The human touch: a meta-analysis of anthropogenic effects on plant-pollinator interaction networks.

Background: Anthropogenic activities significantly impact natural ecosystems, leading to alterations in plant and pollinator diversity and abundance. These changes often result in shifts within interacting communities, potentially reshaping the structure of plant-pollinator interaction networks. Given the escalating human footprint on habitats, evaluating the response of these networks to anthropization is critical for devising effective conservation and management strategies. Methods: We conducted a comprehensive review of the plant-pollinator network literature to assess the impact of anthropization on network structure. We assessed network metrics such as nestedness measure based on overlap and decreasing fills (NODF), network specialization (H2'), connectance (C), and modularity (Q) to understand structural changes. Employing a meta-analytical approach, we examined how anthropization activities, such as deforestation, urbanization, habitat fragmentation, agriculture, intentional fires and livestock farming, affect both plant and pollinator richness. Results: We generated a dataset for various metrics of network structure and 36 effect sizes for the meta-analysis, from 38 articles published between 2010 and 2023. Studies assessing the impact of agriculture and fragmentation were well-represented, comprising 68.4% of all studies, with networks involving interacting insects being the most studied taxa. Agriculture and fragmentation reduce nestedness and increase specialization in plant-pollinator networks, while modularity and connectance are mostly not affected. Although our meta-analysis suggests that anthropization decreases richness for both plants and pollinators, there was substantial heterogeneity in this regard among the evaluated studies. The meta-regression analyses helped us determine that the habitat fragment size where the studies were conducted was the primary variable contributing to such heterogeneity. Conclusions: The analysis of human impacts on plant-pollinator networks showed varied effects worldwide. Responses differed among network metrics, signaling nuanced impacts on structure. Activities like agriculture and fragmentation significantly changed ecosystems, reducing species richness in both pollinators and plants, highlighting network vulnerability. Regional differences stressed the need for tailored conservation. Despite insights, more research is crucial for a complete understanding of these ecological relationships.

Flower visits and pollinator pollen load networks reveal the effects of pollinator sharing on heterospecific pollen transfer in a subalpine plant community.

The mutualistic network of plant-pollinator also involves interspecific pollination caused by pollinator sharing. Plant-pollinator networks are commonly based on flower visit observations, which may not adequately represent the actual pollen transfer between co-flowering plant species. Here, we compared the network structure of plant-pollinator interactions based on flower visits (FV) and pollen loads (PL) on the bodies of pollinators and tested how the degree of pollinator sharing in the two networks affected heterospecific pollen transfer (HPT) between plant species in a subalpine meadow. The FV and PL networks were largely overlapped. PL network included more links than FV network. The positions of plant and pollinator species in the FV and PL networks were positively correlated, indicating that both networks could detect major plant-pollinator interactions. The degree of pollinator sharing, based on either the FV or the PL network, positively influenced the amount of heterospecific pollen transferred between plant species pairs. However, the degree of pollinator sharing had a low overall explanatory power for HPT, and the explanatory powers of the FV and PL networks were similar. Overall, our study highlights the importance of FV and PL for understanding the drivers and outcomes of plant-pollinator interactions, as well as their relevance to HPT.

Pollen nutrition structures bee and plant community interactions.

As bees' main source of protein and lipids, pollen is critical for their development, reproduction, and health. Plant species vary considerably in the macronutrient content of their pollen, and research in bee model systems has established that this variation both modulates performance and guides floral choice. Yet, how variation in pollen chemistry shapes interactions between plants and bees in natural communities is an open question, essential for both understanding the nutritional dynamics of plant-pollinator mutualisms and informing their conservation. To fill this gap, we asked how pollen nutrition (relative protein and lipid content) sampled from 109 co-flowering plant species structured visitation patterns observed among 75 subgenera of pollen-collecting bees in the Great Basin/Eastern Sierra region (USA). We found that the degree of similarity in co-flowering plant species' pollen nutrition predicted similarity among their visitor communities, even after accounting for floral morphology and phylogeny. Consideration of pollen nutrition also shed light on the structure of this interaction network: Bee subgenera and plant genera were arranged into distinct, interconnected groups, delineated by differences in pollen macronutrient values, revealing potential nutritional niches. Importantly, variation in pollen nutrition alone (high in protein, high in lipid, or balanced) did not predict the diversity of bee visitors, indicating that plant species offering complementary pollen nutrition may be equally valuable in supporting bee diversity. Nutritional diversity should thus be a key consideration when selecting plants for habitat restoration, and a nutritionally explicit perspective is needed when considering reward systems involved in the community ecology of pollination.

Manipulating network connectance by altering plant attractiveness.

Background: Mutualistic interactions between plants and their pollinating insects are critical to the maintenance of biodiversity. However, we have yet to demonstrate that we are able to manage the structural properties of these networks for the purposes of pollinator conservation and preserving functional outcomes, such as pollination services. Our objective was to explore the extent of our ability to experimentally increase, decrease, and maintain connectance, a structural attribute that reflects patterns of insect visitation and foraging preferences. Patterns of connectance relate to the stability and function of ecological networks. Methods: We implemented a 2-year field experiment across eight sites in urban Dublin, Ireland, applying four agrochemical treatments to fixed communities of seven flowering plant species in a randomized block design. We spent ~117 h collecting 1,908 flower-visiting insects of 92 species or morphospecies with standardized sampling methods across the 2 years. We hypothesized that the fertilizer treatment would increase, herbicide decrease, and a combination of both maintain the connectance of the network, relative to a control treatment of just water. Results: Our results showed that we were able to successfully increase network connectance with a fertilizer treatment, and maintain network connectance with a combination of fertilizer and herbicide. However, we were not successful in decreasing network connectance with the herbicide treatment. The increase in connectance in the fertilized treatment was due to an increased species richness of visiting insects, rather than changes to their abundance. We also demonstrated that this change was due to an increase in the realized proportion of insect visitor species rather than increased visitation by common, generalist species of floral visitors. Overall, this work suggests that connectance is an attribute of network structure that can be manipulated, with implications for management goals or conservation efforts in these mutualistic communities.

Impacts of land use change on native plant-butterfly interaction networks from central Mexico.

Background: Land use change is a key catalyst of global biodiversity loss and ecosystem degradation. Deforestation and conversion of natural habitats to agricultural or urban areas can profoundly disrupt plant-flower visitor interactions by altering their abundances and distribution. Yet, specific studies analyzing the effects of land use change on the structure of networks of the interactions between particular groups of flower visitors and their plants are still scarce. Here, we aimed to analyze how converting native habitats affects the species composition of butterfly communities and their plants, and whether this, in turn, leads to changes in the structure of interaction networks in the modified habitats. Methods: We performed bi-monthly censuses for a year to record plant-butterfly interactions and assess species diversity across three habitat types, reflecting a land-use change gradient. From original native juniper forest to urban and agricultural zones in central Mexico, one site per land use type was surveyed. Interactions were summarized in matrices on which we calculated network descriptors: connectance, nestedness and modularity. Results: We found highest butterfly diversity in native forest, with the most unique species (i.e., species not shared with the other two sites). Agricultural and urban sites had similar diversity, yet the urban site featured more unique species. The plant species richness was highest in the urban site, and the native forest site had the lowest plant species richness, with most of the plants being unique to this site. Butterfly and plant compositions contrasted most between native forest and modified sites. Network analysis showed differences between sites in the mean number of links and interactions. The urban network surpassed agriculture and native forest networks in links, while the native forest network had more interactions than the agriculture and urban networks. Native plants had more interactions than alien species. All networks exhibited low connectance and significant nestedness and modularity, with the urban network featuring the most modules (i.e., 10 modules). Conclusions: Converting native habitats to urban or agricultural areas reshapes species composition, diversity and interaction network structure for butterfly communities and plants. The urban network showed more links and modules, suggesting intricate urban ecosystems due to diverse species, enhanced resources, and ecological niches encouraging interactions and coexistence. These findings emphasize the impacts of land use change on plant-butterfly interactions and the structure of their interaction networks.

Individual bee foragers are less-efficient transporters of pollen for plants from which they collect the most pollen in their scopae.

PREMISE: Bees provision most of the pollen removed from anthers to their larvae and transport only a small proportion to stigmas, which can negatively affect plant fitness. Though most bee species collect pollen from multiple plant species, we know little about how the efficiency of bees' pollen transport varies among host plant species or how it relates to other aspects of generalist bee foraging behavior that benefit plant fitness, such as specialization on individual foraging bouts. METHODS: We compared the pollen collected and transported by three bee species for 46 co-occurring plant species. Specifically, we compared the relative abundance of pollen taxa in the individual bees' scopae, structures where bees store pollen to provision larvae, with the relative abundance of pollen taxa on the rest of bees' bodies, which is more likely to be transferred to stigmas. RESULTS: Bees carried five times more pollen grains in their scopae than elsewhere on their bodies. Within foraging bouts, bees were relatively specialized in their pollen collection, but transported proportionally less pollen for the host plants on which they specialized. Across foraging bouts, two bee species transported proportionally less pollen for some of their host plants than for others, though differences didn't consistently follow the same trend as at the foraging bout scale. CONCLUSIONS: Our results suggest that foraging-bout specialization, which is known to reduce heterospecific pollen transfer, also results in less-efficient pollen transport. Thus, bee foragers that visit predominantly one plant species may have contrasting effects on that plant's fitness.

Evading tipping points in socio-mutualistic networks via structure mediated optimal strategy.

The threat of large-scale pollinator decline is increasing globally under stress from multiple anthropogenic pressures. Traditional approaches have focused on managing endangered species at an individual level, in which the effect of complex interactions such as mutualism and competition are amiss. Here, we develop a coupled socio-mutualistic network model that captures the change in pollinator dynamics with human conservation opinion in a deteriorating environment. We show that the application of social norm (or conservation) at the pollinator nodes is fit to prevent sudden community collapse in representative networks of varied topology. Whilst primitive strategies have focused on regulating abundance as a mitigation strategy, the role of network structure has been largely overlooked. Here, we develop a novel network structure-mediated conservation strategy to find the optimal set of nodes on which norm implementation successfully prevents community collapse. We find that networks of intermediate nestedness require conservation at a minimum number of nodes to prevent a community collapse. We claim the robustness of the optimal conservation strategy (OCS) after validation on several simulated and empirical networks of varied complexity against a broad range of system parameters. Dynamical analysis of the reduced model shows that incorporating social norms allows the pollinator abundance to grow that would have otherwise crossed a tipping point and undergo extinction. Together, this novel means OCS provides a potential plan of action for conserving plant-pollinator networks bridging the gap between research in mutualistic networks and conservation ecology.

Environmental Effects on Bee Microbiota.

Anthropogenic activities and increased land use, which include industrialization, agriculture and urbanization, directly affect pollinators by changing habitats and floral availability, and indirectly by influencing their microbial composition and diversity. Bees form vital symbioses with their microbiota, relying on microorganisms to perform physiological functions and aid in immunity. As altered environments and climate threaten bees and their microbiota, characterizing the microbiome and its complex relationships with its host offers insights into understanding bee health. This review summarizes the role of sociality in microbiota establishment, as well as examines if such factors result in increased susceptibility to altered microbiota due to environmental changes. We characterize the role of geographic distribution, temperature, precipitation, floral resources, agriculture, and urbanization on bee microbiota. Bee microbiota are affected by altered surroundings regardless of sociality. Solitary bees that predominantly acquire their microbiota through the environment are particularly sensitive to such effects. However, the microbiota of obligately eusocial bees are also impacted by environmental changes despite typically well conserved and socially inherited microbiota. We provide an overview of the role of microbiota in plant-pollinator relationships and how bee microbiota play a larger role in urban ecology, offering microbial connections between animals, humans, and the environment. Understanding bee microbiota presents opportunities for sustainable land use restoration and aiding in wildlife conservation.

The non-random assembly of network motifs in plant-pollinator networks.

Ecological processes leave distinct structural imprints on the species interactions that shape the topology of animal-plant mutualistic networks. Detecting how direct and indirect interactions between animals and plants are organised is not trivial since they go beyond pairwise interactions, but may get blurred when considering global network descriptors. Recent work has shown that the meso-scale, the intermediate level of network complexity between the species and the global network, can capture this important information. The meso-scale describes network subgraphs representing patterns of direct and indirect interactions between a small number of species, and when these network subgraphs differ statistically from a benchmark, they are often referred to as 'network motifs'. Although motifs can capture relevant ecological information of species interactions, they remain overlooked in natural plant-pollinator networks. By exploring 60 empirical plant-pollinator networks from 18 different studies with wide geographical coverage, we show that some network subgraphs are consistently under- or over-represented, suggesting the presence of worldwide network motifs in plant-pollinator networks. In addition, we found a higher proportion of densely connected network subgraphs that, based on previous findings, could reflect that species relative abundances are the main driver shaping the structure of the meso-scale on plant-pollinator communities. Moreover, we found that distinct subgraph positions describing species ecological roles (e.g. generalisation and number of indirect interactions) are occupied by different groups of animal and plant species representing their main life-history strategies (i.e. functional groups). For instance, we found that the functional group of 'bees' was over-represented in subgraph positions with a lower number of indirect interactions in contrast to the rest of floral visitors groups. Finally, we show that the observed functional group combinations within a subgraph cannot be retrieved from their expected probabilities (i.e. joint probability distributions), indicating that plant and floral visitor associations within subgraphs are not random either. Our results highlight the presence of common network motifs in plant-pollinator communities that are formed by a non-random association of plants and floral visitors functional groups.

Conserving diversity in Irish plant-pollinator networks.

Beneficial insects provide valuable services upon which we rely, including pollination. Pollinator conservation is a global priority, and a significant concern in Ireland, where over half of extant bee species have declined significantly in recent decades. As flower-visiting insects rely on flowering plants, one way to conserve and promote pollinator populations is to protect high-quality habitat. We analyzed the structure of insect-flower interactions from multiple habitat categories in a large database of interactions from Ireland. Our primary goals were to compare spatial and temporal variation in Irish network structures, compare Irish networks to published networks from other countries, and provide evidence-based recommendations for pollinator conservation in Ireland by identifying well-visited plant species that may promote high pollinator diversity, abundance, and functional complementarity. Habitat types within Ireland differed substantially: seminatural grasslands had the highest pollinator species richness and largest number of unique pollinator species, while intensively managed habitats exhibited negative asymmetry (more plant than pollinator species). This negative asymmetry is notable because most plant-pollinator networks exhibit a positive asymmetry. Within intensively managed habitats, agricultural and urban habitats differed. Urban habitats had the highest number of non-native plant species while agricultural habitats had the lowest pollinator species richness. We also found Irish networks varied across the growing season, where July had the highest plant and insect species richness. When comparing Irish networks to published networks from other countries, we found Irish networks had a higher ratio of plant species to pollinator species, and that this difference was most evident in agricultural habitats. This ratio means the typical network asymmetry (more pollinator than plant species) was flipped (more plant than pollinator species) in the Irish network. We conclude that conserving seminatural grasslands in Ireland will be an essential component of pollinator conservation and identify thirty-five plant species important for restoring seminatural habitats.

Functional traits of plants and pollinators explain resource overlap between honeybees and wild pollinators.

Managed and wild pollinators often cohabit in both managed and natural ecosystems. The western honeybee, Apis mellifera, is the most widespread managed pollinator species. Due to its density and behaviour, it can potentially influence the foraging activity of wild pollinators, but the strength and direction of this effect are often context-dependent. Here, we observed plant-pollinator interactions in 51 grasslands, and we measured functional traits of both plants and pollinators. Using a multi-model inference approach, we explored the effects of honeybee abundance, temperature, plant functional diversity, and trait similarity between wild pollinators and the honeybee on the resource overlap between wild pollinators and the honeybee. Resource overlap decreased with increasing honeybee abundance only in plant communities with high functional diversity, suggesting a potential diet shift of wild pollinators in areas with a high variability of flower morphologies. Moreover, resource overlap increased with increasing trait similarity between wild pollinators and the honeybee. In particular, central-place foragers of family Apidae with proboscis length similar to the honeybee exhibited the highest resource overlap. Our results underline the importance of promoting functional diversity of plant communities to support wild pollinators in areas with a high density of honeybee hives. Moreover, greater attention should be paid to areas where pollinators possess functional traits similar to the honeybee, as they are expected to be more prone to potential competition with this species.

Pollen-insect interaction meta-networks identify key relationships for conservation in mosaic agricultural landscapes.

Flower visitors use different parts of the landscape through the plants they visit, however these connections vary within and among land uses. Identifying which flower-visiting insects are carrying pollen, and from where in the landscape, can elucidate key pollen-insect interactions and identify the most important sites for maintaining community-level interactions across land uses. We developed a bipartite meta-network, linking pollen-insect interactions with the sites they occur in. We used this to identify which land-use types at the site- and landscape-scale (within 500 m of a site) are most important for conserving pollen-insect interactions. We compared pollen-insect interactions across four different land uses (remnant native forest, avocado orchard, dairy farm, rotational potato crop) within a mosaic agricultural landscape. We sampled insects using flight intercept traps, identified pollen carried on their bodies and quantified distinct pollen-insect interactions that were highly specialized to both natural and modified land uses. We found that sites in crops and dairy farms had higher richness of pollen-insect interactions and higher interaction strength than small forest patches and orchards. Further, many interactions involved pollinator groups such as flies, wasps, and beetles that are often under-represented in pollen-insect network studies, but were often connector species in our networks. These insect groups require greater attention to enable wholistic pollinator community conservation. Pollen samples were dominated by grass (Poaceae) pollen, indicating anemophilous plant species may provide important food resources for pollinators, particularly in modified land uses. Field-scale land use (within 100 m of a site) better predicted pollen-insect interaction richness, uniqueness, and strength than landscape-scale. Thus, management focused at smaller scales may provide more tractable outcomes for conserving or restoring pollen-insect interactions in modified landscapes. For instance, actions aimed at linking high-richness sites with those containing unique (i.e., rare) interactions by enhancing floral corridors along field boundaries and between different land uses may best aid interaction diversity and connectance. The ability to map interactions across sites using a meta-network approach is practical and can inform land-use planning, whereby conservation efforts can be targeted toward areas that host key interactions between plant and pollinator species.

Mutualistic networks: moving closer to a predictive theory.

Plant-animal mutualistic networks sustain terrestrial biodiversity and human food security. Global environmental changes threaten these networks, underscoring the urgency for developing a predictive theory on how networks respond to perturbations. Here, I synthesise theoretical advances towards predicting network structure, dynamics, interaction strengths and responses to perturbations. I find that mathematical models incorporating biological mechanisms of mutualistic interactions provide better predictions of network dynamics. Those mechanisms include trait matching, adaptive foraging, and the dynamic consumption and production of both resources and services provided by mutualisms. Models incorporating species traits better predict the potential structure of networks (fundamental niche), while theory based on the dynamics of species abundances, rewards, foraging preferences and reproductive services can predict the extremely dynamic realised structures of networks, and may successfully predict network responses to perturbations. From a theoretician's standpoint, model development must more realistically represent empirical data on interaction strengths, population dynamics and how these vary with perturbations from global change. From an empiricist's standpoint, theory needs to make specific predictions that can be tested by observation or experiments. Developing models using short-term empirical data allows models to make longer term predictions of community dynamics. As more longer term data become available, rigorous tests of model predictions will improve.

Network modularity influences plant reproduction in a mosaic tropical agroecosystem.

Biodiversity influences ecosystem function, but there is limited understanding of the mechanisms that support this relationship across different land use types in mosaic agroecosystems. Network approaches can help to understand how community structure influences ecosystem function across landscapes; however, in ecology, network analyses have largely focused on species-species interactions. Here, we use bipartite network analysis in a novel way: to link pollinator communities to sites in a tropical agricultural landscape. We used sentinel plants of Brassica rapa to examine how the structure of the community network influences plant reproduction. Diptera was the most common order of flower visitors at every site. Syrphidae visits were the strongest contributor to the number of fertilized pods, while visits by Syrphidae, Hymenoptera and Lepidoptera had the strongest effect on the number of seeds per pod. Sentinel pots at forest sites were visited by more unique species (i.e. species with higher d') than sites in other land uses, and dairy sites had more visitors that were common across the network. Participation coefficients, which indicate how connected a single node is across network modules, were strong predictors of ecosystem function: plant reproduction increased at sites with higher participation coefficients. Flower visitor taxa with higher participation coefficients also had the strongest effect on plant reproduction. Hymenoptera visits were the best predictor for participation coefficients but an Allograpta sp. (Diptera: Syrphidae) was the most influential flower visitor species in the landscape network. A diverse insect community contributed to plant reproduction and connection among nodes in this system. Identifying the 'keystone' flower visitor species and sites that have a strong influence on network structure is a significant step forward to inform conservation priorities and decision-making in diverse agroecosystems.

Climate drives plant-pollinator interactions even along small-scale climate gradients: the case of the Aegean.

Plant-pollinator network structure is the outcome of ecological and evolutionary processes, and although the importance of environmental factors is beyond doubt, our knowledge of how abiotic factors (e.g. climate) shape plant-pollinator networks remains limited. This knowledge gap is critical, as climate change poses a major threat to ecosystems, especially in the Mediterranean. This study focuses on one of the hottest parts of the Mediterranean Basin, the Aegean Archipelago, Greece, and examines how climate affects species richness and network properties (e.g. nestedness, modularity and specialisation) - either directly or indirectly through species richness. We sampled systematically 39 local plant-pollinator networks on eight islands along a north-south climate gradient in the Aegean. All plant-pollinator material used in the analyses was collected in 2012 and identified to species level. Aspects of climate used in the models were expressed as average conditions (mean temperature and annual precipitation) or as seasonal variability (isothermality and temperature seasonality). Structural properties of plant-pollinator networks were found to be strongly associated with species richness, which was in turn affected by climate, implying that pollination network structure is driven indirectly by climate. In addition, climate had a direct effect on network structure, especially on modularity and specialisation. Different aspects of climate affected network properties in different ways. We highlight that even in a relatively narrow latitudinal gradient, such as within the Aegean Sea region, climate constitutes a significant driver of plant-pollinator interactions.

The influence of floral traits on specialization and modularity of plant-pollinator networks in a biodiversity hotspot in the Peruvian Andes.

BACKGROUND AND AIMS: Modularity is a ubiquitous and important structural property of ecological networks which describes the relative strengths of sets of interacting species and gives insights into the dynamics of ecological communities. However, this has rarely been studied in species-rich, tropical plant-pollinator networks. Working in a biodiversity hotspot in the Peruvian Andes we assessed the structure of quantitative plant-pollinator networks in nine valleys, quantifying modularity among networks, defining the topological roles of species and the influence of floral traits on specialization. METHODS: A total of 90 transects were surveyed for plants and pollinators at different altitudes and across different life zones. Quantitative modularity (QuanBiMo) was used to detect modularity and six indices were used to quantify specialization. KEY RESULTS: All networks were highly structured, moderately specialized and significantly modular regardless of size. The strongest hubs were Baccharis plants, Apis mellifera, Bombus funebris and Diptera spp., which were the most ubiquitous and abundant species with the longest phenologies. Species strength showed a strong association with the modular structure of plant-pollinator networks. Hubs and connectors were the most centralized participants in the networks and were ranked highest (high generalization) when quantifying specialization with most indices. However, complementary specialization d' quantified hubs and connectors as moderately specialized. Specialization and topological roles of species were remarkably constant across some sites, but highly variable in others. Networks were dominated by ecologically and functionally generalist plant species with open access flowers which are closely related taxonomically with similar morphology and rewards. Plants associated with hummingbirds had the highest level of complementary specialization and exclusivity in modules (functional specialists) and the longest corollas. CONCLUSIONS: We have demonstrated that the topology of networks in this tropical montane environment was non-random and highly organized. Our findings underline that specialization indices convey different concepts of specialization and hence quantify different aspects, and that measuring specialization requires careful consideration of what defines a specialist.

Beyond neutral and forbidden links: morphological matches and the assembly of mutualistic hawkmoth-plant networks.

A major challenge in evolutionary ecology is to understand how co-evolutionary processes shape patterns of interactions between species at community level. Pollination of flowers with long corolla tubes by long-tongued hawkmoths has been invoked as a showcase model of co-evolution. Recently, optimal foraging models have predicted that there might be a close association between mouthparts' length and the corolla depth of the visited flowers, thus favouring trait convergence and specialization at community level. Here, we assessed whether hawkmoths more frequently pollinate plants with floral tube lengths similar to their proboscis lengths (morphological match hypothesis) against abundance-based processes (neutral hypothesis) and ecological trait mismatches constraints (forbidden links hypothesis), and how these processes structure hawkmoth-plant mutualistic networks from five communities in four biogeographical regions of South America. We found convergence in morphological traits across the five communities and that the distribution of morphological differences between hawkmoths and plants is consistent with expectations under the morphological match hypothesis in three of the five communities. In the two remaining communities, which are ecotones between two distinct biogeographical areas, interactions are better predicted by the neutral hypothesis. Our findings are consistent with the idea that diffuse co-evolution drives the evolution of extremely long proboscises and flower tubes, and highlight the importance of morphological traits, beyond the forbidden links hypothesis, in structuring interactions between mutualistic partners, revealing that the role of niche-based processes can be much more complex than previously known.

Influences of sampling effort on detected patterns and structuring processes of a Neotropical plant-hummingbird network.

Virtually all empirical ecological interaction networks to some extent suffer from undersampling. However, how limitations imposed by sampling incompleteness affect our understanding of ecological networks is still poorly explored, which may hinder further advances in the field. Here, we use a plant-hummingbird network with unprecedented sampling effort (2716 h of focal observations) from the Atlantic Rainforest in Brazil, to investigate how sampling effort affects the description of network structure (i.e. widely used network metrics) and the relative importance of distinct processes (i.e. species abundances vs. traits) in determining the frequency of pairwise interactions. By dividing the network into time slices representing a gradient of sampling effort, we show that quantitative metrics, such as interaction evenness, specialization (H2 '), weighted nestedness (wNODF) and modularity (Q; QuanBiMo algorithm) were less biased by sampling incompleteness than binary metrics. Furthermore, the significance of some network metrics changed along the sampling effort gradient. Nevertheless, the higher importance of traits in structuring the network was apparent even with small sampling effort. Our results (i) warn against using very poorly sampled networks as this may bias our understanding of networks, both their patterns and structuring processes, (ii) encourage the use of quantitative metrics little influenced by sampling when performing spatio-temporal comparisons and (iii) indicate that in networks strongly constrained by species traits, such as plant-hummingbird networks, even small sampling is sufficient to detect their relative importance for the frequencies of interactions. Finally, we argue that similar effects of sampling are expected for other highly specialized subnetworks.

Pollinators visit related plant species across 29 plant-pollinator networks.

Understanding the evolution of specialization in host plant use by pollinators is often complicated by variability in the ecological context of specialization. Flowering communities offer their pollinators varying numbers and proportions of floral resources, and the uniformity observed in these floral resources is, to some degree, due to shared ancestry. Here, we find that pollinators visit related plant species more so than expected by chance throughout 29 plant-pollinator networks of varying sizes, with "clade specialization" increasing with community size. As predicted, less versatile pollinators showed more clade specialization overall. We then asked whether this clade specialization varied with the ratio of pollinator species to plant species such that pollinators were changing their behavior when there was increased competition (and presumably a forced narrowing of the realized niche) by examining pollinators that were present in at least three of the networks. Surprisingly, we found little evidence that variation in clade specialization is caused by pollinator species changing their behavior in different community contexts, suggesting that clade specialization is observed when pollinators are either restricted in their floral choices due to morphological constraints or innate preferences. The resulting pollinator sharing between closely related plant species could result in selection for greater pollinator specialization.

The potential for indirect effects between co-flowering plants via shared pollinators depends on resource abundance, accessibility and relatedness.

Co-flowering plant species commonly share flower visitors, and thus have the potential to influence each other's pollination. In this study we analysed 750 quantitative plant-pollinator networks from 28 studies representing diverse biomes worldwide. We show that the potential for one plant species to influence another indirectly via shared pollinators was greater for plants whose resources were more abundant (higher floral unit number and nectar sugar content) and more accessible. The potential indirect influence was also stronger between phylogenetically closer plant species and was independent of plant geographic origin (native vs. non-native). The positive effect of nectar sugar content and phylogenetic proximity was much more accentuated for bees than for other groups. Consequently, the impact of these factors depends on the pollination mode of plants, e.g. bee or fly pollinated. Our findings may help predict which plant species have the greatest importance in the functioning of plant-pollination networks.