Allopatric divergence can result in the evolution of incomplete reproductive barriers, which are put to the test when species come into secondary contact. North American red-backed voles (, ) form a broad contact zone, presenting an opportunity to investigate outcomes of secondary contact. Using RADseq data for > 200 red-backed vole specimens across three transects of the contact zone (Southeast Alaska, British Columbia, Northwest Territories), we test for evidence of admixture and describe the biogeographic history of each region. We find limited evidence for nuclear admixture, but by analyzing nuclear and mitochondrial RAD loci separately, we detect extensive unidirectional mitochondrial introgression. In British Columbia, the pattern of mitonuclear discordance is consistent with a moving hybrid zone, and suggests the possibility of adaptive mitochondrial introgression. In Southeast Alaska, mitochondrial introgression is localized to a smaller geographic region, and involves a distinct mitochondrial haplotype that is only found in specimens with nuclear genomes. In contrast, we find no evidence for mitochondrial introgression in the Northwest Territories. Together, our results suggest a complex biogeographic history for North American red-backed voles, mediated by the availability of ice-free colonization pathways following the LGM and incomplete barriers to reproduction, leading to different outcomes of secondary contact across the continent.

Latitudinal gradient can influence ecosystem dynamics and species distribution, yet the influence on some aspects, such as intra-species diversity, is less well understood. The large-scale distribution of harbour porpoises () in the Northeast Atlantic, indicates that ecological adaptations within the species might be greater than currently recognised. This study investigates variation in foraging behaviour using long-term passive acoustic monitoring data collected between 2009 and 2023 from Iceland, Sweden, Ireland, and France. In each area, Generalised Additive Models (GAMs) were used to investigate the influence of large-scale environmental variables (water temperature, salinity, primary production, diel phase and daylength) on foraging behaviour. Our results show variability and complexity in foraging and differences in temporal patterns between areas. In the northernmost regions, with the largest variation in daylight, foraging behaviour was related to diel phase, with primarily nocturnal foraging recorded during the year, but with predominantly diurnal foraging in Iceland during late fall. In the southernmost regions, less effect of diel phase on foraging was found. Harbour porpoises in Sweden and Iceland exhibited increased day-round foraging during calving periods, highlighting the role of reproductive energetics in behavioural adaptations, and the complexity and importance of foraging during different seasons. Our findings underscore the influence of environmental drivers in shaping foraging strategies, supporting the concept that harbour porpoises optimise these based on local conditions and prey availability. Using long-term datasets, spanning broad geographical and temporal scales, this study contributes to the wider ecological understanding of animals with extensive latitudinal distributions, highlighting intra-species variance and the need for region-specific conservation and management.

Environmentally mediated transmission is a critical pathway for disease spread, particularly for pathogens with environmental persistence. However, distinguishing which overlapping host movements result in transmission remains difficult, as most mobility models assume homogeneous exposure and overlook key spatial and behavioral factors. We extend previous models of indirect contact to account for variation in host movement behavior that could affect the probability of indirect transmission and examine these effects on transmission dynamics. We considered four models with different assumptions about the influence of movement behavior on indirect contact and transmission probability. The pathogen decay model includes all overlaps occurring within the pathogen's viability window. The high-use area model restricts transmission to frequently visited locations, assuming greater pathogen deposition where hosts repeatedly congregate. The behavioral model filters contacts based on host behaviors relevant to deposition or acquisition. The integrated model incorporates both spatial and behavioral constraints. We applied these models to GPS telemetry data from wild pigs in Florida and simulated disease dynamics using SEIR models with environmental transmission for Influenza A and , representing pathogens with short and long environmental persistence, respectively. Compared to pathogen decay model, total contact numbers decreased by 0.3% in high-use model, 70.3% in behavioral model, and 70.4% in integrated model under the short-lived condition. Edge density declined from 0.60 to 0.50, transitivity from 0.81 to 0.77, and assortativity from 0.43 to 0.17. declined from 2.42 ± 0.47 to 2.06 ± 0.39 (high-use), 1.56 ± 0.42 (behavioral), and 1.35 ± 0.40 (integrated), with a lag of approximately 21 days in peak incidence. The long-lived scenarios, were quantitively similar to those of short-lived scenarios. These results underscore that assumptions about how indirect contact from movement data maps to pathogen transmission significantly influence network configuration and epidemic projections. Our findings highlight the importance of mechanistically defining indirect transmission in disease modeling.

Forest pests and diseases may indirectly drive critical shifts in rates of litter decomposition, ultimately impacting carbon and nutrient cycling. Beech leaf disease (BLD) is a rapidly emerging forest disease affecting American beech and driving significant changes in leaf traits and associated microbial communities-both critical drivers of decomposition. However, the extent to which these BLD-driven changes may alter rates of litter decomposition remains unknown. Here, we explore rates of decomposition in BLD symptomatic and asymptomatic leaf litter using a field-based litterbag experiment. In addition to monitoring rates of decomposition, we also explored differences in the associated fungal communities. We found that BLD-symptomatic litter decomposed more quickly than asymptomatic litter. Furthermore, fungal communities differed in composition across symptomatic and asymptomatic leaf litter, with higher abundances of saprotrophic fungi found on symptomatic litter throughout the decomposition process. Given the dominance of American beech in many forested systems in northeastern North America, these shifts in litter decomposition rates driven by BLD may have important consequences for nutrient and carbon cycling in many deciduous forests.

Dispersal is a key process in the spatial and temporal dynamics of insect populations. Dispersal depends on the flight performance of individual insects, which is influenced by their environment, morphology, and physiological state. Temperature affects flight performance and costs through its effect on the metabolism of ectotherms. It can also induce developmental changes in flight-related traits that influence flight biomechanics and insect size, affecting the resources available for this activity. We thus need to understand how temperature during flight modulates flight performance and costs, but also how morphology affects them. Using flight mills, models were developed to describe how flight performance and costs of individual spruce budworm females varied over a range of temperatures (10°C-35°C). Variations of barometric pressure and morphological traits were also incorporated in these models. Flight propensity dropped below 20°C, and when female mass increased relative to wing area, suggesting that there is a wing load beyond which flight probability decreases. Speed, duration, and distance of flight decreased as temperature deviated from 23°C, while wingbeat frequency increased consistently with temperature. Females with long and broad wings had greater flight endurance. Mass loss and use of water and fuel (lipids and carbohydrates) increased with temperature, while the remaining lipids were not affected. As a result, female budworms allocated a daily energy budget to flight, which was proportional to their mass. Large females also benefited from an energy saving due to their mass during flight. Water loss was proportional to female mass but decreased with distance traveled, reaching hydric quasi-homoeostasis at distances greater than 10 km. Our approach reveals the underlying mechanisms of flight and highlights the factors that influence the landing process after dispersal in the spruce budworm. The relationships presented in this study can help develop more realistic models of dispersal for this boreal forest pest.

Parasites are an abundant and diverse group of organisms that are often excluded from biodiversity surveys, limiting our understanding of host-parasite relationships and parasite diversity. Parasites are dependent on their hosts for survival and parasite populations are at risk if their host populations decline. The aim of our study was to quantify and compare the ectoparasite communities of two sympatric Atlantic Canadian bat species, and . Ectoparasites were collected from bats captured for research throughout Atlantic Canada between 1999 and 2017 during the active season (May-October). The prevalence and mean intensity of infection were calculated for each identified ectoparasite species and generalized linear mixed models were used to assess host differences in infection by the two most abundant ectoparasites. Both bat species hosted ectoparasite communities dominated by the mite and the flea with other ectoparasites being rarely encountered. Despite being the most common ectoparasites of both bat species, our results suggest that infections of these ectoparasites vary between them with prevalence being greater on and prevalence being greater on We suggest these differences in infection burden are related to the social and roosting behaviors of these hosts and the life history of the ectoparasites. Monitoring parasites concurrently with focal species is important for capturing this aspect of biodiversity and for understanding how host-parasite dynamics may be disrupted if hosts undergo drastic demographic changes.

[This corrects the article DOI: 10.1002/ece3.72300.].

Characterizing the complex web of ecological interactions is a central challenge in molecular ecology. Shotgun metagenomics of environmental samples offers a powerful, high-resolution approach, yet its potential for simultaneously generating multiple genomic resources from different trophic levels remains underexplored. This study serves as a proof-of-concept, using deep sequencing of a single, complex sample-the larval food of the stingless bee -to demonstrate the method's capacity to recover genomic information across varying template abundances. We successfully assembled three genomes of different completeness levels: a near-complete bacterial genome (, 2,097,977 bp with 0.002% ambiguous bases), a draft mitochondrial genome (, 15,498-15,549 bp), and a fragmented chloroplast genome (, 130,532 bp with 23.47% ambiguous bases). The assembly quality gradient, observed from complete to fragmented, directly reflects the relative abundance of each DNA template in the environmental sample, demonstrating the method's sensitivity and ecological informativeness. Beyond these genomic resources, the data provided a comprehensive biodiversity profile, revealing DNA from seven major taxonomic groups, including 209 bacterial genera, 123 plant families, and 55 insect taxa. Additionally, genomic comparisons using Average Nucleotide Identity (ANI) and digital DNA-DNA Hybridization (dDDH) analyses suggest that the dominant bacterial strain represents a putative novel species within the genus . This approach simultaneously provided insights into host genetics, food sources, and microbial communities, illustrating the potential of single metagenomic datasets to generate multiple valuable genomic resources for molecular ecology research.

Fruit traits have the potential to influence disperser communities and vice versa. Elevational gradients, where functional traits are known to change rapidly with distance, enable the study of these relationships at local scales. In this pilot study, we examine the trends in three fruit traits related to dispersal by frugivores: size, colour and presentation (i.e., location of displayed fruits on the trunk or on the branches) along a 200-2700 m asl rainforest elevation gradient in Papua New Guinea. We found fruit size to be lower at higher elevations. While specific fruit colours showed few significant elevational patterns, colours typically attributed to attracting avian dispersers were more prevalent at higher elevations. The proportion of ramiflorous species (bearing fruits from branches) increased with elevation. Finally, we use phylogenetic information to test the 'dispersal syndromes' hypothesis: that combinations of fruit traits have evolved in accordance with the preferences and sensory abilities of different frugivore guilds. All fruit traits except presentation showed little evidence of phylogenetic signal but we found fruits displaying colours attributed to attracting mammalian dispersers to be larger than 'bird colour' fruits. We found evidence for the correlated evolution of fruit size and colour, in support of the dispersal syndromes hypothesis. We encourage research at larger scales of time and space to further explore the relationships between fruit traits and frugivores, across elevational and other ecological gradients.

Rewilding captive animals is an important strategy for rehabilitating individuals and ecosystems. Comparing the behaviors of released animals to their wild counterparts enables the evaluation of their adaptive response to new environments, assuming that wild animals are better suited to natural conditions. We examined how movement patterns of captive African savannah elephants () before and after soft release compared with movement patterns of other elephant groups, rehabilitated and wild elephants, in the western Okavango Delta, Botswana. We monitored 12 adult female elephants using GPS collars: six captive elephants, subjected to a three-year phased soft release, two elephants released more than a decade earlier and four wild elephants. We quantified 30-min diurnal and nocturnal distances, cumulative daily distances, daily displacement, and monthly home range sizes across seasonal flood cycles. We analyzed the effects of release, season, time of day, and elephant group on movement metrics, comparing captive elephants before and after release, and with rehabilitated and wild elephants. Before release, captive elephants moved longer diurnal and shorter nocturnal 30-min distances, covered longer cumulative daily distances, and occupied smaller home ranges. After release, these metrics shifted, reducing differences with rehabilitated and wild elephants, although captive elephant home ranges remained significantly smaller. This suggests that captive elephants changed their movement patterns post-release in response to environmental cues. However, even the movement patterns of rehabilitated elephants were not completely similar to those of wild elephants, likely due to sample size, individual variation, or effects of prior taming. These results highlight the critical importance of long-term monitoring of animals since the movement patterns of released animals may take several years to converge with those of wild counterparts.

Population turnover (local extinction and recolonization) is a fundamentally important ecological process but one that is not often (or easily) studied. Even more poorly known are the local and landscape factors that influence population turnover. We leverage a long-term monitoring dataset on a short-lived frog (Blanchard's cricket frog, ) to provide empirical estimates of local extinction and colonization frequency in an occupancy modeling framework that explicitly addresses detection probability. We surveyed 102 aquatic habitats in northwestern Ohio (USA) using 1491 listening surveys from 2004-2008 and 2017-2022. Estimated colonization and extinction rates were very similar over the entire period 2004-2022 (0.1099 (SE = 0.0149) and 0.1014 (SE = 0.0301), respectively). However, colonization and extinction rate estimates were substantially higher in the 2017-2022 interval compared to the 2004-2008 interval. The two most important variables influencing colonization and extinction probabilities were the distance to the nearest "hotspot" (a site that was continuously or nearly continuously occupied throughout our study) and the proximity to a large, interstate highway. The strong importance of "hotspot" proximity suggests that resilient core populations reduce extinction rates and boost colonization rates in nearby populations and generally play an outsized role in overall regional population dynamics. While many other habitat, landscape structure and connectivity variables were not consistently informative, landscape connectivity metrics generally were more powerful than habitat or landscape structure variables. These results highlight the importance of long-term ecological data in understanding the role of episodic processes operating in natural populations.

The Mediterranean ant-ant association between and has fascinated naturalists for long, with a focus on documenting the attraction of workers to trails. Little attention, in contrast, has been put on similar behaviors between other and species. Moreover, neither the adaptive value of mimicry in , which mimics the color patterns of different species, nor its evolution is understood. Data and video records (https://figshare.com/s/b627084f6a7f60e6dc4c) of 2 one-month-long and several short field trips brought insights into this mimicry evolution: We recorded trail following, a behavior hitherto known from four Mediterranean species, from 10 Camponotini species. The percentage of workers to those of on trails was only 2.4%. , , and species were the only trail followers mimicking color patterns. Dealate gynes of and founded colonies next to trails. Of 211 colonies, 79% were associated with . nests putatively associated with had 2.2 times more workers and a 2.2 times higher chance to produce sexuals. major workers defended entrances against by blocking them with their heads, in a similar manner as . At 15 of 19 sites, lacertid lizards were present; some ingested ants but avoided . Several characteristics of concerning its behavior, morphology, colony structure, and nesting site selection are advantageous for the mimicry strategy. Of these, we identified those shared with related non-mimetic species as putative preadaptations. The manifestation of regional color morphs could be the only species-specific mimicry adaptation, while several common characteristics among Mediterranean Camponotini appear as preadaptations to trail following or to mimicry. Trail following facilitates access to food and protection; color mimics likely evolved from non-mimetic trail followers and enjoy reduced predation through Batesian mimicry.

Open terrestrial ecosystems such as savannas have been experiencing marked increases in woody cover driven by shrub encroachment. Despite this widespread pattern, understanding the consequences for faunal communities remains challenging because long-term data are often not available and other structural changes, such as changing tree cover, may confound conclusions on shrub encroachment effects. We used satellite data and surveys of bird communities spanning 22 years to assess vegetation-cover dynamics and its effects on bird communities across the savanna ecosystem of Eswatini. We employed a hierarchical multi-species occupancy model that accounted for imperfect detection to assess changes in species occurrence, richness, and community assemblages. Between 1998 and 2020, shrub cover increased from 16% to 44% and tree cover increased from 17% to 28%. Across 64 species, shrub cover tended to have greater effects on bird occupancy than tree cover, with 34 (53%) species exhibiting positive linear associations with shrub cover and 15 (23.4%) species exhibiting a non-linear response to shrubs, where occupancy peaked at < 50% shrub cover. Shrub cover generated non-linear responses across all four bird diet groups and four of the five nesting groups. Species richness increased over time, with changes in the composition of bird assemblages being driven more by changes in shrub than tree cover. With recent predictions indicating a potential increase in shrub cover that is driven by global factors, community-wide changes in bird communities may intensify. To minimize negative consequences of changes in bird communities, land managers should initiate and expand existing woody cover management regimes in southern African savannas, where shrub cover remains high.

The taxonomy of the scorpion genus is complex due to the considerable increase in newly reported species, their high degree of similarity, and consequently, the great difficulty in their morphological differentiation. Tunisian species are not exempt from this issue, with several references highlighting the need for taxonomic revisions. This study integrates DNA sequence data and morphological assessments to investigate the diversity present in Tunisia and to provide morphological details that facilitate species identification. The results show that most Tunisian specimens are distributed within two clades. One clade comprises four subclades corresponding to Herbst, 1800, C. L. Koch, 1839, Kovařík 2006 and a southern group corresponding probably to Rossi, Tropea & Yağmur, 2013. The second clade represents a new species described in this study as Hajri, Bahri & Harris, . No evidence of Kovařík 2006 have been recorded in the studied samples. Distances between all five species exceed the minimum divergence thresholds for species. The greatest distance was observed between . and the southern group, while the smallest distance was between and . Although the genetic differences revealed considerable divergence of the new group from the four remaining species, the morphological assessment did not identify the same pattern. These five species demonstrate a morphological shape gradient in which and the southern group represent the two extremes, with being the most ornamented and the latter the least. The new species presents an intermediate morphology. The geographic distributions of the five reported species are discussed in this work according to the topography and orography of the region. Additional lineages known from Algeria may also enter the western fringes of Tunisia.

Diaspore (e.g., seed and spore) dispersal is recognized as a key mechanism in plant dynamics, including endozoochory, which can be a risky journey for diaspores. Endozoochory is achieved when diaspores are consumed and may germinate after the mastication, the gut and fecal matrix passage, all representing filters for diaspores. Nevertheless, endozoochory is a highly studied mechanism through numerous methods, notably based on the observation of frugivorous behavior, diaspores retrieved in feces or germination experiments. However, most of those methods consider partially the endozoochorous filters (ingestion, mastication, gut, feces). Hence, the combined effect of the methods and filters consideration may lead to biased inference of endozoochory. In this study, we used a collection of 52 brown bear () feces to highlight five methods inferring endozoochory. Two methods consider the ingestion filter and used metabarcoding of fecal eDNA to identify fleshy fruits (i) or plants during fruiting periods (ii). The third method (iii) was based on the intact propagules retrieved in feces, considering ingestion and mastication filters. Another method (iv) was based on the germination from disaggregated feces, considering up to the gut passage filter. The last method (v) was based on the germination from aggregated feces, considering the four filters. We compared the number of taxa, the community and the plant life forms inferred among methods. We inferred the largest number of taxa in method (iii), but the germination-based methods inferred the most diverse plant life forms. We identify few shared taxa among methods. The metabarcoding-based methods might be an interesting tool as a first approximation of endozoochory while detailing the diet. The method (v) appeared as the most reliable. Overall, we invite future studies to mitigate their interpretations according to the filters of endozoochory considered and plant detectability related to the method used.

[This corrects the article DOI: 10.1002/ece3.72268.].

Tiger () and leopard () are two congener species and wherever they live together are found to compete for space and habitat. The study on habitat suitability and overlap between tigers and leopards in the Banke-Bardia Complex, Nepal, aims to assess the ecological preferences and spatial distribution patterns of these big cat species within the given landscape. For the study the Banke-Bardia Complex was considered as it is one of the priority tiger conservation units. We have gathered the GPS location of tigers and leopards from the camera trap survey. Maximum Entropy (Maxent) is software used to model the suitable habitat of species by using geo-referenced occurrence data and environmental variables. We used area under the receiver-operator curve (AUC) as the threshold-independent method. We found a fair AUC for the model of leopard (0.674+/-0.043) and tiger (0.690+/- 0.012) habitat suitability model. We found 854.15 km and 867.21 km of suitable habitat for common leopard and tiger respectively throughout the study area. We identified 388.16 km of overlapping habitat between the species, which constituted 45.60% of the habitat of leopard and 44.75% of the habitat of tiger. Most of the overlapping habitat was located in the southern part of Banke National Park and western parts of Bardia National Park. The findings from this study highlight the need for inclusive conservation strategies extending beyond core protected areas. Engaging local communities in conservation efforts and promoting sustainable land-use practices can mitigate human-wildlife conflicts and support broader ecological integrity.

has severely invaded mangroves in China. In order to explore the possible belowground interspecific interaction along with its invasion, the rhizosphere effect enhancing the competition of neighboring mangroves was hypothesized. Here, both rhizosphere soil of and bulk soil were collected from the center of marsh and border sites where was adjacent to and , respectively, in both vigorous growth and senescent periods. Soil nutrient properties, rhizospheric low-molecular-weight organic acids (LMWOAs), soil microbiomes, and microbial functional genes were analyzed. Soil total carbon and total nitrogen contents of neighboring mangroves were increased, and its LMWOAs were altered when adjacent to mangroves in both vigorous growth and senescent periods. These changes were significantly correlated with variation in the composition of rhizosphere microbiome. Microbial interkingdom co-occurrence networks were simplified when neighbored mangroves, while network modularity significantly increased. Metagenomics indicated that genes involved in methanogenesis (, , etc.) and nitrogen fixation (, , etc.) were significantly enriched in those neighboring , and genes related to phosphate transporter (, , etc.) were significantly enriched in those neighboring . These results demonstrated that the rhizosphere effect intensified the belowground interspecific competition of adjacent to mangroves by altering root exudates, changing the soil microbial composition, and modulating strategies for core nutrient metabolism.

Fluctuating asymmetry (FA) is often proposed as an early warning indicator of subtle changes in plant functioning. Here, we tested whether leaf FA responds consistently to the alleviation of cold stress in three boreal willow species-, and . We enclosed 10 naturally growing individuals of each species in open-top chambers at budburst and compared their leaf traits to those of unenclosed control plants after leaf development had ceased. All measurements were conducted blind to treatment. Willows in open-top chambers showed a 9% increase in specific leaf area, indicating that the 1°C-2°C warming within chambers affected leaf development. However, neither leaf length nor FA responded significantly to the warming treatment. FA also did not differ among species or individual plants, suggesting that it may reflect statistical noise rather than a reliable biological signal in this context. These findings add to growing concerns that many reported FA responses to environmental change may result from confirmation bias-an issue that can be mitigated by adopting blind measurement protocols.

Species expand their geographic distribution when environmental conditions are favorable or when mutations arise that allow them to live in previously unfavorable conditions. The European wasp spider, , has expanded its range poleward, and populations at the northern edge show higher tolerance to cold and are genetically differentiated from the core populations. We aimed to investigate the degree and limits of plasticity in a recently cold-adapted Estonian population by exposing overwintering juveniles (spiderlings) to three fixed winter regimes over the course of three months. These regimes differed in absolute and relative day and night temperature: cold (5°C day, -15°C night), moderate (5°C day, -5°C night), and warm (15°C day, -5°C night). We expected a differential response to the winter regimes in survival, lipid content, metabolites, and gene expression patterns. The survival probability of the spiderlings decreased over winter by approximately 20% and their lipid content by 28%, with no significant differences between groups. Spiderlings also did not differ in content of saturated and monounsaturated fatty acids per dry weight. However, in spiderlings exposed to the warm winter regime, short-chain omega-3 PUFAs were less abundant (~57%) and long-chain omega-3 PUFAs more abundant (~66%) compared to the other regimes. The gene expression response was low under the cold regime and much higher under the warm regime, as compared to the moderate regime. The affected pathways suggest a more pronounced stress response under warmer winter temperatures. Taken together, our findings demonstrate that spiderlings from a northern population can endure very different winter regimes. However, the observed physiological responses to the warmer regime suggest metabolic costs that may reduce spiderling survival probability after emergence from the egg sac. We conclude that, despite remarkable tolerance to different winter regimes, warmer winters have nuanced effects on spiderling physiology beyond survival probability.

The phenomenon of floral color change (FCC) in angiosperms is frequently associated with pollination events. However, research on triggers, functions, and ecological effects of FCC still remains confined to phenomenological speculation or model-based validation due to the overlap between natural senescence and multiple ecological roles, as well as the difficulties in quantitative analysis of pollen transfer. a typical distylous species characterized by five gradually fading purple spots on its corolla, serves as an excellent paradigmatic system for studying the triggers and adaptability of FCC, owing to its heteromorphic pollen-stigma morphology. Therefore, we investigated the flower syndrome, dynamic changes of spots under controlled pollination, pollinator visiting behavior and pollen deposition mediated by artificial spots. The results showed that emasculating and bagging, as well as inter- or intra-morph pollination, can accelerate or delay the time of spots fading. When artificial spots were added to some spotless older flowers to mimic natural ones, these flowers, whether with natural or artificial spots, attracted significantly more pollinators than those without spots. Additionally, the number of incompatible pollen grains on the stigmas of both L-morph and S-morph flowers increased significantly, while the compatible pollen grain counts remained unchanged. In conclusion, as a typical distylous plant with weak self-incompatibility in the S-morph, FCC in , which is influenced by pollen input-output dynamics, plays a critical role in reducing self-pollen deposition. For the first time, the study systematically evaluated the induction and ecological function of FCC through employing the manual control experiment, quantitative analysis of stigma pollen deposition, and investigation of pollinator visitation behavior in a distylous population.