Cryptic Species Pair Research Articles

Out of taxonomic crypsis: A new trans-arctic cryptic species pair corroborated by phylogenetics and molecular evidence

Cryptic species are a common phenomenon in cosmopolitan marine species. The use of molecular tools has often uncovered cryptic species occupying a fraction of the geographic range of the original morphospecies. Shipworms (Teredinidae) are marine bivalves, living in drift and fixed wood, many of which have a conserved morphology across cosmopolitan distributions. Herein novel and GenBank mitochondrial (cytochrome c oxidase subunit I) and nuclear (18S rRNA) DNA sequences are employed to produce a phylogeny of the Teredinidae and delimit a cryptic species pair in the Psiloteredo megotara complex. The anatomy, biogeography, and ecology of P. megotara, Psiloteredo sp. and Nototeredo edax are compared based on private and historic museum collections and a thorough literature review. Morphological and anatomical characters of P. megotara from the North Atlantic and Psiloteredo sp. from Japan were morphologically indistinguishable, and differ in pallet architecture and soft tissue anatomy from N. edax. The two Psiloteredo species were then delimited as genetically distinct species using four molecular-based methods. Consequently, the Northwest Pacific species, Psiloteredo pentagonalis, first synonymized with N. edax and then with P. megotara, is resurrected. Nototeredo edax, P. megotara and P. pentagonalis are redescribed based upon morphological and molecular characters. Phylogenetic analysis further revealed cryptic species complexes within the cosmopolitan species Bankia carinata and possibly additional cryptic lineages within the cosmopolitan Lyrodus pedicellatus.

Broad‐scale patterns of geographic avoidance between species emerge in the absence of fine‐scale mechanisms of coexistence

AbstractAimThe need to forecast range shifts under future climate change has motivated an increasing interest in better understanding the role of biotic interactions in driving diversity patterns. The contribution of biotic interactions to shaping broad‐scale species distributions is, however, still debated, partly due to the difficulty of detecting their effects. We aim to test whether spatial exclusion between potentially competing species can be detected at the species range scale, and whether this pattern relates to fine‐scale mechanisms of coexistence.LocationWestern Palearctic.MethodsWe develop and evaluate a measure of geographic avoidance that uses outputs of species distribution models to quantify geographic exclusion patterns expected if interspecific competition affects broad‐scale distributions. We apply the measure to 10 Palearctic bat species belonging to four morphologically similar cryptic groups in which competition is likely to occur. We compare outputs to null models based on pairs of virtual species and to expectations based on ecological similarity and fine‐scale coexistence mechanisms. We project changes in range suitability under climate change taking into account effects of geographic avoidance.ResultsValues of geographic avoidance were above null expectations for two cryptic species pairs, suggesting that interspecific competition could have contributed to shaping their broad‐scale distributions. These two pairs showed highest levels of ecological similarity and no trophic or habitat partitioning. Considering the role of competition modified predictions of future range suitability.Main conclusionsOur results support the role of interspecific competition in limiting the geographic ranges of morphologically similar species in the absence of fine‐scale mechanisms of coexistence. This study highlights the importance of incorporating biotic interactions into predictive models of range shifts under climate change, and the need for further integration of community ecology with species distribution models to understand the role of competition in ecology and biogeography.

Bat echolocation plasticity in allopatry: a call for caution in acoustic identification of Pipistrellus sp.

Animals modify their behaviours and interactions in response to changing environments. In bats, environmental adaptations are reflected in echolocation signalling that is used for navigation, foraging and communication. However, the extent and drivers of echolocation plasticity are not fully understood, hindering our identification of bat species with ultrasonic detectors, particularly for cryptic species with similar echolocation calls. We used a combination of DNA barcoding, intensive trapping, roost and emergence surveys and acoustic recording to study a widespread European cryptic species complex (Pipistrellus pipistrellus and Pipistrellus pygmaeus) to investigate whether sibling bat species could exhibit extreme echolocation plasticity in response to certain environmental conditions or behaviours. We found that P. pygmaeus occupied the acoustic niche of their absent congeneric species, producing calls with P. pipistrellus’ characteristic structure and peak frequencies and resulting in false positive acoustic records of that species. Echolocation frequency was significantly affected by the density of bats and by maternity rearing stage, with lower frequency calls emitted when there was a high density of flying bats, and by mothers while juveniles were non-volant. During roost emergence, 29% of calls had peak frequencies typical of P. pipistrellus, with calls as low as 44 kHz, lower than ever documented. We show that automatic and manual call classifiers fail to account for echolocation plasticity, misidentifying P. pygmaeus as P. pipistrellus. Our study raises a vital limitation of using only acoustic sampling in areas with high densities of a single species of a cryptic species pair, with important implications for bat monitoring.Significance statementUltrasonic acoustic detectors are widely used in bat research to establish species inventories and monitor species activity through identification of echolocation calls, enabling new methods to study and understand this elusive understudied group of nocturnal mammals. However, echolocation call signalling in bats is intrinsically different to that of other taxa, serving a main function of navigation and foraging. This study demonstrates an extreme level of plasticity, showing large variation in call frequency and structure in different situations. We showcase the difficulty and limitation in using acoustic sampling alone for bat monitoring and the complications of setting parameters for species identification for manual and automatic call classifiers. Our observations of call frequency variation correlated with density and absence of congenerics provide novel insights of behavioural echolocation plasticity in bats.

Candidate genes involved in cuticular hydrocarbon differentiation between cryptic, parabiotic ant species

Insect cuticular hydrocarbons (CHCs) are highly diverse and have multiple functions, including communication and waterproofing. CHC profiles form species-specific, complex blends of up to 150 compounds. Especially in ants, even closely related species can have largely different profiles, raising the question how CHC differences are mirrored in the regulation of biosynthetic pathways. The neotropical ants Crematogaster levior and Camponotus femoratus both consist of two cryptic species each that are morphologically similar, but express strongly different CHC profiles. This is ideal to study the molecular basis of CHC differences. We thus investigated gene expression differences in fat-body transcriptomes of these ants. Despite common garden conditions, we found several thousand differentially expressed transcripts within each cryptic species pair. Many of these were related to metabolic processes, probably accounting for physiological differences. Moreover, we identified candidate genes from five gene families involved in CHC biosynthesis. By assigning candidate transcripts to orthologs in Drosophila, we inferred which CHCs might be influenced by differential gene expression. Expression of these candidate genes was often mirrored in the CHC profiles. For example, Cr. levior A, which has longer CHCs than its cryptic sister species, had a higher expression of elongases and a lower expression of fatty acyl- CoA reductases. This study is one of the first to identify CHC candidate genes in ants and will provide a basis for further research on the genetic basis of CHC biosynthesis.

Postzygotic isolation drives genomic speciation between highly cryptic Hypocnemis antbirds from Amazonia.

How species evolve reproductive isolation in the species-rich Amazon basin is poorly understood in vertebrates. Here, we sequenced a reference genome and used a genome-wide sample of SNPs to analyze a hybrid zone between two highly cryptic species of Hypocnemis warbling-antbirds-the Rondonia warbling-antbird (H. ochrogyna) and Spix's warbling-antbird (H. striata)-in a headwater region of southern Amazonia. We found that both species commonly hybridize, producing F1 s and a variety of backcrosses with each species but we detected only one F2 -like hybrid. Patterns of heterozygosity, hybrid index, and interchromosomal linkage disequilibrium in hybrid populations closely match expectations under strong postzygotic isolation. Hybrid zone width (15.4 km) was much narrower than expected (211 km) indicating strong selection against hybrids. A remarkably high degree of concordance in cline centers and widths across loci, and a lack of reduced interspecific Fst between populations close to versus far from the contact zone, suggest that genetic incompatibilities have rendered most of the genome immune to introgression. These results support intrinsic postzygotic isolation as a driver of speciation in a moderately young cryptic species pair from the Amazon and suggest that species richness of the Amazon may be grossly underestimated.

Cryptic species of black flies (Diptera: Simuliidae) at high elevations in the Oriental Region: The Simulium vernum species group in Thailand

High-mountain summits in tropical regions are habitat islands that provide refugia for northern taxa and opportunities for speciation. These focal hotspots of biodiversity are also threatened by increasing anthropogenic pressures and changing climatic conditions. The largest species group of black flies, the Simulium vernum group, reaches its southern limit in the mountains of Southeast Asia, providing an opportunity to examine macrogenetic diversification at the southern edge of the group's range. We investigated the polytene chromosomes of two morphologically similar species in the S. vernum group, S. chomthongense Takaoka, Srisuka & Choochote, and S. khunklangense Takaoka & Srisuka, from their type localities near the summit of Doi Inthanon, Thailand's highest mountain. Chromosomal analyses of 112 larvae demonstrated reproductive isolation between S. chomthongense and S. khunklangense and revealed a cryptic species of S. chomthongense. The three species have the same fixed banding sequence in five of the six chromosome arms, but they are distinguished by the successive addition of overlapping inversions in the IIIL arm. The two cytospecies (A and B) of S. chomthongense s. l. are the third example of endemic or nearly endemic cryptic species pairs near the top of Doi Inthanon, suggesting the possibility of sympatric speciation and the importance of conserving high-elevation stream habitats.

A taxonomic monograph of the liphistiid spider genus Heptathela, endemic to Japanese islands.

Among the eight extant genera of primitively segmented spiders, family Liphistiidae, two are confined to East Asian islands, Heptathela Kishida, 1923 and Ryuthela Haupt, 1983. In this paper, a taxonomic revision of the genus Heptathela (Heptathelinae) from Kyushu and Ryukyu archipelago, Japan is provided. This study follows a multi-tier species delimitation strategy within an integrative taxonomic framework that is presented in a parallel paper, in which diagnosable lineages are considered as valid species. There, the initial hypothesis of species diversity (19) based on classical morphological diagnoses is tested with multiple species delimitation methods aimed at resolving conflict in data. This revision follows those analyses that converge on the species diversity of 20, which includes a pair of cryptic species that would have been undetected with morphology alone. After this revision, eight previously described species remain valid, two junior synonyms are proposed, and 12 new Heptathela species are described based on diagnostic evidence. To ease identification and to hint at putative evolutionary units, Heptathela is divided into three groups. The Kyushu group contains H. higoensis Haupt, 1983, H. kikuyai Ono, 1998, H. kimurai (Kishida, 1920), and H. yakushimaensis Ono, 1998; the Amami group contains H. amamiensis Haupt, 1983, H. kanenoi Ono, 1996, H. kojimasp. nov., H. sumiyosp. nov., and H. ukensp. nov.; and the Okinawa group contains H. yanbaruensis Haupt, 1983, H. ahasp. nov., H. gayozansp. nov., H. kubayamasp. nov., H. maesp. nov., H. otohasp. nov., H. shurisp. nov., H. tokashikisp. nov., H. untensp. nov., and H. cryptasp. nov.Heptathela helios Tanikawa & Miyashita, 2014 is not assigned to a species group. A combination of diagnostic tools augments the morphological diagnoses that, in isolation, would be prone to error in morphologically challenging groups of organisms.

Genomic and Mitochondrial Data Identify Different Species Boundaries in Aposematically Polymorphic Eniclases Net-Winged Beetles (Coleoptera: Lycidae).

Species delineation is essential for any evolutionary and biodiversity research, and recent advances in genomic sequencing have made it possible to robustly define species boundaries and detect hidden diversity. Here, we studied 14 species of aposematically colored New Guinean Eniclases (Coleoptera: Lycidae) whose conventional morphology- and single-locus mtDNA-based taxonomy has been contentious. We analyzed mitochondrial and restriction site associated DNA fragments to obtain a phylogenetic hypothesis and compared relationships recovered by the RAD analysis with species limits based on other information. The results show the presence of cryptic diversity and common mitonuclear discordance when over 30% of individuals were incorrectly assigned to species if only mitogenomic markers were considered. Nuclear data falsified the species rank of one species and identified one earlier unrecognized lineage deserving species rank. Further, our analyses demonstrate a highly variable phenotypic differentiation, with several pairs of cryptic species standing in contrast with genetically close but phenotypically highly divergent lineages. We show that morphological and mitogenomic analyses produce reliable information for taxonomy in most cases. Nevertheless, the species boundaries among closely related species should be based on all lines of evidence, including nuclear markers.

Cuticular hydrocarbons as potential mediators of cryptic species divergence in a mutualistic ant association.

Upon advances in sequencing techniques, more and more morphologically identical organisms are identified as cryptic species. Often, mutualistic interactions are proposed as drivers of diversification. Species of the neotropical parabiotic ant association between Crematogaster levior and Camponotus femoratus are known for highly diverse cuticular hydrocarbon (CHC) profiles, which in insects serve as desiccation barrier but also as communication cues. In the present study, we investigated the association of the ants’ CHC profiles with genotypes and morphological traits, and discovered cryptic species pairs in both genera. To assess putative niche differentiation between the cryptic species, we conducted an environmental association study that included various climate variables, canopy cover, and mutualistic plant species. Although mostly sympatric, the two Camponotus species seem to prefer different climate niches. However in the two Crematogaster species, we could not detect any differences in niche preference. The strong differentiation in the CHC profiles may thus suggest a possible role during speciation itself either by inducing assortative mating or by reinforcing sexual selection after the speciation event. We did not detect any further niche differences in the environmental parameters tested. Thus, it remains open how the cryptic species avoid competitive exclusion, with scope for further investigations.

Evolutionary and biogeographic history of the black fly Simulium wayani (Diptera: Simuliidae) on the island of Timor

A recently described species of black fly, Simulium wayani Takaoka and Chen, from the island of Timor was chromosomally mapped to provide insights into its evolutionary and biogeographic history. The morphologically based species status of S. wayani is supported by a suite of fixed chromosomal rearrangements and unique sex chromosomes derived primarily from a large pool of polymorphisms in the S. ornatipes complex in Australia. The banding patterns of its polytene chromosomes indicate that S. wayani is closely related to a pair of homosequential cryptic species (S. norfolkense Dumbleton and S. ornatipes cytoform A2) in the S. ornatipes Skuse complex on mainland Australia; all three species uniquely share the same amplified band in their chromosomal complement. The low level of polymorphism and heterozygosity in S. wayani, relative to Australian populations of the S. ornatipes complex, suggests few colonization events from the larger land mass.

Lyrodus mersinensis sp. nov. (Bivalvia: Teredinidae) another cryptic species in the Lyrodus pedicellatus (Quatrefages, 1849) complex.

New data from barcode index numbers (BINs) and 28S rRNA gene sequences confirm a cryptic species pair in Lyrodus pedicellatus from the eastern Mediterranean and European Atlantic coasts. Therefore, it is paramount to associate the new species to a scientific name for a reliable reference system of biological information. To this end, we describe Lyrodus mersinensis sp. nov., another cryptic species in the L. pedicellatus complex, and redescribe the `true´ L. pedicellatus. Both the description and redescription are based on molecular diagnostic characters obtained from sequences of the mitochondrial cytochrome c oxidase subunit I (COI) and nuclear 28S rRNA genes. The 18S rRNA gene sequences did not yield diagnostic characters to distinguish these species. A morphological diagnosis of pedicellatus-like Lyrodus species is also provided.

From Nature to the Lab: Establishing Drosophila Resources for Evolutionary Genetics

In recent years important tools have been developed in Drosophila to capture with the greatest possible accuracy the variation found in nature. Efforts such as the Drosophila melanogaster Genetic Reference Panel (DGRP) or the Drosophila Synthetic Population Resource Panel (DSRP) allied to the advances in whole-genome sequencing and analysis have propelled to unprecedented level our capacity to dissect the genotype-phenotype map. However, several practical problems arise upstream of these analyses starting with the collection and identification of wild specimens. These problems are dealt with in different ways by each researcher generating solutions not necessarily compatible across laboratories. Here, we provide a systematic coverage of every phase of this process based on our experience, and suggest procedures to maximize and share the generated resources potentiating future applications. We propose a detailed pipeline to guide researchers from collection in the wild to the development of a large array of molecular and genetic resources. We designed a multiplex-PCR that distinguishes sister species D. melanogaster and D. simulans and is diagnostic of the presence/absence of Wolbachia infection. These procedures may extend to other cryptic species pairs and endosymbionts. We developed a standardized protocol to create, replicate and maintain isofemale lines and outbred populations. Finally, we explore the potential of outbred populations across several applications from experimental evolution, to introgression of transgenic constructs or mutant alleles, and genomic analyses. We hope to contribute to the success in developing Drosophila resources for evolutionary genetics studies and facilitate exchanges across laboratories based on a common set of procedures.

Discussion on the taxonomy of African fern Abrodictyum rigidum (Sw.) Ebihara & Dubuisson and description of two new Abrodictyum C.Presl species (Hymenophyllaceae, Polypodiidae) for the Afro-Malagasy region

The taxonomy of the genus Abrodictyum is complex, with many species showing similar gross morphology, especially those supposedly related to A. rigidum. Morphological and morphometric comparisons were performed on Neotropical A. rigidum specimens, African specimens first attributed to A. rigidum and two supposedly new distinct Abrodictyum species from the western Indian Ocean. All species share similar gross morphology and therefore are often confused in the wild and in herbarium collections. Morphological and morphometric analyses have not led us to the discrimination of distinct taxa or morpho-groups, except for one of the two supposedly new species, which appears to be endemic to Madagascar and shows some differences in frond shape, frond division and the indusium. Molecular analyses nevertheless confirm the distinctiveness of the Malagasy endemic new species, and the clustering of African A. rigidum with the second supposedly new species, both new taxa being consequently distinct from true Neotropical A. rigidum. We describe thus two new species: Abrodictyum pseudorigidum sp. nov., which is widespread in tropical Africa and in the western Indian Ocean and forms with true A. rigidum a pair of cryptic species, and Abrodictyum franceae sp. nov. endemic to Madagascar.

Phylogeny of deepwater snappers (Genus Etelis) reveals a cryptic species pair in the Indo-Pacific and Pleistocene invasion of the Atlantic

Evolutionary genetic patterns in shallow coastal fishes are documented with dozens of studies, but corresponding surveys of deepwater fishes (>200m) are scarce. Here we investigate the evolutionary history of deepwater snappers (genus Etelis), comprised of three recognized Indo-Pacific species and one Atlantic congener, by constructing a phylogeny of the genus with two mtDNA loci and two nuclear introns. Further, we apply range-wide Indo-Pacific sampling to test for the presence and distribution of a putative cryptic species pair within E. carbunculus using morphological analyses and mtDNA cytochrome b sequences from 14 locations across the species range (N=1696). These analyses indicate that E. carbunculus is comprised of two distinct, non-interbreeding lineages separated by deep divergence (d=0.081 in cytochrome b). Although these species are morphologically similar, we identified qualitative differences in coloration of the upper-caudal fin tip and the shape of the opercular spine, as well as significant differences in adult body length, body depth, and head length. These two species have overlapping Indo-Pacific distributions, but one species is more widespread across the Indo-Pacific, whereas the other species is documented in the Indian Ocean and Western Central Pacific. The dated Etelis phylogeny places the cryptic species divergence in the Pliocene, indicating that the biogeographic barrier between the Indian and Pacific Oceans played a role in speciation. Based on historic taxonomy and nomenclature, the species more widespread in the Pacific Ocean is E. carbunculus, and the other species is previously undescribed (referred to here as E. sp.). The Atlantic congener E. oculatus has only recently (∼0.5Ma) diverged from E. coruscans in the Indo-Pacific, indicating colonization via southern Africa. The pattern of divergence at the Indo-Pacific barrier, and Pleistocene colonization from the Indian Ocean into the Atlantic, is concordant with patterns observed in shallow coastal fishes, indicating similar drivers of evolutionary processes.

Seven new species of Paleanotus (Annelida: Chrysopetalidae) described from Lizard Island, Great Barrier Reef, and coral reefs of northern Australia and the Indo-Pacific: two cryptic species pairs revealed between western Pacific Ocean and the eastern Indian Ocean.

Morphological investigation into the paleate genus Paleanotus Schmarda 1861 of the family Chrysopetalidae from northern Australian coral reefs, primarily Lizard Island and outlying reefs, included a complex of very small, slender individuals (length < 5 mm). This complex resolved into 7 new species, described herein: Paleanotus inornatus n. sp., P. adornatus n. sp., P. chrysos n. sp., P. aquifolia n. sp., P. latifolia n. sp., P. silus n. sp., and P. silopsis n. sp. A key is provided to the new species and Paleanotus distinguished from Treptopale and Hyalopale, two closely related genera. Diagnostic features of the apical structure and shape of the notochaetal main paleae plus median paleae shape and raised rib pattern, differentiates each species from the other. Gametous states are described. Two cryptic species pairs (Paleanotus silopsis n. sp. and P. silus n. sp.; Paleanotus aquifolia n. sp. and P. latifolia n. sp.) were identified. In each case one species is restricted to either the NE or NW Australian coast. In each pair the most eastern point for the NW Australian species range occurs at Darwin, western Arnhemland, Northern Territory. Additional material for each species pair extends their respective ranges northwards: NW Australia to Thailand, Andaman Sea, eastern Indian Ocean or NE Australia, Great Barrier Reef to the Philippines, western Pacific Ocean. Cryptic morphology and potential genetic diversity is discussed in Paleanotus inornatus n. sp. and P. adornatus n. sp. that possess overlapping widespread distribution patterns across northern Australia and Indo-Pacific reefs. The smallest bodied taxon, Paleanotus chrysos n. sp. is the only species with a Coral Sea range encompassing Lizard Island, Heron Island and New Caledonia.

Why do cryptic species tend not to co-occur? A case study on two cryptic pairs of butterflies.

As cryptic diversity is being discovered, mostly thanks to advances in molecular techniques, it is becoming evident that many of these taxa display parapatric distributions in mainland and that they rarely coexist on islands. Genetic landscapes, haplotype networks and ecological niche modeling analyses were performed for two pairs of non-sister cryptic butterfly species, Aricia agestis-A. cramera and Polyommatus icarus—P. celina (Lycaenidae), to specifically assess non-coexistence on western Mediterranean islands, and to test potential causes producing such chequered distribution patterns. We show that the morphologically and ecologically equivalent pairs of species do not coexist on any of the studied islands, although nearly all islands are colonized by one of them. According to our models, the cryptic pairs displayed marked climatic preferences and ‘precipitation during the driest quarter’ was recovered as the most important climatic determinant. However, neither dispersal capacity, nor climatic or ecological factors fully explain the observed distributions across particular sea straits, and the existence of species interactions resulting in mutual exclusion is suggested as a necessary hypothesis. Given that the studied species are habitat generalists, feeding on virtually unlimited resources, we propose that reproductive interference, together with climatic preferences, sustain density-dependent mechanisms like “founder takes all” and impede coexistence on islands. Chequered distributions among cryptic taxa, both sister and non-sister, are common in butterflies, suggesting that the phenomenon revealed here could be important in determining biodiversity patterns.

Characterizing a hybrid zone between a cryptic species pair of freshwater snails.

Characterizing hybrid zones and their dynamics is a central goal in evolutionary biology, but this is particularly challenging for morphologically cryptic species. The lack of conspicuous divergence between parental types means intermediate hybrid forms often go undetected. We aimed to detect and characterize a suspected hybrid zone between a pair of morphologically cryptic lineages of the freshwater snail, Radix. We sampled Radix from across a contact zone between two mitochondrial lineages (Radix balthica and an undescribed lineage termed 'MOTU3') and detected admixture between two nuclear genotype clusters, which were significantly but not categorically associated with the mitochondrial lineages. Using a model selection approach, we show that the admixture cline is best explained by an interaction between precipitation and temperature gradients over the area, rather than geographic distance. We thus hypothesize that the correlation with climatic gradients suggests environmental selection has played a role in maintaining the hybrid zone. In a 2050 climate change scenario, we furthermore predict an expansion of one of the nuclear clusters and a widening of the hybrid zone as the climate warms and dries.

Multilocus evidence for globally distributed cryptic species and distinct populations across ocean gyres in a mesopelagic copepod.

Zooplanktonic taxa have a greater number of distinct populations and species than might be predicted based on their large population sizes and open-ocean habitat, which lacks obvious physical barriers to dispersal and gene flow. To gain insight into the evolutionary mechanisms driving genetic diversification in zooplankton, we developed eight microsatellite markers to examine the population structure of an abundant, globally distributed mesopelagic copepod, Haloptilus longicornis, at 18 sample sites across the Atlantic and Pacific Oceans (n = 761). When comparing our microsatellite results with those of a prior study that used a mtDNA marker (mtCOII, n = 1059, 43 sample sites), we unexpectedly found evidence for the presence of a cryptic species pair. These species were globally distributed and apparently sympatric, and were separated by relatively weak genetic divergence (reciprocally monophyletic mtCOII lineages 1.6% divergent; microsatellite FST ranging from 0.28 to 0.88 across loci, P < 0.00001). Using both mtDNA and microsatellite data for the most common of the two species (n = 669 for microsatellites, n = 572 for mtDNA), we also found evidence for allopatric barriers to gene flow within species, with distinct populations separated by continental landmasses and equatorial waters in both the Atlantic and Pacific Ocean basins. Our study shows that oceanic barriers to gene flow can act as a mechanism promoting allopatric diversification in holoplanktonic taxa, despite the high potential dispersal abilities and pelagic habitat for these species.

Differentiation of white-footed mice (Peromyscus leucopus) and deer mice (Peromyscus maniculatus) of the Upper Midwest using PCR melt curve analysis

Cryptic species pairs including white-footed mice, Peromyscus leucopus, and deer mice, Peromyscus maniculatus, are difficult to identify using morphological and laboratory techniques. To improve rapid and accurate determination of these species, we developed an assay reliant on differences in PCR-product melt curves (MCA). We differentiated 235 P. maniculatus gracilis and 78 P. leucopus using our assay. We compared this to morphological identification using ear length, which also provided reliable species identification. We find that MCA accurately differentiates these species and provides more reliable identifications than certain morphological measures, therefore making our approach ideal for identification of these species.

Ascidian Mitogenomics: Comparison of Evolutionary Rates in Closely Related Taxa Provides Evidence of Ongoing Speciation Events

Ascidians are a fascinating group of filter-feeding marine chordates characterized by rapid evolution of both sequences and structure of their nuclear and mitochondrial genomes. Moreover, they include several model organisms used to investigate complex biological processes in chordates. To study the evolutionary dynamics of ascidians at short phylogenetic distances, we sequenced 13 new mitogenomes and analyzed them, together with 15 other available mitogenomes, using a novel approach involving detailed whole-mitogenome comparisons of conspecific and congeneric pairs. The evolutionary rate was quite homogeneous at both intraspecific and congeneric level, and the lowest congeneric rates were found in cryptic (morphologically undistinguishable) and in morphologically very similar species pairs. Moreover, congeneric nonsynonymous rates (dN) were up to two orders of magnitude higher than in intraspecies pairs. Overall, a clear-cut gap sets apart conspecific from congeneric pairs. These evolutionary peculiarities allowed easily identifying an extraordinary intraspecific variability in the model ascidian Botryllus schlosseri, where most pairs show a dN value between that observed at intraspecies and congeneric level, yet consistently lower than that of the Ciona intestinalis cryptic species pair. These data suggest ongoing speciation events producing genetically distinct B. schlosseri entities. Remarkably, these ongoing speciation events were undetectable by the cox1 barcode fragment, demonstrating that, at low phylogenetic distances, the whole mitogenome has a higher resolving power than cox1. Our study shows that whole-mitogenome comparative analyses, performed on a suitable sample of congeneric and intraspecies pairs, may allow detecting not only cryptic species but also ongoing speciation events.