Cryptic Sister Species Research Articles

A revision of the genus Wrangelia (Wrangeliaceae, Ceramiales) in Bermuda resolves six new species including W. ryancraigii from the mesophotic zone.

Four species of the genus Wrangelia are presently known from the western Atlantic Ocean: W. argus, W. bicuspidata, W. penicillata, and W. gordoniae, with the first three historically being reported from Bermuda. Morphological and molecular barcode (COI-5P) and phylogenetic analyses used in this study (SSU, LSU, rbcL) indicated eight species groupings of Wrangelia in Bermuda, excluding two of the historically recognized species, retaining only W. argus while adding seven new species, of which six are formally described. What had been historically reported as W. penicillata from Bermuda was shown to be distinct from Mediterranean Sea specimens (type locality) and was shown to be a mixture of W. hesperia sp. nov. and W. incrassata sp. nov. Along with these two, three other new species (W. laxa sp. nov., W. ryancraigii sp. nov., and W. secundiramea sp. nov.) have complete rhizoidal cortication tightly covering axial cells of indeterminate axes below the apices, distinguishing them from the two local incompletely corticated congeners W.argus and W. abscondita sp. nov., the latter a morphologically cryptic sister species with W. bicuspidata from the Caribbean Sea. Only one of the new species, W. ryancraigii, has thus far been observed in the mesophotic zone off the Bermuda platform, and it is morphologically cryptic with the euphotic zone's W. laxa.

Biogeography, speciation and niche evolution of doraditos (Aves: Pseudocolopteryx)

AbstractEcological and geographical factors shape the current distribution of species. Analysing their interplay in a phylogenetic framework is key to understand the historical processes that have shaped the evolution of a group. Here, we modelled the ecological niches and geographic distributions of the five species of doraditos (Pseudocolopteryx spp.) to study their biogeographic histories, niche evolution and speciation process in a phylogenetic framework. Our potential distribution models uncovered novel range‐wide distributional patterns and seasonal movements in the doraditos, where four species are migratory with distinct breeding and non‐breeding distributions, and one (P. sclateri) exhibits a complex spatiotemporal distribution indicating nomadism. Ecological niche pairwise comparisons showed that none of the doraditos have equivalent niches and that niche differences are due to species‐specific habitat preferences. Phylogenetically weighted geographical and ecological analyses showed patterns of allopatric speciation and niche lability in the evolution of doraditos. The divergence of P. sclateri seems tied to its tropical‐to‐temperate wetland specialization. The montane P. acutipennis expanded to human‐modified lowlands following speciation, highlighting the need to control for post‐speciational changes in ecological niche comparisons as done here. In turn, P. dinelliana, P. citreola and P. flaviventris showed essentially allopatric breeding distributions, as a product of environmentally mediated divergence during their speciation processes. The distribution and migration data of the recently diverged cryptic sister species P. citreola and P. flaviventris are consistent with two possible speciation scenarios: peripatric speciation and migration dosing speciation.

Taxonomic revision of Tinocladia (Ectocarpales s.l., Phaeophyceae): merger of Tinocladia with Eudesme and description of E. pseudocrassa sp. nov

ABSTRACT A molecular phylogeny of Tinocladia and Eudesme based on specimens covering a large proportion of the known species was done using mitochondrial cox1 and cox3, chloroplast atpB, psaA, psbA and rbcL genes and 5.8S rDNA and its ITS2 region sequences. The phylogeny revealed a close phylogenetic relationship between the two genera and the occurrence of a cryptic species within the generitype T. crassa. Eudesme species (E. borealis, E. shandongensis and the generitype E. virescens) showed a sister relationship with the northern hemisphere Tinocladia (T. crassa, including a cryptic sister species T. pseudocrassa sp. nov., and T. sanrikuensis), whereas southern hemisphere Tinocladia (T. australis, T. falklandica, T. novae-zelandiae, and a cryptic species from New Zealand) nested in the Eudesme clade. Morphologically, Tinocladia species have been distinguished from Eudesme by their denser medullary filaments and more extensive subcortical layer, but our molecular phylogenetic analyses suggest that these features do not correlate with genetic differences. The cox3 sequence divergence between northern hemisphere Tinocladia and the lineage that includes Eudesme and southern hemisphere Tinocladia was comparable to or smaller than those within selected ectocarpalean genera. We therefore propose to merge Tinocladia with Eudesme and resurrect Eudesme crassa (Suringar) Okamura for T. crassa. Although the two independent lineages of T. crassa do not show marked morphological differences, they are genetically isolated even in sympatric populations. We therefore propose to treat them as independent species. In this paper we neotypify Tinocladia crassa (=Eudesme crassa), describe E. pseudocrassa sp. nov., and determine that the Californian population of E. crassa is an introduction from Northeast Asia. HIGHLIGHTS •A taxonomic revision of Eudesme and Tinocladia, based on genetic and morpho-anatomy, provides evidence to merge Tinocladia with Eudesme. •The finding of cryptic species in New Zealand highlights the need for further taxonomic studies in this region. •The genetic data have revealed the occurrence of a cryptic species within T. crassa in Japan.

A Polyphasic Approach including Whole Genome Sequencing Reveals Paecilomyces paravariotii sp. nov. as a Cryptic Sister Species to P. variotii.

Whole genome sequencing is rapidly increasing phylogenetic resolution across many groups of fungi. To improve sequencing coverage in the genus Paecilomyces (Eurotiales), we report nine new Paecilomyces genomes representing five different species. Phylogenetic comparison between these genomes and those reported previously showed that Paecilomyces paravariotii is a distinct species from its close relative P. variotii. The independence of P. paravariotii is supported by analysis of overall gene identify (via BLAST), differences in secondary metabolism and an inability to form ascomata when paired with a fertile P. variotii strain of opposite mating type. Furthermore, whole genome sequencing resolves the P. formosus clade into three separate species, one of which lacked a valid name that is now provided.

First report of black root rot (Berkeleyomyces rouxiae) on greenhouse hemp (Cannabis sativa) in Kentucky.

Hemp, Cannabis sativa, was reintroduced to the US in 2014 after 50 years of prohibition. Since then, growers have focused primarily on cannabinoid production using female plants. Thus, most modern hemp has been asexually propagated in greenhouses and transplanted into fields. In December 2019, a commercial transplant producer in Fayette County, Kentucky reported 30% dieback on 'Seagull' hemp. Plants were potted into artificial media (unknown origin) immediately upon removal from the mist bench, and symptoms developed approximately two weeks afterward. Scattered plants developed stunting, yellowing, and wilting symptoms and then died within one week of symptom development. Roots had brown to black lesions distributed along roots that progressed into necrosis. Aleuriospores (chlamydospores) were visible under the dissecting microscope. Root pieces were surface sterilized with 10% household bleach for 2 min and then plated onto cut centers of ~1 cm thick sterilized carrot cross section (Williamson 2014). Resulting colonies were single-spored and maintained on PDA plates on laboratory benchtops (23°C, fluorescent lights). Three isolates were selected for pathogenicity testing and identification. Resulting colonies initially appeared white but darkened to black within two days following the development of aleuriospores (chlamydospores). Aleuriospores were dark brown, formed in chains of 4 to 5, and measured 10.00 to 14.27 × 5.62 to 11.23 μm (n=50). Endoconidia were hyaline, cylindrical, and measured 11.63 to 34.10 × 3.95 to 5.58 μm (n=50). To confirm pathogenicity, soil was washed from three 4-week-old hemp plants (proprietary cultivar) and roots were soaked in a 1x106 spore/mL conidial suspension for 5 minutes. In addition, the roots of three control plants were mock inoculated using sterile water. Inoculated and control plants were transplanted into sterile potting media and maintained in separate trays. Inoculated plants developed leaf yellowing and wilting symptoms 14 dai. Roots examined 21 dai had scattered brown lesions throughout and were colonized by the fungus. Aleuriospores and endoconidia isolated from the symptomatic roots were identical to the inoculum. Control plants did not develop symptoms after 21 dai. For molecular analyses, DNA was extracted from an actively growing (5 d) representative isolate (19FY005) on PDA using Quick-DNA™ Fungal/Bacterial Miniprep Kit (Zymo Research, Irvine, CA). Fragments of commonly used diagnostic loci ITS, ACT, RPBII and Mcm7 were amplified as described by de Beer et al (2014), Fourie et al (2015) and Duong et al (2012). Amplicons were sequenced by Sanger sequencing and the consensus sequences were compared with the NCBI GenBank database by BLASTn. ACT amplicon (OK135163) top hit was B. rouxiae (MF967149.1) with 100% similarity, Mcm7 (OK135165) top hit was B. rouxiae (MF967103.1) with 100% similarity, RPBII (OK135166) top hit was B. rouxiae (MF967194.1) with 99.80% similarity, and ITS (OK135164) top hit was B. rouxiae (MF952402.1) with 99.82% similarity. Berkeleyomyces spp (syn Thielaviopsis sp.) is the causal agent of black root rot in field crops such as cotton and tobacco and in ornamentals such as holly and pansy; it is also a common disease in greenhouse production systems. Introduction of a cryptic sister species in 2017 may suggest that reports of black root rot in many crops may be either B. basicola or B. rouxiae, both species are indistinguishable by morphology (Nel et al. 2017). Although both species have wide host ranges with no host specificity, speciation may be important in terms of pathogenicity, host susceptibility, and other cultural factors. As hemp acreage increases across the US, black root rot may become a more prominent disease in greenhouse transplant production systems and in fields.

Cryptic Diversity in Paramecium multimicronucleatum Revealed with a Polyphasic Approach.

Paramecium (Ciliophora) systematics is well studied, and about twenty morphological species have been described. The morphological species may include several genetic species. However, molecular phylogenetic analyses revealed that the species diversity within Paramecium could be even higher and has raised a problem of cryptic species whose statuses remain uncertain. In the present study, we provide the morphological and molecular characterization of two novel Paramecium species. While Paramecium lynni n. sp., although morphologically similar to P. multimicronucleatum, is phylogenetically well separated from all other Paramecium species, Paramecium fokini n. sp. appears to be a cryptic sister species to P. multimicronucleatum. The latter two species can be distinguished only by molecular methods. The number and structure of micronuclei, traditionally utilized to discriminate species in Paramecium, vary not only between but also within each of the three studied species and, thus, cannot be considered a reliable feature for species identification. The geographic distribution of the P. multimicronucleatum and P. fokini n. sp. strains do not show defined patterns, still leaving space for a role of the geographic factor in initial speciation in Paramecium. Future findings of new Paramecium species can be predicted from the molecular data, while morphological characteristics appear to be unstable and overlapping at least in some species.

Genome-wide markers reveal differentiation between and within the cryptic sister species, sunset and vermilion rockfish

The vermilion rockfish complex, which consists of the cryptic sister species vermilion and sunset rockfish, is one of the most valuable recreational fisheries on the U.S. West Coast. These species are currently managed as a single complex, and because of uncertainty surrounding the relative contribution of each species within existing data sources, the stock status of each species is not fully known. A reliable and cost-effective method is needed to disentangle these species that will allow for the development of abundance indices, life history profiles, and catch histories that may potentially support species-specific stock assessments. Using restriction-site associated DNA sequence (RADseq) markers we generated 10,003 polymorphic loci to characterize the vermilion rockfish complex. PCA and Bayesian clustering approaches based on these loci clearly distinguished between sunset and vermilion rockfishes and identified hybrid individuals. These loci included 203 highly differentiated (FST ≥ 0.99) single nucleotide polymorphisms, which we consider candidates in the planned development of a diagnostic assay capable of distinguishing between these cryptic species. In addition to clearly delineating to species, subsets of the interspecific markers allowed for insight into intraspecific differentiation in both species. Population genetic analyses for sunset rockfish identified two weakly divergent genetic groups with similar levels of genetic diversity. Vermilion rockfish, however, were characterized by three distinct genetic groups with much stronger signals of differentiation and significantly different genetic diversities. Collectively, these data will contribute to well-informed, species-specific management strategies to protect this valuable species complex.

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.

Little parallelism in genomic signatures of local adaptation in two sympatric, cryptic sister species.

Species living in sympatry and sharing a similar niche often express parallel phenotypes as a response to similar selection pressures. The degree of parallelism within underlying genomic levels is often unexplored, but can give insight into the mechanisms of natural selection and adaptation. Here, we use multi-dimensional genomic associations to assess the basis of local and climate adaptation in two sympatric, cryptic Crematogaster levior ant species along a climate gradient. Additionally, we investigate the genomic basis of chemical communication in both species. Communication in insects is mainly mediated by cuticular hydrocarbons (CHCs), which also protect against water loss and, hence, are subject to changes via environmental acclimation or adaptation. The combination of environmental and chemical association analyses based on genome-wide Pool-Seq data allowed us to identify single nucleotide polymorphisms (SNPs) associated with climate and with chemical differences. Within species, CHC changes as a response to climate seem to be driven by phenotypic plasticity, since there is no overlap between climate- and CHC-associated SNPs. The only exception is the odorant receptor OR22c, which may be a candidate for population-specific CHC recognition in one of the species. Within both species, climate is significantly correlated with CHC differences, as well as to allele frequency differences. However, associated candidate SNPs, genes and functions are largely species-specific and we find evidence for minimal parallel evolution only on the level of genomic regions (J=0.04). This highlights that even closely related species may follow divergent evolutionary trajectories when expressing similar adaptive phenotypes.

The taxonomic position of Asian Holopedium (Crustacea: Cladocera) confirmed by morphological and genetic analyses

The cladoceran Holopedium gibberum Zaddach, 1855 (Ctenopoda: Holopediidae) was once thought to occur broadly in the northern hemisphere, but its cryptic sister species was recently separated from H. gibberum sensu stricto (s.s.) as a new species, Holopedium glacialis. In East Asia, although “H. gibberum” occurrence has been recorded in many water bodies, the identity of the surveyed populations has rarely been confirmed via molecular analyses. Thus, it is unclear whether it is actually H. gibberum s.s. or H. glacialis that is distributed in East Asia. We used DNA-barcoding techniques to check the taxonomic status of Holopedium samples collected in Japan. We sequenced mitochondrial cytochrome c oxidase subunit 1 (mtCOI) and nuclear 18S ribosomal DNA (nr18S) of Japanese Holopedium and compared the results with those of H. gibberum s.s. collected in Norway and H. glacialis collected in Canada. The mtCOI sequence divergences between Norwegian H. gibberum s.s. and Japanese Holopedium were at most 2.4%, which was within the degree of intraspecific differentiation in cladocerans. Norwegian H. gibberum s.s. and Japanese Holopedium shared identical nr18S haplotypes. Individuals of Canadian H. glacialis were genetically different from those of Japanese Holopedium. We therefore concluded that Japanese Holopedium can be identified as H. gibberum s.s.

Scolopsis igcarensis Mishra, Biswas, Russell, Satpathy amp; Selvanayagam, 2013, a junior synonym of S. vosmeri (Bloch, 1792) (Perciformes: Nemipteridae).

Scolopsis igcarensis Mishra, Biswas, Russell, Satpathy Selvanayagam, 2013 was described from specimens collected from coastal waters of southern India and Sri Lanka. A comparison of recently collected specimens from Bangladesh, initially identified as S. igcarensis, with Scolopsis vosmeri (Bloch, 1792) showed morphological differences between the two species are minor, and that specimens of S. igcarensis in fact represent juvenile and subadult colour forms of S. vosmeri. Underwater and aquarium observations, as well as molecular data based on the COI barcode region, support this conclusion. Accordingly, S. igcarensis is regarded as a junior synonym of S. vosmeri, which is redescribed herein. Phylogenetic analysis of COI barcodes of Scolopsis specimens produced in this study, together with those available from GenBank, indicate S. vosmeri is part of a species complex which includes two additional cryptic sister species that require further taxonomic investigation.

A taxonomic reassessment of Caulerpa (Chlorophyta, Caulerpaceae) in southern Australia, based on tufA and rbcL sequence data

ABSTRACTMolecular characterisation (using tufA and rbcL DNA sequences) of newly collected specimens of the genus Caulerpa in southern Australia identified 24 species and one variety, which, with the addition of four species and one variety recorded previously but not found during the present study, brings the total number of Caulerpa taxa in the region to 28 species and two varieties. Three new species are described based on molecular and morphological data: Caulerpa crispata comb. & stat. nov. (for C. longifolia f. crispata), Caulerpa coppejansii sp. nov., and Caulerpa perplexa sp. nov. Caulerpa coppejansii is a tropical West Pacific species and is included here due to its similarity to the southern Australian Caulerpa sedoides. Caulerpa perplexa is found in Western Australia and is a cryptic sister species of C. parvifolia Harvey from eastern Australia. The name Caulerpa muelleri is reinstated for C. flexilis var. muelleri. A new name, Caulerpa lucasii nom. nov., is proposed for Caulerpa annulata Lucas nom. illeg., a taxon shown to be distinct from Caulerpa hodgkinsoniae. Caulerpa corynephora and Caulerpa fergusonii are newly reported from the region, and a population of the potentially invasive Caulerpa cylindracea is reported from Portland, Victoria. An updated nomenclature is provided and reference tufA sequences are proposed, as well as an updated identification key for southern Australian Caulerpa species.

Morphologically cryptic Amazonian bird species pairs exhibit strong postzygotic reproductive isolation.

We possess limited understanding of how speciation unfolds in the most species-rich region of the planet-the Amazon basin. Hybrid zones provide valuable information on the evolution of reproductive isolation, but few studies of Amazonian vertebrate hybrid zones have rigorously examined the genome-wide underpinnings of reproductive isolation. We used genome-wide genetic datasets to show that two deeply diverged, but morphologically cryptic sister species of forest understorey birds show little evidence for prezygotic reproductive isolation, but substantial postzygotic isolation. Patterns of heterozygosity and hybrid index revealed that hybrid classes with heavily recombined genomes are rare and closely match simulations with high levels of selection against hybrids. Genomic and geographical clines exhibit a remarkable similarity across loci in cline centres, and have exceptionally narrow cline widths, suggesting that postzygotic isolation is driven by genetic incompatibilities at many loci, rather than a few loci of strong effect. We propose Amazonian understorey forest birds speciate slowly via gradual accumulation of postzygotic genetic incompatibilities, with prezygotic barriers playing a less important role. Our results suggest old, cryptic Amazonian taxa classified as subspecies could have substantial postzygotic isolation deserving species recognition and that species richness is likely to be substantially underestimated in Amazonia.

A new genus and species for the globally important, multihost root pathogen Thielaviopsis basicola

The plant pathogenic asexual fungus Thielaviopsis basicola (Ascomycota) causes black root rot on many important agricultural and ornamental plant species. Since its first description in 1850, this species has had a tumultuous taxonomic history, being classified in many different genera. Thus far, DNA‐based techniques have not played a significant role in identification of T. basicola and have been used only to confirm its placement in the Microascales. This investigation reconsidered the phylogenetic placement of T. basicola, using DNA sequence data for six different gene regions. It included 41 isolates identified as T. basicola from 13 geographical locations worldwide. Phylogenetic analyses showed that these isolates grouped in a well‐supported lineage distinct from other genera in the Ceratocystidaceae, here described as Berkeleyomyces gen. nov. The data also provided robust evidence that isolates of T. basicola include a cryptic sister species. As a result, this report provides a new combination as B. basicola comb. nov. and introduces a new species as B. rouxiae sp. nov.

Synalpheus pinkfloydi sp. nov., a new pistol shrimp from the tropical eastern Pacific (Decapoda: Alpheidae).

A new, conspicuously coloured species of the alpheid genus Synalpheus Spence Bate, 1888, is described based on material collected on the Pacific coast of Panama. Synalpheus pinkfloydi sp. nov. is closely related to the western Atlantic S. antillensis Coutière, 1909, the two taxa being transisthmian, cryptic sister species. Both species are characterised by the distal areas of their major and minor chelae coloured in an intense, almost glowing pink-red. The morphological differences between S. pinkfloydi sp. nov. and S. antillensis Coutière, 1909 are subtle, being limited to the slightly different proportions of the merus of both chelipeds, distodorsal armature of the major cheliped merus, relative length of the antennal scaphocerite, and body size. However, they are genetically different with a 10.2% sequence divergence in COI. Based on molecular clock estimates, these transisthmian taxa diverged around 6.8-7.8 mya, i.e. well before the final closure of the Isthmus of Panama 2.5-3 mya.

Population genetic structure and hybridization patterns in the cryptic sister species Chironomus riparius and Chironomus piger across differentially polluted freshwater systems

Chironomids are an integral and functionally important part of many freshwater ecosystems. Yet, to date, there is limited understanding of their microevolutionary processes under chemically polluted natural environments.In this study, we investigated the genetic variation within populations of the ecotoxicological model species Chironomus riparius and its cryptic sister species Chironomus piger at 18 metal-contaminated and reference sites in northwestern Portugal. Microsatellite analysis was conducted on 909 samples to answer if metal contamination affects genetic variation in natural chironomid populations as previously suggested from controlled laboratory experiments.Similarly high levels of genetic diversity and significant but weak genetic substructuring were found across all sites and temporal replicates, with no effects of metal contamination on the genetic variation or species’ abundance, although C. piger tended to be less frequent at highly contaminated sites. Our results indicate that high levels of gene flow and population dynamic processes may overlay potential pollutant effects. At least for our study species, we conclude that the “genetic erosion hypothesis”, which suggests that chemical pollution will reduce genome-wide genetic variability in affected populations, does not hold under natural conditions. Interestingly, our study provides evidence of successful hybridization between the two sister species under natural conditions.

A new cryptic species distinguished from Ischnochiton comptus (Gould, 1859) (Polyplacophora: Ischnochitonidae) in central Honshu, Japan

ABSTRACTIschnochiton boninensis Bergenhayn, 1933 and Ischnochiton comptus (Gould, 1859) are commonly found in central Honshu, Japan. These species are frequently misidentified because of the striking similarity in their overall appearance, and with the slight differences requiring microscopic examination. The present study investigated these distinctions, and I. comptus is further divided into a cryptic pair of species on the basis of morphological and genetic analyses. Genetic analyses revealed that I. comptus and its new cryptic sister species are separated by a relatively large pairwise genetic distance, similar to the distance between either of these and I. boninensis. Morphological observations indicated that only I. boninensis has clear granulation of the central areas of the intermediate valves, and the other two species, including I. comptus, are more similar in their radulae and girdle scales. Despite these similarities, specimens originally identified as I. comptus can be divided into separate species based on molecular and differences in the sculpture of the central area of the intermediate valves. One of these is here described as a new cryptic species distinct from I. comptus.

The more we search, the more we find: discovery of a new lineage and a new species complex in the genus Asparagopsis.

In the past few decades, in the marine realm in particular, the use of molecular tools has led to the discovery of hidden taxonomic diversity, revealing complexes of sister species. A good example is the red algal genus Asparagopsis. The two species (A. armata and A. taxiformis) recognized in this genus have been introduced in many places around the world. Within the nominal species A. taxiformis, previous molecular analyses have uncovered several lineages, suggesting the existence of sister species or subspecies. Although the genus has been well studied in some regions (e.g., the Mediterranean Sea and Hawaii), it remains poorly investigated in others (e.g., South Pacific). Our study mainly focused on these latter areas to clarify lineages and better determine lineage status (i.e., native vs. introduced). A total of 188 specimens were collected from 61 sites, 58 of which had never been sampled before. We sequenced the DNA from samples for three markers and obtained 112 sequences for the chloroplastic RuBisCo spacer, 118 sequences for the nuclear LSU rRNA gene, and 174 for the mitochondrial spacer cox2-3. Phylogenetic analyses using all three markers suggested the existence of two cryptic sister species with the discovery of a new clade within A. armata. This clade was found only in Western Australia, Tasmania and New Zealand, and is thus restricted to a subregional biogeographic unit. We also discovered a new, fifth lineage for A. taxiformis restricted to the South Pacific and Western Australia. Except for this newly described lineage, all other lineages showed a global distribution influenced by introduction events. These results illustrate the difficulty in accurately defining cosmopolitan species. Our findings also highlight the need for targeted (i.e., in poorly studied areas) and geographically extensive sampling efforts when studying taxa that have been introduced globally and that are likely to hide species complexes.

Genomic basis of ecological niche divergence among cryptic sister species of non-biting midges.

BackgroundThere is a lack of understanding the evolutionary forces driving niche segregation of closely related organisms. In addition, pinpointing the genes driving ecological divergence is a key goal in molecular ecology. Here, larval transcriptome sequences obtained by next-generation-sequencing are used to address these issues in a morphologically cryptic sister species pair of non-biting midges (Chironomus riparius and C. piger).ResultsMore than eight thousand orthologous open reading frames were screened for interspecific divergence and intraspecific polymorphisms. Despite a small mean sequence divergence of 1.53% between the sister species, 25.1% of 18,115 observed amino acid substitutions were inferred by α statistics to be driven by positive selection. Applying McDonald-Kreitman tests to 715 alignments of gene orthologues identified eleven (1.5%) genes driven by positive selection.ConclusionsThree candidate genes were identified as potentially responsible for the observed niche segregation concerning nitrite concentration, habitat temperature and water conductivity. Additionally, signs of positive selection in the hydrogen sulfide detoxification pathway were detected, providing a new plausible hypothesis for the species’ ecological differentiation. Finally, a divergently selected, nuclear encoded mitochondrial ribosomal protein may contribute to reproductive isolation due to cytonuclear coevolution.

Fifty Shades of Grey: giving colour to the poorly known Angolan Ashy reed frog (Hyperoliidae: Hyperolius cinereus), with the description of a new species.

Phylogenetic reconstruction using the mitochondrial 16S marker shows that geographically separated populations of the poorly known Hyperolius cinereus (Anura: Hyperoliidae) from Angola form two distinct clades. The description of H. cinereus was originally based on only a single preserved adult male. Fresh material of both sexes allowed a detailed redescription of the species, which is restricted mainly to the south-draining Cunene and Cubango river systems. Bioacoustic and morphological characters, in conjunction with colouration differences, allow the description of a cryptic sister species from Lagoa Carumbo in north-eastern Angola, occurring in the Luele and Lovuo river systems of the Congo drainage basin. Tadpoles, for H. cinereus and the new species, are described.