Objectively delimiting species boundaries remains an important challenge in systematics and becomes urgent when unresolved taxonomy complicates conservation and recovery efforts. We examined species boundaries in the imperiled freshwater mussel genus Cyclonaias (Bivalvia: Unionidae) using morphometrics, molecular phylogenetics, and multispecies coalescent models to help guide pending conservation assessments and legislative decisions. Congruence across multiple lines of evidence indicated that current taxonomy overestimates diversity in the C. pustulosa species complex. The only genetically and morphologically diagnosable species in the C. pustulosa species complex were C. pustulosa and C. succissa and we consider C. aurea , C. houstonensis, C. mortoni , and C. refulgens to be synonyms of C. pustulosa . In contrast, all three species in the C. nodulata complex ( C. necki , C. nodulata , and C. petrina ) were genetically, geographically, and morphologically diagnosable. Our findings have important conservation and management implications, as three nominal species ( C. aurea , C. houstonensis , and C. petrina ) are being considered for protection under the Endangered Species Act.

Inaccurate systematics confound our ability to determine evolutionary processes that have led to the diversification of many taxa. The North American freshwater mussel tribe Lampsilini is one of the better-studied groups in Unionidae, however, many supraspecific relationships between lampsiline genera remain unresolved. Two genera previously hypothesized to be non-monophyletic that have been largely overlooked are Leptodea and Potamilus. We set out to resolve supraspecific relationships in Lampsilini and test the monophyly of Leptodea and Potamilus by integrating molecular, morphological, and life history data. Our molecular matrix consisted of four loci: cytochrome c oxidase subunit 1 (CO1), NADH dehydrogenase subunit 1 (ND1), internal transcribed spacer 1 (ITS1), and 28S ribosomal RNA. Secondly, we performed both traditional and Fourier shape morphometric analyses to evaluate morphological differences and finally, we compared our results with available life history data. Molecular data supported the paraphyly of both Leptodea and Potamilus, but nodal support was insufficient to make any conclusions regarding generic-level assignments at this time. In contrast, inference from our integrative taxonomic assessment depicts significant support for the recognition of a new species, Potamilus streckersoni sp. nov., the Brazos Heelsplitter. Our data show clear separation of three taxonomic entities in the P. ohiensis species complex: P. amphichaenus, P. ohiensis, and P. streckersoni sp. nov.; all molecularly, geographically, and morphologically diagnosable. Our findings have profound implications for unionid taxonomy and will aid stakeholders in establishing effective conservation and management strategies.http://www.zoobank.org/urn:lsid:zoobank.org:pub:502647C0-418B-4CC4-85A8-BD89FC3F674F

From a genomics perspective, bivalves (Mollusca: Bivalvia) have been poorly explored with the exception for those of high economic value. The bivalve order Unionida, or freshwater mussels, has been of interest in recent genomic studies due to their unique mitochondrial biology and peculiar life cycle. However, genomic studies have been hindered by the lack of a high-quality reference genome. Here, I present a genome assembly of Potamilus streckersoni using Pacific Bioscience single-molecule real-time long reads and 10X Genomics-linked read sequencing. Further, I use RNA sequencing from multiple tissue types and life stages to annotate the reference genome. The final assembly was far superior to any previously published freshwater mussel genome and was represented by 2,368 scaffolds (2,472 contigs) and 1,776,755,624 bp, with a scaffold N50 of 2,051,244 bp. A high proportion of the assembly was comprised of repetitive elements (51.03%), aligning with genomic characteristics of other bivalves. The functional annotation returned 52,407 gene models (41,065 protein, 11,342 tRNAs), which was concordant with the estimated number of genes in other freshwater mussel species. This genetic resource, along with future studies developing high-quality genome assemblies and annotations, will be integral toward unraveling the genomic bases of ecologically and evolutionarily important traits in this hyper-diverse group.

Genomic resources across squamate reptiles (lizards and snakes) have lagged behind other vertebrate systems and high-quality reference genomes remain scarce. Of the 23 chromosome-scale reference genomes across the order, only 12 of the ~60 squamate families are represented. Within geckos (infraorder Gekkota), a species-rich clade of lizards, chromosome-level genomes are exceptionally sparse representing only two of the seven extant families. Using the latest advances in genome sequencing and assembly methods, we generated one of the highest quality squamate genomes to date for the leopard gecko, Eublepharis macularius (Eublepharidae). We compared this assembly to the previous, short-read only, E. macularius reference genome published in 2016 and examined potential factors within the assembly influencing contiguity of genome assemblies using PacBio HiFi data. Briefly, the read N50 of the PacBio HiFi reads generated for this study was equal to the contig N50 of the previous E. macularius reference genome at 20.4 kilobases. The HiFi reads were assembled into a total of 132 contigs, which was further scaffolded using HiC data into 75 total sequences representing all 19 chromosomes. We identified 9 of the 19 chromosomal-scaffolds were assembled as a near-single contig, while the other 10 chromosomes were each scaffolded together from multiple contigs. We qualitatively identified that percent repeat content within a chromosome broadly affects its assembly contiguity prior to scaffolding. This genome assembly signifies a new age for squamate genomics where high-quality reference genomes rivaling some of the best vertebrate genome assemblies can be generated for a fraction previous cost estimates. This new E. macularius reference assembly is available on NCBI at JAOPLA010000000.

In 2011, the first high-quality genome assembly of a squamate reptile (lizard or snake) was published for the green anole. Dozens of genome assemblies were subsequently published over the next decade, yet these assemblies were largely inadequate for answering fundamental questions regarding genome evolution in squamates due to their lack of contiguity or annotation. As the “genomics age” was beginning to hit its stride in many organismal study systems, progress in squamates was largely stagnant following the publication of the green anole genome. In fact, zero high-quality (chromosome-level) squamate genomes were published between the years 2012–2017. However, since 2018, an exponential increase in high-quality genome assemblies has materialized with 24 additional high-quality genomes published for species across the squamate tree of life. As the field of squamate genomics is rapidly evolving, we provide a systematic review from an evolutionary genomics perspective. We collated a near-complete list of publicly available squamate genome assemblies from more than half-a-dozen international and third-party repositories and systematically evaluated them with regard to their overall quality, phylogenetic breadth, and usefulness for continuing to provide accurate and efficient insights into genome evolution across squamate reptiles. This review both highlights and catalogs the currently available genomic resources in squamates and their ability to address broader questions in vertebrates, specifically sex chromosome and microchromosome evolution, while addressing why squamates may have received less historical focus and has caused their progress in genomics to lag behind peer taxa.