed with an antisense polygalacturonase gene is used to demonstrate the method. The use of the method to analyze other plants transformed with endogenous plant gene coding regions or promoters is discussed.