Phylogeny and cryptic diversity in geckos (Phyllopezus; Phyllodactylidae; Gekkota) from South America's open biomes

The gecko genus Phyllopezus occurs across South America's open biomes: Cerrado, Seasonally Dry Tropical Forests (SDTF, including Caatinga), and Chaco. We generated a multi-gene dataset and estimated phylogenetic relationships among described Phyllopezus taxa and related species. We included exemplars from both described Phyllopezus pollicaris subspecies, P. p. pollicaris and P. p.przewalskii. Phylogenies from the concatenated data as well as species trees constructed from individual gene trees were largely congruent. All phylogeny reconstruction methods showed Bogertia lutzae as the sister species of Phyllopezus maranjonensis, rendering Phyllopezus paraphyletic. We synonymized the monotypic genus Bogertia with Phyllopezus to maintain a taxonomy that is isomorphic with phylogenetic history. We recovered multiple, deeply divergent, cryptic lineages within P. pollicaris. These cryptic lineages possessed mtDNA distances equivalent to distances among other gekkotan sister taxa. Described P. pollicaris subspecies are not reciprocally monophyletic and current subspecific taxonomy does not accurately reflect evolutionary relationships among cryptic lineages. We highlight the conservation significance of these results in light of the ongoing habitat loss in South America's open biomes.     

Bermuda as an Evolutionary Life Raft for an Ancient Lineage of Endangered Lizards

Oceanic islands are well known for harboring diverse species assemblages and are frequently the basis of research on adaptive radiation and neoendemism. However, a commonly overlooked role of some islands is their function in preserving ancient lineages that have become extinct everywhere else (paleoendemism). The island archipelago of Bermuda is home to a single species of extant terrestrial vertebrate, the endemic skink Plestiodon (formerly Eumeces) longirostris. The presence of this species is surprising because Bermuda is an isolated, relatively young oceanic island approximately 1000 km from the eastern United States. Here, we apply Bayesian phylogenetic analyses using a relaxed molecular clock to demonstrate that the island of Bermuda, although no older than two million years, is home to the only extant representative of one of the earliest mainland North American Plestiodon lineages, which diverged from its closest living relatives 11.5 to 19.8 million years ago. This implies that, within a short geological time frame, mainland North American ancestors of P. longirostris colonized the recently emergent Bermuda and the entire lineage subsequently vanished from the mainland. Thus, our analyses reveal that Bermuda is an example of a “life raft” preserving millions of years of unique evolutionary history, now at the brink of extinction. Threats such as habitat destruction, littering, and non-native species have severely reduced the population size of this highly endangered lizard.     

Detecting cryptic species in sympatry and allopatry: analysis of hidden diversity in Polyommatus (Agrodiaetus) butterflies (Lepidoptera: Lycaenidae)

Modern multilocus molecular techniques are a powerful tool in the detection and analysis of cryptic taxa. However, its shortcoming is that with allopatric populations it reveals phylogenetic lineages, not biological species. The increasing power of coalescent multilocus analysis leads to the situation in which nearly every geographically isolated or semi‐isolated population can be identified as a lineage and therefore raised to species rank. It leads to artificial taxonomic inflation and as a consequence creates an unnecessary burden on the conservation of biodiversity. To solve this problem, we suggest combining modern lineage delimitation techniques with the biological species concept. We discuss several explicit principles on how genetic markers can be used to detect cryptic entities that have properties of biological species (i.e. of actually or potentially reproductively isolated taxa). Using these principles we rearranged the taxonomy of the butterfly species close to Polyommatus (Agrodiaetus) ripartii. The subgenus Agrodiaetus is a model system in evolutionary research, but its taxonomy is poorly elaborated because, as a rule, most of its species are morphologically poorly differentiated. The taxon P. (A.) valiabadi has been supposed to be one of the few exceptions from this rule due to its accurately distinguishable wing pattern. We discovered that in fact traditionally recognized P. valiabadi is a triplet of cryptic species, strongly differentiated by their karyotypes and mitochondrial haplotypes.     

The Role of Integrative Taxonomy in the Conservation Management of Cryptic Species: The Taxonomic Status of Endangered Earless Dragons (Agamidae: Tympanocryptis) in the Grasslands of Queensland,Australia

Molecular phylogenetics is increasingly highlighting the prevalence of cryptic species, where morphologically similar organisms have long independent evolutionary histories. When such cryptic species are known to be declining in numbers and are at risk of extinction due to a range of threatening processes, the disjunction between molecular systematics research and conservation policy becomes a significant problem. We investigate the taxonomic status of Tympanocryptis populations in Queensland, which have previously been assigned to T. tetraporophora, using three species delimitation approaches. The taxonomic uncertainties in this species-group are of particular importance in the Darling Downs Earless Dragon (T. cf. tetraporophora), which is ranked as an endangered ‘species’ of high priority for conservation by the Queensland Department of Environment and Heritage Protection. We undertook a morphological study, integrated with a comprehensive genetic study and species delimitation analyses, to investigate the species status of populations in the region. Phylogenetic analyses of two gene regions (mtDNA: ND2; nuclear: RAG1) revealed high levels of genetic divergence between populations, indicating isolation over long evolutionary time frames, and strongly supporting two independent evolutionary lineages in southeastern Queensland, from the Darling Downs, and a third in the Gulf Region of northern Queensland. Of the three species delimitation protocols used, we found integrative taxonomy the most applicable to this cryptic species complex. Our study demonstrates the utility of integrative taxonomy as a species delimitation approach in cryptic complexes of species with conservation significance, where limited numbers of specimens are available.     

Delimitation despite discordance: Evaluating the species limits of a confounding species complex in the face of mitonuclear discordance

The delimitation of species is an essential pursuit of biology, and proper taxonomies are crucial for the assessment and conservation management of organismal diversity. However, delimiting species can be hindered by a number of factors including highly conserved morphologies (e.g., cryptic species), differences in criteria of species concepts, lineages being in the early stages of the speciation or divergence process, and discordance between gene topologies (e.g., mitonuclear discordance). Here we use a taxonomically confounded species complex of toads in Central America that exhibits extensive mitonuclear discordance to test delimitation hypotheses. Our investigation integrates mitochondrial sequences, nuclear SNPs, morphology, and macroecological data to determine which taxonomy best explains the divergence and evolutionary relationships among these toads. We found that a three species taxonomy following the distributions of the nuclear SNP haplotypes offers the best explanation of the species in this complex based off of the integrated data types. Due to the taxonomic instability of this group, we also discuss conservation concerns in the face of improper taxonomic delimitation. Our study provides an empirical and integrative hypothesis testing framework to assess species delimitation hypotheses in the face of cryptic morphology and mitonuclear discordance and highlights the importance that a stable taxonomy has over conservation‐related actions.     

Mitogenomic insights into a recently described and rarely observed killer whale morphotype

Identifying evolutionary divergent taxonomic units, e.g. species and subspecies, is important for conservation and evolutionary biology. The ‘type D’ killer whale, Orcinus orca, is a rarely observed morphotype with a pelagic, circumpolar subantarctic distribution, making dedicated research and therefore taxonomic study extremely difficult to date. In this study, we used DNA target enrichment hybridisation capture coupled to high throughput sequencing, to obtain the first DNA sequence from the only known museum specimen of this recently described morphotype. The high coverage, complete mitogenome sequence was compared to a previously published global dataset of 139 individuals, indicating that this type is highly divergent to all previously genetically sequenced killer whale forms. The estimated divergence time (390,000 years ago) from its most recent common ancestor with other extant killer whale lineages was the second oldest split within the killer whale phylogeny. This study provides the first genetic support of type D potentially being a distinct subspecies or species of killer whale, although further samples are needed to identify whether there is monophyly of mitogenome sequences and whether nuclear DNA also indicates reproductive isolation. These findings also highlight the value of natural history museum collections and new technologies to investigate the taxonomy of rare, cryptic or difficult to access species.     

Systematic revision of mountain dragons (Reptilia: Agamidae: Diploderma) in China,with descriptions of six new species and discussion on their conservation

Cryptic diversity not only introduces confusion to taxonomic studies, but it also poses major challenges to conservation and environmental legislation. One such troubling group are the Mountain Dragons in the genus Diploderma in Southwest China. Previous studies have suggested that the genus contains considerable cryptic diversity, particularly in the D. flaviceps complex. Owing to taxonomic confusion, micro‐endemic lineages are still neglected by the Chinese wildlife protection laws, despite their urgent conservation needs. Combining multivariate morphological and multi‐locus phylogenetic data, we provide the first integrative systematic revision of the genus Diploderma. Specifically, we confirm that the six examined populations of D. cf. flaviceps from the upper Jinsha and Yalong River Valleys in Sichuan and Yunnan Provinces represent six cryptic, undescribed lineages, and we describe each of them as a new species. With the updated taxonomy and distribution information, we discuss the taxonomy of the D. flaviceps complex in Southwest China, provide an updated diagnostic key along with distributional ranges for all species of the genus, and discuss some of the suspicious records of other congeners in China. Lastly, we evaluate the IUCN status of each of the six new species and highlight the major challenges for Diploderma conservation in China due to delayed environmental legislation and misleading conservation assessments.     

‘Old-forest fungi’ are not always what they seem: the case of Antrodia crassa

Putatively old-forest dependent fungi have acquired a distinct position in biodiversity conservation, acting both as management targets and indicators for other nature values in the forest. Establishing old-forest dependency, however, requires rigorous sampling, taxonomy and identification approaches as exemplified here by a case study on Antrodia crassa. This rarely recorded polypore is considered indicative of old conifer forests in Northern Europe. We demonstrate that the material labelled as A. crassa includes not only frequent misidentifications, but it represents two genetically and ecologically distinct cryptic lineages with partly overlapping distributions. A balanced survey of different successional stages in Estonia indicated that one of those lineages prefers deadwood-rich disturbed habitats, notably clear-cuts, which have been previously undersampled. Because similar problems with cryptic lineages and biased surveys are common in mycological research, lists of ‘old-forest fungi’ should be considered as high-priority working hypotheses for taxonomic and ecological research, rather than well-established management tools.     

Non‐monophyly of Bostrychia simpliciuscula (Ceramiales,Rhodophyta): Multiple species with very similar morphologies,a revised taxonomy of cryptic species

The discovery of a plethora of cryptic species in many algal groups has led to speculation as to the causes of this observation and has affected taxonomy, with reluctance to give names to species that look identical. While this is defensible for monophyletic cryptic species complexes, both our understanding of similar morphologies (crypsis) and nomenclature is challenged when we encounter non‐monophyletic ‘cryptic’ species. Bostrychia simpliciuscula is a wide‐ranging species in which multiple cryptic species are known. Our increased sampling shows that this species consists of four lineages that do not form a clade, but lineages are sister to species with different morphologies. Careful morphological examination shows that characters, especially branched monosiphonous laterals and rhizoid morphology in haptera, are able to distinguish these four lineages into two groups, that are still not monophyletic. The similar morphologies in these lineages could be due to convergence, but not developmental constraints or lack of time to diverge morphologically; or possibly maintenance of a generalized body plan. These lineages appear to have specific biogeographic patterns and these will be used to propose a new taxonomy. B. simpliciuscula is now confined to the tropics. Another of these lineages matches a previously described species, B. tenuissima, that was synonymized with B. simpliciuscula and is from cold temperate Australasia, and is resurrected. Another lineage is found in Japan in which a previous name is also available, B. hamana‐tokidae; the last lineage is found in central New South Wales, morphologically it resembles B. tenuissima, with which it overlaps in distribution around Sydney, and is named as a new species, B. kingii sp. nov.     

Systematics of the grey mullets (Teleostei: Mugiliformes: Mugilidae): molecular phylogenetic evidence challenges two centuries of morphology-based taxonomy

The family Mugilidae comprises mainly coastal marine species that are widely distributed in all tropical, subtropical and temperate seas. Mugilid species are generally considered to be ecologically important and they are a major food resource for human populations in certain parts of the world. The taxonomy and systematics of the Mugilidae are still much debated and based primarily on morphological characters. In this study, we provide the first comprehensive molecular systematic account of the Mugilidae using phylogenetic analyses of nucleotide sequence variation at three mitochondrial loci (16S rRNA, cytochrome oxidase I, and cytochrome b) for 257 individuals from 55 currently recognized species. The study covers all 20 mugilid genera currently recognized as being valid. The family comprises seven major lineages that radiated early on from the ancestor to all current forms. All genera that were represented by two species or more, except Cestraeus, turned out to be paraphyletic or polyphyletic. Thus, the present phylogenetic results generally disagree with the current taxonomy at the genus level and imply that the anatomical characters used for the systematics of the Mugilidae may be poorly informative phylogenetically. The present results should provide a sound basis for a taxonomic revision of the mugilid genera. A proportion of the species with large distribution ranges (including Moolgarda seheli, Mugil cephalus and M. curema) appear to consist of cryptic species, thus warranting further taxonomic and genetic work at the infra-generic level.     

Possible links between phenotypic variability,habitats and connectivity in the killifish Aphaniops stoliczkanus in Northeast Oman

There is a significant gap in our knowledge of the intraspecific morphological variability in freshwater fish, although such data are crucial for understanding species diversity. Here we use the killifish Aphaniops stoliczkanus (Day, 1872; Aphaniidae: Cyprinodontiformes), which is a widespread but poorly known freshwater species in the Middle East, to investigate variability in morphological traits within and between its populations. As otolith morphology is known to evolve on ecological timescales and can signal the presence of cryptic lineages, a special focus lies on otolith variability. Based on samples from six populations in northern Oman, we found that variation in pigmentation, disparities in body shape and otolith variability can be associated with distinctive environmental conditions. The unique otolith shape of A. stoliczkanus from a hot sulphuric spring (Nakhal) suggests that a cryptic lineage may have emerged there. Our new data can serve as a benchmark for future studies on the diversity of Aphaniops and other Aphaniidae and help to clarify whether cryptic diversity is present in some lineages. Moreover, our data can serve as an actualistic model for studies on fossil fishes, in which morphological characters provide the only accessible data source for taxonomic and phylogenetic interpretations.     

Genome wide AFLP markers support cryptic species in Coniophora (Boletales)

Numerous fungal morphospecies include cryptic species that routinely are detected by sequencing a few unlinked DNA loci. However, whether the patterns observed by multi-locus sequencing are compatible with genome wide data, such as amplified fragment length polymorphisms (AFLPs), is not well known for fungi. In this study we compared the ability of three DNA loci and AFLP data to discern between cryptic fungal lineages in the three morphospecies Coniophora olivacea, Coniophora arida, and Coniophora puteana. The sequences and the AFLP data were highly congruent in delimiting the morphotaxa into multiple cryptic species. However, while the DNA sequences indicated introgression or hybridization between some of the cryptic lineages the AFLP data did not. We conclude that as few as three polymorphic DNA loci was sufficient to recognize cryptic lineages within the studied Coniophora taxa. However, based on analyses of a few (three) sequenced loci the hybridization could not easily be distinguished from incomplete lineage sorting. Hence, great caution should be taken when concluding about hybridization based on data from just a few loci.     

Alpine lichen diversity in an isolated sky island in the Colorado Plateau,USA—Insight from an integrative biodiversity inventory

Lichens are major components of high altitude/latitude ecosystems. However, accurately characterizing their biodiversity is challenging because these regions and habitats are often underexplored, there are numerous poorly known taxonomic groups, and morphological variation in extreme environments can yield conflicting interpretations. Using an iterative taxonomic approach based on over 800 specimens and incorporating both traditional morphology‐based identifications and information from the standard fungal DNA barcoding marker, we compiled a voucher‐based inventory of biodiversity of lichen‐forming fungi in a geographically limited and vulnerable alpine community in an isolated sky island in the Colorado Plateau, USA—the La Sal Mountains. We used the newly proposed Assemble Species by Automatic Partitioning (ASAP) approach to empirically delimit candidate species‐level lineages from family‐level multiple sequence alignments. Specimens comprising DNA‐based candidate species were evaluated using traditional taxonomically diagnostic phenotypic characters to identify specimens to integrative species hypotheses and link these, where possible, to currently described species. Despite the limited alpine habitat (ca. 3,250 ha), we document the most diverse alpine lichen community known to date from the southern Rocky Mountains, with up to 240 candidate species/species‐level lineages of lichen‐forming fungi. 139 species were inferred using integrative taxonomy, plus an additional 52 candidate species within 29 different putative species complexes. Over 68% of sequences could not be assigned to species‐level rank with statistical confidence, corroborating the limited utility of current sequence repositories for species‐level DNA barcoding of lichen‐forming fungi. By integrating vouchered specimens, DNA sequence data, and photographic documentation, we provide an important baseline of lichen‐forming fungal diversity for the limited alpine habitat in the Colorado Plateau. These data provide an important resource for subsequent research in the ecology and evolution of lichens alpine habitats, including DNA barcodes for most putative species/species‐level lineages occurring in the La Sal Mountains, and vouchered collections representing any potentially undescribed species that can be used for future taxonomic studies.     

COI barcoding of Nebelid testate amoebae (Amoebozoa: Arcellinida): extensive cryptic diversity and redefinition of the Hyalospheniidae Schultze

We used Cytochrome Oxidase Subunit 1 (COI) to assess the phylogenetic relationships and taxonomy of Nebela sensu stricto and similar taxa (Nebela group, Arcellinida) in order to clarify the taxonomic validity of morphological characters. The COI data not only successfully separated all studied morphospecies but also revealed the existence of several potential cryptic species. The taxonomic implications of the results are: (1) Genus Nebela is paraphyletic and will need to be split into at least two monophyletic assemblages when taxon sampling is further expanded. (2) Genus Quadrulella, one of the few arcellinid genera building its shell from self-secreted siliceous elements, and the mixotrophic Hyalosphenia papilio branch within the Nebela group in agreement with the general morphology of their shell and the presence of an organic rim around the aperture (synapomorphy for Hyalospheniidae). We thus synonymise Hyalospheniidae and Nebelidae. Hyalospheniidae takes precedence and now includes Hyalosphenia, Quadrulella (previously in the Lesquereusiidae) and all Nebelidae with the exception of Argynnia and Physochila. Leptochlamys is Arcellinida incertae sedis. We describe a new genus Padaungiella Lara et Todorov and a new species Nebela meisterfeldi n. sp. Heger et Mitchell and revise the taxonomic position (and rank) of several taxa. These results show that the traditional morphology-based taxonomy underestimates the diversity within the Nebela group, and that phylogenetic relationships are best inferred from shell shape rather than from the material used to build the shell.     

Mating trials validate the use of DNA barcoding to reveal cryptic speciation of a marine bryozoan taxon

Despite increasing threats to the marine environment, only a fraction of the biodiversity of the oceans has been described, owing in part to the widespread occurrence of cryptic species. DNA-based barcoding through screening of an orthologous reference gene has been proposed as a powerful tool to uncover biological diversity in the face of dwindling taxonomic expertise and the limitations of traditional species identification. Although DNA barcoding should be particularly useful in the sea, given the prevalence of marine cryptic species, the link between taxa identified through DNA barcodes and reproductively isolated taxa (biological species) has rarely been explicitly tested. Here, we use an integrated framework comparing breeding compatibility, morphology and mitochondrial (cytochrome c oxidase 1) and nuclear (elongation factor-1-alpha) DNA sequence variation among globally distributed samples of the cosmopolitan marine bryozoan Celleporella hyalina (L.). Our results reveal that C. hyalina comprises numerous deep, mostly allopatric, genetic lineages that are reproductively isolated, yet share very similar morphology, indicating rampant cryptic speciation. The close correspondence between genetic lineages and reproductively isolated taxa in the context of minimal morphological change suggests that DNA barcoding will play a leading role in uncovering the hidden biodiversity of the oceans and that the sole use of morphologically based taxonomy would grossly underestimate the number of marine species.     

The relative roles of adaptation and phylogeny in determination of larval traits in diversifying anuran lineages

I measured phenotypic traits important to the fitness of larval anurans to assess the relative roles of ancestral trait value and selective regime in determining present-day phenotypes. The positions of 14 species from three taxonomic families and three different habitats in a phenotypic space defined by 19 traits provided measures of taxonomic and ecological similarity. The distribution of phenotypic distances among species revealed that neither taxonomy nor habitat overwhelmingly determined phenotype. There appear to be multiple ways in which anurans can exploit pond types. However, the direction of phenotypic movement was not random from one species to the next. Independent contrasts revealed significant correlations in the evolution of traits that were consistent among lineages. These correlations reflected well-known trade-offs that result from functional relationships among the constituent traits. Although there is no simple pattern in the distribution of mean phenotypes across environments and lineages, the pattern of the evolutionary trajectories that created that distribution is consistent with a predictive theory of multivariate evolution.     

Integrative taxonomy and conservation of cryptic beetles in the Mediterranean region (Hydrophilidae)

Arribas, P., Andújar, C., Sánchez‐Fernández, D., Abellán, P. & Millán, A. (2012). Integrative taxonomy and conservation of cryptic beetles in the Mediterranean region (Hydrophilidae). —Zoologica Scripta, 00, 000–000. Because biodiversity inventory forms the basis for the effective conservation of species and habitats, there is an imperative need to discover and describe new species. A significant part of presently unknown biodiversity is constituted by cryptic species complexes, where traditional taxonomy usually fails due to a lack of clear taxonomic characters in the external structures of specimens. Integrative taxonomy offers a powerful tool to shed light on this part of encrypted biodiversity, combining multiple operational criteria in an evolutionary context in order to delineate species boundaries. The present study used an integrative approach to explore the species boundaries in a water beetle complex (Enochrus falcarius species complex) inhabiting saline streams, a rare and threatened habitat across the Mediterranean region. First, hypotheses about the candidate species on the basis of phylogenetic analyses and biogeographical information were proposed. Second, lineage divergence was evaluated between candidate species using (i) molecular cluster delimitation, (ii) morphometry (both linear body morphometrics and pronotum outlines) and (iii) ecological niche similarity estimates. We found divergence between candidate species on the basis of molecular, biogeographical and niche information, and consequently, four species were delimited within the E. falcarius complex (i.e. Enochrus jesusarribasi sp. n., Enochrus blazquezae sp. n., Enochrus risii sp. n. and Enochrus falcarius), despite the fact that they showed high morphological similarity. Enochrus falcarius, as previously considered, had not been proposed to be of conservational concern, because until now, it had been regarded as a single broadly distributed species in the Mediterranean region. However, the four entities here delimited within this species complex displayed characteristics that categorised them as vulnerable taxa. Hence, these results show how applying integrative taxonomy approaches and rapid vulnerability assessments to lineages from threatened habitats with the potential to comprise cryptic diversity could become a fundamental tool for biodiversity conservation, driving the discovery of cryptic species and consequently the modification of previous, inadequately evaluated vulnerability categorisations.     

The niche of higher plants: evidence for phylogenetic conservatism.

A species' ecological niche depends on the species' adaptations to its present habitat, but also on the legacy from its ancestors. Most authors argue that such a phylogenetic niche conservatism is of minor importance, although no quantitative analyses across a major taxon is available. Higher plants from central Europe offer a unique opportunity for such an exercise, as the niche positions along various environmental gradients are available for most species. We quantified niche conservatism by two approaches. First, we used a phylogenetic tree and quantified the degree of retention of niches across the tree. Depending on the gradient, the values ranged from 0.43 to 0.22. This was significantly greater than the null expectation. Second, we used a taxonomy and quantified the amount of variance among species that could be explained at higher taxonomic levels. The values ranged from 25 to 72%. Again, this was significantly higher than the null expectation. Thus, both approaches indicated a clear niche conservatism. The distribution of conservatism across taxonomic levels differed considerably among environmental gradients. The differences among environmental gradients could be correlated with the palaeoenvironmental conditions during the radiation of the phylogenetic lineages. Thus, niche conservatism among extant plant species may reflect the opportunities of their ancestors during their diversification.