Geographic variation in oviposition choice of a leaf beetle: the relationship between host plant ranking, specificity, and motivation

The degree of adaptation of herbivorous insects to their local flora is an important component of the evolutionary processes that lead to host plant specialization in insects. In this study we investigated geographic variations in the oviposition preference of the leaf beetle Oreina elongata Suffrian (Coleoptera: Chrysomelidae: Chrysolini) in relation to differences in host plant specialization, in the field. We focused on the mechanisms of host choice and asked whether potential differences among populations are due to variations in host plant ranking and/or host plant specificity. We performed a combination of simultaneous choice and sequential no‐choice experiments with two of the major host plants of the beetle [Cirsium spinosissimum (L.) and Adenostyles alliariae (Gouan) (Asteraceae)]. The results suggested that spatial variation in host plant specialization has resulted in differences between populations in some aspects of the oviposition choice of O. elongata, while other aspects seem unaffected. We found no variation in host plant ranking among populations, as estimated in simultaneous choice tests. In contrast, the sequential no‐choice test indicated that host plant specificity was lower in a population that never encountered the highest ranked plant in the field. This finding agreed with our expectations, and we discuss our results in relation to the commonly used hierarchical threshold model. The results suggested that the mechanism for the differences in specificity is the variation among populations in the general motivation to oviposit, rather than quantitative differences in relative preference for the two hosts. We stress that it is essential to establish which of the two mechanisms is most important, as it will affect the probability of evolutionary change in host plant ranking.     

Testing the relationship between intraspecific competition and individual specialization across both behavior and diet

Individual specialization within populations is increasingly recognized as important in both ecology and evolution, but researchers working on intraspecific variation in behavior and diet infrequently interact. This may be because individual specialization on diet and behavior was historically difficult to investigate simultaneously on the same individuals. However, approaches like stable isotope analysis that allow hindcasting past field diets for laboratory organisms may provide opportunities to unite these areas of inquiry. Here, we tested the role of intraspecific competition on individual specialization through analysis of both behavior and diet simultaneously. We focused on intraspecific competition as a mechanism that might drive individual specialization of both diet and behavior. We conducted this study in Vilas County, Wisconsin, United States (US), using rusty crayfish Faxonius rusticus from six lakes across a relative abundance gradient. We conducted six assays to measure individual specialization of behavior and used stable isotope analysis to measure individual specialization of diet. We then related both measures of individual specialization to relative abundance of F. rusticus using linear and quadratic models. We found a unimodal relationship between intraspecific competition and individual specialization of diet for F. rusticus, likely because some preferred resources are unavailable to specialize on at the highest densities of this well‐studied crayfish invader. Conversely, we found greater support for a linear relationship between individual specialization of behavior and intraspecific competition, perhaps because specialization by behavior is not inherently resource‐limited. Our results show that dietary and behavioral specialization may exhibit different responses to increased intraspecific competition, and demonstrate a potential technique that can be used to investigate individual specialization of diet and behavior simultaneously for the same individuals and populations.     

How intraspecific variation in seed‐dispersing animals matters for plants

Seed dispersal by animals is a complex phenomenon, characterized by multiple mechanisms and variable outcomes. Most researchers approach this complexity by analysing context‐dependency in seed dispersal and investigating extrinsic factors that might influence interactions between plants and seed dispersers. Intrinsic traits of seed dispersers provide an alternative way of making sense of the enormous variation in seed fates. I review causes of intraspecific variability in frugivorous and granivorous animals, discuss their effects on seed dispersal, and outline likely consequences for plant populations and communities. Sources of individual variation in seed‐dispersing animals include sexual dimorphism, changes associated with growth and ageing, individual specialization, and animal personalities. Sexual dimorphism of seed‐dispersing animals influences seed fate through diverse mechanisms that range from effects caused by sex‐specific differences in body size, to influences of male versus female cognitive functions. These differences affect the type of seed treatment (e.g. dispersal versus predation), the number of dispersed seeds, distance of seed dispersal, and likelihood that seeds are left in favourable sites for seeds or seedlings. The best‐documented consequences of individual differences associated with growth and ageing involve quantity of dispersed seeds and the quality of seed treatment in the mouth and gut. Individual specialization on different resources affects the number of dispersed plant species, and therefore the connectivity and architecture of seed‐dispersal networks. Animal personalities might play an important role in shaping interactions between plants and dispersers of their seeds, yet their potential in this regard remains overlooked. In general, intraspecific variation in seed‐dispersing animals often influences plants through effects of these individual differences on the movement ecology of the dispersers. Two conditions are necessary for individual variation to exert a strong influence on seed dispersal. First, the individual differences in traits should translate into differences in crucial characteristics of seed dispersal. Second, individual variation is more likely to be important when the proportions of particular types of individuals fluctuate strongly in a population or vary across space; when proportions are static, it is less likely that intraspecific differences will be responsible for changes in the dynamics and outcomes of plant–animal interactions. In conclusion, focusing on variation among foraging animals rather than on species averages might bring new, mechanistic insights to the phenomenon of seed dispersal. While this shift in perspective is unlikely to replace the traditional approach (based on the assumption that all important variation occurs among species), it provides a complementary alternative to decipher the enormous variation observed in animal‐mediated seed dispersal.     

Comparative phylogeography of five bumblebees: impact of range fragmentation,range size and diet specialization

We characterised the phylogeographic patterns displayed by five species of bumblebees with largely overlapping ranges in Eurasia, but different levels of range fragmentation, range size and food specialization. Genetic variation across the range of each species was explored by using sequence variation of a total of 368 specimens at one mitochondrial and two nuclear DNA fragments (total of ~2380 bp). Comparing patterns of genetic variation across species allowed us to investigate whether diet specialization, relative range size and/or fragmentation, impact phylogeographic patterns in bumblebees. As expected, stronger fragmentations of the species range are associated with a stronger overall geographic differentiation. Furthermore, diet specialization appears to increase population structure at the landscape level, presumably due to the less widespread and more heterogeneously distributed food resources. Conversely, no clear association was highlighted between diet specialization or overall range size and genetic diversity. Surprisingly, the two generalist and co‐distributed species investigated, B. pratorum and B. hortorum, displayed widely divergent patterns in terms of genetic diversity and population structure. We suggest these differences are best explained by contrasting responses to past climate changes, possibly involving different glacial refuges. Overall, our results are compatible with a combined impact of two interacting parameters on intraspecific genetic variation: environment disturbances (presumably related to past climate changes) and features specific to the organism, such as diet specialization. They thus further highlight the challenge of dissociating both parameters in phylogeographic studies.     

Intraspecific difference among herbivore lineages and their host‐plant specialization drive the strength of trophic cascades

Trophic cascades – the indirect effect of predators on non‐adjacent lower trophic levels – are important drivers of the structure and dynamics of ecological communities. However, the influence of intraspecific trait variation on the strength of trophic cascade remains largely unexplored, which limits our understanding of the mechanisms underlying ecological networks. Here we experimentally investigated how intraspecific difference among herbivore lineages specialized on different host plants influences trophic cascade strength in a terrestrial tri‐trophic system. We found that the occurrence and strength of the trophic cascade are strongly influenced by herbivores’ lineage and host‐plant specialization but are not associated with density‐dependent effects mediated by the growth rate of herbivore populations. Our findings stress the importance of intraspecific heterogeneities and evolutionary specialization as drivers of trophic cascade strength and underline that intraspecific variation should not be overlooked to decipher the joint influence of evolutionary and ecological factors on the functioning of multi‐trophic interactions.     

Local ecological divergence of two closely related stag beetles based on genetic,morphological, and environmental analyses

The process of phenotypic adaptation to the environments is widely recognized. However, comprehensive studies integrating phylogenetic, phenotypic, and ecological approaches to assess this process are scarce. Our study aims to assess whether local adaptation may explain intraspecific differentiation by quantifying multidimensional differences among populations in closely related lucanid species, Platycerus delicatulus and Platycerus kawadai, which are endemic saproxylic beetles in Japan. First, we determined intraspecific analysis units based on nuclear and mitochondrial gene analyses of Platycerus delicatulus and Platycerus kawadai under sympatric and allopatric conditions. Then, we compared differences in morphology and environmental niche between populations (analysis units) within species. We examined the relationship between morphology and environmental niche via geographic distance. P. kawadai was subdivided into the “No introgression” and “Introgression” populations based on mitochondrial COI gene – nuclear ITS region discordance. P. delicatulus was subdivided into “Allopatric” and “Sympatric” populations. Body length differed significantly among the populations of each species. For P. delicatulus, character displacement was suggested. For P. kawadai, the morphological difference was likely caused by geographic distance or genetic divergence rather than environmental differences. The finding showed that the observed mitochondrial–nuclear discordance is likely due to historical mitochondrial introgression following a range of expansion. Our results show that morphological variation among populations of P. delicatulus and P. kawadai reflects an ecological adaptation process based on interspecific interactions, geographic distance, or genetic divergence. Our results will deepen understanding of ecological specialization processes across the distribution and adaptation of species in natural systems.     

Genetic Diversity and Evolution of Chinese Traditional Medicinal Fungus Polyporus umbellatus (Polyporales,Basidiomycota)

Background

Polyporus umbellatus is an important medicinal fungus distributed throughout most area of China. Its wide distribution may have resulted in substantial intraspecific genetic diversity for the fungus, potentially creating variation in its medical value. To date, we know little about the intraspecific genetic diversity of P. umbellatus.

Methodology/Principal Findings

The objective of this research was to assess genetic differences of P. umbellatus from geographically diverse regions of China based on nrDNA ITS and 28S rRNA (LSU, large subunit) sequences. Significant sequence variations in the ITS and LSU sequences were detected. All sclerotial samples were clustered into four clades based on phylogenetic analysis of ITS, LSU and a combined data set of both regions. Heterogeneity of ITS and LSU sequences was detected in 5 and 7 samples respectively. All clone sequences clustered into the same clade except for one LSU clone sequences (from Henan province) which clustered into two clades (Clade I and Clade II). Significant genetic divergence in P. umbellatus was observed and the genetic diversification was greater among sclerotial samples from Shaanxi, Henan and Gansu provinces than among other provinces. Polymorphism of ITS and LSU sequences indicated that in China, P. umbellatus may spread from a center (Shaanxi, Henan and Gansu province) to other regions.

Conclusions/Significance

We found sclerotial samples of P. umbellatus contained levels of intraspecific genetic diversity. These findings suggested that P. umbellatus populations in Shaanxi, Henan and Gansu are important resources of genetic diversity and should be conserved accordingly.     

Gene-Swapping Mediates Host Specificity among Symbiotic Bacteria in a Beneficial Symbiosis

Environmentally acquired beneficial associations are comprised of a wide variety of symbiotic species that vary both genetically and phenotypically, and therefore have differential colonization abilities, even when symbionts are of the same species. Strain variation is common among conspecific hosts, where subtle differences can lead to competitive exclusion between closely related strains. One example where symbiont specificity is observed is in the sepiolid squid-Vibrio mutualism, where competitive dominance exists among V. fischeri isolates due to subtle genetic differences between strains. Although key symbiotic loci are responsible for the establishment of this association, the genetic mechanisms that dictate strain specificity are not fully understood. We examined several symbiotic loci (lux-bioluminescence, pil = pili, and msh-mannose sensitive hemagglutinin) from mutualistic V. fischeri strains isolated from two geographically distinct squid host species (Euprymna tasmanica-Australia and E. scolopes-Hawaii) to determine whether slight genetic differences regulated host specificity. Through colonization studies performed in naïve squid hatchlings from both hosts, we found that all loci examined are important for specificity and host recognition. Complementation of null mutations in non-native V. fischeri with loci from the native V. fischeri caused a gain in fitness, resulting in competitive dominance in the non-native host. The competitive ability of these symbiotic loci depended upon the locus tested and the specific squid species in which colonization was measured. Our results demonstrate that multiple bacterial genetic elements can determine V. fischeri strain specificity between two closely related squid hosts, indicating how important genetic variation is for regulating conspecific beneficial interactions that are acquired from the environment.     

Specificity of assemblage,not fungal partner species,explains mycorrhizal partnerships of mycoheterotrophic Burmannia plants

Mycoheterotrophic plants (MHPs) growing on arbuscular mycorrhizal fungi (AMF) usually maintain specialized mycorrhizal associations. The level of specificity varies between MHPs, although it remains largely unknown whether interactions with mycorrhizal fungi differ by plant lineage, species, and/or by population. Here, we investigate the mycorrhizal interactions among Burmannia species (Burmanniaceae) with different trophic modes using high-throughput DNA sequencing. We characterized the inter- and intraspecific dynamics of the fungal communities by assessing the composition and diversity of fungi among sites. We found that fully mycoheterotrophic species are more specialized in their fungal associations than chlorophyllous species, and that this specialization possibly results from the gradual loss of some fungal groups. In particular, although many fungal species were shared by different Burmannia species, fully MHP species typically host species-specific fungal assemblages, suggesting that they have a preference for the selected fungi. Although no apparent cophylogenetic relationship was detected between fungi and plants, we observe that evolutionarily closely related plants tend to have a greater proportion of shared or closely related fungal partners. Our findings suggest a host preference and specialization toward fungal assemblages in Burmannia, improving understanding of interactions between MHPs and fungi.Subject terms: Fungi, Plant sciences, Evolution     

Effects of Hybridization and Evolutionary Constraints on Secondary Metabolites: The Genetic Architecture of Phenylpropanoids in European Populus Species

The mechanisms responsible for the origin, maintenance and evolution of plant secondary metabolite diversity remain largely unknown. Decades of phenotypic studies suggest hybridization as a key player in generating chemical diversity in plants. Knowledge of the genetic architecture and selective constraints of phytochemical traits is key to understanding the effects of hybridization on plant chemical diversity and ecological interactions. Using the European Populus species P. alba (White poplar) and P. tremula (European aspen) and their hybrids as a model, we examined levels of inter- and intraspecific variation, heritabilities, phenotypic correlations, and the genetic architecture of 38 compounds of the phenylpropanoid pathway measured by liquid chromatography and mass spectrometry (UHPLC-MS). We detected 41 quantitative trait loci (QTL) for chlorogenic acids, salicinoids and flavonoids by genetic mapping in natural hybrid crosses. We show that these three branches of the phenylpropanoid pathway exhibit different geographic patterns of variation, heritabilities, and genetic architectures, and that they are affected differently by hybridization and evolutionary constraints. Flavonoid abundances present high species specificity, clear geographic structure, and strong genetic determination, contrary to salicinoids and chlorogenic acids. Salicinoids, which represent important defence compounds in Salicaceae, exhibited pronounced genetic correlations on the QTL map. Our results suggest that interspecific phytochemical differentiation is concentrated in downstream sections of the phenylpropanoid pathway. In particular, our data point to glycosyltransferase enzymes as likely targets of rapid evolution and interspecific differentiation in the ‘model forest tree’ Populus.     

Inter‐ and intraspecific variation in mycotoxin tolerance: A study of four Drosophila species

Many mycophagous Drosophila species have adapted to tolerate high concentrations of mycotoxins, an ability not reported in any other eukaryotes. Although an association between mycophagy and mycotoxin tolerance has been established in many Drosophila species, the genetic mechanisms of the tolerance are unknown. This study presents the inter‐ and intraspecific variation in the mycotoxin tolerance trait. We studied the mycotoxin tolerance in four Drosophila species from four separate clades within the immigrans‐tripunctata radiation from two distinct locations. The effect of mycotoxin treatment on 20 isofemale lines per species was studied using seven gross phenotypes: survival to pupation, survival to eclosion, development time to pupation and eclosion, thorax length, fecundity, and longevity. We observed interspecific variation among four species, with D. falleni being the most tolerant, followed by D. recens, D. neotestacea, and D. tripunctata, in that order. The results also revealed geographical variation and intraspecific genetic variation in mycotoxin tolerance. This report provides the foundation for further delineating the genetic mechanisms of the mycotoxin tolerance trait.     

The evolution of wing color in Colias butterflies: heritability,sex linkage,and population divergence

We investigated the genetic background of intraspecific variation in wing color across an elevational gradient in the butterfly Colias philodice eriphyle. The degree of wing melanization was an accelerating function of elevation, and differences in wing melanization persisted in a common environment. Full-sibling analysis and parent-offspring regression yielded consistent, moderate to high heritabilities for the degree of wing melanization. The breeding experiments also demonstrated that wing melanization is strongly sex linked. Because traits that differentiate sister species also tend to be sex linked, our results suggest that the genetic mechanisms underlying intraspecific differences in wing melanization are not fundamentally different from those that have been shown to differentiate sister species.     

Variabilité biochimique et génétique de Sphaeroma ghigii,S. hookeri et S. serratum. Variations intraspécifiques

Electrophoresis on polyacrylamide gel of enzymes (esterases, phosphatases) and total proteins of Sphaeroma ghigii, S. hookeri and S. serratum shows the existence of important intraspecific variability. Esterases, alkaline phosphatases and major low ionic strength extractable proteins also demonstrate quantitative and qualitative differences between individuals; these differences are the expression of protein and enzyme polymorphism. Some of these variations represent sexual differences which are an expression of a biochemical sexual dimorphism that appears particularly in esterases and phosphatases. Among males, this variability leads to the delineation of several biochemical phenotypes and gives an insight into the genetic variability of populations based on the protein composition of organisms. Taking into consideration the ecological characteristics of the various species of genus Sphaeroma, this polymorphism provides a source of information on ecophysiological relations at the level of proteins and enzymes, that may lead to a better understanding of intraspecific variability.     

The role of food quality in clonal succession in Daphnia: an experimental test

A high genetic variation and recurrent changes in the genetic structure have been found in many pelagic populations. However, evidence that directly links these changes to differences in the ecological performance of particular genotypes is scarce. We hypothesized that within Daphnia, the specialization of clones occurring in a particular season to the food quality specific for that time of the year is responsible for the observed changes in the genetic structure of a population. This hypothesis was tested by comparing the fitness of spring and summer clones of the Daphnia longispina group, given food of biochemical quality relevant to these seasons. We identified significant intraspecific differences between clones of Daphnia that are specific for particular seasons, but there was no evidence that clones are adapted to the food quality available at the respective times of year. Summer clones reproduce at smaller size, and have a lower juvenile specific growth rate as compared to spring clones, irrespective of food quality. Spring clones invest more energy in somatic growth at the cost of reproduction, whereas summer clones invest more energy in reproduction at the cost of somatic growth. On the basis of the observed differences between spring and summer clones in their patterns of energy allocation, we suggest that other factors, most likely predation, are the major forces driving phenotypic and genetic diversity in the investigated Daphnia population of a large lake.     

Mechanisms of the adaptation of the Kildin cod <Emphasis Type="Italic">Gadus morhua kildinensis</Emphasis> Derjugin, 1920 (Pisces: Gadidae) to the specific conditions of Lake Mogilnoye

The mechanisms of the adaptation of the Kildin cod to the conditions of a meromictic lake were analyzed based on the study of morphological, biological, and genetic characteristics. Significant differences between the Kildin cod and Atlantic cod have been found in their morphological and genetic characteristics. The results of an X-ray microanalysis of otoliths, which were conducted for the first time, confirm that the distribution range of the Kildin cod in Lake Mogilnoye also extends to brackish waters. It has been shown that the formation of the reproductively isolated lacustrine population of Kildin cod is accompanied by specialization in a number of characteristics (body color, feeding strategy, behavior, etc.) and by a decrease in genetic diversity.     

Intraspecific plant–soil feedback and intraspecific overyielding in Arabidopsis thaliana

Understanding the mechanisms of community coexistence and ecosystem functioning may help to counteract the current biodiversity loss and its potentially harmful consequences. In recent years, plant–soil feedback that can, for example, be caused by below‐ground microorganisms has been suggested to play a role in maintaining plant coexistence and to be a potential driver of the positive relationship between plant diversity and ecosystem functioning. Most of the studies addressing these topics have focused on the species level. However, in addition to interspecific interactions, intraspecific interactions might be important for the structure of natural communities. Here, we examine intraspecific coexistence and intraspecific diversity effects using 10 natural accessions of the model species Arabidopsis thaliana (L.) Heynh. We assessed morphological intraspecific diversity by measuring several above‐ and below‐ground traits. We performed a plant–soil feedback experiment that was based on these trait differences between the accessions in order to determine whether A. thaliana experiences feedback at intraspecific level as a result of trait differences. We also experimentally tested the diversity–productivity relationship at intraspecific level. We found strong differences in above‐ and below‐ground traits between the A. thaliana accessions. Overall, plant–soil feedback occurred at intraspecific level. However, accessions differed in the direction and strength of this feedback: Some accessions grew better on their own soils, some on soils from other accessions. Furthermore, we found positive diversity effects within A. thaliana: Accession mixtures produced a higher total above‐ground biomass than accession monocultures. Differences between accessions in their feedback response could not be explained by morphological traits. Therefore, we suggest that they might have been caused by accession‐specific accumulated soil communities, by root exudates, or by accession‐specific resource use based on genetic differences that are not expressed in morphological traits. Synthesis. Our results provide some of the first evidence for intraspecific plant–soil feedback and intraspecific overyielding. These findings may have wider implications for the maintenance of variation within species and the importance of this variation for ecosystem functioning. Our results highlight the need for an increased focus on intraspecific processes in plant diversity research to fully understand the mechanisms of coexistence and ecosystem functioning.     

Diversity and succession of pelagic microorganism communities in a newly restored Illinois River floodplain lake

Stenotopic specialization to a fragmented habitat promotes the evolution of genetic structure. It is not yet clear whether small-scale population structure generally translates into large-scale intraspecific divergence. In the present survey of mitochondrial genetic structure in the Lake Tanganyika endemic Altolamprologus (Teleostei, Cichlidae), a rock-dwelling cichlid genus comprising A. compressiceps and A. calvus, habitat-induced population fragmentation contrasts with weak phylogeographic structure and recent divergence among genetic clades. Low rates of dispersal, perhaps along gastropod shell beds that connect patches of rocky habitat, and periodic secondary contact during lake level fluctuations are apparently sufficient to maintain genetic connectivity within each of the two Altolamprologus species. The picture of genetic cohesion was interrupted by a single highly divergent haplotype clade in A. compressiceps restricted to the northern part of the lake. Comparisons between mitochondrial and nuclear phylogenetic reconstructions suggested that the divergent mitochondrial clade originated from ancient interspecific introgression. Finally, ‘isolation-with-migration’ models indicated that divergence between the two Altolamprologus species was recent (67–142 KYA) and proceeded with little if any gene flow. As in other rock-dwelling cichlids, recent population expansions were inferred in both Altolamprologus species, which may be connected with drastic lake level fluctuations.     

Phylogeny and Diagnostic Markers of Six Tetranychus Species (Acarina: Tetranychidae) in Korea Based on the Mitochondrial Cytochrome Oxidase Subunit I

To understand the evolutionary relationship among six Tetranychus species in Korea, 598 nucleotides of cytochrome oxidase subunit I (COI) were sequenced and analyzed. The sequences were extremely rich in A+T (77% on average). The nucleotide differences in pairwise comparison among species ranged from 6.7% to 14.9%. Nucleotide substitutions based on all pairwise comparison showed 55% of transversions. The largest genetic distance was found between species T. viennensis and T. piercei. The inferred phylogenetic tree indicates that T. urticae, T. cinnabarinus and T. kanzawai are clustered together and showing sister-group relationship with T. truncatus at the highest bootstrap support. T. viennensis showed the largest genetic distance from all other species. T. piercei, a species recently found in Korea, was also distinct from the group composed of four species and also from T. viennensis. The restriction sites of Alu I, Dde I and Sau 3A were searched in the sequences of COI. The restriction fragment length polymorphisms (RFLPs) in COI genes and intraspecific variabilities of three restriction sites in diverse colonies of each species collected from different locations and various host plants were shown.     

Different ecological conditions support individual specialization in closely related,ecologically similar species

Individual generalist predators often have more specialized diets than their populations do. Individual specialization (IS) is influenced by ecological opportunity, intraspecific competition, and interspecific competition, although the effects of these parameters are inconsistent across studies. We investigated IS in five species of frogs and toads, Anaxyrus americanus, A. fowleri, Lithobates catesbeianus, L. clamitans, and L. sphenocephalus. We used the natural history and ecology of each species to predict which parameters would influence IS. Our predictions were supported for some species but not others. We predicted IS would be positively influenced by resource diversity in all species, but this prediction held for only three species, with the relationship significant in A. fowleri and L. catesbeianus and marginally significant in A. americanus. We also predicted that interspecific competition would have a negative relationship with IS in L. clamitans because L. catesbeianus is competitively superior to L. clamitans and likely to suppress its foraging options. This prediction was upheld. Finally, we predicted that IS in A. americanus, A. fowleri, and L. clamitans would be influenced by intraspecific competition. However, IS was not influenced by intraspecific competition in any species, a surprising result given that intraspecific competition has traditionally been assumed to be the ecological parameter with the strongest effects on IS. Many previous studies did not simultaneously consider all three ecological parameters, which may have increased the apparent importance of intraspecific competition for IS. Our results revealed that the ecological parameters affected IS differently even across closely related and ecologically similar species, and demonstrated that these differences are sometimes predictable based on natural history. This study also suggests that sympatric ecological speciation based on IS may be rare because the ecological parameters driving IS are inconsistent across species, and the strength of their effects on intraspecific diet variation varies in space.