Rapid protocol for visualization of rust fungi structures using fluorochrome Uvitex 2B

Background

Histological examination using fluorochromes is one of the standard methods for observation of microorganisms in tissues and other compartments. In the study of fungi, especially those that cannot be cultured in axenic media such as biotrophic fungi, histological examination of processes associated with the fungal growth, differentiation, infection and other cellular functions can lead to the better understanding of host-parasite interactions. Fluorescence microscopy coupled with Fluorochrome Uvitex 2B have been extensively utilized to study rust fungi structures and host-pathogen interactions. In this study, we report development of a rapid staining protocol of the rust fungus Puccinia triticina using fluorochrome Uvitex 2B. The newly developed rapid procedure was compared with a standard staining technique to observe in planta fungal infection structures development during the wheat—Puccinia triticina interaction.

Results

While significantly reducing the time for staining, the rapid protocol described here was equally efficient or better compared to standard procedure in detecting fungal infection structures using Uvitex 2B. In the rapid staining procedure, pre-heating of the stain increased efficiency to detect all the infection structures including haustoria with highly reduced background noise from plant tissue.

Conclusion

This staining process described here is simple and quick. It can be completed in 4 h, which is of 6 times faster than the standard Uvitex 2B staining procedure.

     

Dry-washes determine gene flow and genetic diversity in a common desert shrub

Context

In deserts, many plant species exhibit a patchy spatial distribution within a harsh habitat matrix, where the likelihood of propagule dispersal among patches is uncertain, but may be promoted by landscape corridors or dispersal vectors.

Objectives

We examine the connectivity of a representative desert plant species (Acacia (Senegalia) greggii), and the ability of three major factors (animal dispersal agents, water flow along dry-washes, and climate) to facilitate dispersal within four watersheds in the Mojave National Preserve.

Methods

We genotyped 323 individuals sampled across 22 one-hectare sites using ten nuclear microsatellite markers.

Results

A hierarchical AMOVA revealed no significant differentiation among watersheds (F _RT = 0.00, P > 0.10), and very little genetic structure among all sites (F _ST = 0.03, P < 0.001), indicating regional connectivity. Mantel tests indicated distance along dry-washes best explained genetic distance between sites (r = 0.47, P < 0.05) when compared to Euclidean distance (P > 0.05), a distance measure based on rodent dispersal (P > 0.05), and a distance measure avoiding inhospitable climate (P > 0.05). An AIC comparison of generalized linear models found that within site genetic diversity (H _E and allelic richness) and average relatedness were best explained by slope (which increases seed dispersal potential via water flow) and area of the upstream watershed (which determines the number of potential seed donors), rather than plant density or habitat suitability.

Conclusions

Together, these findings indicate that dry-washes are key landscape features that enhance dispersal and regional connectivity in this patchy desert plant.

     

Connectivity analysis as a tool for assessing restoration success

Context

Methods for measuring restoration success that include functional connectivity between species’ populations are rare in landscape ecology and restoration practices. We developed an approach that analyzes connectivity between populations of target species and their dispersal probabilities to assess restoration success based on easily accessible input data. Applying this method to landscape development scenarios can help optimize restoration planning.

Objectives

We developed an assessment for restoration success and restoration planning based on functional connectivity between species’ populations and spatially explicit scenarios. The method was used in a case study to test its applicability.

Methods

Based on data on available habitat, species’ occurrence and dispersal ranges, connectivity metrics and dispersal probabilities for target species are calculated using the software Conefor Sensinode. The metrics are calculated for scenarios that reflect possible changes in the landscape to provide a basis for future restoration planning. We applied this approach to floodplain meadows along the Upper Rhine for four plant species and three future scenarios.

Results

In the case study, habitats of the target species were poorly connected. Peucedanum officinale and Sanguisorba officinalis were more successful in recolonizing new habitats than Iris spuria and Serratula tinctoria. The scenarios showed that restoration of species-rich grassland was beneficial for dispersal of the target species. As expected in the agriculturally dominated study area, restoration of former arable land significantly increased dispersal probabilities.

Conclusions

In the case study, the developed approach was easily applicable and provided reasonable results. Its implementation will be helpful in decision-making for future restoration planning.

     

How landscape,pollen intake and pollen quality affect colony growth in <Emphasis Type="Italic">Bombus terrestris</Emphasis>

Context

Abundance and diversity of bumblebees have been declining over the past decades. To successfully conserve bumblebee populations, we need to understand how landscape characteristics affect the quantity and quality of floral resources collected by colonies and subsequently colony performance.

Objectives

We therefore investigated how amount and composition of pollen collected by buff-tailed bumblebee Bombus terrestris colonies was affected by the surrounding landscape (i.e. the proportion of forest, urban, semi-natural habitats) and how they were related to colony growth.

Methods

Thirty B. terrestris colonies were placed at grassland sites differing in surrounding landscape. Colonies were established in spring when availability of flowering plants was highest, and their weight gain was monitored for 1 month. We additionally recorded the quantity and compared plant taxonomic composition and nutritional quality (i.e. amino acid composition) of pollen stored.

Results

Bumblebee colonies varied little in the pollen spectra collected despite differences in surrounding landscape composition. They collected on average 80 % of pollen from woody plants, with 34 % belonging to the genus Acer. Early colony growth positively correlated with total amount of woody pollen and protein collected and decreased with increasing proportions of semi-natural habitats and total amino acid concentrations.

Conclusions

Our results suggest that woody plant species represent highly important pollen sources for the generalist forager B. terrestris early in the season. We further show that colony growth of B. terrestris is predominantly affected by the quantity, not quality, of forage, indicating that several abundant plant species flowering throughout the bumblebees’ foraging season may cover the colonies’ nutritional needs.

     

Are landscapes more than the sum of their patches?

Context

The species–area relationship (SAR) is the most ubiquitous scaling relationship in ecology, yet we still do not know how different aspects of scale affect this relationship. Scale is defined by grain, extent, and focus. Focus here pertains to whether patches or landscapes are used to derive SARs.

Objective

To explore whether altering the focal scale influences the resulting SAR. If the SAR is scale-invariant, patch-based and landscape-based SARs should be congruent.

Methods

I fit a power-law function (S = cA ^z) to arthropod data obtained from an experimental landscape system, in which habitat amount and configuration (clumped vs. fragmented) of red clover (Trifolium pratense) varied among plots (256 m²). The scaling coefficient (z) was compared among patch-based and landscape-based SARs for congruence.

Results

Patches gained species at a faster rate than landscapes (z = 0.37 vs. 0.26, respectively), producing domains of incongruity in the SAR. Landscape richness (S _L) was greater than patch richness (S _P) below 30 % habitat, but S _P > S _L above 60 % habitat. Landscape configuration contributed to this incongruity below 30 % habitat (fragmented S _L > clumped S _L), but landscape context (whether the largest patch was embedded in a fragmented or clumped landscape) was important above 60 % habitat for understanding the SAR in this domain.

Conclusions

Landscape configuration exerts both direct (<30 % habitat) and indirect (>60 % habitat) effects on the SAR. Because patch-based and landscape-based SARs may not be congruent, we should exercise care when extrapolating from patches to landscapes to make inferences about the effects of habitat loss and fragmentation on species richness.

     

Disentangling the relative effect of light pollution,impervious surfaces and intensive agriculture on bat activity with a national-scale monitoring program

Context

Light pollution is a global change affecting a major proportion of global land surface. Although the impacts of Artificial Light At Night (ALAN) have been documented locally for many taxa, the extent of effect of ALAN at a landscape scale on biodiversity is unknown.

Objectives

We characterized the landscape-scale impacts of ALAN on 4 insectivorous bat species Pipistrellus pipistrellus, Pipistrellus kuhlii, Eptesicus serotinus, Nyctalus leisleri, and compared the extent of their effects to other major land-use pressures.

Methods

We used a French national-scale monitoring program recording bat activity among 2-km car transect surveys, and extracted landscape characteristics around transects with satellite and land cover layers. For each species, we performed multi-model averaging at 4 landscape scales (from 200 to 1000 m buffers around transects) to compare the relative effects of the average radiance, the proportion of impervious surface and the proportion of intensive agriculture.

Results

For all species, ALAN had a stronger negative effect than impervious surface at the 4 landscape scales tested. This effect was weaker than the effect of intensive agriculture. The negative effect of ALAN was significant for P. pipistrellus, P. kuhlii and E. serotinus, but not for N. leisleri. The effect of impervious surface varied among species while intensive agriculture had a significant negative effect on the 4 species.

Conclusion

Our results highlight the need to consider the impacts of ALAN on biodiversity in land-use planning and suggest that using only impervious surface as a proxy for urbanization may lead to underestimated impacts on biodiversity.

     

Determinants of spread in an urban landscape by an introduced lizard

Context

Urban landscapes are a mixture of built structures, human-altered vegetation, and remnant semi-natural areas. The spatial arrangement of abiotic and biotic conditions resulting from urbanization doubtless influences the establishment and spread of non-native species in a city.

Objectives

We investigated the effects of habitat structure, thermal microclimates, and species coexistence on the spread of a non-native lizard (Anolis cristatellus) in the Miami metropolitan area of South Florida (USA).

Methods

We used transect surveys to estimate lizard occurrence and abundance on trees and to measure vegetation characteristics, and we assessed forest cover and impervious surface using GIS. We sampled lizard body temperatures, habitat use, and relative abundance at multiple sites.

Results

At least one of five Anolis species occupied 79 % of the 1035 trees surveyed in primarily residential areas, and non-native A. cristatellus occupied 25 % of trees. Presence and abundance of A. cristatellus were strongly associated with forest patches, dense vegetation, and high canopy cover, which produced cooler microclimates suitable for this species. Presence of A. cristatellus was negatively associated with the ecologically similar non-native A. sagrei, resulting in reduced abundance and a shift in perch use of A. cristatellus.

Conclusions

The limited spread of A. cristatellus in Miami over 35 years is due to the patchy, low-density distribution of wooded habitat, which limits dispersal by diffusion. The presence of congeners may also limit spread. Open habitats—some parks, yards and roadsides—contain few if any A. cristatellus, and colonization of isolated forest habitat appears to depend on human-mediated dispersal.

     

Methanol fixation of plant tissue for Scanning Electron Microscopy improves preservation of tissue morphology and dimensions

Background

It is well known that preparation of biological (plant and animal) tissues for Scanning Electron Microscopy (SEM) by chemical fixation and critical point drying results in shrinkage of tissues, often by up to 20-30%, depending on the tissue type and fixation protocol used. We sought to identify a protocol that would preserve tissue size and morphology better than standard chemical fixatives and dehydration regimes. We compared a range of processing techniques by quantifying changes in tissue size and recording details of surface morphology using leaf tissues from three commonly studied species; Arabidopsis thaliana, barley and cotton.

Results

All processing protocols altered tissue dimensions. Methanol fixation and dehydration, followed by a further short (1 h) dehydration step in ethanol and critical point drying (which was based on a previously published method), preserved tissue dimensions most consistently of all protocols tested, although it did cause 8% shrinkage in all three species. This protocol was also best for preservation of surface morphology in all three species. We outline a recommended protocol and advise that the method is best trialled for different tissues, especially thicker or larger samples.

Conclusions

This study shows that simultaneous fixation and dehydration in methanol followed by ethanol results in better preservation of dimensions and morphology of critical point dried plant tissues than other fixation and dehydration procedures. It is a quick and simple method, and requires standard SEM preparation equipment.

     

Surface metrics: scaling relationships and downscaling behavior

Context

Considerable research has examined scale effects for patch-based metrics with the ultimate goal of predicting values at finer resolutions (i.e., downscaling), but results have been inconsistent. Surface metrics have been suggested as an alternative to patch-based metrics, although far less is known about their scaling relationships and downscaling potential. If successful, downscaling would enable integration of disparate datasets and comparison of landscapes using different resolution datasets.

Objectives

(1) Determine how surface metrics scale as resolution changes and how consistent those scaling relationships are across landscapes. (2) Test whether these scaling relationships can be accurately downscaled to predict metric values for finer resolutions.

Methods

Various scaling functions were fit to 16 surface metrics computed for multiple resolutions for a set of landscapes. Best-fitting functions were then extrapolated to test downscaling behavior (i.e., predict metric value for a finer resolution) for an independent set of validation landscapes. Relative error was assessed between the predicted and true values to determine downscaling robustness.

Results

Seven surface metrics (Sa, Sq, S10z, Sdq, Sds, Sdr, Srwi) fit consistently well (R² > 0.99) with a 3rd order polynomial or power law. Of those, the scaling functions for Sa, Sq, and S10z were able to predict metric values at a finer resolution within 5 %. Three metrics, (Ssk, Sku, Sfd) were also notable in terms of fit and downscaling.

Conclusions

Many metrics exhibit consistent scaling relations across resolution, and several are able to accurately predict values at finer resolutions. However, prediction accuracy is likely related to the amount of information lost during aggregation.

     

Temporal variability in potential connectivity of <Emphasis Type="Italic">Vallisneria americana</Emphasis> in the Chesapeake Bay

Context

Submersed aquatic vegetation (SAV) performs water quality enhancing functions that are critical to the overall health of estuaries such as the Chesapeake Bay. However, eutrophication and sedimentation have decimated the Bay’s SAV population to a fraction of its historical coverage. Understanding the spatial distribution of and connectedness among patches is important for assessing the dynamics and health of the remaining SAV population.

Objectives

We seek to explore the distribution of SAV patches and patterns of potential connectivity in the Chesapeake Bay through time.

Methods

We assess critical distances, from complete patch isolation to connection of all patches, in a merged composite coverage map that represents the sum of all probable Vallisneria americana containing patches between 1984 and 2010 and in coverage maps for individual years within that timeframe for which complete survey data are available.

Results

We have three key findings: First, the amount of SAV coverage in any given year is much smaller than the total recently occupied acreage. Second, the vast majority of patches of SAV that are within the tolerances of V. americana are ephemeral, being observed in only 1 or 2 years out of 26 years. Third, this high patch turnover results in highly variable connectivity from year to year, dependent on dispersal distance and patch arrangement.

Conclusions

Most of the connectivity thresholds are beyond reasonable dispersal distances for V. americana. If the high turnover in habitat occupancy is due to marginal water quality, relatively small improvements could greatly increase V. americana growth and persistence.

     

Elephants,termites and mound thermoregulation in a progressively warmer world

Context

With global change, microclimates become important refuges for temperature-sensitive, range-restricted organisms. In African savannas, woody vegetation on Macrotermes mounds create widely-dispersed microclimates significantly cooler than the surrounding matrix, which buffer against elevated temperatures at the finer scale of mounds, allowing species to persist at the landscape scale. Termite colonies cultivate symbiotic fungi to digest lignin, but the fungi require temperatures between 29 and 32 °C, which termites strive to maintain. Mound-associated vegetation is a hot-spot for elephant herbivory, so removal of woody species cover by elephants could influence mound-associated microclimates, impacting temperature regulation by termites.

Objectives

We explored the interaction between two prominent ecosystem engineers (termites and elephants) to ascertain whether elephant removal of mound woody cover affects (1) external mound-associated microclimate and (2) internal mound temperature.

Methods

We surveyed 44 mounds from three sites in Kruger National Park, South Africa, during an El Niño/Southern Oscillation-induced drought and heatwave, recording whether sub-canopy, external, mound-surface and internal mound temperatures varied with vegetation removal by elephant.

Results

Elephant damage to mound-associated vegetation reduces the fine-scale microclimate effect provided by vegetation on Macrotermes mounds. Despite this, termites were able to regulate internal mound temperatures, whereas internal temperatures of abandoned mounds increased with elevated surface temperatures.

Conclusions

Termites can persist despite loss of mound-associated microclimates, but the loss likely increases energetic costs of mound thermoregulation. Since mound vegetation buffers against drought, loss of widely-dispersed, fine-scale microclimates could increase as megaherbivores remain constrained to protected areas, impacting climate-sensitive organisms and ecosystem function at a range of scales.

     

Landscape genetics of the foundational salt marsh plant species black needlerush (<Emphasis Type="Italic">Juncus roemerianus</Emphasis> Scheele) across the northeastern Gulf of Mexico

Context

Common species important for ecosystem restoration stand to lose as much genetic diversity from anthropogenic habitat fragmentation and climate change as rare species, but are rarely studied. Salt marshes, valuable ecosystems in widespread decline due to human development, are dominated by the foundational plant species black needlerush (Juncus roemerianus Scheele) in the northeastern Gulf of Mexico.

Objectives

We assessed genetic patterns in J. roemerianus by measuring genetic and genotypic diversity, and characterizing population structure. We examined population connectivity by delineating possible dispersal corridors, and identified landscape factors influencing population connectivity.

Methods

A panel of 19 microsatellite markers was used to genotype 576 samples from ten sites across the northeastern Gulf of Mexico from the Grand Bay National Estuarine Research Reserve (NERR) to the Apalachicola NERR. Genetic distances (F_ST and D_ch) were used in a least cost transect analysis (LCTA) within a hierarchical model selection framework.

Results

Genetic and genotypic diversity results were higher than expected based on life history literature, and samples structured into two large, admixed genetic clusters across the study area, indicating sexual reproduction may not be as rare as predicted in this clonal macrophyte. Digitized coastal transects buffered by 500 m may represent possible dispersal corridors, and developed land may significantly impede population connectivity in J. roemerianus.

Conclusions

Results have important implications for coastal restoration and management that seek to preserve adaptive potential by sustaining natural levels of genetic diversity and conserving population connectivity. Our methodology could be applied to other common, widespread and understudied species.

     

Effect of connectivity and habitat availability on the occurrence of the Chestnut-throated Huet-Huet (<Emphasis Type="Italic">Pteroptochos castaneus,</Emphasis> Rhinocryptidae) in fragmented landscapes of central Chile

Context

Although small isolated habitat patches may not be able to maintain a minimum viable population, small patches that are structurally isolated may be functionally connected if individuals can cross the gaps between them, in which case, their areas could be added to form a larger habitat patch, eventually surpassing the size threshold for holding a viable population.

Objectives

We studied whether models based on the size and isolation of habitat patches could be used to predict the distribution of the Chestnut-throated Huet-Huet (Pteroptochos castaneus) in fragmented landscapes of the coastal range of the Maule region, central Chile.

Methods

We selected seven 10,000-ha landscapes (8.4–70.7% forest cover). For each habitat patch we made 18 predictions of the presence of the species based on the combination of two thresholds: three critical patch sizes for maintaining a viable population (62.5, 125 and 250 ha) and six critical isolation distances between patches (0, 10, 50, 100, 150 and 200 m). We used playbacks in 59 sampling points to estimate the species’ presence/absence. We used logistic regressions to test whether the output of the patch-matrix models could explain part of the variation in the presence of Pteroptochos castaneus.

Results

The best predictions for the presence of P. castaneus were obtained with the most conservative scenarios (125–250 ha to 0–10 m), including a positive effect of the understory cover and a lack of effect of the forest type (native or exotic).

Conclusions

Our findings suggest that the long term persistence of P. castaneus may depend on the existence of large and/or very connected forest tracts.

     

How do field margins contribute to the functional connectivity of insect-pollinated plants?

Context

Habitat fragmentation generates a loss of functional connectivity detrimental to the persistence of biodiversity. The French agricultural intensification initiated in the 1950s has caused a decline in field margins.

Objectives

As field margins may facilitate species dispersal while providing socio-economic benefits, it is of interest to assess their contribution to the functional connectivity of insect-pollinated plants in agro-ecosystems. This will help develop appropriate management strategies mitigating fragmentation.

Methods

We addressed this issue by studying the links between landscape structure and the patterns of abundance and pollen dispersal (using fluorescent dye particles) for two contrasted insect-pollinated plants occurring in field margins (Crepis sancta and Euphorbia serrata). We investigated the influence of field margins quality and of the surrounding matrix on pollen dispersal and compared the relevance of the least-cost algorithm with a straight-line approach to depict pollinators’ movements.

Results

The influence of landscape structure on plant abundance is species and scale-specific. Pollen dispersal decreases with distance from the source. For E. serrata, it was preferentially dispersed via field margins, confirming the relevance of the least-cost algorithm, while C. sancta dispersal followed a straight-line.

Conclusions

Euphorbia serrata, which grows strictly on field margins with a greater dispersal ability and a more diversified pollinator guild than C. sancta, is less affected by land-use changes. Our study demonstrates the contrasting contributions of field margins to pollen dispersal as they may act as functional corridors favouring pollinators’ movement depending on the species of interest.

     

Fine-scale wetland features mediate vector and climate-dependent macroscale patterns in human West Nile virus incidence

Context

Mosquito-borne diseases in the United States are notorious for their seemingly erratic temporal and spatial outbreak patterns in humans. Identifying linkages between fine-scale phenomenon and macroscale outbreak patterns can improve the understanding of these complex socio–ecological systems.

Objectives

Using West Nile virus (WNV) as a model system, we examine whether fine-scale wetland characteristics—wetland size, connectivity, and inundation regime—have an important effect on macroscale human disease outbreak in the northeastern and midwestern United States.

Methods

We take a spatially explicit GIS-based approach, derive new macroscale wetland metrics based on fine-scale wetland characteristics, and use county-level annual human WNV incidence data over 11 years to evaluate linkages between the fine and macroscales (1.6 million km²).

Results

In regions dominated by the Culex tarsalis WNV vector, U.S. counties with a small average wetland size had more than 100 % higher human WNV incidence than counties with a large average wetland size. In regions dominated by the Cx. pipiens and Cx. restuans vectors, a low proportional area of connected wetland was associated with at least 50 % higher human WNV incidence than a high proportional area of connected wetland. Finally, Cx. tarsalis-dominated counties with a high proportional area of semi-permanent wetland that were experiencing drought conditions had over 300 % higher annual WNV incidence than drought-affected counties with a low proportional area of semi-permanent wetland.

Conclusions

Our results suggest that phenomena occurring at the individual wetland scale may aggregate to influence macroscale human WNV outbreak patterns and may be mediated by the interplay of other factors such as vector species-specific traits and climate.

     

Microhabitat conditions and landscape pattern explain nocturnal rodent activity,but not seed removal,in burned and unburned lodgepole pine forests

Context

Disturbances create spatial variation in environments that may influence animal foraging. Granivory by rodents can influence seed supply and thus plant establishment. However, effects of disturbance patterns on rodent seed removal in western North American conifer forests are generally unknown.

Objectives

We conducted a study in lodgepole pine (Pinus contorta var. latifolia) forests of Greater Yellowstone (Wyoming, USA) to answer: (1) How do seed removal and rodent activity vary between recently burned and adjacent unburned forests and with distance from fire perimeter? (2) Which microhabitat conditions explain variability in seed removal and rodent activity?

Methods

One or two years after wildfires, we established transects (n = 23) with four stations each: at 10 and 40 m from the fire perimeter in both burned and unburned forest. At stations, we deployed trays with lodgepole pine seeds and cameras pointed at trays for 28 days and quantified habitat structure and seed abundance.

Results

Seed removal, which averaged 85%, and diurnal rodent activity did not differ between burned and unburned forests or with distance from the fire perimeter; however, nocturnal rodent activity was lower in burned forests. Seed removal and diurnal rodent activity were not associated with any microhabitat conditions we measured. However, nocturnal rodent activity was associated with microhabitat in both burned and unburned forests.

Conclusions

High seed removal rates suggested that rodent foraging was not reduced by high-severity wildfire. If observed seed removal represents natural conditions, post-dispersal seed predation could influence post-fire recruitment of a widespread foundation tree species.

     

Contrasting impacts of invasive plants and human-altered landscape context on nest survival and brood parasitism of a grassland bird

Context

Humans have altered grasslands in recent decades through crop conversion, woody encroachment, and plant invasions. Concurrently, grassland birds have experienced range-wide declines. Studies have reported effects of plant invasions and land conversion on nest ecology, but few have assessed relative impacts of these changes.

Objectives

We compared impacts of invasive plants and landscape context on nest survival of a grassland songbird, the dickcissel (Spiza americana). We also compared effects on parasitism by brown-headed cowbirds (Molothrus ater) and tested whether parasitism affects survival.

Methods

From 2013–2016, we monitored 477 dickcissel nests. We measured nest-site vegetation (including woody plants, tall fescue Schedonorus arundinaceous, and other invasive grasses) and measured landscape context at broad scales.

Results

Nest survival declined with increasing tall fescue cover at nest sites, and parasitism was more common at nests with greater fescue and woody cover. Some evidence suggested a negative effect of row-crop cover within 1000 m on nest survival, but no landscape patterns unambiguously affected survival. Woodland cover and wooded-edge prevalence were associated with reduced parasitism risk. Parasitized nests had smaller clutches, failed more frequently, and produced fewer fledglings than non-parasitized nests.

Conclusions

Determining the impacts of invasive plants and other anthropogenic changes on grassland birds will aid in prioritizing management to improve habitat quality. Our results indicate that optimizing landscape context around habitats may not affect dickcissel nest survival strongly, except perhaps through effects on parasitism. In contrast, controlling tall fescue and shrubs within grasslands could benefit birds by increasing nest success and reducing parasitism.

     

Transient co-expression with three <Emphasis Type="Italic">O</Emphasis>-glycosylation enzymes allows production of GalNAc-<Emphasis Type="Italic">O</Emphasis>-glycosylated Granulocyte-Colony Stimulating Factor in <Emphasis Type="Italic">N. benthamiana</Emphasis>

Background

Expression of economically relevant proteins in alternative expression platforms, especially plant expression platforms, has gained significant interest in recent years. A special interest in working with plants as bioreactors for the production of pharmaceutical proteins is related to low production costs, product safety and quality. Among the different properties that plants can also offer for the production of recombinant proteins, protein glycosylation is crucial since it may have an impact on pharmaceutical functionality and/or stability.

Results

The pharmaceutical glycoprotein human Granulocyte-Colony Stimulating Factor was transiently expressed in Nicotiana benthamiana plants and subjected to mammalian-specific mucin-type O-glycosylation by co-expressing the pharmaceutical protein together with the glycosylation machinery responsible for such post-translational modification.

Conclusions

The pharmaceutical glycoprotein human Granulocyte-Colony Stimulating Factor can be expressed in N. benthamiana plants via agroinfiltration with its native mammalian-specific mucin-type O-glycosylation.

     

Quantifying spatiotemporal drivers of environmental heterogeneity in Kruger National Park,South Africa

Context

Environmental heterogeneity is considered an important mechanism of biodiversity. How environmental heterogeneity is characterised by the compositional, structural and functional variation of biotic and abiotic components is a central research theme in conservation.

Objectives

We explore how environmental heterogeneity relates to the underlying physical landscape template and how that relationship changes over space and time. We examine how, in some areas, environmental heterogeneity may also be driven by dynamic ecological processes, and how this relates to patterns of plant species richness.

Method

We use local geographically weighted regression to spatially partition environmental heterogeneity, measured as Landsat spectral variance, into the portion explained by stable physical landscape properties (R²) and the portion unexplained (1?R²) which we term landscape complexity. We explore how this relationship varies spatially and temporally as a function of dynamic ecological processes such as rainfall and season in Kruger National Park, as well as plant species richness at landscape scales.

Results

The significance and direction of relationships varied over space and time and as a function of rainfall and season. R² values generally decreased in higher rainfall summer months and revealed patterns describing the importance of known stable factors relative to unknown dynamic factors. Landscape complexity (1?R²) explained over 70 % of variation in species richness.

Conclusions

Rainfall and seasonality are important drivers of environmental heterogeneity. The spatial arrangement and magnitude of model agreement helped disentangle the relative influence of the physical landscape template on environmental heterogeneity. Given the high correlation with species richness, landscape complexity provides complementary guidance to biodiversity research and monitoring prioritization.

     

Scale dependence of felid predation risk: identifying predictors of livestock kills by tiger and leopard in Bhutan

Context

Livestock predation by tiger and leopard in Bhutan is a major threat to the conservation of these felids. Conflict mitigation planning would benefit from an improved understanding of the spatial pattern of livestock kills by the two predators.

Objectives

We aimed to identify the landscape features that predict livestock kills by tiger and leopard throughout Bhutan. Our goals were to: (1) identify the predictors that have the largest influence in determining livestock kills, (2) assess the influence of scale across the different predictors evaluated and identify the scale at which each was most important.

Methods

We used livestock kills obtained from compensation records of tiger (n = 326) and leopard (n = 377) across Bhutan between 2003 and 2012 to run predation risk models with MaxEnt algorithm, using a multi-scale modeling approach (1, 2, 4, 8 and 16 km).

Results

Human-presence (density of settlements and roads) and land-cover (percentage of tree cover and meadow patches) were the main variables contributing to livestock kills by both species. Livestock kills were likely driven by a trade-off between livestock density and predator ecology, and the balance of this trade-off varied with scale. Risk maps revealed different hotspots for tiger and leopard kills, and analysis showed both species preferentially killed equids over other livestock types.

Conclusions

Our results highlight the importance of evaluating scale when investigating the spatial attributes of livestock kills by tiger and leopard. Our findings provide guidance for reducing conflict between humans and large felids throughout the country.