Taxonomy as a hypothesis: testing the status of the Bermuda buckeye butterfly Junonia coenia bergi (Lepidoptera: Nymphalidae)

Species determination and definition in eukaryotes have traditionally been based on morphology, with little focus on genetic differentiation. Molecular methods allow for the independent assessment of morphology‐based taxonomic hypotheses. Three criteria used to define a full species for taxonomic purposes are morphological distinction, formation of a monophyletic lineage, and reproductive isolation. Junonia butterflies (Nymphalidae) are becoming an important experimental model system, but the taxonomy of many New World Junonia species is unclear. One of these species is J. coenia, which contains the subspecies J. coenia coenia, J. coenia grisea and J. coenia bergi. Previous studies suggest that J. coenia grisea may meet the criteria for full species status. Therefore, we evaluated the geographically isolated and rarely studied Bermuda buckeye butterfly J. coenia bergi to determine if it was similarly distinct. Physical examination of specimens and phylogenetic and population genetic analyses of mitochondrial cytochrome c oxidase subunit I, nuclear wingless, and complete mitochondrial genome sequences suggest that while J. coenia bergi is smaller in body size than many Junonia and has distinctive ventral hindwing colouration, it does not form a monophyletic lineage and shows indications of continued gene flow with North American mainland J. coenia coenia populations. Thus, J. coenia bergi does not meet the criteria for full species designation, but geographic isolation, morphological distinctiveness, and cultural importance suggest that it remain recognized as a subspecies of J. coenia. Similar analyses will be useful for addressing further taxonomic questions in Junonia and other taxa, especially where morphology‐based taxonomic determinations are ambiguous.     

Speciation in North American Junonia from a genomic perspective

Delineating species boundaries in phylogenetic groups undergoing recent radiation is a daunting challenge akin to discretizing continuity. Here, we propose a general approach exemplified by American butterflies from the genus Junonia Hübner notorious for the variety of similar phenotypes, ease of hybridization, and the lack of consensus about their classification. We obtain whole-genome shotgun sequences of about 200 specimens. We reason that discreteness emerges from continuity by means of a small number of key players, and search for the proteins that diverged markedly between sympatric populations of different species, while keeping low polymorphism within these species. Being 0.25% of the total number, these three dozen ‘speciation’ proteins indeed partition pairs of Junonia populations into two clusters with a prominent break in between, while all proteins taken together fail to reveal this discontinuity. Populations with larger divergence from each other, comparable to that between two sympatric species, form the first cluster and correspond to different species. The other cluster is characterized by smaller divergence, similar to that between allopatric populations of the same species and comprise conspecific pairs. Using this method, we conclude that J. genoveva (Cramer), J. litoralis Brévignon, J. evarete (Cramer), and J. divaricata C. & R. Felder are restricted to South America. We find that six species of Junonia are present in the United States, one of which is new: Junonia stemosa Grishin, sp.n. (i), found in south Texas and phenotypically closest to J. nigrosuffusa W. Barnes & McDunnough (ii) in its dark appearance. In the pale nudum of the antennal club, these two species resemble J. zonalis C. & R. Felder (iii) from Florida and the Caribbean Islands. The pair of sister species, J. grisea Austin & J. Emmel (iv) and J. coenia Hübner (v), represent the classic west/east U.S.A. split. The mangrove feeder (as caterpillar), dark nudum J. neildi Brévignon (vi) enters south Texas as a new subspecies Junonia neildi varia Grishin ssp.n. characterized by more extensive hybridization with and introgression from J. coenia, and, as a consequence, more variable wing patterns compared with the nominal J. n. neildi in Florida. Furthermore, a new mangrove-feeding species from the Pacific Coast of Mexico is described as Junonia pacoma Grishin sp.n. Finally, genomic analysis suggests that J. nigrosuffusa may be a hybrid species formed by the ancestors of J. grisea and J. stemosa sp.n. This published work has been registered on Zoobank, http://zoobank.org/urn:lsid:zoobank.org:act:C6E70C96-463D-4E6A-95CC-B0384B0EEEBA .     

Relative susceptibility to predation of two species of caterpillar on plantain

Summary The relative susceptibility to predators of a cryptic generalist caterpillar (Spilosoma congrua: Arctiidae) and a non-cryptic specialist (Junonia coenia: Nymphalidae) using the same hostplant species (Plantago lanceolata) was examined. In a laboratory experiment using predatory stinkbugs (Podisus maculiventris), more Junonia caterpillars than Spilosoma caterpillars were killed (70% vs. 16%). This result was a consequence of the Spilosoma spending some time under cover, moving frequently, feeding on leaves while under or adjacent to them, and spending little time on the leaves. In a field experiment using predatory wasps (Polistes fuscatus), the wasps found 7 times as many of the Junonia as the Spilosoma, and overall 6 times as many Junonia were killed as Spilosoma. Initially, 71% of the Junonia caterpillars encountered by wasps were killed, but by the fourth day of the test, only 22% of the Junonia encountered by wasps were killed. Over three full days of observations, a constant 50% of the Spilosoma caterpillars encountered by the wasps per day were killed. For the Junonia, evasion of predators rested on passive chemical defense. For the Spilosoma, evasion depended on being unapparent, speedy movement between feeding and resting sites and, if found, on fleeing immediately and quickly. These results indicate that Spilosoma caterpillars, by way of cryptic and escape behaviors, can be less susceptible to insect predators than Junonia caterpillars.     

Genetic population structure of buckeye butterflies (Junonia) from Argentina

There are nine named species of buckeye butterflies (genus Junonia Hübner) in the Western Hemisphere. There is considerable geographic variation within Junonia species, and possible ongoing hybridization between species, suggesting that Junonia may be a ring species, but also making this a very difficult group to define taxonomically. We tried to determine whether two forms of Junonia from Argentina – conventionally referred to as Junonia genoveva hilaris C. & R. Felder, the light buckeye butterfly, and Junonia evarete flirtea (Fabricius), the dark buckeye butterfly – were genetically distinct species or simply colour forms of a single species using morphological characters, mitochondrial cytochrome oxidase I (COI) DNA barcodes, nuclear wingless (wg) locus DNA sequences, and anonymous nuclear Randomly Amplified Fingerprints (RAF). Phylogenetic analysis of COI identified two distinct mitochondrial haplotypes that differ by about 4% sequence divergence; one confined to light‐coloured Junonia specimens and one shared between some light‐coloured Junonia and all of dark‐coloured Junonia specimens. Analysis of nuclear wingless sequences revealed 32 alleles among 22 Junonia specimens and showed significant genetic differentiation between light‐coloured and dark‐coloured Junonia. Analysis of RAF genotypes suggests that there are actually three genetically distinct Junonia populations in Argentina: two with light wing coloration, and one with dark wing coloration. Genetic evidence of recent hybridization among these populations was also observed, consistent with the ring species hypothesis. Careful comparisons of morphological characters between Argentinian Junonia and Junonia species from elsewhere in South America suggests that the two light‐coloured populations correspond to J. genoveva and either a genetically disparate population of the same species or an undescribed cryptic Junonia species, The dark‐coloured population may correspond to J. wahlbergi Brévignon. Our data suggest that COI DNA barcodes by themselves are not very useful for studying Junonia taxonomy, population structure or evolution.     

Presence of predatory wasps and stinkbugs alters foraging behavior of cryptic and non-cryptic caterpillars on plantain (Plantago lanceolata)

We examined the foraging patterns of two species of caterpillar (Junonia coenia: Nymphalidae and Spilosoma congrua: Arctiidae) that contrast in feeding specialization and crypticity on plantain (Plantago lanceolata) in the absence and presence of two different insect predators [stinkbugs, Podisus maculiventris (Pentatomidae) and wasps, Polistes fuscatus (Vespidae)]. Junonia larvae were quite apparent to human observers, feeding on upper leaf surfaces during daylight, whereas Spilosoma larvae were relatively cryptic, often hiding under leaves and in soil crevices during daylight. In the presence of either predator species, the non-cryptic Junonia caterpillars more quickly left the plant on which they were initially placed and were less apparent than Junonia larvae not exposed to predators. The presence of predators had no detectable influence on where the caterpillars occurred on the plants (new, intermediate-aged or mature leaves, or reproductive stalks). Surprisingly, the predators influenced the behavior of the inherently cryptic Spilosoma: the apparency of these larvae at night increased when wasps had access to the plots during the day. Survivorship of the non-cryptic Junonia was less than 12% when stinkbugs were present compared to 60% in their absence. Although the presence of wasps resulted in a lower relative growth rate for the non-cryptic Junonia larvae, the indirect effect of predators on reduction in survivorship due to alterations in prey growth rate through behavioral changes was less than 3%. After taking into account the decline in caterpillars per plot through predation, we found that both the amount of leaves eaten and the proportion of plants eaten were altered on plots with predators present, which suggests that the caterpillars' increased consumption countered increased maintenance costs due to the presence of predators. Overall, our results indicate that hostplant size, level of predation and type of predator can influence the degree to which these caterpillars react to the presence of insect predators. In contrast, degree of inherent feeding specialization and cryptic behavior seemed to have little effect on the expression of reactive behaviors of these caterpillars to predators.     

Larval feeding experience influences adult predator acceptance of chemically defended prey

Generalist predatory paper wasps, Polistes dominulus, experience plant secondary defensive compounds as developing larvae through their herbivorous lepidopteran caterpillar prey and as adults through attacking caterpillars while foraging. We evaluated the role that larval and early adult experience with unpalatable prey plays in subsequent foraging choices by adult wasps. For periods of two or four weeks, caged wasps were raised exclusively on caterpillars of either unpalatable Buckeye, Junonia coenia, that sequester substantial levels of iridoid glycosides (IG) or on Painted Lady, Vanessa cardui, that contain very low levels of IG. Wasps were then allowed to forage on both caterpillar species simultaneously. Patterns of prey capture differed significantly based on previous prey experience. Regardless of previous feeding experience, adult wasps overwhelmingly preferred to take Vanessa. Yet Junonia-experienced wasps continued to attack and take back to the nest over 50% more Junonia than did Vanessa-experienced wasps. The longer the wasps' larval experience with Junonia, the more likely they were to capture Junonia caterpillars. However, the life stage at which the wasps experienced Junonia was also influential as young adult experience with the unpalatable prey was more of a deterrent than was experience strictly as larvae for Junonia-experienced wasps. The results demonstrate that, in these predators, previous experience with deterrent chemicals during their larval development alters patterns of prey acceptability to the adult insects.     

A tale of two haplotype groups: evaluating the New World Junonia ring species hypothesis using the distribution of divergent COI haplotypes

The New World Junonia butterflies are a possible ring species with a circum‐Caribbean distribution. Previous reports suggest a steady transition between North and South American forms in Mesoamerica, but in Cuba the forms were thought to co‐exist without interbreeding representing the overlapping ends of the ring. Three criteria establish the existence of a ring species: a ring‐shaped geographic distribution, gene flow among intervening forms and genetic isolation in the region of range overlap. We evaluated mitochondrial cytochrome oxidase I haplotypes in Junonia from nine species in the Western Hemisphere to test the Junonia ring species hypothesis. Junonia species are generally not monophyletic with respect to COI haplotypes, which are shared across species. However, two major COI haplotype groups exist. Group A predominates in South America, and Group B predominates in North and Central America. Therefore, COI haplotypes can be used to assess the degree of genetic influence a population receives from each continent. Junonia shows a ring‐shaped distribution around the Caribbean, and evidence is consistent with gene flow among forms of Junonia, including those from Mesoamerica. However, we detected no discontinuity in gene flow in Cuba or elsewhere in the Caribbean consistent with genetic isolation in the region of overlap. Although sampling is still very limited in the critical region, the only remaining possibility for a circum‐Caribbean discontinuity in gene flow is at the Isthmus of Panama, where there may be a transition from 98% Group B haplotypes in Costa Rica to 85–100% Group A haplotypes in South America.     

Distribution,host plant affiliation,phenology, and phylogeny of walnut‐infesting Rhagoletis flies (Diptera: Tephritidae) in Mexico

Many taxa of Nearctic origin have diversified in the subtropical highlands of Mexico. In particular, flies in the genus Rhagoletis have undergone episodes of isolation and gene flow during Pleistocene glaciations and post‐glacial times that have produced lineage differentiation and reproductive isolation. To reach a better understanding of the phylogeography of the genus Rhagoletis, a host plant survey of the walnut‐infesting Rhagoletis suavis species group was conducted across sixteen states comprising 34 different collecting sites in Mexico over a 9‐year period. Five species of Juglans were found to be infested by three species of walnut‐attacking Rhagoletis flies. Several species of parasitoids were also recovered from collections, but in contrast to their walnut fly hosts, they revealed little evidence for host or geographic subdivision. There was no consistent difference in mean eclosion time between walnut fly species or populations associated with different host walnuts in Mexico, unlike the case for other Rhagoletis species, in which allochronic isolation arising from variation in diapause timing is a major ecological adaptation, reproductively isolating flies. We compare the distribution of R. suavis flies in Mexico with those of other Rhagoletis species attacking hawthorns and cherries, and discuss its implications for population divergence and speciation. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110 , 765–779.     

Phylogenetic relationships and historical biogeography of tribes and genera in the subfamily Nymphalinae (Lepidoptera: Nymphalidae)

We infer for the first time the phylogenetic relationships of genera and tribes in the ecologically and evolutionarily well‐studied subfamily Nymphalinae using DNA sequence data from three genes: 1450 bp of cytochrome oxidase subunit I (COI) (in the mitochondrial genome), 1077 bp of elongation factor 1‐alpha (EF1‐α) and 400–403 bp of wingless (both in the nuclear genome). We explore the influence of each gene region on the support given to each node of the most parsimonious tree derived from a combined analysis of all three genes using Partitioned Bremer Support. We also explore the influence of assuming equal weights for all characters in the combined analysis by investigating the stability of clades to different transition/transversion weighting schemes. We find many strongly supported and stable clades in the Nymphalinae. We are also able to identify ‘rogue’ taxa whose positions are weakly supported (the different gene regions are in conflict with each other) and unstable. Our main conclusions are: (1) the tribe Coeini as currently constituted is untenable, and Smyrna, Colobura and Tigridia are part of Nymphalini; (2) ‘Kallimini’ is paraphyletic with regard to Melitaeini and should be split into three tribes: Kallimini s.s., Junoniini and Victorinini; (3) Junoniini, Victorinini, Melitaeini and the newly circumscribed Nymphalini are strongly supported monophyletic groups, and (4) Precis and Junonia are not synonymous or even sister groups. The species Junonia coenia, a model system in developmental biology, clearly belongs in the genus Junonia. A dispersal‐vicariance analysis suggests that dispersal has had a major effect on the distributions of extant species, and three biotic regions are identified as being centres of diversification of three major clades: the Palaearctic for the Nymphalis‐group, the Afrotropics for Junoniini and the Nearctic for Melitaeini. © 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 86 , 227–251.     

Phylogeography of a Patagonian lizard and frog: Congruent signature of southern glacial refuges

Pleistocene glaciations produced significant increases in continental ice cover in polar and mid‐latitude temperate areas, sea‐level declines and shifts and reshuffling of biomes, all of which promote either isolation, coalescence or fragmentation in the distribution of land biota. If populations of several taxa have been co‐distributed for a prolonged time, and if the periods between perturbation or vicariance processes have been more or less stable, it is expected that divergence patterns of closely related and ecologically similar species will be congruent because of their similar biological and demographic characteristics. Based on this premise, we analysed the phylogeographic structure (cytochrome b) of Liolaemus pictus and Batrachyla leptopus, two widely co‐distributed lizard and frog species, respectively, in the Chiloé Archipelago of southern Chile, to decipher their genetic structure in response to a common climatic and environmental history. Haplotype network analysis and Bayesian inference suggest an evolutionary pattern of genetic diversity for the two species that is consistent with the Quaternary glacial history of southern Chile, and suggests a complex phylogeographic history in the Liolaemus and Batrachyla species. High‐divergence levels among haplotypes in some island populations of the archipelago also suggest genetic connectivity between putative refuges from Chiloé Island and the mainland along the exposed continental shelf during sea level minima associated with the most recent Quaternary glaciations. Our results are consistent with our hypothesis that two species have responded to parallel historical events in which the historical process during the last glacial maximum (approximately 41°S) has been sufficient to influence their phylogeographic structure.     

Biogeographical research on European species of the lichen genus Physconia

Abstract Aim The (world) distribution of the European species of Physconia Poelt (Lichenes: Physciaceae) [except Ph. distorta (With.) J. R. Laundon and Ph. americana Essl.] was investigated. The study was complemented by data on the ecological behaviour of the species. Location The location of the study is the whole world with a local emphasis on Europe. Methods The geographical distribution of the lichen species under consideration was investigated from collection data of herbaria and field studies in a databank. Additional data were taken from existing literature and critically considered. Distribution maps were created with ArcView GIS. The distribution patterns are expressed as three‐dimensional areal formulas, regarding zonal distribution, altitudinal range and oceanity of the species distributions, using the method of Meusel et al. (1965 ). Results The distribution patterns of the ten Physconia taxa considered are represented in detailed maps and expressed as three‐dimensional areal formulas. The ecological behaviour was discussed. Physconia muscigena and Ph. perisidiosa are species of bipolar distribution type, the other species are holarctic types. Physconia detersa and Ph. enteroxantha occur in both Eurasia and North America, the remaining species (Ph.grisea ssp. grisea, Ph. grisea ssp. lilacina, Ph. petraea, Ph. servitii, Ph. subpulverulenta and Ph.venusta) are pure Old world taxa with a much more limited distribution.The majority of the European Physconia taxa are distributed meridionally to submeridionally and occur preferentially in suboceanic areas. Main conclusions Some of the species under consideration have a wide distribution, nevertheless they show clear affinities to ecological conditions that influence their distribution patterns. A number of species show not only zonal and regional restrictions, but also affinities to the western climatic type. The Mediterranean is the centre of diversity of the genus Physconia in Europe.     

CHROMOSOME NUMBERS IN UMBELLIFERAE. III

Chromosome numbers are reported for 156 collections representing 100 taxa of Umbelliferae. Approximately two thirds of the collections are from Mexico, Central and South America and indicate a high percentage of polyploid species in certain genera found in this area. Chromosome numbers for plants belonging to 78 taxa are published here for the first time, previously published chromosome numbers are verified for 18 taxa and chromosome numbers differing from those previously published are reported in seven instances. No chromosome counts have been previously published for nine of the genera included here. Further aneuploidy and polyploidy were found in Eryngium, and Lomatium columbianum has been found to be a high polyploid with 2n = 14x. Every chromosome count is referable to a cited herbarium specimen.     

New notharctine primate fossils from the early Eocene of New Mexico and southern Wyoming and the phylogeny of Notharctinae

Previously undescribed notharctine primate fossils are reported from the early Eocene San Jose Formation, San Juan Basin, New Mexico, and the early Eocene Wasatch Formation, southern Wyoming. These collections include the most complete specimens yet discovered of the poorly known species Copelemur tutus and Copelemur praetutus; the first upper dentitions of Cantius angulatus and Cantius frugivorus from the type area of these taxa; and fossils attributable to two new notharctine species, Copelemur australotutus and Smilodectes gingerichi. These new fossils reveal that current ideas concerning notharctine phylogeny are incorrect. Two major, monophyletic clades are apparent within the subfamily: the tribe Copelemurini, consisting of the genera Copelemur and Smilodectes, and the tribe Notharctini, comprising the genera Cantius, Pelycodus, and Notharctus. Analysis of the paleobiogeographic distribution of the Copelemurini indicates that this clade was limited to more southerly regions of western North America during early Eocene time. Northward migration of more tropical habitats during the late Wasatchian and early Bridgerian in western North America, associated with an overall climatic warming trend through the early and middle Eocene, appears to have allowed several mammalian taxa, including Smilodectes, to extend their ranges northward during this time interval. Such taxa thus possess diachronous distributions and have been partly responsible for the long-standing confusion regarding the biostratigraphic correlation of early Eocene faunas from New Mexico with those from Wyoming. Based on several taxa which are also known from the Wasatchian of Wyoming, the age of the San Jose Formation appears to be middle Wasatchian.     

Dietary specialization and the effects of plant species on potential multitrophic interactions of three species of nymphaline caterpillars

The diet breadth of insect herbivores influences their response to variation in plant quality, and these bitrophic interactions have implications for the higher‐level trophic interactions between herbivores and their natural enemies. In this comparative study, we examined the role of host plant species and plant secondary chemistry on the potential interactions between three species of nymphaline caterpillars and their natural enemies. The caterpillar species (all Lepidoptera: Nymphalidae) varied in their degree of specialization: the buckeye, Junonia coenia Hübner, is a specialist on plants that contain iridoid glycosides (IGs); the white peacock, Anartia jatrophae L., feeds on plants in five families, some of which contain IGs and some of which do not; and the painted lady, Vanessa cardui L., is a generalist, feeding on plants in at least 15 families. Each species was reared on leaves of an introduced host plant, Plantago lanceolata L. (Plantaginaceae), which produces two IGs, aucubin and catalpol, and on another plant species that is a common host plant. These alternate host plants were Plantago major L. (Plantaginaceae) for J. coenia, Bacopa monnieri (L.) Pennell (Plantaginaceae) for A. jatrophae, and Malva parviflora L. (Malvaceae) for V. cardui. We examined growth, sequestration, and immune response of these caterpillars on the different host plant species. Junonia coenia developed more rapidly and sequestered higher IG concentrations when reared on P. lanceolata, whereas both other species grew more slowly on P. lanceolata. Host plant did not influence immune response of J. coenia or A. jatrophae, whereas V. cardui immune response was weaker when reared on P. lanceolata. Junonia coenia was most efficient at IG sequestration and A. jatrophae was least efficient, when all three species were reared on P. lanceolata. These results indicate that diet breadth may play an important role in structuring tritrophic interactions, and this role should be further explored.     

Arion transsylvanus (Mollusca,Pulmonata, Arionidae): rediscovery of a cryptic species

Jordaens, K., Pinceel, J., Van Houtte, N., Breugelmans, K. & Backeljau, T. (2010). Arion transsylvanus (Mollusca, Pulmonata, Arionidae): rediscovery of a cryptic species. —Zoologica Scripta, 39, 343–362. Cryptic species are abundant among invertebrates and are often hard to recognise. Molecular markers are an extremely useful tool to delineate cryptic taxa, although they should be applied with caution because different genes and techniques may yield different outcomes. We illustrate how cross‐validation by molecular and morphological data can be applied to optimise taxonomic interpretations when cryptic species are involved. This is performed for the terrestrial slug Arion subfuscus species complex which represents a historical ‘taxonomic garbage can’. Gonad morphology, allozymes and mtDNA data consistently showed that slugs from Romania and a location in E Poland represent a strongly differentiated taxon within this complex. These slugs are therefore formally redescribed and assigned to A. transsylvanus Simroth 1885 ; a forgotten nominal taxon from Transylvania. Diagnostic characters, including DNA sequences for the mitochondrial 16S rDNA are presented. Animals with the morphology of A. brunneus Lehmann 1862 , a nominal taxon which has also been reported from Romania, have the gonad type, allozyme alleles and 16S rDNA haplotypes of either A. fuscus, A. subfuscus or A. transsylvanus. Therefore, A. brunneus is regarded as a colour morph shared by several species and hence, A. transsylvanus is probably the only A. subfuscus‐like species in the Romanian Carpathians.     

Phylogeography and cryptic variation within the Lacerta viridis complex (Lacertidae, Reptilia)

It is well known that the current genetic pattern of many European species has been highly influenced by climatic changes during the Pleistocene. While there are many well known vertebrate examples, knowledge about squamate reptiles is sparse. To obtain more data, a range‐wide sampling of Lacerta viridis was conducted and phylogenetic relations within the L. viridis complex were analysed using an mtDNA fragment encompassing part of cytochrome b, the adjacent tRNA genes and the noncoding control region. Most genetic divergence was found in the south of the distribution range. The Carpathian Basin and the regions north of the Carpathians and Alps are inhabited by the same mitochondrial lineage, corresponding to Lacerta viridis viridis. Three distinct lineages occurred in the south‐eastern Balkans — corresponding to L. v. viridis, L. v. meridionalis, L. v. guentherpetersi— as well as a fourth lineage for which no subspecies name is available. This distribution pattern suggests a rapid range expansion of L. v. viridis after the Holocene warming, leading to a colonization of the northern part of the species range. An unexpected finding was that a highly distinct genetic lineage occurs along the western Balkan coast. Phylogenetic analyses (Bayesian, maximum likelihood, maximum parsimony) suggested that this west Balkan lineage could represent the sister taxon of Lacerta bilineata. Due to the morphological similarity of taxa within the L. viridis complex this cryptic taxon was previously assigned to L. v. viridis. The distribution pattern of several parapatric, in part highly, distinct genetic lineages suggested the existence of several refuges in close proximity on the southern Balkans. Within L. bilineata sensu stricto a generally similar pattern emerged, with a high genetic diversity on the Apennine peninsula, arguing for two distinct refuges there, and a low genetic diversity in the northern part of the range. Close to the south‐eastern Alps, three distinct lineages (L. b. bilineata, L. v. viridis, west Balkan taxon) occurred within close proximity. We suggest that the west Balkan lineage represents an early offshoot of L. bilineata that was isolated during a previous Pleistocene glacial from the more western L. bilineata populations, which survived in refuges on the Apennine peninsula.     

Evolutionary drivers of phylogeographical diversity in the highlands of Mexico: a case study of the Crotalus triseriatus species group of montane rattlesnakes

Aim To assess the genealogical relationships of widespread montane rattlesnakes in the Crotalus triseriatus species group and to clarify the role of Late Neogene mountain building and Pleistocene pine–oak forest fragmentation in driving the diversification of Mexican highland taxa. Location Highlands of mainland Mexico and the south‐western United States (Texas, New Mexico, and Arizona). Methods A synthesis of inferences was used to address several associated questions about the biogeography of the Mexican highlands and the evolutionary drivers of phylogeographical diversity in co‐distributed taxa. We combined extensive range‐wide sampling (130 individuals representing five putative species) and mixed‐model phylogenetic analyses of 2408 base pairs of mitochondrial DNA to estimate genealogical relationships and divergence times within the C. triseriatus species group. We then assessed the tempo of diversification using a maximum likelihood framework based on the birth–death process. Estimated times of divergences provided a probabilistic temporal component and questioned whether diversification rates have remained constant or varied over time. Finally, we looked for phylogeographical patterns in other co‐distributed taxa. Results We identified eight major lineages within the C. triseriatus group, and inferred strong correspondence between maternal and geographic history within most lineages. At least one cryptic species was detected. Relationships among lineages were generally congruent with previous molecular studies, with differences largely attributable to our expanded taxonomic and geographic sampling. Estimated divergences between most major lineages occurred in the Late Miocene and Pliocene. Phylogeographical structure within each lineage appeared to have been generated primarily during the Pleistocene. Although the scale of genetic diversity recognized affected estimated rates of diversification, rates appeared to have been constant through time. Main conclusions The biogeographical history of the C. triseriatus group implies a dynamic history for the highlands of Mexico. The Neogene formation of the Transvolcanic Belt appears responsible for structuring geographic diversity among major lineages. Pleistocene glacial–interglacial climatic cycles and resultant expansions and contractions of the Mexican pine–oak forest appear to have driven widespread divergences within lineages. Climatic change, paired with the complex topography of Mexico, probably produced a myriad of species‐specific responses in co‐distributed Mexican highland taxa. The high degree of genetic differentiation recovered in our study and others suggests that the Mexican highlands may contain considerably more diversity than currently recognized.     

Effects of prey quality on social wasps when given a choice of prey

The effect of prey quality on foraging behavior and colony demographics of the social wasp Polistes fuscatus was examined by providing a choice between non‐toxic prey (Manduca sexta caterpillars) and sublethally toxic prey (Junonia coenia caterpillars), and then comparing the performance of these colonies to others given only the non‐toxic prey. In the choice, one of two types of Manduca were used: those fed an artificial diet or those fed plantain (Plantago lanceolata), which contains iridoid glycosides (IGs) that Junonia coenia store but which Manduca does not. Despite the negative correlation between the number of Junonia prey used and number of adult offspring produced, when a surplus of non‐toxic prey was available, the wasps did not completely avoid the toxic prey. However, they were more discriminating when the choice was between Manduca fed an artificial diet and Junonia fed plantain vs. when both prey species had eaten the plantain. Because the wasps had a choice of prey types and had a surplus of prey on about one‐third of the days, the wasps were able to take enough non‐toxic prey to avoid some of the negative consequences of IGs. For example, the total number of wasp offspring per nest was not affected, but mean weight of female offspring per colony was less for colonies given both prey types eating plantain, compared to that for colonies fed only non‐toxic prey, or those given a choice of non‐toxic prey vs. toxic prey. In addition, compared to the control (only non‐toxic prey), the proportion of males produced was less in the treatment that provided a clear contrast between non‐toxic and toxic prey. Why these wasps did not avoid the toxic prey is discussed.     

Glacial refuges and cryptic speciation in a Southern Palearctic tiger beetle (Coleoptera: Cicindelidae)

Pleistocene glaciations had a determining role for shaping the current distribution and diversity of organisms, especially in the Palearctic region. In this work, we carry out a phylogeographic analysis of Iberian and two Eastern European populations of the tiger beetle Calomera littoralis (Fabricius, 1787) in order to infer the processes that may have affected their evolutionary history. According to our results, the genetic diversity of central Iberian C. littoralis populations is very low. The haplotype networks also suggest that these populations experienced a genetic bottleneck in the past, possibly related to the last glacial maxima, similar to that observed in other cicindelid taxa. These results highlight the remarkable dispersal capacity of this species, being able to move freely from one locality to another, despite the relatively long distances of sub-optimal habitat that separates them. The genetic data of central Iberian populations contrast with those of the Eastern European populations, with higher genetic diversity and no hints of any past bottleneck. This can be explained by the different characteristics of both (Iberian and Pontic) glacial refuges. The high degree of genetic differentiation between the three C. littoralis clades, and the inclusion of C. lunulata between them, suggests that the three analysed populations could be considered as different cryptic species. In that case, C. littoralis may correspond to a species complex that is still undergoing a process of speciation, similar to that observed in Cicindela campestris.