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1.

Background

Rust fungi are obligate pathogens with multiple life stages often including different spore types and multiple plant hosts. While individual rust pathogens are often associated with specific plants, a wide range of plant species are infected with rust fungi. To study the interactions between these important pathogenic fungi and their host plants, one must be able to differentiate fungal tissue from plant tissue. This can be accomplished using the In situ hybridization (ISH) protocol described here.

Results

To validate reproducibility using the ISH protocol, samples of Chrysanthemum × morifolium infected with Puccinia horiana, Gladiolus × hortulanus infected with Uromyces transversalis and Glycine max infected with Phakopsora pachyrhizi were tested alongside uninfected leaf tissue samples. The results of these tests show that this technique clearly distinguishes between rust pathogens and their respective host plant tissues.

Conclusions

This ISH protocol is applicable to rust fungi and potentially other plant pathogenic fungi as well. It has been shown here that this protocol can be applied to pathogens from different genera of rust fungi with no background staining of plant tissue. We encourage the use of this protocol for the study of plant pathogenic fungi in paraffin embedded sections of host plant tissue.
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2.

Background

Scanning electron microscopy (SEM) has been used for high-resolution imaging of plant cell surfaces for many decades. Most SEM imaging employs the secondary electron detector under high vacuum to provide pseudo-3D images of plant organs and especially of surface structures such as trichomes and stomatal guard cells; these samples generally have to be metal-coated to avoid charging artefacts. Variable pressure-SEM allows examination of uncoated tissues, and provides a flexible range of options for imaging, either with a secondary electron detector or backscattered electron detector. In one application, we used the backscattered electron detector under low vacuum conditions to collect images of uncoated barley leaf tissue followed by simple quantification of cell areas.

Results

Here, we outline methods for backscattered electron imaging of a variety of plant tissues with particular focus on collecting images for quantification of cell size and shape. We demonstrate the advantages of this technique over other methods to obtain high contrast cell outlines, and define a set of parameters for imaging Arabidopsis thaliana leaf epidermal cells together with a simple image analysis protocol. We also show how to vary parameters such as accelerating voltage and chamber pressure to optimise imaging in a range of other plant tissues.

Conclusions

Backscattered electron imaging of uncoated plant tissue allows acquisition of images showing details of plant morphology together with images of high contrast cell outlines suitable for semi-automated image analysis. The method is easily adaptable to many types of tissue and suitable for any laboratory with standard SEM preparation equipment and a variable-pressure-SEM or tabletop SEM.
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3.

Context

Despite decades of research, there is an intense debate about the consistency of the hump-shaped pattern describing the relationship between diversity and disturbance as predicted by the intermediate disturbance hypothesis (IDH). Previous meta-analyses have not explicitly considered interactive effects of disturbance frequency and intensity of disturbance on plant species diversity in terrestrial landscapes.

Objective

We conducted meta-analyses to test the applicability of IDH by simultaneously examining the relationship between species richness, disturbance frequency (quantified as time since last disturbance as originally proposed) and intensity of disturbance in forest landscapes.

Methods

The effects of disturbance frequency, intensity, and their interaction on species richness was evaluated using a mixed-effects model.

Results

We found that species richness peaks at intermediate frequency after both high and intermediate disturbance intensities, but the richness-frequency relationship differed between intensity classes.

Conclusions

Our study highlights the need to measure multiple disturbance components that could help reconcile conflicting empirical results on the effect of disturbance on plant species diversity.
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4.

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.
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5.
6.

Context

One of the key challenges for landscape planners is to reframe the meaning of ecosystem services. In this context, alternative concepts such as ecologies have potential to complement ecosystem services when applied to human–nature relationships in changing landscapes.

Objectives

The objectives of this article are: (1) to review how landscape planners use major critical approaches to translate the meaning of ecosystem services and (2) to introduce why ecologies provides helpful insights to complement ecosystem services.

Methods

A conceptual framework examines how landscape planners use critique to reframe the meaning of ecosystem services. This framework is then revised as a scenario to reframe the meanings of ecologies and ecosystem services.

Results

Landscape planners use three critical approaches to reframe the meaning of ecosystem services to advance the understanding of human–nature relationships in changing landscapes. Yet, they identify some important issues and gaps that emerge when it is applied. These issues and gaps are part of the rationale for why landscape planning is at a crossroads with ecosystem services. This rationale is then extended to create a scenario for why a revised conceptual framework is needed for landscape planners to reframe the meanings of ecologies and ecosystem services.

Conclusion

The translational challenge of ecologies and ecosystem services is an example of the key role that landscape planners play in developing a deeper understanding of human–nature relationships.
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7.

Background

The compressional modulus of elasticity is an important mechanical property for understanding stalk lodging, but this property is rarely available for thin-walled plant stems such as maize and sorghum because excised tissue samples from these plants are highly susceptible to buckling. The purpose of this study was to develop a testing protocol that provides accurate and reliable measurements of the compressive modulus of elasticity of the rind of pith-filled plant stems. The general approach was to relying upon standard methods and practices as much as possible, while developing new techniques as necessary.

Results

Two methods were developed for measuring the compressional modulus of elasticity of pith-filled node–node specimens. Both methods had an average repeatability of ± 4%. The use of natural plant morphology and architecture was used to avoid buckling failure. Both methods relied up on spherical compression platens to accommodate inaccuracies in sample preparation. The effect of sample position within the test fixture was quantified to ensure that sample placement did not introduce systematic errors.

Conclusions

Reliable measurements of the compressive modulus of elasticity of pith-filled plant stems can be performed using the testing protocols presented in this study. Recommendations for future studies were also provided.
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8.

Context

Multiple ecological drivers generate spatial patterns in species’ distributions. Changes to natural disturbance regimes can place early successional habitat specialists at an increased risk of extinction by altering landscape patterns of habitat suitability.

Objectives

We developed a series of hypotheses to evaluate the effects of landscape structure, fire history, and site-level habitat quality on site occupancy by an early successional specialist, the eastern chestnut mouse (Pseudomys gracilicaudatus).

Methods

We obtained eight years of monitoring data from 26 sites in recently burned heathland in southeast Australia. We used generalised linear models to determine which explanatory variables were related to occupancy. We also explored predictability in patterns of small mammal species co-occurrence.

Results

Landscape structure (patch area, landscape heterogeneity) was strongly related to site occupancy. Site occupancy was associated with dead shrubs in the understory and rock cover on ground layer, but was not directly influenced by recent or historical fire. Contrary to contemporary ecological theory, we found no predictable species associations in our early successional community.

Conclusions

We recommend surveys take account of landscape configuration and proximity to suitable habitat for optimal results. Fire regimes expected to promote eastern chestnut mouse population growth should encourage the retention of critical habitat features rather than be based on temporal rates of successional stages. For management to adequately account for post-disturbance patterns in early successional communities, a species-by-species, multi-scaled approach to research is necessary.
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9.

Context

Competitive interactions potentially play an important role in structuring bird communities. It is unclear how differences in functional traits influence the niche dimensions of highly mobile waterbird species, particularly when they co-exist in spatiotemporally heterogeneous communities.

Objectives

We investigated the inter-relationships between waterbird trait groupings (movement, dietary and foraging habitat) and environmental variable groupings (rainfall, land cover, vegetation structure and water quality). Specifically, we tested whether the scale of environmental variables filtered movement traits and whether these traits operated in conjunction with dietary and foraging habitat traits to form distinct ecological niches in waterbirds.

Methods

We conducted waterbird and environmental variable surveys in 60 sites, sampled seven times each at bimonthly intervals, in KwaZulu-Natal, South Africa. Trait-environment relationships were tested using a combination of RLQ and fourth-corner analyses.

Results

Several significant trait-environment relationships emerged in bivariate correlations and multivariate ordination space. Movement traits correlated with the scale of environmental variables; migrant and nomadic species responded to broad scale environmental variables. Vegetation structure and land cover were particularly important in explaining the abundance of species foraging in emergent vegetation. Three groups emerged along a gradient in multivariate ordination space providing evidence for ecological niche separation of waterbirds with different movement traits.

Conclusions

Our findings suggest that the scale of landscape resources can act as a filter of movement traits, and that in conjunction with dietary and foraging traits, waterbirds with different movement traits occupy distinct ecological niches.
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10.
11.

Context

Species are expected to shift their distributions in response to global environmental changes and additional protected areas are needed to encompass the corresponding changes in the distributions of their habitats. Conservation policies are likely to become obsolete unless they integrate the potential impacts of climate and land-use change on biodiversity.

Objectives

We identify conservation priority areas for current and future projected distributions of Iberian bird species. We then investigate the extent to which global change informed priority areas are: (i) covered by existing protected area networks (national protected areas and Natura 2000); (ii) threatened by agricultural or urban land-use changes.

Methods

We use outputs of species distributions models fitted with climatic data as inputs in spatial prioritization tools to identify conservation priority areas for 168 bird species. We use projections of land-use change to then discriminate between threatened and non-threatened priority areas.

Results

19% of the priority areas for birds are covered by national protected areas and 23% are covered by Natura 2000 sites. The spatial mismatch between protected area networks and priority areas for birds is projected to increase with climate change. But there are opportunities to improve the protection of birds under climate change, as half of the priority areas are currently neither protected nor in conflict with urban or agricultural land-uses.

Conclusions

We identify critical areas for bird conservation both under current and climate change conditions, and propose that they could guide the establishment of new conservation areas across the Iberian Peninsula complementing existing protected areas.
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12.

Context

Seagrass landscapes vary substantially in extent and pattern, resulting from depth zonation and hydrodynamic stress gradients and may exhibit threshold behavior in response to changes in physical drivers. Seagrass landscapes persist in a delicate balance between processes of disturbance and recovery and therefore may exhibit behavior typical of classic critical systems.

Objectives

Examine how hydrodynamic drivers and physical setting influence seagrass landscape composition and configuration. Determine if seagrass patch size distributions typify patterns observed for critical systems.

Methods

We used landscape metrics to quantify the spatial configuration of seagrass and then modeled the response of these metrics to wave energy, tidal current speed, and water depth at 62 estuarine sites in North Carolina, USA. Seagrass landscapes were representative of cover types observed in the estuary generated by wave energy.

Results

Percent cover, patch size, and number of patches all declined with increasing wave energy. Threshold behavior occurred at wave energy change points between 675–774 J m?1. Seagrass landscapes differed in spatial configuration and physical setting, above and below change points. There was moderate support for a power law relationship for patch size distribution across a wide range of seagrass landscape cover and wave energy.

Conclusions

With weather extremes on the rise, much of this estuarine seagrass will be exposed to increased wave energy. Where seagrass exists just below the wave energy change points, increases in wave energy could tip those habitats into a new stable state of lower cover resulting in less cover overall in the estuary.
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13.

Context

Hoverflies are often used as bio-indicators for ecosystem conservation, but only few studies have actually investigated the key factors explaining their richness in woodlands.

Objectives

In a fragmented landscape in southwest France, we investigated the joint effects of woodland area, structural heterogeneity, connectivity and history on the species richness of forest-specialist hoverflies, and whether there was a time lag in the response of hoverflies to habitat changes, and tested the effect of spatiotemporal changes.

Methods

Current species richness was sampled in 48 woodlands using 99 Malaise traps. Structural variables were derived from a rapid habitat assessment protocol. Old maps and aerial photographs were used to extract past and present spatial patterns of the woodlands since 1850. Relationships between species richness and explanatory variables were explored using generalized linear models.

Results

We show that current habitat area, connectivity, historical continuity and the average density of tree-microhabitats explained 35 % of variation in species richness. Species richness was affected differently by changes in patch area between 1979 and 2010, depending on woodland connectivity. In isolated woodlands, extinction debt and colonization credit were revealed, showing that even several decades are not sufficient for hoverflies to adapt to landscape-scale habitat conditions.

Conclusions

These findings emphasise the importance of maintaining connectedness between woodlands, which facilitates the dispersion in a changing landscape. Our results also highlight the benefits of using a change-oriented approach to explain the current distribution patterns of species, especially when several spatial processes act jointly.
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14.

Context

Interactions among disturbances, climate, and vegetation influence landscape patterns and ecosystem processes. Climate changes, exotic invasions, beetle outbreaks, altered fire regimes, and human activities may interact to produce landscapes that appear and function beyond historical analogs.

Objectives

We used the mechanistic ecosystem-fire process model FireBGCv2 to model interactions of wildland fire, mountain pine beetle (Dendroctonus ponderosae), and white pine blister rust (Cronartium ribicola) under current and future climates, across three diverse study areas.

Methods

We assessed changes in tree basal area as a measure of landscape response over a 300-year simulation period for the Crown of the Continent in north-central Montana, East Fork of the Bitterroot River in western Montana, and Yellowstone Central Plateau in western Wyoming, USA.

Results

Interacting disturbances reduced overall basal area via increased tree mortality of host species. Wildfire decreased basal area more than beetles or rust, and disturbance interactions modeled under future climate significantly altered landscape basal area as compared with no-disturbance and current climate scenarios. Responses varied among landscapes depending on species composition, sensitivity to fire, and pathogen and beetle suitability and susceptibility.

Conclusions

Understanding disturbance interactions is critical for managing landscapes because forest responses to wildfires, pathogens, and beetle attacks may offset or exacerbate climate influences, with consequences for wildlife, carbon, and biodiversity.
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15.

Context

Connectivity models for animal movement frequently use resistance surfaces, but rarely incorporate actual movement data and multiple scale drivers of landscape resistance.

Objectives

Using GPS data, we developed a multi-scale model of landscape resistance for tiger (Panthera tigris) dispersal in central India and evaluated the performance, interpretation and predictions against single scale models.

Methods

Six dispersing tiger paths were subjected to a path level analysis with conditional logistic regression to parameterize a resistance surface. We evaluated for 21 scales of available habitat and selected the best scale for each variable. We derived a scale-optimized multivariate path selection function and predicted landscape resistance across the landscape.

Results

The tigers preferred to move along areas with forest cover at relatively high elevations along the ridges with rugged topography at broad scale, while avoiding areas with agriculture-village matrix at fine scale. We found that the scale that was most supported by Akaike’s information criterion was not always the scale that maximized the magnitude (effect size) of the relationship. Further, the multi-scale optimized model differed substantially from the single scale models in terms of variable importance, magnitude of coefficients and predictions of connectivity.

Conclusions

Our results demonstrate that the variables in landscape resistance models produce markedly different predictions of population connectivity depending on the scales of analyses and interpretation. Thus, scale optimization in parameterization is critical for appropriate inferences and sound management strategies.
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16.

Context

In a global context of erosion of biodiversity, the current environmental policy in Europe is oriented towards the creation and the preservation of ecological networks for wildlife. However, most of the management guidelines arose from a structural landscape diagnostic without truly taking into consideration species’ needs.

Objectives

We tested whether and how landscape elements influence the functional connectivity of landscapes for a forest specialist species, the European pine marten (Martes martes), in Northeastern France.

Methods

We collected pine marten scats and tissues from 13 evenly distributed study sites across the whole study area in order to test several types of barriers such as highways, waterways, and open agricultural fields. We crossed the results of several methods: spatial autocorrelation analysis, causal modelling framework, and clustering methods.

Results

The study indicates significant genetic differentiation among the sampling sites. A signal of isolation by distance was detected but disappeared after partialling out landscape or barrier resistance. The only model that was fully supported by causal modelling was the one identifying waterways as the main driver of genetic differentiation. Moreover, clustering analyses indicated the presence of genetic clusters, suggesting that pine marten spatial genetic pattern could be explained by the presence of waterways but also by their reluctance to cross open fields.

Conclusions

The current ecological network could thus be improved by increasing permeability of waterways, in particular navigation canals, and by maintaining and restoring forested corridors in agricultural plains.
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17.

Context

Traditionally soils have not received much attention in urban planning. For this, tools are needed that can both be understood both by soil scientists and urban planners.

Purpose

The purpose of this paper is to enhance the role of soil knowledge in urban planning practice, through the following objectives: (1) identifying the role soil plays in recent urban plans; (2) analysing the ecosystem services and indicators used in soil science in an urban context; and (3) inferring the main challenges and opportunities to integrate soil into urban planning.

Methods

Seven urban plans and reports of world cities that include sustainability goals were analysed using text-mining and qualitative analysis, with a critical view on the inclusion of soil-related concepts. Secondly, the contribution of soil science to urban planning was assessed with an overview of case studies in the past decade that focus on soil-related ecosystem services in urban context.

Results

The results show an overall weak attention to soil and soil-related ecosystem services in the implementation and monitoring phases of urban plans. The majority of soil science case studies uses a haphazard approach to measure ecosystem service indicators which may not capture the ecosystem services appropriately and hence lack relevance for urban planning.

Conclusions

Even though the most urban plans assessed recognize soil as a key resource, most of them fail to integrate indicators to measure or monitor soil-related functions. There is a need to develop soil-related ecosystem services that can be easily integrated and understood by other fields.
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18.

Context

In heterogeneous landscapes, habitat complementation is a key process underlying the distribution of mobile species able to exploit non-substitutable resources over large home ranges. For instance, insectivorous bats need to forage in a diversity of habitat patches offering varied compositions and structures within forest landscape mosaics to fulfill their life cycle requirements.

Objectives

We aimed at analyzing the effects of forest structure and composition measured at the stand and landscape scales on bat species richness, abundance and community composition in pine plantation forests of south-western France.

Methods

We sampled bat communities at different periods of the summer season using automatic ultrasound recorders along a tree composition gradient from pine monocultures to pure oak stands. We analyzed bat species activity (as a proxy for bat abundance) and species richness with linear mixed models. Distance-based constrained ordinations were used to partition the spatio-temporal variation in bat communities.

Results

Deciduous tree cover increased bat activity and modified community composition at both stand and landscape scales. Changes in bat communities were mostly driven by landscape-scale variables while bat activity responded more to stand-scale predictors.

Conclusions

The maintenance of deciduous trees at both stand and landscape scales is likely critical for bat communities living in fast-growing conifer plantations, by increasing the availability and diversity of prey and roosting sites. Our study suggests that bats respond to forest composition at both stand and landscape scales in mosaic plantation landscapes, mainly through a resource complementation process.
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19.

Context

Regime shifts are well known for driving penetrating ecological change, yet we do not recognise the consequences of these shifts much beyond species diversity and productivity. Sound represents a multidimensional space that carries decision-making information needed for some dispersing species to locate resources and evaluate their quantity and quality.

Objectives

Here we assessed the effect of regime shifts on marine soundscapes, which we propose has the potential function of strengthening the positive or negative feedbacks that mediate ecosystem shifts.

Methods

We tested whether biologically relevant cues are altered by regime shifts in kelp forests and seagrass systems and how specific such shifted soundscapes are to the type of driver; i.e. local pollution (eutrophication) vs. global change (ocean acidification).

Results

Here, we not only provide the first evidence for regime-shifted soundscapes, but also reveal that the modified cues of shifted ecosystems are similar regardless of spatial scale and type of environmental driver. Importantly, biological sounds can act as functional cues for orientation by dispersing larvae, and observed shifts in soundscape loudness may alter this function.

Conclusions

These results open the question as to whether shifted soundscapes provide a functional role in mediating the positive or negative feedbacks that govern the arrival of species associated with driving change or stasis in ecosystem state.
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20.

Context

There are few detailed data for short-term (≤?monthly) fluctuations in flowering and nectar availability at relatively large spatial scales. Such information is critical for understanding the governors of variation in flowering and for the management of floral resources assisting the persistence of nectar consumers in landscapes.

Objectives

To obtain monthly measurements of patterns of nectar availability in a 314,400 ha region, and to relate these patterns to potential environmental predictors.

Methods

Flowering was measured at 83 sites in natural vegetation and in eight domestic gardens in subtropical, eastern Australia. A nectar-availability index was developed was based on nectarivore visitation rates and plant-specific flowering patterns. Spatial–temporal patterns were related to environmental variables using boosted regression trees.

Results

The large between-year variation was due mostly to irregular flowering by several eucalypt species. There was a ‘lean season’ in the austral spring (August–September). Coastal vegetation was an important source of nectar for much of the year, including the lean season. Gardens produced prolific nectar throughout the year, peaking in August–October.

Conclusions

Nectar availability was most closely associated with primary productivity over the previous 12 months, average annual solar radiation, topographic wetness, and rainfall over the previous 6 months, although some relationships seemed counter-intuitive. There were large differences in nectar availabilities among broad vegetation types (especially rainforests vs. sclerophyllous forests), which partially accounted for the unintuitive results.
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