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At least 109 records · Page 6Linked to original sources

Optimization of wetland environmental DNA metabarcoding protocols for Great Lakes region herpetofauna

Many species of reptiles and amphibians (herpetofauna) rely on wetlands that are being degraded and lost at a high rate. Characterization of herpetofauna diversity in different wetland types may help guide conservation strategies. However, traditional survey methods often involve sampling within small temporal windows, and the gear deployed may be taxonomically biased, thus, they may fail to accurately characterize species presence/absence and diversity. In contrast, environmental (e)DNA metabarcoding has been shown to effectively survey entire aquatic communities and can provide a useful complement to traditional surveys. The objective of this study was to design and optimize eDNA sampling and laboratory protocols for wetland herpetofauna. Protocols evaluated included different water sampling approaches (point versus transect sampling), seasonality of sampling, and choice of metabarcoding marker (mitochondrial 12S versus 16S rDNA). Samples collected from 10 sites across southern Michigan detected 17 amphibian and five reptile species, including four species of conservation concern ( Ambystoma texanum , Clemmys guttata , Rana palustris , and Sternotherus odoratus ). We observed no difference in the number of species detected between point and transect samples ( p = 0.70), but point sampling required less time ( p = 0.03) and allowed significantly larger volumes of water to be filtered ( p = 1.13e-5). No difference in species richness was observed between the 12S and 16S mitochondrial DNA markers ( p = 0.96). However, a greater number of taxa were identifiable at the species level when using the 16S locus. There was also a significant difference in the number of species detected between early and late summer sampling periods (more species detected in the earlier period; p = 6.31e-6), and some species were only found in the early or late sampling period. Sampling during multiple periods to fully characterize species composition, the use of point sampling, and the 16S mtDNA marker for herpetofauna eDNA metabarcoding studies may increase efficiency and reliability of results.

Environmental DNA

Hybridization and the phylogenetic relationship between polecats and domestic ferrets in Britain

Ferrets ( Mustela furo ) were domesticated from polecats ( M. putorius, M. eversmannii ) over 2000 years ago. Following their introduction to Britain, they escaped and hybridized with native European polecats ( M. putorius ). Native polecats declined to the point of near extinction prior to World War I, but have recently begun to expand from a Welsh refugium. Concern has arisen as to the extent of polecat/ferret introgression, and in particular, whether the expanding population is of mainly hybrid origin. Therefore, mitochondrial DNA sequencing was used to investigate polecat genetic diversity in Britain. Two geographically distinct lineages were found, where one may be ancestral to the British polecat, and the other to the domestic ferret. The ancestral distribution of each lineage, or assortative mating is sufficient to explain the observed pattern. A further comparison between the distribution of the polecat phenotype and mitochondrial haplotype implies that the current population expansion may be mediated by dispersing male polecats hybridizing with female feral ferrets. However, the wild source of the ferret remains obscure. Relatively recent speciation from European mink ( M. lutreola ) and black-footed ferrets ( M. nigripes ), and/or the effects of hybridization result in an unresolved molecular phylogeny.

Biological Conservation

Mitochondrial and nuclear genetic relationships of deer ( Odocoileus spp.) in western North America

Odocoileus hemionus (mule deer and black-tailed deer) and Odocoileus virginanus (white-tailed deer) are sympatric in western North America and are characterized by distinct morphology, behavior, and allozyme allele frequencies. However, there is discordance among nuclear and mitochondrial genetic relationships, as mule deer ( O . h . hemionus ) and white-tailed deer have similar mitochondrial DNA (mtDNA) which is very different from that of black-tailed deer ( O . h . columbianus , O . h . sitkensis ). I expanded previous studies to clarify the genetic relationships of these groups by determining mtDNA haplotype and allozyme genotypes for 667 deer from several locations in northwestern North America. Different mtDNA haplotypes in mule deer, black-tailed deer, and white-tailed deer indicate that mitochondrial gene flow is restricted. Allozyme allele frequencies indicate that there is also restriction of nuclear gene flow between O . virginianus and O . hemionus , and to a lesser extent between mule deer and black-tailed deer. There is a low level of introgressive hybridization of mtDNA from mule deer and black-tailed deer into white-tailed deer populations and considerable interbreeding of mule deer and black-tailed deer in a contact zone. The discordance of mitochondrial and nuclear genomes is apparent only if mtDNA sequence divergences, and not haplotype frequencies, are considered.

Canadian Journal of Zoology

Changes in genetic diversity and differentiation in Red‐cockaded woodpeckers (Dryobates borealis) over the past century

Red‐cockaded woodpeckers (RCW; Dryobates borealis ) declined after human activities reduced their fire‐maintained pine ecosystem to <3% of its historical range in the southeastern United States and degraded remaining habitat. An estimated 1.6 million RCW cooperative breeding groups declined to about 3,500 groups with no more than 10,000 birds by 1978. Management has increased RCW population abundances since they were at their lowest in the 1990s. However, no range‐wide study has been undertaken since then to investigate the impacts of this massive bottleneck or infer the effects of conservation management and recent demographic recoveries. We used mitochondrial DNA sequences (mtDNA) and nine nuclear microsatellite loci to determine if range‐wide demographic declines resulted in changes to genetic structure and diversity in RCW by comparing samples collected before 1970 (mtDNA data only), between 1992 and 1995 (mtDNA and microsatellites), and between 2010 and 2014 (mtDNA and microsatellites). We show that genetic diversity has been lost as detected by a reduction in the number of mitochondrial haplotypes. This reduction was apparent in comparisons of pre‐1970 mtDNA data with data from the 1992–1995 and 2010–2014 time points, with no change between the latter two time points in mtDNA and microsatellite analyses. The mtDNA data also revealed increases in range‐wide genetic differentiation, with a genetically panmictic population present throughout the southeastern United States in the pre‐1970s data and subsequent development of genetic structure that has remained unchanged since the 1990s. Genetic structure was also uncovered with the microsatellite data, which like the mtDNA data showed little change between the 1992–1995 and 2010–2014 data sets. Temporal haplotype networks revealed a consistent, star‐like phylogeny, suggesting that despite the overall loss of haplotypes, no phylogenetically distinct mtDNA lineages were lost when the population declined. Our results may suggest that management during the last two decades has prevented additional losses of genetic diversity.

Alabama, Arkansas, Florida, Georgia, Kentucky, Lou

Importance of tributary streams for rainbow trout reproduction: insights from a small stream in Georgia and a bi-genomic approach

Tributaries of tailwater fisheries in the southeastern USA have been used for spawning by stocked rainbow trout (Oncorhynchus mykiss), but their importance may have been underestimated using traditional fish survey methods such as electrofishing and redd counts. We used a bi-genomic approach, mitochondrial DNA sequences and nuclear microsatellite loci, to estimate the number of spawning adults in one small tributary (Cabin Creek) of the Chattahoochee River, Georgia, where rainbow trout are known to spawn and have successful recruitment. We extracted and analysed DNA from seven mature male rainbow trout and four juveniles that were captured in February 2006 in Cabin Creek and from 24 young-of-year (YOY) trout that were captured in April 2006. From these samples, we estimated that 24 individuals were spawning to produce the amount of genetic variation observed in the juveniles and YOY, although none of the mature males we sampled were indicated as sires. Analysis of the mitochondrial D-loop region identified four distinct haplotypes, suggesting that individuals representing four maternal lineages contributed to the offspring. Our analyses indicated that many more adults were spawning in this system than previously estimated with direct count methods and provided insight into rainbow trout spawning behavior.

River Research and Applications

Metabarcoding assays for the detection of freshwater mussels (Unionida) with environmental DNA

Freshwater mussels of the order Unionida are a widely distributed taxon that are important in maintaining freshwater ecosystems and are also highly imperiled throughout the world. Monitoring of mussel populations with environmental DNA (eDNA) is an attractive alternative to traditional methods because it is noninvasive and requires less labor and taxonomic knowledge from field personnel. We developed eDNA metabarcoding assays specific to freshwater mussels and tested them at six sites in the Clinch River, located in the southeastern United States. Our objective was to determine the utility of eDNA metabarcoding for future monitoring of mussel populations and restoration efforts in this watershed. Two metabarcoding assays that target the mitochondrial DNA regions of the cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit (ND1) genes were developed and tested. Our assays appear to be order specific, amplifying members from the two families found in North America, Unionidae and Margaritiferidae, while not amplifying nontarget fish or other bivalve species. From the field collected samples, our assays together detected 19 species, eight of which are listed as federally endangered. The assays also detected 42%, 58%, and 54% of the species identified by recent quantitative visual mussel surveys at three sampling sites. Increased sampling effort by processing a greater water volume or number of samples will likely increase species detections. These eDNA metabarcoding assays may enable enhanced monitoring of freshwater mussel assemblages and subsequently inform conservation efforts.

Virginia

Genetic diversity in a morphologically conservative invasive taxon: Multiple introductions of swamp eels to the southeastern United States

Genetic analysis of introduced populations, especially in morphologically conservative taxa, can clarify introduction histories, identify management units and source populations, provide a more realistic estimate of the frequency of successful invasion, and suggest strategies for preventing further introductions. In the last 7 years, populations of swamp eels, referred to the Asian genus Monopterus (Family Synbranchidae) on the basis of external morphology, have been discovered in aquatic habitats near Atlanta, Georgia; Tampa, Florida; North Miami, Florida; and most recently in close proximity to Everglades National Park in Homestead, Florida. Swamp eels are large predators capable of dispersal over land and have the potential to disrupt already threatened ecosystems. We analyzed mitochondrial DNA sequences from four known populations in the continental United States and samples from Malaysia, Indonesia, Vietnam, and two locations in China to determine introduction histories, source populations, genetic diversity, and relationships among populations. Our results indicate that there have been at least three independent introductions of genetically distinct forms. Introduced populations in close proximity (separated by <40 km) are genetically distinct. The level of sequence difference among introduced populations reaches levels seen among sister families of teleost fishes for the same region of the mitochondrial genome. These genetically distinct introduced populations in all likelihood represent at least two and possibly three species. Regardless of species status, these genetically distinct lineages may be expected to vary in ecological or life-history traits, representing different potential threats to the ecosystems where they have been introduced. Given the success of swamp eels in invading many habitats around the world, further study of these eels is warranted to elucidate the characteristics of successful invaders and invasions.

Conservation Biology

Population status and population genetics of northern leopard frogs in Arizona

Increasing isolation of populations by habitat fragmentation threatens the persistence of many species, both from stochastic loss of small isolated populations, and from inbreeding effects in populations that have become genetically isolated. In the southwestern United States, amphibian habitat is naturally patchy in occurrence because of the prevailing aridity of the region. Streams, rivers, and other wetlands are important both as habitat and as corridors that connect populations. However, populations of some species have become more fragmented and isolated by habitat degradation and loss. Northern leopard frogs (Rana pipiens) have experienced serious declines in the Southwest. We conducted an extensive survey across the known range of northern leopard frogs in Arizona to determine the current distribution and abundance of the species. From a range that once spanned much of the northern and central part of the State, northern leopard frogs have been reduced to three or four widely separated populations, near Lyman Lake in east-central Arizona, in the Stoneman Lake area south of Flagstaff, along Truxton Wash near Peach Springs, and a population of uncertain extent on Navajo Nation lands. The Lyman Lake and Truxton Wash populations are small and extremely isolated. The Stoneman Lake population, however, is an extensive metapopulation spread across several stream drainages, including numerous ponds, wetlands, and artificial tanks. This is the only population in Arizona that is increasing in extent and numbers, but there is concern about the apparent introduction of nonnative genetic stock from eastern North America into this area. We analyzed genetic diversity within and genetic divergence among populations of northern leopard frogs, across both extant and recently extirpated populations in Arizona. We also analyzed mitochondrial DNA to place these populations into a larger phylogenetic framework and to determine whether any populations contained genetic material not native to the region. We found a high level of genetic divergence among the population centers (Lyman Lake, Stoneman Lake, Truxton Wash), and low genetic diversity in the small populations at Lyman Lake and Truxton. The extensive population in the Stoneman Lake area had high genetic diversity and relatively high gene flow among ponds and tanks across the entire extent of the area. However, this population also contained a mitochondrial haplotype from northern leopard frogs from the northeastern United States or southeastern Canada, probably representing the introduction of released pets or laboratory animals. These eastern frogs were extensively distributed through this population, and probably contributed to its high genetic diversity. Genetic diversity in the outlying populations such as Truxton Wash, East Buckskin Tank, and Hess Tank was low and showed signs of recent bottlenecks. However, supplementing genetic diversity in these native populations with artificial gene flow from the Stoneman Lake area may only be advisable in extreme situations for which there are no other alternatives. Until the nature and effects of genetic mixing of eastern and western genetic stocks of northern leopard frogs are better understood, the long-term persistence of the species in the Southwest may be best served by retaining as much genetic integrity of remaining native populations as possible.

Arizona

Molecular and phenotypic diversity in Chionactis occipitalis (Western Shovel-nosed Snake), with emphasis on the status of C. o. klauberi (Tucson Shovel-nosed Snake).

Chionactis occipitalis (Western Shovel-nosed Snake) is a small colubrid snake inhabiting the arid regions of the Mojave, Sonoran, and Colorado deserts. Morphological assessments of taxonomy currently recognize four subspecies. However, these taxonomic proposals were largely based on weak morphological differentiation and inadequate geographic sampling. Our goal was to explore evolutionary relationships and boundaries among subspecies of C. occipitalis, with particular focus on individuals within the known range of C. o. klauberi (Tucson Shovel-nosed snake). Population sizes and range for C. o. klauberi have declined over the last 25 years due to habitat alteration and loss prompting a petition to list this subspecies as endangered. We examined the phylogeography, population structure, and subspecific taxonomy of C. occipitalis across its geographic range with genetic analysis of 1100 bases of mitochondrial DNA sequence and reanalysis of 14 morphological characters from 1543 museum specimens. We estimated the species gene phylogeny from 81 snakes using Bayesian inference and explored possible factors influencing genetic variation using landscape genetic analyses. Phylogenetic and population genetic analyses reveal genetic isolation and independent evolutionary trajectories for two primary clades. Our data indicate that diversification between these clades has developed as a result of both historical vicariance and environmental isolating mechanisms. Thus these two clades likely comprise 'evolutionary significant units' (ESUs). Neither molecular nor morphological data are concordant with the traditional C. occipitalis subspecies taxonomy. Mitochondrial sequences suggest specimens recognized as C. o. klauberi are embedded in a larger geographic clade whose range has expanded from western Arizona populations, and these data are concordant with clinal longitudinal variation in morphology. ?? 2007 Springer Science+Business Media B.V.

Conservation Genetics

Conservation genetics of imperiled striped whipsnake in Washington

Conservation of wide-ranging species is aided by population genetic information that provides insights into adaptive potential, population size, interpopulation connectivity, and even extinction risk in portions of a species range. The Striped Whipsnake (Masticophis taeniatus) occurs across 11 western U.S. states and into Mexico but has experienced population declines in parts of its range, particularly in the state of Washington. We analyzed nuclear and mitochondrial DNA extracted from 192 shed skins, 63 muscle tissue samples, and one mouth swab to assess local genetic diversity and differentiation within and between the last known whipsnake populations in Washington. We then placed that information in a regional context to better understand levels of differentiation and diversity among whipsnake populations in the northwestern portion of the range of the species. Microsatellite data analyses indicated that there was comparable genetic diversity between the two extant Washington populations, but gene flow may be somewhat limited. We found moderate to high levels of genetic differentiation among states across all markers, including five microsatellites, two nuclear genes, and two mitochondrial genes. Pairwise state-level comparisons and dendrograms suggested that Washington whipsnakes are most closely related to those in Oregon, and distinct from Idaho, Nevada, and Utah, approximately following an isolation by distance model. We conclude that Washington populations of whipsnakes have experienced recent isolating events, but they have yet to lose genetic diversity. The longevity and high vagility of the species may provide opportunity for conservation of whipsnakes in the state as long as shrubland habitat is available

Washington

A nuclear DNA perspective on delineating evolutionarily significant lineages in polyploids: the case of the endangered shortnose sturgeon ( Acipenser brevirostrum )

The shortnose sturgeon, Acipenser brevirostrum , oft considered a phylogenetic relic, is listed as an “endangered species threatened with extinction” in the US and “Vulnerable” on the IUCN Red List. Effective conservation of A. brevirostrum depends on understanding its diversity and evolutionary processes, yet challenges associated with the polyploid nature of its nuclear genome have heretofore limited population genetic analysis to maternally inherited haploid characters. We developed a suite of polysomic microsatellite DNA markers and characterized a sample of 561 shortnose sturgeon collected from major extant populations along the North American Atlantic coast. The 181 alleles observed at 11 loci were scored as binary loci and the data were subjected to multivariate ordination, Bayesian clustering, hierarchical partitioning of variance, and among-population distance metric tests. The methods uncovered moderately high levels of gene diversity suggesting population structuring across and within three metapopulations (Northeast, Mid-Atlantic, and Southeast) that encompass seven demographically discrete and evolutionarily distinct lineages. The predicted groups are consistent with previously described behavioral patterns, especially dispersal and migration, supporting the interpretation that A. brevirostrum exhibit adaptive differences based on watershed. Combined with results of prior genetic (mitochondrial DNA) and behavioral studies, the current work suggests that dispersal is an important factor in maintaining genetic diversity in A. brevirostrum and that the basic unit for conservation management is arguably the local population.

PLoS ONE

DNA Sequencing confirms Tundra Bean Goose (Anser serrirostris serrirostris) occurrence in the Mississippi Alluvial Valley in Arkansas, USA

—First sighting records of rare occurrences may become increasingly important for recognizing changes in distribution, changes in migratory strategies, or increases in hybridization. We focumented the first record of a Tundra Bean Goose in the Mississippi Alluvial Valley, the outlet and historic floodplain for much of North America and one of the most important waterfowl wintering areas on the continent. We also document the first genetically confirmed record in the contiguous USA. Bean Goose (Anser fabalis and A. serrirostris) occurrences in North America are rare, especially outside of Alaska. On 24 January 2018, a Tundra Bean Goose (A. s. serrirostris) was harvested by a hunter in a winter-flooded rice field in Desha County, Arkansas, USA, near Dumas. The goose was mixed with a flock of 50 Greater White-Fronted Geese (A. albifrons). Because this individual was legally, albeit accidentally shot, we had the rare and exciting opportunity to obtain morphometric measurements and biological samples. As a result, we were able to verify the species and subspecies through genetic and morphological analysis. We determined the goose was an adult female Tundra Bean Goose, and mitochondrial DNA control region sequence data indicated this specimen was the subspecies A. s. serrirostris.

Arkansas

Distribution of tiger salamanders in northern Sonora, Mexico: Comparison of sampling methods and possible implications for an endangered subspecies

Many aquatic species in the arid USA-Mexico borderlands region are imperiled, but limited information on distributions and threats often hinders management. To provide information on the distribution of the Western Tiger Salamander ( Ambystoma mavortium ), including the USA-federally endangered Sonoran Tiger Salamander ( Ambystoma mavortium stebbinsi ), we used traditional (seines, dip-nets) and modern (environmental DNA [eDNA]) methods to sample 91 waterbodies in northern Sonora, Mexico, during 2015-2018. The endemic Sonoran Tiger Salamander is threatened by introgressive hybridization and potential replacement by another sub-species of the Western Tiger Salamander, the non-native Barred Tiger Salamander ( A. m. mavortium ). Based on occupancy models that accounted for imperfect detection, eDNA sampling provided a similar detection probability (0.82 [95% CI: 0.56-0.94]) as seining (0.83 [0.46-0.96]) and much higher detection than dip-netting (0.09 [0.02-0.23]). Volume of water filtered had little effect on detection, possibly because turbid sites had greater densities of salamanders. Salamanders were estimated to occur at 51 sites in 3 river drainages in Sonora. These results indicate tiger salamanders are much more widespread in northern Sonora than previously documented, perhaps aided by changes in land and water management practices. However, because the two subspecies of salamanders cannot be reliably distinguished based on morphology or eDNA methods that are based on mitochondrial DNA, we are uncertain if we detected only native genotypes or if we documented recent invasion of the area by the non-native sub-species. Thus, there is an urgent need for methods to reliably distinguish the subspecies so managers can identify appropriate interventions.

Sonora

Phylogeographic implications for release of critically endangered manatee calves rescued in Northeast Brazil

1. The Antillean manatee (Trichechus manatus manatus), a subspecies of the West Indian manatee, is a large-bodied marine mammal found in fresh, brackish, and marine habitats throughout the Caribbean Islands and Central and South America. Antillean manatees in Brazil are classified as critically endangered, with a census size of approximately 500 individuals. The population in the Northeast region of Brazil is suspected to have approximately 300 manatees and is threatened by habitat alteration and incidental entanglement in fishing gear. 2. A high incidence of dependent calf strandings have been identified near areas of altered critical manatee habitat. The majority of the calves are neonates, discovered alive, with no potential mothers nearby. These calves typically require human intervention to survive. Since 1989 the calves have been rescued (N=67), rehabilitated, and released (N=25) to supplement the small wild manatee population. The rescued calves, and those born in captivity, are typically, not released to their rescue location, mainly for logistical reasons. Therefore, phylogeographic analyses can help to identify related populations and appropriate release sites. 3. Here, mitochondrial DNA analyses identified low haplotype (h=0.08) and nucleotide (π=0.0026) genetic diversity in three closely related haplotypes. All three haplotypes (M01, M03, and a previously unidentified haplotype, M04) were found in the northern portion of the region, while only a single haplotype (M01) was represented in the south. This suggests the presence of two genetic groups with a central mixing zone. Release of rehabilitated calves to unrelated populations may result in genetic swamping of locally adapted alleles or genotypes, limiting the evolutionary potential of the population. 4. The small population size coupled with low genetic diversity indicates that the Northeast Brazil manatee population is susceptible to inbreeding depression and possible local extinction. Further conservation measures incorporating genetic information could be beneficial to the critically endangered Brazilian manatee population.

Aquatic Conservation: Marine and Freshwater Ecosys

Where east meets west: Phylogeography of the high Arctic North American brant goose

Genetic variation in Arctic species is often influenced by vicariance during the Pleistocene, as ice sheets fragmented the landscape and displaced populations to low- and high-latitude refugia. The formation of secondary contact or suture zones during periods of ice sheet retraction has important consequences on genetic diversity by facilitating genetic connectivity between formerly isolated populations. Brant geese ( Branta bernicla ) are a maritime migratory waterfowl (Anseriformes) species that almost exclusively uses coastal habitats. Within North America, brant geese are characterized by two phenotypically distinct subspecies that utilize disjunct breeding and wintering areas in the northern Pacific and Atlantic. In the Western High Arctic of Canada, brant geese consist of individuals with an intermediate phenotype that are rarely observed nesting outside this region. We examined the genetic structure of brant geese populations from each subspecies and areas consisting of intermediate phenotypes using mitochondrial DNA (mtDNA) control region sequence data and microsatellite loci. We found a strong east–west partition in both marker types consistent with refugial populations. Within subspecies, structure was also observed at mtDNA while microsatellite data suggested the presence of only two distinct genetic clusters. The Western High Arctic (WHA) appears to be a secondary contact zone for both Atlantic and Pacific lineages as mtDNA and nuclear genotypes were assigned to both subspecies, and admixed individuals were observed in this region. The mtDNA sequence data outside WHA suggests no or very restricted intermixing between Atlantic and Pacific wintering populations which is consistent with published banding and telemetry data. Our study indicates that, although brant geese in the WHA are not a genetically distinct lineage, this region may act as a reservoir of genetic diversity and may be an area of high conservation value given the potential of low reproductive output in this species.

Ecology and Evolution

Hybridization among Arctic white-headed gulls (Larus spp.) obscures the genetic legacy of the Pleistocene

We studied the influence of glacial oscillations on the genetic structure of seven species of white-headed gull that breed at high latitudes ( Larus argentatus, L. canus, L. glaucescens, L. glaucoides, L. hyperboreus, L. schistisagus, and L. thayeri ). We evaluated localities hypothesized as ice-free areas or glacial refugia in other Arctic vertebrates using molecular data from 11 microsatellite loci, mitochondrial DNA (mtDNA) control region, and six nuclear introns for 32 populations across the Holarctic. Moderate levels of genetic structure were observed for microsatellites ( F ST = 0.129), introns ( Φ ST = 0.185), and mtDNA control region ( Φ ST = 0.461), with among-group variation maximized when populations were grouped based on subspecific classification. Two haplotype and at least two allele groups were observed across all loci. However, no haplotype/allele group was composed solely of individuals of a single species, a pattern consistent with recent divergence. Furthermore, northernmost populations were not well differentiated and among-group variation was maximized when L. argentatus and L. hyberboreus populations were grouped by locality rather than species, indicating recent hybridization. Four populations are located in putative Pleistocene glacial refugia and had larger t estimates than the other 28 populations. However, we were unable to substantiate these putative refugia using coalescent theory, as all populations had genetic signatures of stability based on mtDNA. The extent of haplotype and allele sharing among Arctic white-headed gull species is noteworthy. Studies of other Arctic taxa have generally revealed species-specific clusters as well as genetic structure within species, usually correlated with geography. Aspects of white-headed gull behavioral biology, such as colonization ability and propensity to hybridize, as well as their recent evolutionary history, have likely played a large role in the limited genetic structure observed.

Ecology and Evolution

Genetic differentiation and inferred dynamics of a hybrid zone between Northern Spotted Owls (Strix occidentalis caurina) and California Spotted Owls (S. o. occidentalis) in northern California

Genetic differentiation among Spotted Owl ( Strix occidentalis ) subspecies has been established in prior studies. These investigations also provided evidence for introgression and hybridization among taxa but were limited by a lack of samples from geographic regions where subspecies came into close contact. We analyzed new sets of samples from Northern Spotted Owls (NSO: S. o. caurina ) and California Spotted Owls (CSO: S. o. occidentalis ) in northern California using mitochondrial DNA sequences (mtDNA) and 10 nuclear microsatellite loci to obtain a clearer depiction of genetic differentiation and hybridization in the region. Our analyses revealed that a NSO population close to the northern edge of the CSO range in northern California (the NSO Contact Zone population) is highly differentiated relative to other NSO populations throughout the remainder of their range. Phylogenetic analyses identified a unique lineage of mtDNA in the NSO Contact Zone, and Bayesian clustering analyses of the microsatellite data identified the Contact Zone as a third distinct population that is differentiated from CSO and NSO found in the remainder of the subspecies' range. Hybridization between NSO and CSO was readily detected in the NSO Contact Zone, with over 50% of individuals showing evidence of hybrid ancestry. Hybridization was also identified among 14% of CSO samples, which were dispersed across the subspecies' range in the Sierra Nevada Mountains. The asymmetry of hybridization suggested that the hybrid zone may be dynamic and moving. Although evidence of hybridization existed, we identified no F1 generation hybrid individuals. We instead found evidence for F2 or backcrossed individuals among our samples. The absence of F1 hybrids may indicate that (1) our 10 microsatellites were unable to distinguish hybrid types, (2) primary interactions between subspecies are occurring elsewhere on the landscape, or (3) dispersal between the subspecies' ranges is reduced relative to historical levels, potentially as a consequence of recent regional fires.

California

Genetic analyses reveal cryptic introgression in secretive marsh bird populations

Hybridization is common in bird populations but can be challenging for management, especially if one of the two parent species is of greater conservation concern than the other. King rails (Rallus elegans) and clapper rails (R. crepitans) are two marsh bird species with similar morphologies, behaviors, and overlapping distributions. The two species are found along a salinity gradient with the king rail in freshwater marshes and the clapper in estuarine marshes. However, this separation is not absolute; they are occasionally sympatric, and there are reports of interbreeding. In Virginia, USA, both king and clapper rails are identified by the state as Species of Greater Conservation Need, although clappers are thought to be more abundant and king rails have a higher priority ranking. We used a mitochondrial DNA marker and 13 diagnostic nuclear single nucleotide polymorphisms (SNPs) to identify species, classify the degree of introgression, and explore the evolutionary history of introgression in two putative clapper rail focal populations along a salinity gradient in coastal Virginia. Genetic analyses revealed cryptic introgression with site-specific rates of admixture. We identified a pattern of introgression where clapper rail alleles predominate in brackish marshes. These results suggest clapper rails may be displacing king rails in Virginia coastal waterways, most likely as a result of ecological selection. As introgression can result in various outcomes from outbreeding depression to local adaptation, continued monitoring of these populations would allow further exploration of hybrid fitness and inform conservation management.

Georgia, New Jersey, North Carolina, Rhode Island,