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Results for “Mitochondrial DNA”

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At least 271 records · Page 15Linked to original sources

USGS2006

Non-invasive method to obtain DNA from freshwater mussels (Bivalvia: Unionidae)

To determine whether DNA could be isolated from tissues obtained by brush-swabbing the mantle, viscera and foot, mantle-clips and swabbed cells were obtained from eight Quadrula pustulosa (Lea, 1831). DNA yields from clips and swabbings were 447.0 and 975.3 ??g/??L, respectively. Furthermore, comparisons of sequences from the ND-1 mitochondrial gene region showed a 100% sequence agreement of DNA from cells obtained by clips and swabs. To determine the number of swabs needed to obtain adequate yields of DNA for analyses, the visceras and feet of 5 Q. pustulosa each were successively swabbed 2, 4 and 6 times. DNA yields from the 2, 4 and 6 swabbed mussel groups were 399.4, 833.8 and 852.6 ng/??L, respectively. ND-1 sequences from the lowest yield still provided 846-901 bp for the ND-1 region. Nevertheless, to ensure adequate DNA yield from cell samples obtained by swabbing, we recommend that 4 swab-strokes of the viscera and foot be obtained. The use of integumental swabbing for collection of cells for determination of genetic relationships among freshwater mussels is noninvasive, when compared with tissue collection by mantle-clipping. Therefore, its use is recommended for freshwater mussels, especially state-protected or federally listed mussel species.

Journal of Shellfish Research↗

USGS2012

Development of a real-time PCR assay for detection of planktonic red king crab ( Paralithodes camtschaticus (Tilesius 1815)) larvae

The Alaskan red king crab ( Paralithodes camtschaticus ) fishery was once one of the most economically important single-species fisheries in the world, but is currently depressed. This fishery would benefit from improved stock assessment capabilities. Larval crab distribution is patchy temporally and spatially, requiring extensive sampling efforts to locate and track larval dispersal. Large-scale plankton surveys are generally cost prohibitive because of the effort required for collection and the time and taxonomic expertise required to sort samples to identify plankton individually via light microscopy. Here, we report the development of primers and a dual-labeled probe for use in a DNA-based real-time polymerase chain reaction assay targeting the red king crab, mitochondrial gene cytochrome oxidase I for the detection of red king crab larvae DNA in plankton samples. The assay allows identification of plankton samples containing crab larvae DNA and provides an estimate of DNA copy number present in a sample without sorting the plankton sample visually. The assay was tested on DNA extracted from whole red king crab larvae and plankton samples seeded with whole larvae, and it detected DNA copies equivalent to 1/10,000th of a larva and 1 crab larva/5mL sieved plankton, respectively. The real-time polymerase chain reaction assay can be used to screen plankton samples for larvae in a fraction of the time required for traditional microscopial methods, which offers advantages for stock assessment methodologies for red king crab as well as a rapid and reliable method to assess abundance of red king crab larvae as needed to improve the understanding of life history and population processes, including larval population dynamics.

Journal of Shellfish Research↗

USGS2020

An evaluation of noninvasive sampling techniques for Malayan sun bears

Traditional mark–recapture studies to estimate abundance and trends of Malayan sun bear ( Helarctos malayanus ) populations are impeded by logistics of live-trapping wild individuals. The development of noninvasive sampling techniques for monitoring sun bear populations is therefore crucial for targeted conservation action. Sun bears have short fur, and conventional hair-snagging devices are ineffective. Moreover, scats are rapidly decomposed by the warm, humid environment, as well as by invertebrates. In combination with camera-sampling, we tested 2 designs of hair traps ( n = 45) in situ at Tabin Wildlife Reserve in Sabah, Malaysia, during April–October 2017, to obtain hair samples from wild sun bears. We also deployed 4 types of hair traps in rainforest enclosures with captive sun bears to evaluate hair-capture success and the effects of weathering, lure, and adhesive on polymerase chain reaction (PCR) amplification success. Wild adult male sun bears displayed back-rubbing behavior at hair traps and 6 individuals were identified based on unique chest marks. We collected 30 hair samples from wild sun bears, including 15 chest mark images of 6 individuals over 1,260 trap-nights. We detected adult males at hair traps more frequently than females and subadults. We obtained 39 hair samples in the captive trials. Extracted DNA from hair roots successfully amplified with mitochondrial (wild bears: 95%; captive bears: 97%) and microsatellite primers (wild bears: 100%; captive bears 87%). Adhesive and lure type did not affect PCR amplification, but weathering reduced amplification of microsatellite loci. This study is the first successful attempt to obtain genetic samples from wild sun bears using inexpensive, readily available materials such as duct tape, polybutyl glue, and locally sourced lures. The quality of genetic material from these genetic samples should be suitable for studies of population size and gene flow.

Sabah↗

USGS2007

A genetic assessment of the recovery units for the mojave population of the desert tortoise, Gopherus agassizii

In the 1994 Recovery Plan for the Mojave population of the desert tortoise, Gopherus agassizii, the US Fish and Wildlife Service established 6 recovery units by using the best available data on habitat use, behavior, morphology, and genetics. To further assess the validity of the recovery units, we analyzed genetic data by using mitochondrial deoxyribonucleic acid (mtDNA) sequences and nuclear DNA microsatellites. In total, 125 desert tortoises were sampled for mtDNA and 628 for microsatellites from 31 study sites, representing all recovery units and desert regions throughout the Mojave Desert in California and Utah, and the Colorado Desert of California. The mtDNA revealed a great divergence between the Mojave populations west of the Colorado River and those occurring east of the river in the Sonoran Desert of Arizona. Some divergence also occurred between northern and southern populations within the Mojave population. The microsatellites indicated a low frequency of private alleles and a significant correlation between genetic and geographic distance among 31 sample sites, which was consistent with an isolation-by-distance population structure. Regional genetic differentiation was complementary to the recovery units in the Recovery Plan. Most allelic frequencies in the recovery units differed. An assignment test correctly placed most individuals to their recovery unit of origin. Of the 6 recovery units, the Northeastern and the Upper Virgin River units showed the greatest differentiation; these units may have been relatively more isolated than other areas and should be managed accordingly. The Western Mojave Recovery Unit, by using the new genetic data, was redefined along regional boundaries into the Western Mojave, Central Mojave, and Southern Mojave recovery units. Large-scale translocations of tortoises and habitat disturbance throughout the 20th century may have contributed to the observed patterns of regional similarity. ?? 2007 Chelonian Research Foundation.

California↗

USGS2019

Drift of larval darters (Family Percidae) in the upper Roanoke River basin, USA, characterized using phenotypic and DNA barcoding markers

Larval fish ecology is poorly characterized because sampling is difficult and tools for phenotypically identifying larvae are poorly developed. While DNA barcoding can help address the latter problem, ‘universal’ primers do not work for all fish species. The Roanoke River in the southeastern United States includes seven darters (Family Percide: Tribe Etheostomatini). We made 393 collections of larval fishes in 2015 and 2018, examined darter larvae for morphometric and pigmentation traits, developed PCR primers amplifying darter DNA, and evaluated three gear types for collecting larval darters. Amplified DNA sequences for 1351 larvae matched archived mitochondrial cytochrome oxidase I sequences for darters occurring in the ecosystem. Larval darters were classified to genus with 100% accuracy using the ratio of pectoral fin length to body length; however, identification to species using morphometrics alone was subject to a misclassification rate of 11.8%, which can be resolved by considering pigmentation patterns. Gear-types varied considerably in their capture efficacy for larval darters; most Percina larvae were collected in drift nets. Larval Percina species appeared in the drift before Etheostoma species in both study years. Application of molecular genetic and phenotypic tools to larval fish identification can advance understanding of larval darter ecology.

North Carolina, Virginia↗

USGS2012

Puerto Rico and Florida manatees represent genetically distinct groups

The West Indian manatee ( Trichechus manatus ) populations in Florida ( T. m. latirostris ) and Puerto Rico ( T. m. manatus ) are considered distinct subspecies and are listed together as endangered under the United States Endangered Species Act. Sustained management and conservation efforts for the Florida subspecies have led to the suggested reclassification of the species to a threatened or delisted status. However, the two populations are geographically distant, morphologically distinct, and habitat degradation and boat strikes continue to threaten the Puerto Rico population. Here, 15 microsatellite markers and mitochondrial control region sequences were used to determine the relatedness of the two populations and investigate the genetic diversity and phylogeographic organization of the Puerto Rico population. Highly divergent allele frequencies were identified between Florida and Puerto Rico using microsatellite (F ST = 0.16; R ST = 0.12 (P < 0.001)) and mitochondrial (F ST = 0.66; Φ ST = 0.50 (P < 0.001)) DNA. Microsatellite Bayesian cluster analyses detected two populations (K = 2) and no admixture or recent migrants between Florida (q = 0.99) and Puerto Rico (q = 0.98). The microsatellite genetic diversity values in Puerto Rico (H E = 0.45; N A = 3.9), were similar, but lower than those previously identified in Florida (H E = 0.48, N A = 4.8). Within Puerto Rico, the mitochondrial genetic diversity values (π = 0.001; h = 0.49) were slightly lower than those previously reported (π = 0.002; h = 0.54) and strong phylogeographic structure was identified (F ST global = 0.82; Φ ST global = 0.78 (P < 0.001)). The genetic division with Florida, low diversity, small population size (N = 250), and distinct threats and habitat emphasize the need for separate protections in Puerto Rico. Conservation efforts including threat mitigation, migration corridors, and protection of subpopulations could lead to improved genetic variation in the endangered Puerto Rico manatee population.

Florida↗

USGS2025

Development of PCR blocking primers enabling DNA metabarcoding analysis of dietary composition in hematophagous sea lamprey

Conventional dietary assessments are challenging in hematophagous species, particularly in sea lamprey ( Petromyzon marinus ). However, recent technological developments and molecular approaches have provided an attractive alternative through the use of DNA metabarcoding. While DNA metabarcoding has been used for dietary analyses in numerous species, including lampreys, applications of universal primers that detect a diverse set of prey items can be limited by the amplification of predator DNA. In this study, we designed and tested eight blocking primers designed to suppress the amplification of sea lamprey DNA with vertebrate-universal primers targeting the mitochondrial 12S rRNA gene. This approach allowed for the use of a single marker to amplify a taxonomically diverse suite of host species, in contrast to previous studies that used multiple taxon-specific primer pairs (e.g., Salmonidae, Cyprinidae, and Catostomidae). Candidate blocking primers evaluated in this study differed in base pair length, end sequence modification, and purification method. Samples with different sea lamprey-to-host DNA ratios were subjected to multiple detection methods including gel electrophoresis, quantitative PCR, and DNA metabarcoding to assess the ability of each blocking primer to selectively suppress amplification of the sea lamprey 12S gene region. All blocking primers tested performed well and demonstrated high effectiveness, suppressing sea lamprey reads by > 99.9% in mock communities and improving host DNA sequence recovery across various sample types, including wild-caught lamprey. Results show that the blocking primers evaluated can facilitate molecular diet analysis in sea lamprey, allowing the amplification of a taxonomically diverse range of host fish species with universal primers.

Great Lakes↗

USGS2018

Detecting southern California’s white sharks with environmental DNA

To improve ability to detect white sharks without the need for tags, or visual census, we developed a species-specific environmental DNA (eDNA) assay that targets a 163 bp fragment of the white shark ( Carcharodon carcharias ) mitochondrial cytochrome B gene on a digital droplet PCR (ddPCR) platform. We used this marker to detect white shark DNA in 250 ml water samples taken from across two sites in Santa Barbara, California (United States) frequented by juvenile white sharks. We did not detect white shark DNA in samples from two neighboring sites where sharks are presumably absent, suggesting that eDNA can indicate nearby white sharks. This marker development, testing, and opportunistic application in a region with known distributions of white sharks indicates that eDNA could be developed further to monitor white sharks, thereby informing conservation planning and public safety. With the potential increase in white shark populations due to decades of protection, there is a need for fishery independent methods for assessing white shark distributions, and eDNA may provide an ideal, non-intrusive tool for coastal assessments.

California↗

USGS2019

Discriminating among Pacific salmon, Rainbow Trout, and Atlantic Salmon species using common genetic screening methods

The five most common species of Pacific salmon, Rainbow Trout (steelhead) Oncorhynchus spp., and Atlantic Salmon Salmo salar intermingle in the North Pacific Ocean and its freshwater tributaries. Efficient morphological methods for distinguishing among these species are sometimes limited by condition of the specimen (degraded or missing morphology), life history stage, or training of the observer. Researchers have successfully applied various genetic methods to distinguish among these species when morphological analyses are not possible, but they cannot easily incorporate these methods into standard fish and wildlife population monitoring analysis workflows. Here we test five 5′–3′ exonuclease (TaqMan) assays developed from mitochondrial genes and provide novel methods that take advantage of TaqMan output to distinguish among these species. We found that combinations of as few as two of the five assays were adequate to distinguish all species. TaqMan chemistry is designed to interrogate a single nucleotide locus. We also explore the basis for the variation in the observed scatter plot distributions (variation in florescent signals) and show that this variation is due to nucleotide diversity in and near the probe site. Because the SNPs underlying the assays developed here are all physically close to one another along the mitochondrial genome, the potential exists to develop a single DNA sequence-based assay to discriminate among salmon species. This single assay can be added to a genotyping-by-sequencing panel to identify and exclude nontarget species from analyses.

Journal of Fish and Wildlife Management↗

USGS2003

A quantitative evaluation of two methods for preserving hair samples

Hair samples are an increasingly important DNA source for wildlife studies, yet optimal storage methods and DNA degradation rates have not been rigorously evaluated. We tested amplification success rates over a one-year storage period for DNA extracted from brown bear (Ursus arctos) hair samples preserved using silica desiccation and -20C freezing. For three nuclear DNA microsatellites, success rates decreased significantly after a six-month time point, regardless of storage method. For a 1000 bp mitochondrial fragment, a similar decrease occurred after a two-week time point. Minimizing delays between collection and DNA extraction will maximize success rates for hair-based noninvasive genetic sampling projects.

Molecular Ecology Notes↗

USGS2016

Genetic diversity and population structure in the threatened Oregon silverspot butterfly ( Speyeria zerene hippolyta ) in western Oregon and northwestern California— Implications for future translocations and the establishment of new populations

Executive Summary We present results of population genetic analyses performed on Oregon silverspot butterflies (OSB; Speyeria zerene hippolyta ) in western Oregon and northwestern California. We used DNA sequences from a 561-base pair region of the mitochondrial cytochrome oxidase subunit I (COI) gene for a dataset comprised of 112 S. z. hippolyta and 32 S. z. gloriosa individuals collected at 9 locations in western Oregon and northwestern California. The most pertinent findings thus far are summarized as follows: Among OSB populations, genetic diversity is lowest at Mount Hebo and highest at Rock Creek and Bray Point. Of the 32 haplotypes detected in OSB, only 2 were shared among populations (1 shared by Mount Hebo, Cascade Head, Bray Point, and Rock Creek, and 1 shared by Rock Creek and Lake Earl). The remaining 30 haplotypes were identified in individual populations, highlighting the strong differentiation among sites. It is unclear if the shared haplotypes represent widespread, naturally occurring genetic variation or if allele sharing among populations is due to translocation history. Using full siblings of individuals that were released at Rock Creek and Bray Point in 2012 as comparison standards, the analyses suggest that 54 percent of the sampled individuals from Bray Point were naturally recruited into the population and were not originating from the 2012 release of captive reared individuals. Likewise, 33 percent of the analyzed individuals from Rock Creek were naturally recruited. Both of these estimates may be underestimates if the shared alleles that we identified among populations are naturally occurring and not a product of the 2012 translocations. The results suggest that there are about 12–13 COI haplotypes in the Mount Hebo population. The U.S. Fish and Wildlife Service anticipates using Mount Hebo as the source of individuals when establishing new populations in the future. Nonlinear regression models based on a series of rarefaction analyses suggest that progeny from 12, 37, 109, and 326 female individuals would be required to respectively capture 25, 50, 75, and 90 percent of the allelic diversity from Mount Hebo. Phylogenetic analyses identified two different haplotype groups, but the two groups did not correspond to the different subspecies used in the analysis. One group included 22 S. z. hippolyta haplotypes and 7 haplotypes identified in S. z. gloriosa . The second group included eight haplotypes from S. z. hippolyta, three haplotypes from S. z. gloriosa , and one haplotype that was detected in both subspecies.

California, Oregon↗

USGS2001

Cytochrome b sequences in black-crowned night-herons (Nycticorax nycticorax) from heronries exposed to genotoxic contaminants

DNA sequence analysis of a 215 base-pair region of the mitochondrial cytochrome b gene was used to examine genetic variation and search for evidence of an increased mutation rate in black-crowned night-herons. We examined five populations exposed to environmental contamination (primarily PAHs and PCBs) and one reference population from the eastern U.S. There was no evidence of a high mutation rate even within populations previously shown to exhibit increased variation in DNA content among somatic cells as a result of petroleum exposure. Three haplotypes were observed among 99 individuals. The low level of variability could be evidence for a genetic bottleneck, or that cytochrome b is too conservative for use in population genetic studies of this species. With the exception of one population from Louisiana, pair-wise Phi st estimates were very low, indicative of little population structure and potentially high rates of effective migration among populations.

Ecotoxicology↗

USGS2012

Concurrent speciation in the eastern woodland salamanders (Genus Plethodon):DNA sequences of the complete albumin nuclear and partialmitochondrial 12s genes

Salamanders of the North American plethodontid genus Plethodon are important model organisms in a variety of studies that depend on a phylogenetic framework (e.g., chemical communication, ecological competition, life histories, hybridization, and speciation), and consequently their systematics has been intensively investigated over several decades. Nevertheless, we lack a synthesis of relationships among the species. In the analyses reported here we use new DNA sequence data from the complete nuclear albumin gene (1818 bp) and the 12s mitochondrial gene (355 bp), as well as published data for four other genes (Wiens et al., 2006), up to a total of 6989 bp, to infer relationships. We relate these results to past systematic work based on morphology, allozymes, and DNA sequences. Although basal relationships show a strong consensus across studies, many terminal relationships remain in flux despite substantial sequencing and other molecular and morphological studies. This systematic instability appears to be a consequence of contemporaneous bursts of speciation in the late Miocene and Pliocene, yielding many closely related extant species in each of the four eastern species groups. Therefore we conclude that many relationships are likely to remain poorly resolved in the face of additional sequencing efforts. On the other hand, the current classification of the 45 eastern species into four species groups is supported. The Plethodon cinereus group (10 species) is the sister group to the clade comprising the other three groups, but these latter groups (Plethodon glutinosus [28 species], Plethodon welleri [5 species], and Plethodon wehrlei [2 species]) probably diverged from each other at approximately the same time.

Molecular Phylogenetics and Evolution↗

USGS2011

Sperm quality assessments for endangered razorback suckers Xyrauchen texanus

Flow cytometry (FCM) and computer-assisted sperm motion analysis (CASA) methods were developed and validated for use with endangered razorback suckers Xyrauchen texanus collected ( n =64) during the 2006 spawning season. Sperm motility could be activated within osmolality ranges noted during milt collections (here 167–343 mOsm/kg). We hypothesized that sperm quality of milt collected into isoosmotic (302 mOsm/kg) or hyperosmotic (500 mOsm/kg) Hanks' balanced salt solution would not differ. Pre-freeze viabilities were similar between osmolalities (79%±6 (S.E.M.) and 76%±7); however, post-thaw values were greater in hyperosmotic buffer (27%±3 and 12%±2; P =0.0065), as was mitochondrial membrane potential (33%±4 and 13%±2; P =0.0048). Visual estimates of pre-freeze motility correlated with total ( r =0.7589; range 23–82%) and progressive motility ( r =0.7449) by CASA and were associated with greater viability ( r =0.5985; P <0.0001). Count (FCM) was negatively correlated with post-thaw viability ( r =-0.83; P =0.0116) and mitochondrial function ( r =-0.91; P =0.0016). By FCM-based assessments of DNA integrity, whereby increased fluorochrome binding indicated more fragmentation, higher levels were negatively correlated with count ( r =-0.77; P <0.0001) and pre-freeze viabilities ( r =-0.66; P =0.0004). Fragmentation was higher in isotonic buffer ( P =0.0234). To increase reproductive capacity of natural populations, the strategy and protocols developed can serve as a template for use with other imperiled fish species, biomonitoring, and genome banking.

Reproduction↗

USGS2013

Population genetics and evaluation of genetic evidence for subspecies in the Semipalmated Sandpiper (Calidris pusilla)

Semipalmated Sandpipers ( Calidris pusilla ) are among the most common North American shorebirds. Breeding in Arctic North America, this species displays regional differences in migratory pathways and possesses longitudinal bill length variation. Previous investigations suggested that genetic structure may occur within Semipalmated Sandpipers and that three subspecies corresponding to western, central, and eastern breeding groups exist. In this study, mitochondrial control region sequences and nuclear microsatellite loci were used to analyze DNA of birds (microsatellites: n = 120; mtDNA: n = 114) sampled from seven North American locations. Analyses designed to quantify genetic structure and diversity patterns, evaluate genetic evidence for population size changes, and determine if genetic data support the existence of Semipalmated Sandpiper subspecies were performed. Genetic structure based only on the mtDNA data was observed, whereas the microsatellite loci provided no evidence of genetic differentiation. Differentiation among locations and regions reflected allele frequency differences rather than separate phylogenetic groups, and similar levels of genetic diversity were noted. Combined, the two data sets provided no evidence to support the existence of subspecies and were not useful for determining migratory connectivity between breeding sites and wintering grounds. Birds from western and central groups displayed signatures of population expansions, whereas the eastern group was more consistent with a stable overall population. Results of this analysis suggest that the eastern group was the source of individuals that colonized the central and western regions currently utilized by Semipalmated Sandpipers.

Waterbirds↗

USGS2021

Population genomics of free-ranging Great Plains white-tailed and mule deer reflects a long history of interspecific hybridization

Hybridization is a natural process at species-range boundaries that may variably promote the speciation process or break down species barriers but minimally will influence management outcomes of distinct populations. White-tailed deer ( Odocoileus virginianus ) and mule deer ( Odocoileus hemionus ) have broad and overlapping distributions in North America and a recognized capacity for interspecific hybridization. In response to contemporary environmental change to any of one or multiple still-unknown factors, mule deer range is contracting westward accompanied by a westward expansion of white-tailed deer, leading to increasing interactions, opportunities for gene flow, and associated conservation implications. To quantify genetic diversity, phylogenomic structure, and dynamics of hybridization in sympatric populations of white-tailed and mule deer, we used mitochondrial cytochrome b data coupled with SNP loci discovered with double-digest restriction site-associated DNA sequencing. We recovered 25,018 SNPs across 92 deer samples from both species, collected from two regions of western Kansas. Eight individuals with unambiguous external morphology representing both species were of hybrid origin (8.7%), and represented the product of multi-generational backcrossing. Mitochondrial data showed both ancient and recent directional discordance with morphological species assignments, reflecting a legacy of mule deer males mating with white-tailed deer females. Mule deer had lower genetic diversity than white-tailed deer, and both mitochondrial and nuclear data suggest contemporary mule deer effective population decline. Landscape genetic analyses show relative isolation between the two study regions for white-tailed deer, but greater connectivity among mule deer, with predominant movement from north to south. Collectively, our results suggest a long history of gene flow between these species in the Great Plains and hint at evolutionary processes that purge incompatible functional genomic elements as a result of hybridization. Surviving hybrids evidently may be reproductive, but with unknown consequences for the future integrity of these species, population trajectories, or relative susceptibility to emerging pathogens.

Kansas↗

USGS2008

An introduced and a native vertebrate hybridize to form a genetic bridge to a second native species

The genetic impacts of hybridization between native and introduced species are of considerable conservation concern, while the possibility of reticulate evolution affects our basic understanding of how species arise and shapes how we use genetic data to understand evolutionary diversification. By using mitochondrial NADH dehydrogenase subunit 2 (ND2) sequences and 467 amplified fragment-length polymorphism nuclear DNA markers, we show that the introduced white sucker (Catostomus commersoni) has hybridized with two species native to the Colorado River Basin - the flannelmouth sucker (Catostomus latipinnis) and the bluehead sucker (Catostomus discobolus). Hybrids between the flannelmouth sucker and white sucker have facilitated introgression between the two native species, previously isolated by reproductive barriers, such that individuals exist with contributions from all three genomes. Most hybrids had the mitochondrial haplotype of the introduced white sucker, emphasizing its pivotal role in this three-way hybridization. Our findings highlight how introduced species can threaten the genetic integrity of not only one species but also multiple previously reproductively isolated species. Furthermore, this complex three-way reticulate (as opposed to strictly bifurcating) evolution suggests that seeking examples in other vertebrate systems might be productive. Although the present study involved an introduced species, similar patterns of hybridization could result from natural processes, including stream capture or geological formations (e.g., the Bering land bridge). ?? 2008 by The National Academy of Sciences of the USA.

Proceedings of the National Academy of Sciences of↗

USGS2020

Novel molecular resources to facilitate future genetics research on freshwater mussels (Bivalvia: Unionidae)

Molecular data have been an integral tool in the resolution of the evolutionary relationships and systematics of freshwater mussels, despite the limited number of nuclear markers available for Sanger sequencing. To facilitate future studies, we evaluated the phylogenetic informativeness of loci from the recently published anchored hybrid enrichment (AHE) probe set Unioverse and developed novel Sanger primer sets to amplify two protein-coding nuclear loci with high net phylogenetic informativeness scores: fem-1 homolog C (FEM1) and UbiA prenyltransferase domain-containing protein 1 (UbiA). We report the methods used for marker development, along with the primer sequences and optimized PCR and thermal cycling conditions. To demonstrate the utility of these markers, we provide haplotype networks, DNA alignments, and summary statistics regarding the sequence variation for the two protein-coding nuclear loci (FEM1 and UbiA). Additionally, we compare the DNA sequence variation of FEM1 and UbiA to three loci commonly used in freshwater mussel genetic studies: the mitochondrial genes cytochrome c oxidase subunit 1 (CO1) and NADH dehydrogenase subunit 1 (ND1), and the nuclear internal transcribed spacer 1 (ITS1). All five loci distinguish among the three focal species (Potamilus fragilis, Potamilus inflatus, and Potamilus purpuratus), and the sequence variation was highest for ND1, followed by CO1, ITS1, UbiA, and FEM1, respectively. The newly developed Sanger PCR primers and methodologies for extracting additional loci from AHE probe sets have great potential to facilitate molecular investigations targeting supraspecific relationships in freshwater mussels, but may be of limited utility at shallow taxonomic scales.

Data↗