Evidence for divergence between sympatric stone charr and Dolly Varden along unique environmental gradients in Kamchatka

Here, we describe the biological and ecological differences between partially anadromous Dolly Varden and riverine stone charr distributed in the Kamchatka River middle course. Endemic stone charr, being the ambush predator, is defined by the accelerated growth and prolonged lifespan, robust body, and large mouth, as well as specific marble coloration since an early age. The significant restriction in gene flow between the stone charr and benthos-eating Dolly Varden was supported by an allelic distribution of eight microsatellite loci. Herewith, the identity of mitochondrial DNA control region and intron sequences of growth hormone and metallothionein genes confirms a relatively recent diversification of the groups within the single basin. We suggest the key role of the specific environment for reproduction isolation of stone charr. It occupies coniferous taiga zone with dark-colored water, whereas Dolly Varden spawns uppercourse in the zone of stunted, windblown forest. Conifer litter decomposition products could drive natural selection of stone charr via metabolism acceleration and resistance to toxicants. The metabolic difference could promote piscivory and development of the specific morphological peculiarities of the stone charr. Therefore, we present the first evidence in favor of trophic-based sympatric diversification in a river basin at high latitudes.     

Analysis of character divergence along environmental gradients and other covariates

Character displacement is typically identified by comparing phenotypic differences in sympatry and allopatry. Recently, however, Goldberg and Lande (2006) pointed out that when phenotypic characters vary along an environmental gradient, the standard approach may fail to identify sympatric character divergence. Here we present a general analytical procedure for identifying sympatric character divergence while accounting for phenotypic changes that covary with environmental variables. Our approach uses residual randomization from a generalized linear model, and allows the statistical comparison of sympatric phenotypic divergence to allopatric phenotypic divergence while accounting for phenotypic variation along a gradient. Through simulation we demonstrate that our approach correctly identifies patterns of sympatric character divergence when they are present, and does not identify such patterns when they are not. Our analytical approach complements and extends the suggestions of Goldberg and Lande (2006), by allowing a full statistical assessment of the varied patterns of character displacement along environmental gradients, or while accounting for other covariates and sources of variation.     

Sphagnum distribution patterns along environmental gradients in Bulgaria

Abstract

The distribution patterns of 18 Sphagnum species along base-richness and altitudinal gradients were studied in Bulgarian treeless wetlands which are noteworthy because of the edge-of-range occurrence of many mire species including Sphagnum. Of 483 spring and mire sites studied, 202 samples contained some Sphagnum species. The most common species were S. subsecundum (n=85), S. platyphyllum (46), S. contortum (41), S. teres (40) and S. capillifolium (26). The significance of Sphagnum responses to environmental gradients was tested by comparing generalized additive models against the null model. Many Sphagnum species displayed a significant response to the altitudinal gradient. Several species were clearly linked to low or to high altitudes, but the realized niche of other species was wide with respect to altitude. Most species significantly responded to water pH, both above and below the timberline. The same result was obtained for water conductivity below the timberline, whereas only a few species had a significant response to conductivity above the timberline. The highest water conductivity under which Sphagnum species occurred was 280 μS cm^?1. Sphagnum contortum was the species occupying the mires with the highest mineral content. On the contrary, Sphagnum warnstorfii, one of the most calcitolerant species in many regions of Europe, often occurred in extremely mineral-poor mires above the timberline. Some other Sphagnum species growing in mineral-rich mires below the timberline, also inhabited extremely mineral-poor mires above the timberline. This could be explained by adaptation to local conditions during long-term isolation on mineral-poor bedrock or by changed competition pressure.     

Speciation and the evolution of dispersal along environmental gradients

We analyze the joint evolution of an ecological character and of dispersal distance in asexual and sexual populations inhabiting an environmental gradient. Several interesting phenomena resulting from the evolutionary interplay of these characters are revealed. First, asexual and sexual populations exhibit two analogous evolutionary regimes, in which either speciation in the ecological character occurs in conjunction with evolution of short-range dispersal, or dispersal distance remains high and speciation does not occur. Second, transitions between these two regimes qualitatively differ between asexual and sexual populations, with the former showing speciation with long-range dispersal and the latter showing no speciation with short-range dispersal. Third, a phenotypic gradient following the environmental gradient occurs only in the last case, i.e., for non-speciating sexual populations evolving towards short-range dispersal. Fourth, the transition between the evolutionary regimes of long-range dispersal with no speciation and short-range dispersal with speciation is typically abrupt, mediated by a positive feedback between incipient speciation and the evolution of short-range dispersal. Fifth, even though the model of sexual evolution analyzed here does not permit assortative mating preferences, speciation occurs for a surprisingly wide range of conditions. This illustrates that dispersal evolution is a powerful alternative to preference evolution in enabling spatially distributed sexual populations to respond to frequency-dependent disruptive selection.     

The influence of habitat boundaries on evolutionary branching along environmental gradients

It is well known that habitat boundaries affect ecological dynamics, but their influence on evolutionary dynamics is less well understood. Here, we study the effects of different kinds of boundaries on evolutionary branching in clonal populations along environmental gradients by systematically analyzing individual-based stochastic models in small- and large-range systems, as well as their large-population-size limits through deterministic approximations. Specifically, we examine four prototypical kinds of boundaries: impermeable boundaries at which individuals stop (“stopping”), or from which they continue back into the interior as if bouncing back mechanically (“reflecting”), or that let them abort the dispersal attempt, return to their original position and try a different direction (“reprising”), and semipermeable boundaries that can be crossed without hindrance, but do not allow the crossing individual to return (“absorbing”).We find that boundary conditions shape branching patterns only in small-range systems, where stopping boundaries generate disruptive selection for a wide range of parameters, whereas absorbing boundaries always generate stabilizing selection. Reflecting and reprising boundaries generate disruptive selection at low individual mobilities, and stabilizing selection at high mobilities. To further analyze these findings, we introduce a simple approximation of the invasion fitness in a mobile population, which predicts the observed outcome. The effect of stochasticity on evolutionary outcomes is small even in small populations: stochasticity causes random branch extinctions at steeper slopes and higher mobilities. In large-range systems, frequency-dependent interactions alone induce evolutionary branching for almost all parameters and independent of boundary conditions.     

Disentangling diversity patterns in sandy beaches along environmental gradients

Species richness in sandy beaches is strongly affected by concurrent variations in morphodynamics and salinity. However, as in other ecosystems, different groups of species may exhibit contrasting patterns in response to these environmental variables, which would be obscured if only aggregate richness is considered. Deconstructing biodiversity, i.e. considering richness patterns separately for different groups of species according to their taxonomic affiliation, dispersal mode or mobility, could provide a more complete understanding about factors that drive species richness patterns. This study analyzed macroscale variations in species richness at 16 Uruguayan sandy beaches with different morphodynamics, distributed along the estuarine gradient generated by the Rio de la Plata over a 2 year period. Species richness estimates were deconstructed to discriminate among taxonomic groups, supralittoral and intertidal forms, and groups with different feeding habits and development modes. Species richness was lowest at intermediate salinities, increasing towards oceanic and inner estuarine conditions, mainly following the patterns shown for intertidal forms. Moreover, there was a differential tolerance to salinity changes according to the habitat occupied and development mode, which determines the degree of sensitivity of faunal groups to osmotic stress. Generalized (additive and linear) mixed models showed a clear increase of species richness towards dissipative beaches. All taxonomic categories exhibited the same trend, even though responses to grain size and beach slope were less marked for crustaceans and insects than for molluscs or polychaetes. However, supralittoral crustaceans exhibited the opposite trend. Feeding groups decreased from dissipative to reflective systems, deposit feeders being virtually absent in the latter. This deconstructive approach highlights the relevance of life history strategies in structuring communities, highlighting the relative importance that salinity and morphodynamic gradients have on macroscale diversity patterns in sandy beaches.     

Algal community patterns in Slovenian bogs along environmental gradients

In 2005 and 2006, epiphyton samples were collected from seven lowland and montane peat bogs in Slovenia. Water temperature, pH, conductivity, dissolved oxygen and saturation were measured at the same time. Diatoms, desmids and Cyanobacteria were the most abundand groups in species number. Canonical Correspondence Analysis (CCA) was carried out on Cyanobacteria, diatom and desmid flora composition. This analysis showed that shading was the most important parameter in Cyanobacteria distribution and bedrock the most important one in that of diatoms and desmids. Cluster analyses were carried out based on the Cyanobacteria, diatom and desmid data. The Cyanobacteria and diatom data separated sites, whereas the desmid data revealed a temporal aspect.     

Macrozoobenthos patterns along environmental gradients and hydrological connectivity of oxbow lakes

Hydrological connectivity and the frequency and intensity of floods are the key factors determining the structure of macroinvertebrates inhabiting wetland ecosystems in river valleys. In 2007, water and macroinvertebrate samples were collected on four occasions in the middle course of the S?upia River and in five oxbow lakes (Northern Poland) to determine the hydrological relations in a regulated lowland river environment marked by a moderate climate. The water bodies selected for the study featured different types of connections with the main river valley: two of them were completely cut off from the valley, one was connected via a single branch, one featured a forced-flow connection through drainage pipes, and one was connected by a system of drainage channels. Macroinvertebrates, mostly Chironomidae larvae, were predominant in the eutrophic waters of the river. The prevalent macroinvertebrates found in the eutrophicated oxbow lakes isolated from the river were Chironomidae larvae and Crustacea (mainly Asellus aquaticus). In unobstructed oxbow lakes, the main component of benthic fauna was Crustacea, while Ephemeroptera were found mostly in the water body connected to the river via a drainage channel. A canonical correspondence analysis (CCA) showed that hydrological connectivity was the main factor responsible for the structure of invertebrate populations, followed by the physical and chemical parameters of the local environment. A non-conformance analysis revealed that hydrological connectivity enhanced invertebrate abundance and biological diversity, while the overall abundance was marked by unimodal distribution. The developed general model indicates that in the group of measured environmental variables, nitrite concentrations were highly correlated with Shannon diversity and invertebrate composition, while sulphate levels were closely associated with invertebrate abundance in the waters of the analyzed ecosystems.     

The evolution of conditional dispersal and reproductive isolation along environmental gradients

Dispersal modulates gene flow throughout a population's spatial range. Gene flow affects adaptation at local spatial scales, and consequently impacts the evolution of reproductive isolation. A recent theoretical investigation has demonstrated that local adaptation along an environmental gradient, facilitated by the evolution of limited dispersal, can lead to parapatric speciation even in the absence of assortative mating. This and other studies assumed unconditional dispersal, so individuals start dispersing without regard to local environmental conditions. However, many species disperse conditionally; their propensity to disperse is contingent upon environmental cues, such as the degree of local crowding or the availability of suitable mates. Here, we use an individual-based model in continuous space to investigate by numerical simulation the relationship between the evolution of threshold-based conditional dispersal and parapatric speciation driven by frequency-dependent competition along environmental gradients. We find that, as with unconditional dispersal, parapatric speciation occurs under a broad range of conditions when reproduction is asexual, and under a more restricted range of conditions when reproduction is sexual. In both the asexual and sexual cases, the evolution of conditional dispersal is strongly influenced by the slope of the environmental gradient: shallow environmental gradients result in low dispersal thresholds and high dispersal distances, while steep environmental gradients result in high dispersal thresholds and low dispersal distances. The latter, however, remain higher than under unconditional dispersal, thus undermining isolation by distance, and hindering speciation in sexual populations. Consequently, the speciation of sexual populations under conditional dispersal is triggered by a steeper gradient than under unconditional dispersal. Enhancing the disruptiveness of frequency-dependent selection, more box-shaped competition kernels dramatically lower the speciation-enabling slope of the environmental gradient.     

Relationships of marsh seed banks to vegetation patterns along environmental gradients

• 1 The relationship of the seed bank to the vegetation of a freshwater marsh was studied along gradients of water depth and soil organic matter content. Characters examined included standing crop, seedling density, and species composition, distribution and richness.
• 2 The seed bank differed from the vegetation in that only nine of twenty-seven species were present in both, abundant seed-bank species were uncommon as adults, and adults showed different distributions along a gradient of soil organic matter content whereas their seeds were most abundant in soils with high organic matter.
• 3 The seed bank resembled the vegetation in that separate multivariate analyses of the communities revealed that variation in the species composition of each was significantly correlated with water depth and soil organic matter content. Further, species richness in both communities decreased with water depth and increased with soil organic matter content. Lastly, the standing crop of the vegetation and the number of seedlings both decreased with water depth and increased with soil organic matter.
• 4 Consideration of spatial patterns and environmental gradients revealed more similarities between vegetation and seed banks than were obtained by comparing species lists. The results suggest that artificial stimulation of seed bank germination for management purposes will not produce vegetation changes as large as those suggested by differences in species lists.

     

Contrasting patterns of local adaptation along climatic gradients between a sympatric parasitic and autotrophic tree species

Sympatric tree species are subject to similar climatic drivers, posing a question as to whether they display comparable adaptive responses. However, no study has explicitly examined local adaptation of co‐occurring parasitic and autotrophic plant species to the abiotic environment. Here we test the hypotheses that a generalist parasitic tree would display a weaker signal of selection and that genomic variation would associate with fewer climatic variables (particularly precipitation) but have similar spatial patterns to a sympatric autotrophic tree species. To test these hypotheses, we collected samples from 17 sites across the range of two tree species, the hemiparasite Nuytsia floribunda (n = 264) and sympatric autotroph Melaleuca rhaphiophylla (n = 272). We obtained 5,531 high‐quality genome‐wide single nucleotide polymorphisms (SNPs) for M. rhaphiophylla and 6,727 SNPs for N. floribunda using DArTseq genome scan technology. Population differentiation and environmental association approaches were used to identify signals of selection. Generalized dissimilarly modelling was used to detect climatic and spatial patterns of local adaptation across climatic gradients. Overall, 322 SNPs were identified as putatively adaptive for the autotroph, while only 57 SNPs were identified for the parasitic species. We found genomic variation to associate with different sets of bioclimatic variables for each species, with precipitation relatively less important for the parasite. Spatial patterns of predicted adaptive variability were different and indicate that co‐occurring species with disparate life history traits may not respond equally to selective pressures (i.e., temperature and precipitation). Together, these findings provide insight into local adaptation of sympatric parasitic and autotrophic tree species to abiotic environments.     

Grassland composition,structure, and diversity patterns along major environmental gradients in the Central Tien Shan

What species and traits signal vegetation types along prominent environmental gradients in the Central Tien Shan and what are the corresponding diversity patterns? Vegetation was sampled at 41 sites throughout the Kyrgyz Republic using quadrats stratified throughout a 1,000-m² sample area. Relationships among major environmental gradients, vegetation structure, and species composition were explored with nonmetric multidimensional scaling. Species distributions were examined to characterize phytogeographic patterns. Seven vegetation types ranging from desert grassland to meadow steppe were identified with cluster analysis, ordered primarily along elevation/mean annual temperature gradients. Four arid grassland types were distinguished, ranging mainly from 900 to 1,700 m elevation, and characterized by co-dominance of grasses and forbs with secondary dominance by shrubs. Annual and biennial forbs equaled perennial forbs in total importance. Grasses include C3 and C4 species. Three montane grassland types were recognized and characterized by co-dominance of perennial C3 grasses and forbs. Transition to montane steppe occurred from 1,500 to 1,900 m and is correlated with absence of C4 grasses and dominance of Festuca valesiaca. Highest diversity was found at intermediate elevations, from 1,800 to 2,600 m, in meadow steppe habitats. Forty-six percent of 580 identified species are Middle Asian endemics and remaining species primarily have distributions including Eastern Europe, the Caucasus, and western Siberia. Although grassland degradation from overgrazing has been chronic throughout the region, grasslands are widespread throughout the Kyrgyz Republic and many, particularly mid-elevation meadow steppes, retain high levels of native species diversity.     

Adaptive divergence along environmental gradients in a climate‐change‐sensitive mammal

In the face of predicted climate change, a broader understanding of biotic responses to varying environments has become increasingly important within the context of biodiversity conservation. Local adaptation is one potential option, yet remarkably few studies have harnessed genomic tools to evaluate the efficacy of this response within natural populations. Here, we show evidence of selection driving divergence of a climate‐change‐sensitive mammal, the American pika (Ochotona princeps), distributed along elevation gradients at its northern range margin in the Coast Mountains of British Columbia (BC), Canada. We employed amplified‐fragment‐length‐polymorphism‐based genomic scans to conduct genomewide searches for candidate loci among populations inhabiting varying environments from sea level to 1500 m. Using several independent approaches to outlier locus detection, we identified 68 candidate loci putatively under selection (out of a total 1509 screened), 15 of which displayed significant associations with environmental variables including annual precipitation and maximum summer temperature. These candidate loci may represent important targets for predicting pika responses to climate change and informing novel approaches to wildlife conservation in a changing world.     

Biological traits of stream macroinvertebrates from a semi-arid catchment: patterns along complex environmental gradients

1. The relationships between biological traits of macroinvertebrates and environmental characteristics were investigated in streams with contrasting physical, chemical or landscape level attributes. We used an ordination technique, RLQ analysis, which links an environmental table (R) with traits table (Q) through an abundance table (L) to investigate the relationship between habitat characteristics and biological traits.
2. A major environmental axis explaining the distribution of species and their distinctive biological features was obtained. This axis included variables of anthropogenic pressure (agricultural and urban uses) and natural variability (climatic and geologic) that are strongly intercorrelated in the study area, with a clear spatial component.
3. The attributes of species from frequently disturbed systems (small size, multivoltinism, diapause, ovoviviparity, etc.) were associated with semi-arid areas whereas traits common in more stable and favourable environments (large body size, semi-voltinism, isolated eggs, etc.) were found in upland forested areas.
4. The natural climatic variation was proposed as a disturbance axis of a theoretical habitat templet (driven by the intense hydrological disturbances typical of semi-arid streams), while anthropogenic pressure (mainly intensive agriculture) and high salinity, a natural consequence of geology, was proposed as an adversity axis. Different life-histories associated with contrasting environmental features were superimposed in this habitat templet.
5. The ecological–evolutionary scenario in which stream macroinvertebrates have evolved and by which their communities are organized, is closely linked to disturbance, environmental harshness and human pressure.     

Widespread phenotypic and genetic divergence along altitudinal gradients in animals

Altitudinal gradients offer valuable study systems to investigate how adaptive genetic diversity is distributed within and between natural populations and which factors promote or prevent adaptive differentiation. The environmental clines along altitudinal gradients tend to be steep relative to the dispersal distance of many organisms, providing an opportunity to study the joint effects of divergent natural selection and gene flow. Temperature is one variable showing consistent altitudinal changes, and altitudinal gradients can therefore provide spatial surrogates for some of the changes anticipated under climate change. Here, we investigate the extent and patterns of adaptive divergence in animal populations along altitudinal gradients by surveying the literature for (i) studies on phenotypic variation assessed under common garden or reciprocal transplant designs and (ii) studies looking for signatures of divergent selection at the molecular level. Phenotypic data show that significant between‐population differences are common and taxonomically widespread, involving traits such as mass, wing size, tolerance to thermal extremes and melanization. Several lines of evidence suggest that some of the observed differences are adaptively relevant, but rigorous tests of local adaptation or the link between specific phenotypes and fitness are sorely lacking. Evidence for a role of altitudinal adaptation also exists for a number of candidate genes, most prominently haemoglobin, and for anonymous molecular markers. Novel genomic approaches may provide valuable tools for studying adaptive diversity, also in species that are not amenable to experimentation.     

Biodiversity–productivity relationship in ponds: Community and metacommunity patterns along time and environmental gradients

Primary production correlates with diversity in various ways. These patterns may result from the interaction of various mechanisms related to the environmental context and the spatial and temporal scale of analysis. However, empirical evidence on diversity‐productivity patterns typically considers single temporal and spatial scales, and does not include the effect of environmental variables. In a metacommunity of macrophytes in ephemeral ponds, we analysed the diversity‐productivity relationship patterns in the field, the importance of the environmental variables of pond size and heterogeneity on such relationship, and the variation of these patterns at local (community level) and landscape scales (metacommunity level) across 52 ponds on twelve occasions, over five years (2005–2009). Combining all sampling dates, there were 377 ponds and 1954 sample‐unit observations. Vegetation biomass was used as a proxy for productivity, and biodiversity was represented by species richness, evenness, and their interaction. Environmental variables comprised pond area, depth and internal heterogeneity. Productivity and species richness were not directly related at the metacommunity level, and were positively related at the community level. Taking environmental variables into account revealed positive species richness‐productivity relationships at the metacommunity level and positive quadratic relationships at the community level. Productivity showed both positive and negative linear and nonlinear relationships with the size and heterogeneity of ponds. We found a weak relationship between productivity and evenness. The identity of variables associated with productivity changed between spatial scales and through time. The pattern of relationships between productivity and diversity depends on spatial scale and environmental context, and changes idiosyncratically through time within the same ecosystem. Thus, the diversity‐productivity relationship is not only a property of the study system, but also a consequence of environmental variations and the temporal and spatial scale of analysis.     

Life strategies, dominance patterns and mechanisms promoting species coexistence in phytoplankton communities along complex environmental gradients

This paper analyses the life strategies, the dominance patterns and the diversity in phytoplankton communities in large and deep lakes. The study was carried out on the largest Italian Lake (Lake Garda) from 1995 to 2000. Different statistical analyses were applied. For phytoplankton the time variable represents a complex environmental gradient driving annual succession; this gradient was made explicit by the application of PCA analyses to the environmental data. The use of Non Metric Multi Dimensional Scaling applied to Bray-Curtis dissimilarity matrices revealed an ordered and cyclic development of phytoplankton every year; the Bray-Curtis index, calculated between pairs of chronologically contiguous samples, was also used as a measure of the community change rate (β_t) over the temporal succession. A significant relationship between β_t and the complex environmental gradient was assessed. Finally, for every phytoplankton species, the optimum conditions for growth and the realised niches were determined. The positioning of the species on the complex environmental gradient, and the contemporaneous application of cluster analysis based on the different specific environmental optima, highlighted primarily the existence of two groups at the extreme of the complex environmental gradient. The first group included the large late winter/spring diatoms, which developed during high water turbulence and strong physical control, high nutrient concentrations, low light conditions and reduced competition. The second group was composed by many heterogeneous summer species characterised by the ability to contrast losses by grazing and sinking in stratified and stable conditions, and the ability of tolerating nutrient deficiency. A third group of species developed during environmental conditions in the middle of the two previous extremes. These included the three master species Mougeotia sp., Fragilaria crotonensis and Planktothrix rubescens/agardhii. The endogenous and exogenous mechanisms promoting species coexistence are discussed, along with the applicability of competitive and equilibrium/non-equilibrium theories to phytoplankton dynamics.     

Limiting similarity and functional diversity along environmental gradients

Recent developments in community models emphasize the importance of incorporating stochastic processes (e.g. ecological drift) in models of niche‐structured community assembly. We constructed a finite, spatially explicit, lottery model to simulate the distribution of species in a one‐dimensional landscape with an underlying gradient in environmental conditions. Our framework combines the potential for ecological drift with environmentally‐mediated competition for space in a heterogeneous environment. We examined the influence of niche breadth, dispersal distances, community size (total number of individuals) and the breadth of the environmental gradient on levels of species and functional trait diversity (i.e. differences in niche optima). Three novel results emerge from this model: (1) niche differences between adjacent species (e.g. limiting similarity) increase in smaller communities, because of the interaction of competitive effects and finite population sizes; (2) immigration from a regional species pool, stochasticity and niche‐assembly generate a bimodal distribution of species residence times (‘transient’ and ‘resident’) under a heterogeneous environment; and (3) the magnitude of environmental heterogeneity has a U‐shaped effect on diversity, because of shifts in species richness of resident vs. transient species. These predictions illustrate the potential importance of stochastic (although not necessarily neutral) processes in community assembly.     

Comparing species interaction networks along environmental gradients

Knowledge of species composition and their interactions, in the form of interaction networks, is required to understand processes shaping their distribution over time and space. As such, comparing ecological networks along environmental gradients represents a promising new research avenue to understand the organization of life. Variation in the position and intensity of links within networks along environmental gradients may be driven by turnover in species composition, by variation in species abundances and by abiotic influences on species interactions. While investigating changes in species composition has a long tradition, so far only a limited number of studies have examined changes in species interactions between networks, often with differing approaches. Here, we review studies investigating variation in network structures along environmental gradients, highlighting how methodological decisions about standardization can influence their conclusions. Due to their complexity, variation among ecological networks is frequently studied using properties that summarize the distribution or topology of interactions such as number of links, connectance, or modularity. These properties can either be compared directly or using a procedure of standardization. While measures of network structure can be directly related to changes along environmental gradients, standardization is frequently used to facilitate interpretation of variation in network properties by controlling for some co‐variables, or via null models. Null models allow comparing the deviation of empirical networks from random expectations and are expected to provide a more mechanistic understanding of the factors shaping ecological networks when they are coupled with functional traits. As an illustration, we compare approaches to quantify the role of trait matching in driving the structure of plant–hummingbird mutualistic networks, i.e. a direct comparison, standardized by null models and hypothesis‐based metaweb. Overall, our analysis warns against a comparison of studies that rely on distinct forms of standardization, as they are likely to highlight different signals. Fostering a better understanding of the analytical tools available and the signal they detect will help produce deeper insights into how and why ecological networks vary along environmental gradients.