DNA barcodes for the pipefish genus Corythoichthys (Actinopterygii: Syngnathiformes) from the Indian Ocean provide insights into cryptic diversity.

The syngnathiform genus Corythoichthys comprises a group of taxonomically complex, tail-brooding (Syngnathinae) pipefishes widely distributed in the Indo-Pacific region. Due to the presence of overlapping interspecific morphological characters, reliable taxonomic information on Corythoichthys is still lacking. Using 52 CO1 sequences, including seven newly generated, a phylogenetic analysis was carried out to understand the genetic diversity, distribution and 'species groups' within the genus Corythoichthys. Species delimitation using Automatic Barcode Gap Discovery (ABGD) analysis confirmed the presence of 13 species which include 'species-complexes' previously considered as a single taxon. Our results revealed the presence of three species groups, 'C. amplexus', 'C. conspicillatus' and 'C. haematopterus' and four unidentified/undescribed species in the wider Indo-Pacific realm. Interestingly, 60 sequences and a mitogenome identified as Corythoichthys in GenBank are misidentified at the genus level. Based on our findings, we suggest that the taxonomy and systematics of Corythoichthys need to be re-examined and validated using integrative methods, and care should be taken while selecting specimens for genetic studies.

Aenigmachannidae, a new family of snakehead fishes (Teleostei: Channoidei) from subterranean waters of South India.

Pronounced organism-wide morphological stasis in evolution has resulted in taxa with unusually high numbers of primitive characters. These 'living fossils' hold a prominent role for our understanding of the diversification of the group in question. Here we provide the first detailed osteological analysis of Aenigmachanna gollum based on high-resolution nano-CT scans and one cleared and stained specimen of this recently described snakehead fish from subterranean waters of Kerala in South India. In addition to a number of derived and unique features, Aenigmachanna has several characters that exhibit putatively primitive conditions not encountered in the family Channidae. Our morphological analysis provides evidence for the phylogenetic position of Aenigmachanna as the sister group to Channidae. Molecular analyses further emphasize the uniqueness of Aenigmachanna and indicate that it is a separate lineage of snakeheads, estimated to have split from its sister group at least 34 or 109 million years ago depending on the fossil calibration employed. This may indicate that Aenigmachanna is a Gondwanan lineage, which has survived break-up of the supercontinent, with India separating from Africa at around 120 mya. The surprising morphological disparity of Aenigmachanna from members of the Channidae lead us to erect a new family of snakehead fishes, Aenigmachannidae, sister group to Channidae, to accommodate these unique snakehead fishes.

Phylogeny of the hillstream loach genus Mesonoemacheilus reveals widespread diversification through ancient drainage connections in the Western Ghats Biodiversity Hotspot.

Rivers draining the Western Ghats (WG) mountain ranges in peninsular India harbor an exceptionally diverse, unique and evolutionarily distinct assemblage of lower vertebrates with high levels of endemism, attributed to their evolution and potentially long history of isolation during the Late Cretaceous or Early Tertiary. A molecular phylogeny of hillstream loaches of the genus Mesonoemacheilus endemic to the WG revealed the presence of four clades which we designate as 'species groups'. A consensus of various species delimitation methods indicates the likelihood of 'at least' seven more undescribed species within Mesonoemacheilus. Molecular clock analysis dates the basal clade around 38 mya in the Paleogene, and subsequent diversification in the Neogene and Quaternary periods resulting in the current genetic diversity. Biogeographic analysis suggests that vicariance events which separated the rivers on either side of the two geological barriers/gaps, the Palghat and Shencottah, in the Neogene, as well as range contractions and cladogenetic events contributed to the current patterns of diversity and distribution of this genus. Our results also provide preliminary indications on the interconnections and faunal exchange between historical river drainages in the WG region.

Resolving the taxonomic enigma of the iconic game fish, the hump-backed mahseer from the Western Ghats biodiversity hotspot, India.

Growing to lengths and weights exceeding 1.5 m and 45 kg, the hump-backed mahseer fish of the Western Ghats biodiversity hotspot, India, is an iconic, mega-faunal species that is globally recognized as a premier freshwater game fish. Despite reports of their high extinction risk, conservation approaches are currently constrained by their lack of valid taxonomic identity. Using an integrative approach, incorporating morphology, molecular analysis and historical photographs, this fish can now be revealed to be conspecific with Tor remadevii, a species lacking a common name, that was initially, but poorly, described in 2007 from the River Pambar, a tributary of the River Cauvery in Kerala. Currently known to be endemic and restricted to the River Cauvery basin in the Western Ghats, T. remadevii is distinguished from congeners by its prominent hump originating above the pre-opercle and extending to the origin of the dorsal fin, a well-developed mandible resulting in a terminal or slightly superior mouth position, and the dorsal orientation of the eyes. While body colouration varies (silver, bronze, greenish) and is not considered a reliable diagnostic character, orange coloration of the caudal fin (sometimes extending to all fins) is considered a consistent characteristic. Having been first brought to the attention of the scientific community in 1849, and the recreational angling (game fishing) community in 1873, it has taken over 150 years to finally provide this iconic fish with a valid scientific name. This taxonomic clarity should now assist development and delivery of urgent conservation actions commensurate with their extinction risk.

Hypselobarbus bicolor, a new species of large barb (Teleostei: Cyprinidae) from the Western Ghats of India.

There has been much confusion around the identity of Hypselobarbus pulchellus, an endemic large barb of the Western Ghats. Recent works highlighted the fact that it is likely that two similar-looking species have been considered as H. pulchellus. A genetic analysis helps differentiate the two forms: viz., H. pulchellus and a new species, here described as H. bicolor sp. nov. Hypselobarbus bicolor can be distinguished from all other species of Hypselobarbus based on a combination of the following characters: two pairs of barbels, lateral line complete, with 34-36+1-2 pored scales, ½6/1/3½ scales in the transverse row; 11-12 predorsal scales; last simple dorsal ray weak, dorsal fin with 9½ branched rays, gill rakers short and simple, 4-6+11-12 on first gill arch; proximal end of fifth ceratobranchial slightly curved, with 5+3+2 teeth; overall coloration dark grey with an ill-defined silver- or bronze-coloured band running along length of body, two scales high, on lateral-line scale-row and row above it; fins devoid of any markings; and small, closely-placed tubercles restricted to the tip of the snout in large males.

Positive Selection Linked with Generation of Novel Mammalian Dentition Patterns.

A diverse group of genes are involved in the tooth development of mammals. Several studies, focused mainly on mice and rats, have provided a detailed depiction of the processes coordinating tooth formation and shape. Here we surveyed 236 tooth-associated genes in 39 mammalian genomes and tested for signatures of selection to assess patterns of molecular adaptation in genes regulating mammalian dentition. Of the 236 genes, 31 (∼13.1%) showed strong signatures of positive selection that may be responsible for the phenotypic diversity observed in mammalian dentition. Mammalian-specific tooth-associated genes had accelerated mutation rates compared with older genes found across all vertebrates. More recently evolved genes had fewer interactions (either genetic or physical), were associated with fewer Gene Ontology terms and had faster evolutionary rates compared with older genes. The introns of these positively selected genes also exhibited accelerated evolutionary rates, which may reflect additional adaptive pressure in the intronic regions that are associated with regulatory processes that influence tooth-gene networks. The positively selected genes were mainly involved in processes like mineralization and structural organization of tooth specific tissues such as enamel and dentin. Of the 236 analyzed genes, 12 mammalian-specific genes (younger genes) provided insights on diversification of mammalian teeth as they have higher evolutionary rates and exhibit different expression profiles compared with older genes. Our results suggest that the evolution and development of mammalian dentition occurred in part through positive selection acting on genes that previously had other functions.

Horabagrus melanosoma: a junior synonym of Horabagrus brachysoma (Teleostei: Horabagridae).

Horabagrus melanosoma was described from West Venpala in the lower reaches of the Manimala River, in the state of Kerala, India. It was distinguished from its nearest congener, H. brachysoma based on a combination of characters including darker body colour, shorter pelvic fin and greater number of anal fin rays. Examination of the type material revealed significant morphometric and meristic discrepancies with the original description. Based on multivariate morphometric, and genetic analysis of topotypical specimens, we propose that H. melanosoma should be treated as a junior synonym of H. brachysoma.

EASER: Ensembl Easy Sequence Retriever.

The rapid advances in genome sequencing technologies have increased the pace at which biological sequence databases are becoming available to the broad scientific community. Thus, obtaining and preparing an appropriate sequence dataset is a crucial first step for all types of genomic analyses. Here, we present a script that can widely facilitate the easy, fast, and effortless downloading and preparation of a proper biological sequence dataset for various genomics studies. This script retrieves Ensembl defined genomic features, associated with a given Ensembl identifier. Coding (CDS) and genomic sequences can be easily retrieved based on a selected relationship from a set of relationship types, either considering all available organisms or a user specified subset of organisms. The script is very user-friendly and by default starts with an interactive mode if no command-line options are specified.

Morphological and genetic evidence for multiple evolutionary distinct lineages in the endangered and commercially exploited red lined torpedo barbs endemic to the Western Ghats of India.

Red lined torpedo barbs (RLTBS) (Cyprinidae: Puntius) endemic to the Western Ghats Hotspot of India, are popular and highly priced freshwater aquarium fishes. Two decades of indiscriminate exploitation for the pet trade, restricted range, fragmented populations and continuing decline in quality of habitats has resulted in their 'Endangered' listing. Here, we tested whether the isolated RLTB populations demonstrated considerable variation qualifying to be considered as distinct conservation targets. Multivariate morphometric analysis using 24 size-adjusted characters delineated all allopatric populations. Similarly, the species-tree highlighted a phylogeny with 12 distinct RLTB lineages corresponding to each of the different riverine populations. However, coalescence-based methods using mitochondrial DNA markers identified only eight evolutionarily distinct lineages. Divergence time analysis points to recent separation of the populations, owing to the geographical isolation, more than 5 million years ago, after the lineages were split into two ancestral stocks in the Paleocene, on north and south of a major geographical gap in the Western Ghats. Our results revealing the existence of eight evolutionarily distinct RLTB lineages calls for the re-determination of conservation targets for these cryptic and endangered taxa.

The phylogenetic position of Lepidopygopsis typus (Teleostei: Cyprinidae), a monotypic freshwater fish endemic to the Western Ghats of India.

Lepidopygopsis, known as the peninsular-Indian hill trout, is a monotypic genus endemic to the Periyar stream-reservoir system, in the Western Ghats. Due to the morphological similarity of its only species, L. typus, with the Himalayan schizothoracine fishes, it was considered to be a relict species and a classic example of disjunct distribution. Using mitochondrial and nuclear gene sequence datasets, we show that L. typus is not allied to the schizothoracine fishes. Phylogenetic hypothesis-testing unequivocally supports a scenario in which L. typus and a clade comprising various genera of Asian and African barbins such as Tor Gonoproktopterus, Kosswigobarbus and Varicorhinus are sister groups. Based on our results, we suggest that the sheath of tile-like scales covering the anal-fin base of schizothoracine fishes and Lepidopygopsis typus could be a symplesiomorphy or a homoplasy.

Fish lateral line innovation: insights into the evolutionary genomic dynamics of a unique mechanosensory organ.

The mechanosensory lateral line, found only in fishes and amphibians, is an important sense organ associated with aquatic life. Lateral line patterns differ among teleost, the most diverse vertebrate taxa, hypothetically in response to selective pressures from different aquatic habitats. In this article, we conduct evolutionary genomic analyses of 34 genes associated with lateral line system development in teleosts to elucidate the significance of contrasting evolutionary rates and changes in the protein coding sequences. We find that duplicated copies of these genes are preferentially retained in the teleost genomes and that episodic events of positive selection have occurred in 22 of the 30 postduplication branches. In general, teleost genes evolved at a faster rate relative to their tetrapod counterparts, and the mutation rates of 26 of the 34 genes differed among teleosts and tetrapods. We conclude that following whole genome duplication, evolutionary rates and episodic events of positive selection on the lateral line system development genes might have been one of the factors favoring the subsequent adaptive radiation of teleosts into diverse habitats. These results provide the foundation for further detailed explorations into lateral line system genes and the evolution of diverse phenotypes and adaptations.

Adaptive evolution of the Retinoid X receptor in vertebrates.

Retinoid X receptors (RXR) are transcription factors with important roles in development, reproduction, homeostasis, and cell differentiation. Different types of vertebrate RXRs (α (RXRA), ß (RXRB) and γ (RXRG)) have arisen from multiple duplication events. The adaptive evolution mechanism that has preserved duplicate RXR paralogs, as well as their role in development and adaptation, is thus far unknown. In this work, we have investigated different aspects of vertebrate RXR evolution. Codon based tests of positive selection identified that RXR was under significant positive selection immediately after the whole genome duplications in vertebrates. Amino acid based rate shift analysis also revealed significant rate shifts immediately after the whole genome duplications and functional divergence between all the pairs of RXRs. However, the extant RXR genes are highly conserved, particularly the helix involved in dimerization and the DNA-binding domain, but positively selected sites can nevertheless be found in domains for RXR regulation.

Unraveling a 146 years old taxonomic puzzle: validation of Malabar snakehead, species-status and its relevance for channid systematics and evolution.

BACKGROUND: The Malabar snakehead Channa diplogramma is one of the most enigmatic and least understood species within the family Channidae, which comprise one of the most important groups of freshwater food fish in tropical Asia. Since its description from peninsular India in 1865, it has remained a taxonomic puzzle with many researchers questioning its validity, based on its striking similarity with the South East Asian C. micropeltes. In this study, we assessed the identity of the Malabar snakehead, C. diplogramma, using morphological and molecular genetic analyses, and also evaluated its phylogenetic relationships and evolutionary biogeography. METHODOLOGY/PRINCIPAL FINDINGS: The morphometric and meristic analysis provided conclusive evidence to separate C. diplogramma and C. micropeltes as two distinct species. Number of caudal fin rays, lateral line scales, scales below lateral line; total vertebrae, pre-anal length and body depth were the most prominent characters that can be used to differentiate both the species. Channa diplogramma also shows several ontogenic color phases during its life history, which is shared with C. micropeltes. Finally, the genetic distance between both species for the partial mitochondrial 16S rRNA and COI sequences is also well above the intra-specific genetic distances of any other channid species compared in this study. CONCLUSIONS/SIGNIFICANCE: The current distribution of C. diplogramma and C. micropeltes is best explained by vicariance. The significant variation in the key taxonomic characters and the results of the molecular marker analysis points towards an allopatric speciation event or vicariant divergence from a common ancestor, which molecular data suggests to have occurred as early as 21.76 million years ago. The resurrection of C. diplogramma from the synonymy of C. micropeltes has hence been confirmed 146 years after its initial description and 134 years after it was synonymised, establishing it is an endemic species of peninsular India and prioritizing its conservation value.