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Results for “Mitochondrial DNA”

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USGS2002

Molecular analysis of a 11 700-year-old rodent midden from the Atacama Desert, Chile

DNA was extracted from an 11 700-year-old rodent midden from the Atacama Desert, Chile and the chloroplast and animal mitochondrial DNA (mtDNA) gene sequences were analysed to investigate the floral environment surrounding the midden, and the identity of the midden agent. The plant sequences, together with the macroscopic identifications, suggest the presence of 13 plant families and three orders that no longer exist today at the midden locality, and thus point to a much more diverse and humid climate 11 700 years ago. The mtDNA sequences suggest the presence of at least four different vertebrates, which have been putatively identified as a camelid (vicuna), two rodents (Phyllotis and Abrocoma), and a cardinal bird (Passeriformes). To identify the midden agent, DNA was extracted from pooled faecal pellets, three small overlapping fragments of the mitochondrial cytochrome b gene were amplified and multiple clones were sequenced. These results were analysed along with complete cytochrome b sequences for several modern Phyllotis species to place the midden sequence phylogenetically. The results identified the midden agent as belonging to an ancestral P. limatus. Today, P. limatus is not found at the midden locality but it can be found 100 km to the north, indicating at least a small range shift. The more extensive sampling of modern Phyllotis reinforces the suggestion that P. limatus is recently derived from a peripheral isolate.

Molecular Ecology↗

USGS2020

Resolving species boundaries in the critically imperiled freshwater mussel species, Fusconaia mitchelli (Bivalvia: Unionidae)

Species are a fundamental unit of biology, and defining accurate species boundaries is integral to effective conservation and management of imperiled taxa. Freshwater mussels (Bivalvia: Unionidae) are among the most imperiled groups of organisms in North America, yet species boundaries remain uncertain for many taxa. The False Spike, Fusconaia mitchelli (Simpson in Dall, 1895), is a freshwater mussel considered to be endemic to central Texas (Brazos, Colorado, and Guadalupe drainages). Recent research revealed significant intraspecific genetic variation between geographically separated populations of F . mitchelli , which could be indicative of speciation; however, small sample sizes for several of the populations precluded formal taxonomic revision. Here, we increase taxon sampling and use multilocus DNA sequence data and traditional morphometrics to re‐evaluate species boundaries in F . mitchelli . We sequenced three loci: the protein‐coding mitochondrial DNA genes cytochrome c oxidase subunit 1 and NADH dehydrogenase 1 , and the nuclear internal transcribed spacer 1 . Phylogenetic analyses depicted deep genetic divergence between F . mitchelli in the Guadalupe and those in the Brazos and Colorado drainages, which was further supported by available biogeographic information. Morphometric analyses and coalescent‐based species delimitation models integrating both DNA sequence and morphological data provided strong support for the divergence observed between the two geographically isolated clades of F . mitchelli . Based on these results, we revise taxonomy accordingly by elevating the junior synonym Fusconaia iheringi (Wright, 1898) to represent the Brazos and Colorado populations and restrict the distribution of F . mitchelli to the Guadalupe River drainage. Our findings may impact pending management decisions to protect F . mitchelli under the U.S. Endangered Species Act.

Journal of Zoological Systematics and Evolutionary↗

USGS2023

Development of a quantitative PCR assay for detecting northwest salamander (ambystoma gracile) in environmental DNA samples

We developed a primer and probe based quantitative PCR assay for use with environmental DNA to detect Northwest salamander ( Ambystoma gracile ), a species endemic to the temperate Pacific coastal region of North America. The assay targets a region in the mitochondrial DNA (mtDNA) D-loop gene. Tests of the assay were performed in silico (using the NCBI BLAST tool), in vitro (using DNA extracted from A. gracile and related species) and in situ (water samples collected from a lake with a known population of A. gracile ). This assay will be useful in efforts to monitor the distribution and occupancy of this species.

Conservation Genetics Resources↗

USGS2014

The history of sturgeon in the Baltic Sea

Aim Migrants of the Atlantic sturgeon, Acipenser oxyrinchus, from North America are thought to have founded the Baltic sturgeon population during the Little Ice Age around 1200 years ago, replacing the European sturgeon, Acipenser sturio. To test this hypothesis and to further elucidate the colonization of the Baltic Sea by A. oxyrinchus, we carried out DNA analyses of ancient and contemporary populations of both species. Location We analysed DNA from 188 specimens of sturgeons collected from archaeological sites and museums in Poland and of 225 contemporary specimens from North American and European populations. Methods Several mitochondrial DNA fragments were sequenced and eight microsatellite loci were genotyped for species identification, polymorphism and population structure analyses. Approximate Bayesian computation was used to estimate when the Baltic Sea was colonized. Results Of 125 ancient sturgeon specimens from the Baltic Sea, only four were classified as A. sturio, the remainder being A. oxyrinchus oxyrinchus. The ancient A. o. oxyrinchus population over two different time periods was highly polymorphic and genetically distant from contemporary populations of this taxon. The time of entry into the Baltic Sea was estimated to be 4000–5000 years ago. We also detected introgression of A. sturio into the A. o. oxyrinchus gene pool, caused by a prior hybridization event. Main conclusions For the past 2000 years at least, A. o. oxyrinchus has been the dominant sturgeon in the Baltic Sea, indicating a much earlier origin than previously suggested. The most similar extant sturgeon populations to the extinct Baltic stock are those from the St John and St Lawrence rivers in Canada. These populations should be considered the best source of breeding material for the ongoing sturgeon restitution programmes in Poland and Germany.

Journal of Biogeography↗

USGS2007

Polymorphic microsatellite DNA markers for the Florida manatee (Trichechus manatus latirostris)

Florida manatees (Trichechus manatus latirostris) are marine mammals that inhabit the coastal waters and rivers of the southeastern USA, primarily Florida. Previous studies have shown that Florida manatees have low mitochondrial DNA variability, suggesting that nuclear DNA loci are necessary for discriminatory analyses. Here we report 10 polymorphic microsatellite loci with an average of 4.2 alleles per locus, and average heterozygosity of 50.1%. These loci have been developed for use in population studies, parentage assignment, and individual identification. ?? 2007 Blackwell Publishing Ltd.

Molecular Ecology Notes↗

USGS2009

Genetic variation in domestic reindeer and wild caribou in Alaska

Reindeer were introduced into Alaska 100 years ago and have been maintained as semidomestic livestock. They have had contact with wild caribou herds, including deliberate cross-breeding and mixing in the wild. Reindeer have considerable potential as a domestic animal for meat or velvet antler production, and wild caribou are important to subsistence and sport hunters. Our objective was to quantify the genetic relationships of reindeer and caribou in Alaska. We identified allelic variation among five herds of wild caribou and three herds of reindeer with DNA sequencing and restriction enzymes for three loci: a DQA locus of the major histocompatibility complex (Rata-DQA1), k-casein and the D-loop of mitochondrial DNA. These loci are of interest because of their potential influence on domestic animal performance and the fitness of wild populations. There is considerable genetic variation in reindeer and caribou for all three loci, including five, three and six alleles for DQA, k-casein and D-loop respectively. Most alleles occur in both reindeer and caribou, which may be the result of recent common ancestry or genetic introgression in either direction. However, allele frequencies differ considerably between reindeer and caribou, which suggests that gene flow has been limited.

Alaska↗

USGS1998

DNA and allozyme markers provide concordant estimates of population differentiation: Analyses of U.S. and Canadian populations of Yukon River fall-run chum salmon ( Oncorhynchus keta )

Although the number of genetic markers available for fisheries research has steadily increased in recent years, there is limited information on their relative utility. In this study, we compared the performance of different "classes" of genetic markers (mitochondrial DNA (mtDNA), nuclear DNA (nDNA), and allozymes) in terms of estimating levels and partitioning of genetic variation and of the relative accuracy and precision in estimating population allocations to mixed-stock fisheries. Individuals from eight populations of fall-run chum salmon ( Oncorhynchus keta ) from the Yukon River in Alaska and Canada were assayed at 25 loci. Significant differences in mitochondrial haplotype and nuclear allele frequencies were observed among five drainages. Populations from the U.S.-Canada border region were not clearly distinguishable based on multilocus allele frequencies. Although estimates of total genetic diversities were higher for the DNA loci ( H t = 0.592 and h = 0.647 for nDNA and mtDNA, respectively) compared with protein allozymes ( H t = 0.250), estimates of the extent of population differentiation were highly concordant across marker classes (mean theta = 0.010, 0.011, and 0.016 for allozymes, nDNA, and mtDNA, respectively). Simulations of mixed-stock fisheries composed of varying contributions of U.S. and Canadian populations revealed a consistent bias for overallocation of Canadian stocks when expected Canadian contributions varied from 0 to 40%, due primarily to misallocations among genetically similar border populations. No single marker class is superior for differentiating populations of this species at the spatial scale examined.

Yukon River↗

USGS2000

Asymmetric hybridization and introgression between pink salmon and chinook salmon in the Laurentian Great Lakes

Among Pacific salmon collected in the St. Marys River, five natural hybrids of pink salmon Oncorhynchus gorbuscha and chinook salmon Oncorhynchus tshawytscha and one suspected backcross have been detected using morphologic, meristic, and color evidence. One allozyme (LDH, l-lactate dehydrogenase from muscle) and one nuclear DNA locus (growth hormone) for which species-specific fixed differences exist were analyzed to detect additional hybrids and to determine if introgression had occurred. Restriction fragment length polymorphism of mitochondrial DNA (mtDNA) was used to identify the maternal parent of each hybrid. Evidence of introgression was found among the five previously identified hybrids. All hybrid specimens had chinook salmon mtDNA, indicating that hybridization between chinook salmon and pink salmon in the St. Marys River is asymmetric and perhaps unidirectional. Ecological, physiological, and sexual selection forces may contribute to this asymmetric hybridization. Introgression between these highly differentiated species has implications for management, systematics, and conservation of Pacific salmon.

Transactions of the American Fisheries Society↗

USGS2012

Population divergence and gene flow in an endangered and highly mobile seabird

Seabirds are highly vagile and can disperse up to thousands of kilometers, making it difficult to identify the factors that promote isolation between populations. The endemic Hawaiian petrel (Pterodroma sandwichensis) is one such species. Today it is endangered, and known to breed only on the islands of Hawaii, Maui, Lanai and Kauai. Historical records indicate that a large population formerly bred on Molokai as well, but this population has recently been extirpated. Given the great dispersal potential of these petrels, it remains unclear if populations are genetically distinct and which factors may contribute to isolation between them. We sampled petrels from across their range, including individuals from the presumably extirpated Molokai population. We sequenced 524 bp of mitochondrial DNA, 741 bp from three nuclear introns, and genotyped 18 microsatellite loci in order to examine the patterns of divergence in this species and to investigate the potential underlying mechanisms. Both mitochondrial and nuclear data sets indicated significant genetic differentiation among all modern populations, but no differentiation was found between historic samples from Molokai and modern birds from Lanai. Population-specific nonbreeding distribution and strong natal philopatry may reduce gene flow between populations. However, the lack of population structure between extirpated Molokai birds and modern birds on Lanai indicates that there was substantial gene flow between these populations and that petrels may be able to overcome barriers to dispersal prior to complete extirpation. Hawaiian petrel populations could be considered distinct management units, however, the dwindling population on Hawaii may require translocation to prevent extirpation in the near future.

Hawaii↗

USGS1998

Molecular contributions to conservation

Recent advances in molecular technology have opened a new chapter in species conservation efforts, as well as population biology. DNA sequencing, MHC (major histocompatibility complex), minisatellite, microsatellite, and RAPD (random amplified polymorphic DNA) procedures allow for identification of parentage, more distant relatives, founders to new populations, unidentified individuals, population structure, effective population size, population-specific markers, etc. PCR (polymerase chain reaction) amplification of mitochondrial DNA, nuclear DNA, ribosomal DNA, chloroplast DNA, and other systems provide for more sophisticated analyses of metapopulation structure, hybridization events, and delineation of species, subspecies, and races, all of which aid in setting species recovery priorities. Each technique can be powerful in its own right but is most credible when used in conjunction with other molecular techniques and, most importantly, with ecological and demographic data collected from the field. Surprisingly few taxa of concern have been assayed with any molecular technique. Thus, rather than showcasing exhaustive details from a few well-known examples, this paper attempts to present a broad range of cases in which molecular techniques have been used to provide insight into conservation efforts.

Ecology↗

USGS2020

Genomic identification of intergeneric hybrids in New World wood-warblers (Aves: Parulidae)

The documentation of hybrids between distantly related taxa can illustrate an initial step to explain how genes might move between species that do not exhibit complete reproductive isolation. In birds, some of the most phylogenetically distant hybrid combinations occur between genera. Traditionally, morphological and plumage characters have been used to assign the identity of the parental species of a putative hybrid, although recently, nuclear introns also have been used. Here, we demonstrate how high-throughput short-read DNA sequence data can be used to identify the parentage of a putative intergeneric hybrid, in this case between a blue-winged warbler ( Vermivora cyanoptera ) and a cerulean warbler ( Setophaga cerulea ). This hybrid had mitochondrial DNA of a cerulean warbler, indicating the maternal parent. For hundreds of single nucleotide polymorphisms within six regions of the nuclear genome that differentiate blue-winged warblers and golden-winged warblers ( Vermivora chrysoptera ), the hybrid had roughly equal ancestry assignment to blue-winged and cerulean warblers, suggesting a blue-winged warbler as the paternal parent species and demonstrating that this was a first generation (F 1 ) hybrid between these species. Unlike other recently characterized intergeneric warbler hybrids, this individual hybrid learned to song match its maternal parent species, suggesting that it might have been the result of an extra-pair mating and raised in a cerulean warbler nest.

Biological Journal of the Linnean Society↗

USGS2020

Testing environmental DNA from wolf snow tracks for species, sex, and individual identification

Monitoring elusive, relatively low-density, large predators, such as the grey wolf (Canis lupus), has often been accomplished by live-capture and radiocollaring. Increasingly, non-invasive methods are considered best practice whenever it is possible to use them. Recently, environmental DNA (eDNA) deposited in snow tracks was demonstrated as useful for identifying lynx (Lynx canadensis), fisher (Pekania pennanti), wolverine (Gulo gulo), sika deer (Cervus nippon), red fox (Vulpes vulpes), and the Japanese marten (Martes melampus) to species level using mitochondrial DNA (mtDNA) markers. We tested whether eDNA from fresh wolf snow tracks collected in the Superior National Forest, Minnesota, USA during winter 2019 could be used to identify species, sex, and individual. Seven of the 8 snow track samples were successfully identified to “wolf-dog” species using mtDNA, with alleles amplifying in 5 of the samples at 1 or both of loci u250 and FH2096 in the allele range for wolves for this population. None yielded enough high-quality DNA to obtain genotypes to determine individual or sex. We recommend additional field trials to determine the minimum number of tracks required per individual to obtain sufficient, high-quality eDNA, as well as collecting associated urine or blood (from estrus) when possible. If individual wolves could be identified and sexed by the eDNA in their snow tracks, researchers should be able to determine population and family or group metrics with greater precision and less effort than typically required when conducting winter scat or hair-based genetic field studies of low density, elusive carnivores. Furthermore, this method would have applications to other areas of research and management, such as hunting quota determinations, validation of field methods and, in particular regards to wolves and other predators, livestock depredation issues. Overall, this technique holds significant future promise as field and laboratory methods are further refined for greater precision and optimized regarding the varying collection, filtration, and extraction protocols for different species in various environments.

Minnesota↗

USGS2020

High site fidelity does not equate to population genetic structure for common goldeneye and Barrow's goldeneye in North America

Delineation of population structure provides valuable information for conservation and management of species, as levels of demographic and genetic connectivity not only affect population dynamics but also have important implications for adaptability and resiliency of populations and species. Here, we measure population genetic structure and connectivity across the ranges of two sister species of sea ducks: Barrow's goldeneye Bucephala islandica and common goldeneye B. clangula . We use two different marker types: 7–8 nuclear microsatellite loci assayed across 229 samples and 3678 double digest restriction‐site associated DNA sequencing (ddRAD‐seq) loci assayed across 61 samples. First, both datasets found no evidence of genetic structure within common or Barrow's goldeneye, including between North American and European samples of common goldeneye. These results are in contrast with previous mitochondrial DNA, band recovery and telemetry data which suggest that goldeneyes are structured across their range. We posit that the discordance between autosomal genetic markers and other data types suggests that males, possibly subadult males, may be maintaining genetic connectivity across each species' respective ranges. Next, although mate choice consequences resulting from inter‐specific brood parasitism was hypothesized to cause some level of gene flow between goldeneye species, we only identified a single F1 hybrid with no further evidence of contemporary or historical gene flow. Despite ddRAD‐seq demographic analyses which recovered an optimum evolutionary model of split‐with‐migration (i.e. secondary contact), estimates of gene flow were <<1 migrant per generation in both directions. Together, we conclude that either strong ecological barriers or assortative mating are likely playing a role in preventing further backcrossing. Finally, demographic analyses estimated a relatively deep divergence time between Barrow's goldeneye and common goldeneye of ~1.6 million years before present and suggests that the genomes of both species have been under similar evolutionary constraints.

Journal of Avian Biology↗

USGS2016

Ancient DNA reveals substantial genetic diversity in the California Condor ( Gymnogyps californianus ) prior to a population bottleneck

Critically endangered species that have undergone severe population bottlenecks often have little remaining genetic variation, making it difficult to reconstruct population histories to apply in reintroduction and recovery strategies. By using ancient DNA techniques, it is possible to combine genetic evidence from the historical population with contemporary samples to provide a more complete picture of a species' genetic variation across its historical range and through time. Applying this approach, we examined changes in the mitochondrial DNA (mtDNA) control region (526 base pairs) of the endangered California Condor ( Gymnogyps californianus ). Results showed a >80% reduction in unique haplotypes over the past 2 centuries. We found no spatial sorting of haplotypes in the historical population; the periphery of the range contained haplotypes that were common throughout the historical range. Direct examination of mtDNA from California Condor museum specimens provided a new window into historical population connectivity and genetic diversity showing: (1) a substantial loss of haplotypes, which is consistent with the hypothesis that condors were relatively abundant in the nineteenth century, but declined rapidly as a result of human-caused mortality; and (2) no evidence of historical population segregation, meaning that the available genetic data offer no cause to avoid releasing condors in unoccupied portions of their historical range.

The Condor↗

USGS2024

Genetic evidence for the presence of wild-caught sturgeons in commercial markets in Georgia

Sturgeons (Family: Acipenseridae) are among the most endangered taxa worldwide. Significant resources have been invested into the conservation of global sturgeon populations, including the development of commercial aquaculture programs. These programs are intended to improve conservation outcomes by reducing the harvest of wild populations while still meeting commercial demand for sturgeon products. However, there is growing concern that commercial aquaculture programs may contribute to wild population declines through continued, illegal harvest and the escape and/or release of captive individuals into wild environments. These concerns may be particularly acute in the country of Georgia which, despite its small territory and altered landscape, is a globally significant hotspot for sturgeon diversity. In order to understand the potential threat of captive culture on wild sturgeon populations in Georgia, we used mitochondrial DNA sequencing and microsatellite analyses to identify the species and origin of sturgeons encountered in commercial settings. Microsatellite analyses showed significant differentiation between wild and commercial Russian sturgeon populations and highlighted the potential for wild-caught individuals to be present in coastal markets in Georgia. The analyses of mitochondrial haplotypes also suggested that commercial markets may contain sturgeon species that are not native to the region. Overall, our results suggest that wild sturgeon populations may still be exploited to support captive aquaculture programs and commercial sales.

Diversity↗

USGS2017

Phylogeny, diet, and habitat of an extinct ground sloth from Cuchillo Curá, Neuquén Province, southwest Argentina

Advancements in ancient DNA analyses now permit comparative molecular and morphological studies of extinct animal dung commonly preserved in caves of semiarid regions. These new techniques are showcased using a unique dung deposit preserved in a late glacial vizcacha ( Lagidium sp.) midden from a limestone cave in southwestern Argentina (38.5° S). Phylogenetic analyses of the mitochondrial DNA show that the dung originated from a small ground sloth species not yet represented by skeletal material in the region, and not closely related to any of the four previously sequenced extinct and extant sloth species. Analyses of pollen and plant cuticles, as well as analyses of the chloroplast DNA, show that the Cuchillo Curá ground sloth browsed on many of the same herb, grass, and shrub genera common at the site today, and that its habitat was treeless Patagonian scrub-steppe. We envision a day when molecular analyses are used routinely to supplement morphological identifications and possibly to provide a time-lapse view of molecular diversification.

Quaternary Research↗

USGS2006

Use of buccal swabs for sampling DNA from nestling and adult birds

We evaluated the feasibility and efficiency of using swabs to collect buccal epithelial cells from small (2‐ to 13‐ g ) birds as a source of DNA for genetic studies. We used commercially available buccal swab kits to collect samples from 42 adult and 39 nestling (4‐ to 8‐day‐old) black‐capped chickadees (Poecile atricapillus) and from 6 4‐day‐old nestling boreal chickadees (P. hudsonica). We compared DNA from buccal epithelial samples to that from blood samples from the same individuals. We extracted sufficient quantities of DNA for analysis from all buccal samples, and samples remained viable even after being stored in original plastic sampling tubes at room temperature for up to 18 months. Yields were equivalent whether extracted using the proprietary quick‐extraction solution provided with buccal swab kits or using a salt‐extraction process with inexpensive reagents. Yields of DNA from buccal samples were consistently lower than those from blood samples, but quantities were sufficient for all analyses. Assignment of sex, based on DNA extracted from paired buccal and blood samples, was identical for all 87 birds. We found no difference in the genotypes obtained from buccal and blood samples for 12 individuals tested using 5 microsatellite loci and found perfect concordance in sequencing of an 823‐base‐pair segment within the control region of mitochondrial DNA for 7 individuals tested. Use of buccal swabs is highly recommended as a rapid, noninvasive technique for sampling avian genomic DNA, especially for extremely young altricial nestlings or small‐bodied adults, or for any birds for which blood sampling may be impossible or stressful.

Wildlife Society Bulletin↗

USGS2023

Dead-end hollow fiber ultrafiltration capture of environmental DNA for freshwater mussel (Unionidae) species detection with metabarcoding

Insufficient water sample volumes can be a limiting factor for detecting species with environmental DNA (eDNA) from aquatic habitats. We compared detections of freshwater mussel (Unionidae) communities using large water sample volumes and dead-end hollow fiber ultrafiltration (D-HFUF or DEUF) with traditional eDNA filtration methods that use relatively small water sample volumes. Unionid species were detected in approximately 50-L D-HFUF eDNA samples with two mitochondrial DNA metabarcoding markers (COI and ND1) and compared to species detection results from eDNA captured from commonly used 1-L samples filtered with polyethersulfone (PES) filters at three lotic sites in Georgia and Missouri. Of the 431,560 COI and 1,035,472 ND1 reads from all environmental samples of both filter types that passed quality control, 95% (410,755 reads) of COI reads and 85% (883,472 reads) of ND1 reads were assigned to a unionid species. Nineteen different freshwater mussel species were detected across all D-HFUF samples, and 11 species were detected across all PES samples. Reads assigned to the genus Elliptio could not be resolved beyond the genus level with either marker. From D-HFUF samples, 15 and 16 mussel species were detected with the COI and ND1 markers, respectively. From PES samples, nine and seven species were detected with the COI and ND1 markers, respectively. More mussel species were detected at each site in D-HFUF samples than in PES samples regardless of whether results from both markers were combined or evaluated separately. Our results demonstrate the merit of further exploration and optimization of D-HFUF for capturing eDNA from high-volume water samples to facilitate detection of unionids and likely other aquatic organisms.

Environmental DNA↗