Phylogenomic analysis of Syngnathidae reveals novel relationships, origins of endemic diversity and variable diversification rates.

BACKGROUND: Seahorses, seadragons, pygmy pipehorses, and pipefishes (Syngnathidae, Syngnathiformes) are among the most recognizable groups of fishes because of their derived morphology, unusual life history, and worldwide distribution. Despite previous phylogenetic studies and recent new species descriptions of syngnathids, the evolutionary relationships among several major groups within this family remain unresolved. RESULTS: Here, we provide a reconstruction of syngnathid phylogeny based on genome-wide sampling of 1314 ultraconserved elements (UCEs) and expanded taxon sampling to assess the current taxonomy and as a basis for macroevolutionary insights. We sequenced a total of 244 new specimens across 117 species and combined with published UCE data for a total of 183 species of Syngnathidae, about 62% of the described species diversity, to compile the most data-rich phylogeny to date. We estimated divergence times using 14 syngnathiform fossils, including nine fossils with newly proposed phylogenetic affinities, to better characterize current and historical biogeographical patterns, and to reconstruct diversification through time. We present a phylogenetic hypothesis that is well-supported and provides several notable insights into syngnathid evolution. We found nine non-monophyletic genera, evidence for seven cryptic species, five potentially invalid synonyms, and identified a novel sister group to the seahorses, the Indo-Pacific pipefishes Halicampus macrorhynchus and H. punctatus. In addition, the morphologically distinct southwest Pacific seahorse Hippocampus jugumus was recovered as the sister to all other non-pygmy seahorses. As found in many other groups, a high proportion of syngnathid lineages appear to have originated in the Central Indo-Pacific and subsequently dispersed to adjoining regions. Conversely, we also found an unusually high subsequent return of lineages from southern Australasia to the Central Indo-Pacific. Diversification rates rose abruptly during the Middle Miocene Climate Transition and peaked after the closure of the Tethys Sea. CONCLUSIONS: Our results reveal a previously underappreciated diversity of syngnathid lineages. The observed biogeographic patterns suggest a significant role of the southern Australasian region as a source and sink of lineages. Shifts in diversification rates imply possible links to declining global temperatures, the separation of the Atlantic and Pacific faunas, and the environmental changes associated with these events.

Stigmatopora harastii, a new species of pipefish in facultative associations with finger sponges and red algae from New South Wales, Australia (Teleostei, Syngnathidae).

A new species of pipefish, Stigmatopora harastii sp. nov., is described based on the male holotype and two female paratypes, 136.3-145.5 mm SL, collected from red algae (sp.?) at 12 meters depth in Botany Bay, New South Wales (NSW), Australia. The new taxon shares morphological synapomorphies with the previously described members of Stigmatopora, including principle body ridges, fin placement, slender tail, and absence of a caudal fin. It is morphologically and meristically similar to Stigmatopora nigra, including snout length and shape, dorsal-fin origin on 6th-7th trunk ring, and lateral trunk ridge terminating on the first tail ring. Stigmatopora harastii sp. nov. is distinguished from its congeners, however, by characters of the head and first trunk ring, distinct sexual dimorphic markings on sides and venter of anterior trunk rings, and red background coloration in life. The new taxon can be further differentiated by genetic divergence in the mitochondrial COI gene (uncorrected p-distances of 9.8%, 10.1%, 10.7%, and 14.6%, from S. argus, S. macropterygia, S. narinosa, and S. nigra, respectively). The type locality is characterised by semi-exposed deep-water sandy areas interspersed with boulders, flat reefs, and an absence of seagrass beds, in which S. harastii has been observed living in facultative associations with a finger sponge and red algae at depths of 10-25 meters, compared to the shallow coastal and estuarine habitats preferred by the fucoid algae and seagrass-associating members of Stigmatopora. Stigmatopora harastii sp. nov. represents the fourth species of Stigmatopora recorded in temperate southern Australia.

Hippocampus nalu, a new species of pygmy seahorse from South Africa, and the first record of a pygmy seahorse from the Indian Ocean (Teleostei, Syngnathidae).

A new species and the first confirmed record of a true pygmy seahorse from Africa, Hippocampus nalu sp. nov., is herein described on the basis of two specimens, 18.9-22 mm SL, collected from flat sandy coral reef at 14-17 meters depth from Sodwana Bay, South Africa. The new taxon shares morphological synapomorphies with the previously described central Indo-Pacific pygmy seahorses, H. colemani, H. japapigu, H. pontohi, and H. satomiae, and H. waleananus, including diminutive size, twelve trunk rings, prominent cleithral ring and supracleithrum, spines on the fifth and twelfth superior and lateral trunk ridges, respectively, and prominent wing-like protrusions present on the first and/or second superior trunk rings posterior to the head. Hippocampus nalu sp. nov. is primarily distinguished from its pygmy seahorse congeners by highly distinct spine morphology along the anterior segments of the superior trunk ridge. Comparative molecular analysis reveals that the new species demonstrates significant genetic divergence in the mitochondrial COI gene from the morphologically similar H. japapigu and H. pontohi (estimated uncorrected p-distances of 16.3% and 15.2%, respectively). Hippocampus nalu sp. nov. represents the eighth member of the pygmy seahorse clade to be described from the Indo-Pacific, the first confirmed record from the African continent and the Indian Ocean, and an extension of more than 8000 km beyond the previously known range of pygmy seahorses from the Central and Western Indo-Pacific.

Hippocampuswhitei Bleeker, 1855, a senior synonym of the southern Queensland seahorse H.procerus Kuiter, 2001: molecular and morphological evidence (Teleostei, Syngnathidae).

The taxonomic status of the seahorse Hippocampusprocerus Kuiter, 2001, type locality Hervey Bay, QLD, Australia, was re-examined based on its strong morphological similarity and geographical proximity to its congener H.whitei Bleeker, 1855, a species recorded in ten estuaries of New South Wales, Australia. Kuiter (2001) distinguished H.procerus from H.whitei by a taller coronet, marginally lower meristics, and spinier physiognomy. Meristic, morphometric, and key diagnostic morphological character comparisons from vouchered specimens of the two purported species collected from Sydney Harbour, Nelson Bay, Port Stephens, NSW and Hervey Bay, Bundaberg, and Moreton Bay, QLD did not show diagnostic differences to support species-level classification of H.procerus. Furthermore, partial mitochondrial COI sequence data from specimens sampled from known geographical distributions in NSW and Southport, QLD failed to discriminate between populations as a result of shared haplotypes, and revealed an average intraspecific divergence of 0.002%. Hippocampusprocerus is hereby placed in the synonymy of H.whitei; a redescription is provided, with a revised record of its range across eastern Australia.

Hippocampusjapapigu, a new species of pygmy seahorse from Japan, with a redescription of H.pontohi (Teleostei, Syngnathidae).

The pygmy seahorse Hippocampusjapapigusp. n. is described based on three specimens, 13.9-16.3 mm SL, collected from a mixed soft coral and algae reef at 11 m depth at Hachijo-jima Island, Izu Islands, Japan. The new taxon shares morphological synapomorphies with the previously described central Indo-Pacific pygmy seahorses, H.colemani, H.pontohi, H.satomiae, and H.waleananus, including extremely small size, 12 trunk rings, strongly raised continuous cleithral ring, snout spine, large spine on the eighth lateral and fifth and 12 superior trunk ridges, respectively, and unusual wing-like-protrusions immediately posterior to the head. Hippocampusjapapigusp. n. can be distinguished from all congeners by the following combination of features in the anterodorsal area of the trunk: bilaterally paired wing-like protrusions formed by a single pair of large, truncate spines projecting dorsolaterad on the first superior trunk ridge, followed by a unique elevated dorsal ridge formed by triangular bony mounds dorsally on the second to fourth superior trunk ridges. In contrast, H.pontohi possesses a pair of large truncate spines projecting strongly laterad on both the first and second superior trunk ridges followed by flat surfaces dorsally on the third and fourth superior trunk rings. The new species can be further differentiated by genetic divergence from H.pontohi (an uncorrected p-distance of 10.1% in the mitochondrial COI gene) and a striking reticulated white and brown lattice pattern on the head, trunk, and tail. Hippocampusjapapigusp. n. represents the fifth species of pygmy seahorse recorded in Japan.

Molecular phylogeny and patterns of diversification in syngnathid fishes.

The family Syngnathidae is a large and diverse clade of morphologically unique bony fishes, with 57 genera and 300 described species of seahorses, pipefishes, pipehorses, and seadragons. They primarily inhabit shallow coastal waters in temperate and tropical oceans, and are characterized by a fused jaw, male brooding, and extraordinary crypsis. Phylogenetic relationships within the Syngnathidae remain poorly resolved due to lack of generic taxon sampling, few diagnostic morphological characters, and limited molecular data. The phylogenetic placement of the threatened, commercially exploited seahorses remains a topic of intense interest, with conflicting topologies based on morphology and predominantly mitochondrial genetic data. In this study, we integrate eight nuclear and mitochondrial markers and 17 morphological characters to investigate the phylogenetic structure of the family Syngnathidae at the generic level. We include 91 syngnathid species representing 48 of the 57 recognized genera, all major ocean basins, and a broad array of temperate and tropical habitats including rocky and coral reefs, sand and silt, mangroves, seagrass beds, estuaries, and rivers. Maximum likelihood and Bayesian analyses of 5160bp from eight loci produced high congruence among alternate topologies, defining well-supported and sometimes novel clades. We present a hypothesis that confirms a deep phylogenetic split between lineages with trunk- or tail-brood pouch placement, and provides significant new insights into the morphological evolution and biogeography of this highly derived fish clade. Based on the fundamental division between lineages - the tail brooding "Urophori" and the trunk brooding "Gastrophori" - we propose a revision of Syngnathidae classification into only two subfamilies: the Nerophinae and the Syngnathinae. We find support for distinct principal clades within the trunk-brooders and tail-brooders, the latter of which include seahorses, seadragons, independent lineages of pipehorses, and clades that originated in southern Australia and the Western Atlantic. We suggest the seahorse genus Hippocampus is of Indo-Pacific origin and its sister clade is an unexpected grouping of several morphologically disparate Indo-Pacific genera, including the Pacific pygmy pipehorses. Taxonomic revision is required for multiple genera, particularly to reflect deep evolutionary splits in nominal lineages from the Atlantic versus the Indo-Pacific.