Evolution of diverse host infection mechanisms delineates an adaptive radiation of lampsiline freshwater mussels centered on their larval ecology.

North American watersheds contain a high diversity of freshwater mussels (Unionoida). During the long-lived, benthic phase of their life cycle, up to 40 species can co-occur in a single riffle and there is typically little evidence for major differences in their feeding ecology or microhabitat partitioning. In contrast, their brief parasitic larval phase involves the infection of a wide diversity of fish hosts and female mussels have evolved a spectrum of adaptations for infecting host fish with their offspring. Many species use a passive broadcast strategy: placing high numbers of larvae in the water column and relying on chance encounters with potential hosts. Many other species, including most members of the Lampsilini, have a proactive strategy that entails the use of prey-mimetic lures to change the behavior of the hosts, i.e., eliciting a feeding response through which they become infected. Two main lure types are collectively produced: mantle tissue lures (on the female's body) and brood lures, containing infective larvae, that are released into the external environment. In this study, we used a phylogenomic approach (ddRAD-seq) to place the diversity of infection strategies used by 54 North American lampsiline mussels into an evolutionary context. Ancestral state reconstruction recovered evidence for the early evolution of mantle lures in this clade, with brood lures and broadcast infection strategies both being independently derived twice. The most common infection strategy, occurring in our largest ingroup clade, is a mixed one in which mimetic mantle lures are apparently the predominant infection mechanism, but gravid females also release simple, non-mimetic brood lures at the end of the season. This mixed infection strategy clade shows some evidence of an increase in diversification rate and most members use centrarchids (Micropterus & Lepomis spp.) as their predominant fish hosts. Broad linkage between infection strategies and predominant fish host genera is also seen in other lampsiline clades: worm-like mantle lures of Toxolasma spp. with sunfish (Lepomis spp.); insect larvae-like brood lures (Ptychobranchus spp.), or mantle lures (Medionidus spp., Obovaria spp.), or mantle lures combined with host capture (Epioblasma spp.) with a spectrum of darter (Etheostoma & Percina spp.) and sculpin (Cottus spp.) hosts, and tethered brood lures (Hamiota spp.) with bass (Micropterus spp.). Our phylogenetic results confirm that discrete lampsiline mussel clades exhibit considerable specialization in the primary fish host clades their larvae parasitize, and in the host infection strategies they employ to do so. They are also consistent with the hypothesis that larval resource partitioning of fish hosts is an important factor in maintaining species diversity in mussel assemblages. We conclude that, taking their larval ecology and host-infection mechanisms into account, lampsiline mussels may be legitimately viewed as an adaptive radiation.

Millimeter-sized smart sensors reveal that a solar refuge protects tree snail Partula hyalina from extirpation.

Pacific Island land snails are highly endangered due in part to misguided biological control programs employing the alien predator Euglandina rosea. Its victims include the fabled Society Island partulid tree snail fauna, but a few members have avoided extirpation in the wild, including the distinctly white-shelled Partula hyalina. High albedo shell coloration can facilitate land snail survival in open, sunlit environments and we hypothesized that P. hyalina has a solar refuge from the predator. We developed a 2.2 × 4.8 × 2.4 mm smart solar sensor to test this hypothesis and found that extant P. hyalina populations on Tahiti are restricted to forest edge habitats, where they are routinely exposed to significantly higher solar radiation levels than those endured by the predator. Long-term survival of this species on Tahiti may require proactive conservation of its forest edge solar refugia and our study demonstrates the utility of miniaturized smart sensors in invertebrate ecology and conservation.

Phylogenomic analyses confirm a novel invasive North American Corbicula (Bivalvia: Cyrenidae) lineage.

The genus Corbicula consists of estuarine or freshwater clams native to temperate/tropical regions of Asia, Africa, and Australia that collectively encompass both sexual species and clonal (androgenetic) lineages. The latter have become globally invasive in freshwater systems and they represent some of the most successful aquatic invasive lineages. Previous studies have documented four invasive clonal lineages, Forms A, B, C, and Rlc, with varying known distributions. Form A (R in Europe) occurs globally, Form B is found solely in North America, mainly the western United States, Form C (S in Europe) occurs both in European watersheds and in South America, and Rlc is known from Europe. A putative fifth invasive morph, Form D, was recently described in the New World from the Illinois River (Great Lakes watershed), where it occurs in sympatry with Forms A and B. An initial study showed Form D to be conchologically distinct: possessing rust-colored rays and white nacre with purple teeth. However, its genetic distinctiveness using standard molecular markers (mitochondrial cytochrome c oxidase subunit I and nuclear ribosomal 28S RNA) was ambiguous. To resolve this issue, we performed a phylogenomic analysis using 1,699-30,027 nuclear genomic loci collected via the next generation double digested restriction-site associated DNA sequencing method. Our results confirmed Form D to be a distinct invasive New World lineage with a population genomic profile consistent with clonality. A majority (7/9) of the phylogenomic analyses recovered the four New World invasive Corbicula lineages (Forms A, B, C, and D) as members of a clonal clade, sister to the non-clonal Lake Biwa (Japan) endemic, Corbicula sandai. The age of the clonal clade was estimated at 1.49 million years (my; ± 0.401-2.955 my) whereas the estimated ages of the four invasive lineage crown clades ranged from 0.27 to 0.44 my. We recovered very little evidence of nuclear genomic admixture among the four invasive lineages in our study populations. In contrast, 2/6 C. sandai individuals displayed partial nuclear genomic Structure assignments with multiple invasive clonal lineages. These results provide new insights into the origin and maintenance of clonality in this complex system.

Deconstructing an infamous extinction crisis: Survival of Partula species on Moorea and Tahiti.

Eleven of eighteen Society Island Partula species endemic to the Windward Island subgroup (Moorea and Tahiti) have been extirpated by an ill-advised biological control program. The conservation status of this critically endangered tree snail radiation is of considerable import, but is clouded by taxonomic uncertainty due to the extensive lack of congruence among species designations, diagnostic morphologies, and molecular markers. Using a combination of museum, captive, and remnant wild snails, we obtained the first high-resolution nuclear genomic perspective of the evolutionary relationships and survival of fourteen Windward Island Partula species, totaling 93 specimens. We analyzed ~1,607-28,194 nuclear genomic loci collected with the double digest restriction-site associated sequencing method. Results from phylogenomic trees, species estimation, and population assignment tests yielded monophyly of the Windward Island subgroup. Within this group, two well-supported clades encompassing five species complexes were recovered. Clade 1 was restricted to Tahiti and contained two species complexes: "P. affinis" (three species) and "P. otaheitana" (five species). Clade 2 occurred on Moorea and on Tahiti and consisted of three species complexes: one Tahitian, "P. clara/P. hyalina"; the other two, "P. taeniata" (three species) and "P. suturalis" (six species), Moorean. Our genomic results largely corroborated previous mitochondrial DNA survival estimates for Moorea and Tahiti, with all five species complexes having members surviving in captivity and/or as remnant wild populations, although the details vary in each case. Continued, proactive conservation and management may yet ensure a phylogenetically representative survival of the fabled Partula species of Moorea and Tahiti.

Moorean and Tahitian Partula tree snail survival after a mass extinction: New genomic insights using museum specimens.

Natural history museum collections provide a biodiversity window into the past and are of particular importance to the study of extinction-impacted clades such as the Pacific Island tree snail family Partulidae. Deliberate introduction of the predatory rosy wolf snail Euglandina rosea in the late 20th century led to the extinction/extirpation of 55/61 Society Island Partulidae species. In this study, we phylogenomically investigated the inter-relationships of the three surviving Society Island valley Partula species: P. taeniata (Moorea), P. clara and P. hyalina (Tahiti). All three formed a distinct clade in earlier mitochondrial phylogenies. Using Next Generation Sequencing (NGS) double digested Restriction Associated DNA sequencing (ddRADseq), we found that 46-year-old lyophilized museum specimens produced similar numbers of reads, sequencing depth, and loci as 10-year old ethanol-preserved collections. Phylogenomic trees indicated that Tahitian P. clara and P. hyalina are the result of a single founding lineage from Moorea, contrasting previous mitochondrial results and clarifying the enigmatic taxonomic status of P. c. incrassa. Our study highlights the utility and viability of NGS techniques for museum specimens and their increased resolution of evolutionary patterns. Sampling will be expanded to include the remaining Society Island partulid taxa to further explore the evolutionary history of this radiation.

A population genetic window into the past and future of the walleye Sander vitreus: relation to historic walleye and the extinct "blue pike" S. v. "glaucus".

BACKGROUND: Conserving genetic diversity and local adaptations are management priorities for wild populations of exploited species, which increasingly are subject to climate change, habitat loss, and pollution. These constitute growing concerns for the walleye Sander vitreus, an ecologically and economically valuable North American temperate fish with large Laurentian Great Lakes' fisheries. This study compares genetic diversity and divergence patterns across its widespread native range using mitochondrial (mt) DNA control region sequences and nine nuclear DNA microsatellite (µsat) loci, examining historic and contemporary influences. We analyze the genetic and morphological characters of a putative endemic variant- "blue pike" S. v. "glaucus" -described from Lakes Erie and Ontario, which became extinct. Walleye with turquoise-colored mucus also are evaluated, since some have questioned whether these are related to the "blue pike". RESULTS: Walleye populations are distinguished by considerable genetic divergence (mean FST mtDNA = 0.32 ± 0.01, µsat = 0.13 ± 0.00) and substantial diversity across their range (mean heterozygosity mtDNA = 0.53 ± 0.02, µsat = 0.68 ± 0.03). Southern populations markedly differ, possessing unique haplotypes and alleles, especially the Ohio/New River population that houses the oldest haplotype and has the most pronounced divergence. Northern formerly glaciated populations have greatest diversity in Lake Erie (mean heterozygosity mtDNA = 0.79 ± 0.00, µsat = 0.72 ± 0.01). Genetic diversity was much less in the historic Lake Erie samples from 1923-1949 (mean heterozygosity mtDNA = 0.05 ± 0.01, µsat = 0.47 ± 0.06) than today. The historic "blue pike" had no unique haplotypes/alleles and there is no evidence that it comprised a separate taxon from walleye. Turquoise mucus walleye also show no genetic differentiation from other sympatric walleye and no correspondence to the "blue pike". CONCLUSIONS: Contemporary walleye populations possess high levels of genetic diversity and divergence, despite habitat degradation and exploitation. Genetic and previously published tagging data indicate that natal homing and spawning site philopatry led to population structure. Population patterns were shaped by climate change and drainage connections, with northern ones tracing to post-glacial recolonization. Southerly populations possess unique alleles and may provide an important genetic reservoir. Allelic frequencies of Lake Erie walleye from ~70-90 years ago significantly differed from those today, suggesting population recovery after extensive habitat loss, pollution, and exploitation. The historic "blue pike" is indistinguishable from walleye, indicating that taxonomic designation is not warranted.

Systematics of the greenside darter Etheostoma blennioides complex: consensus from nuclear and mitochondrial DNA sequences.

The phylogenetic systematic relationships of the enigmatic greenside darter Etheostoma blennioides complex are analysed using sequences from the mitochondrial (mt) DNA cytochrome b gene and nuclear S7 ribosomal protein intron 1 from putative members of the complex, close relatives, and outgroups (totaling 421 individuals). We compare results from Bayesian and maximum likelihood analysis approaches and a variety of rooting and taxon inclusion scenarios, and include all putative subspecies and intergrade taxa for a new comprehensive analysis. Results reveal that nuclear and mtDNA data congruently, under all scenarios and approaches tested, define a highly-supported restricted greenside darter complex comprising three putative subspecies: E. b. blennioides, E. b. pholidotum, and part of E. b. newmanii (excepting those from the Tennessee/Hiwassee River clade). Within this redefined E. blennioides, only a single putative subspecies -E. b. blennioides - is monophyletic in the mtDNA trees, and none are monophyletic in the nuclear DNA trees. Nuclear and mtDNA results support E. gutselli as a separate species and suggest that the Tennessee/Hiwassee River clade of "E. b. newmanii" also may constitute a separate species (provisionally "E. newmanii"), with neither being a part of our redefined E. blennioides complex. The nuclear DNA trees depict the two as highly-supported divergent clades, but the mtDNA results group them together as a single clade, indicating introgression. Future study with greater sample sizes in the southern watersheds, coupling morphological analyses with additional nuclear gene phylogenies, is recommended to further investigate the relationships within the greenside darter complex.

Molecular, morphological, and biogeographic resolution of cryptic taxa in the Greenside Darter Etheostoma blennioides complex.

The systematic identity and genetic divergence of cryptic taxa and morphological subspecies in the Greenside Darter Etheostoma blennioides complex are analyzed from mitochondrial and nuclear DNA sequence data, along with morphological characters. We sequenced the mtDNA cytochrome b gene and control region and is the nuclear S7 intron 1 for 345 Greenside Darters from 19 locations across their distribution including areas of sympatry and allopatry, in comparison to putative sister species and relatives. Results provide the first genetic evidence that E. gutselli is a separate species and is the sister species of E. blennius, which together with E. rupestre comprise the sister group to the Greenside Darter complex; separating approximately 4.0 mya. MtDNA results show that the complex comprises 6 clades and supports only the morphological subspecies Etheostoma blennioides blennioides, distinguished by theta(ST)=0.94, approximately 1.7 my, scale counts, and ventral squamation. The former E. b. pholidotum and E. b. newmanii are polyphyletic and are invalid taxa, together comprising 5 differentiated clades that diverged approximately 0.90-1.7 mya. Nuclear DNA results recover some of the mtDNA clades, which are distinguished morphologically by subtle meristic count differences. Populations of E. b. blennioides genetically diverge, with diversity increasing to the southwest; attributed to restricted gene flow and genetic isolation with geographic distance. Samples of the former E. b. pholidotum from the Great Lakes/Wabash River clade are less divergent, with diversity increasing to the southwest, reflecting allopatric fragmentation and isolation by distance.