Population genetic structure and ancestry of steelhead/rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) at the extreme southern edge of their range in North America

Steelhead (Oncorhynchus mykiss) populations have declined dramatically in many parts of their range in North America, most critically in Southern California, where these anadromous trout are now classified as ‘Endangered’ under the United States Endangered Species Act. The widespread introduction of hatchery rainbow trout, the domesticated freshwater resident form of the species O. mykiss, is one factor threatening the long-term persistence of native steelhead and other trout populations. To identify where native fish of coastal steelhead lineage remained, we performed a population genetic analysis of microsatellite and SNP genotypes from O. mykiss populations at the extreme southern end of their range in Southern California, USA and Baja California, Mexico. In the northern part of this region, nearly all populations appeared to be primarily descendants of native coastal steelhead. However, in the southern, more urbanized part of this region, the majority of the sampled populations were derived primarily from hatchery trout, indicating either complete replacement of native fish or a strong signal of introgression overlaying native ancestry. Nevertheless, these genetically introgressed populations represent potentially critical genetic resources for the continued persistence of viable networks of O. mykiss populations, given the limited native ancestry uncovered in this region and the importance of genetic variation in adaptation. This study elucidates the geographic distribution of native trout populations in this region, and serves as a baseline for evaluating the impacts of hatchery trout on native O. mykiss populations and the success of steelhead conservation and recovery efforts.     

Temporal Comparisons of Genetic Diversity in Lake Michigan Steelhead, Oncorhynchus mykiss, Populations: Effects of Hatchery Supplementation

Steelhead, Oncorhynchus mykiss, were first introduced into the Great Lakes in the late 1800s. Subsequently, natural recruitment across the Lake Michigan basin has been regularly supplemented by primarily one hatchery strain. Recently, multiple strains derived from locations across the species native range along the west coast of the United States have also been stocked by different management agencies. Prior to 1983, hatchery supplementation of Lake Michigan steelhead populations in Michigan was largely unsuccessful due to low smolting rates of small (<120 mm) hatchery yearlings (estimated survival 0.01%). Accordingly, contributions of hatchery fish to historical adult spawning runs in Michigan tributaries were low (0–30%) across six major drainages. Large yearlings of different hatchery strains (>150 mm) have been stocked exclusively since 1983, increasing estimates of survival to smolting (90%). Consequently, the proportion of hatchery adults in spawning runs increased to 13–79%. We examined the effects of changes in stocking practices on straying rates of hatchery steelhead and to temporal changes in levels of genetic diversity and relationships among populations. We used microsatellite loci to estimate allele frequencies for six populations sampled for two time periods (1983–1984 and 1998–1999). Measures of inter-population divergence (mean F_ST) were not significant for either time period. However, spatial genetic relationships among historical and contemporary populations were significantly correlated with geographic distance; a result not expected if gene flow (natural straying) among populations was mediated solely by hatchery supplementation. Increased numbers of alleles in spawning adults from populations can be attributed to alleles specific to recently introduced hatchery strains.     

Are captive tortoises a reservoir for conservation? An assessment of genealogical affiliation of captive Gopherus agassizii to local, wild populations

The conservation of tortoises poses a unique situation because several threatened species are commonly kept as pets within their native ranges. Thus, there is potential for captive populations to be a reservoir for repatriation efforts. We assess the utility of captive populations of the threatened Agassiz’s desert tortoise (Gopherus agassizii) for recovery efforts based on genetic affinity to local areas. We collected samples from 130 captive desert tortoises from three desert communities: two in California (Ridgecrest and Joshua Tree) and the Desert Tortoise Conservation Center (Las Vegas) in Nevada. We tested all samples for 25 short tandem repeats and sequenced 1,109 bp of the mitochondrial genome. We compared captive genotypes to a database of 1,258 Gopherus samples, including 657 wild caught G. agassizii spanning the full range of the species. We conducted population assignment tests to determine the genetic origins of the captive individuals. For our total sample set, only 44 % of captive individuals were assigned to local populations based on genetic units derived from the reference database. One individual from Joshua Tree, California, was identified as being a Morafka’s desert tortoise, G. morafkai, a cryptic species which is not native to the Mojave Desert. Our data suggest that captive desert tortoises kept within the native range of G. agassizii cannot be presumed to have a genealogical affiliation to wild tortoises in their geographic proximity. Precautions should be taken before considering the release of captive tortoises into the wild as a management tool for recovery.     

Population genetics of steelhead (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) in the Klamath River

An analysis of population structure and genetic diversity was conducted on samples of Oncorhynchus mykiss (steelhead/rainbow trout) from 33 sites in the Klamath–Trinity River basin. Genotype data from 16 microsatellite loci in almost 1,700 fish revealed genetic differentiation between most sampled locations. Two pairs of samples from the same sites in different years were not significantly different, indicating stability of population structure, at least on a short time scale. Most sampling sites were genetically distinct from all other sampling sites, and there was an evidence of geographic structure within the Klamath–Trinity River basin, although populations from tributaries within the watershed (e.g. Salmon River, Scott River, Clear Creek) did not always constitute distinct genetic lineages. Population structure was evident using phylogeographic trees, isolation by distance analyses and individual assignment tests, which all found a relationship between geographic and genetic distance. Populations in the lower Klamath region, below the confluence with the Trinity River, consistently clustered together in phylogeographic analyses and had patterns of genetic diversity that suggest reduced gene flow between these sites and sites above the confluence. Finally, in an analysis that included data from other coastal California rivers, the populations closest to the mouth of the Klamath River appeared intermediate between populations from adjacent watersheds and the lineage formed by the other populations in the Klamath–Trinity basin.     

Population genetic structure and ancestry of Oncorhynchus mykiss populations above and below dams in south-central California

Genetic analyses of coastal Oncorhynchus mykiss, commonly known as steelhead/rainbow trout, at the southern extreme of their geographic range in California are used to evaluate ancestry and genetic relationships of populations both above and below large dams. Juvenile fish from 20 locations and strains of rainbow trout commonly planted in reservoirs in the five study basins were evaluated at 24 microsatellite loci. Phylogeographic trees and analysis of molecular variance demonstrated that populations within a basin, both above and below dams, were generally each other’s closest relatives. Absence of hatchery fish or their progeny in the tributaries above dams indicates that they are not commonly spawning and that above-barrier fish are descended from coastal steelhead trapped at dam construction. Finally, no genetic basis was found for the division of populations from this region into two distinct biological groups, contrary to current classification under the US and California Endangered Species Acts.     

Genetic variation and population structure in desert bighorn sheep: implications for conservation

Bighorn sheep populations experienced a drastic reduction in both distribution and abundance until the advent of modern wildlife management, where improving viability of extant populations and translocating animals into historical habitat range have been the most important management policies. The fact that subspecies relationships among bighorn are ambiguous,together with the importance of selecting appropriate source stock and the expense of translocation projects, makes an understanding of subspecies relationships and genetic variation, within and between populations, important for the management and conservation of this species. In this study, genetic variation in 279 bighorn sheep from 13 study sites in Arizona, California, New Mexico and Alberta, Canada were examined by analyzing ten microsatellite loci to determine interpopulation differentiation and relationships between closely related taxa. All populations contained a substantial amount of genetic variation. Genetic differences between populations were large and roughly proportional to geographic distance. The significance of this to desert subspecies relationships and management is discussed.     

Isolation of 15 single nucleotide polymorphisms from coastal steelhead, Oncorhynchus mykiss (Salmonidae)

We describe 15 single nucleotide polymorphisms (SNPs) isolated in coastal California populations of steelhead (Oncorhynchus mykiss). SNP loci were developed using a 'gene-targeted' approach, which involved the development of primers from functional genes in O. mykiss that were deposited in GenBank or in the published literature. These markers show a wide range of variability in three coastal steelhead populations, and will be useful in population genetic studies and in pedigree reconstruction. Potential applications include evaluation of population structure, introgression between native and hatchery trout, and evaluating reproductive success.     

Islands as refugia of <Emphasis Type="Italic">Trifolium repens</Emphasis> genetic diversity

Island populations are often thought to be more susceptible to the loss of genetic diversity as a consequence of limited population size and genetic drift, greater susceptibility to detrimental stochastic events and low levels of immigration. However the geographic isolation of islands may create refuges for native crop species whose genetic diversity is threatened from the genetic erosion occurring in mainland areas as a result of crop-wild gene flow and genetic swamping. Many UK islands remain uncharacterised in terms of plant genetic diversity. In this study we compared the genetic diversity of mainland populations and landraces of Trifolium repens with wild populations collected from the islands surrounding the UK, including the island of Hirta in the St Kildan archipelago. Individuals from St Kilda represent a unique conservation resource, with populations both highly differentiated from UK mainland populations and genetically distinct from cultivated varieties, whilst able to retain diversity through limited human influence on the islands. In contrast, there is relative genetic similarity of wild UK populations to cultivated forms highlighted in mainland populations, but with geographic barriers preventing complete homogenisation of the mainland UK genepool. We underline the need for conservation priorities to include common species that are threatened by gene flow from cultivation, and draw attention to the potential of islands to preserve natural levels of genetic diversity.     

Genetic diversity and population parameters of sea otters, Enhydra lutris, before fur trade extirpation from 1741-1911

All existing sea otter, Enhydra lutris, populations have suffered at least one historic population bottleneck stemming from the fur trade extirpations of the eighteenth and nineteenth centuries. We examined genetic variation, gene flow, and population structure at five microsatellite loci in samples from five pre-fur trade populations throughout the sea otter's historical range: California, Oregon, Washington, Alaska, and Russia. We then compared those values to genetic diversity and population structure found within five modern sea otter populations throughout their current range: California, Prince William Sound, Amchitka Island, Southeast Alaska and Washington. We found twice the genetic diversity in the pre-fur trade populations when compared to modern sea otters, a level of diversity that was similar to levels that are found in other mammal populations that have not experienced population bottlenecks. Even with the significant loss in genetic diversity modern sea otters have retained historical structure. There was greater gene flow before extirpation than that found among modern sea otter populations but the difference was not statistically significant. The most dramatic effect of pre fur trade population extirpation was the loss of genetic diversity. For long term conservation of these populations increasing gene flow and the maintenance of remnant genetic diversity should be encouraged.     

Genetic and phenotypic variation among geographically isolated populations of the globally threatened Dupont's lark Chersophilus duponti

Identifying genetically and phenotypically distinct populations of threatened species is critical if we are to delineate appropriate plans for their conservation. We conducted an integrated analysis of population genetic structure, historical demographic events, current gene flow (all based on mtDNA sequences) and morphological variation of three geographically separated groups of populations of Dupont's lark Chersophilus duponti, located in the Iberian Peninsula (three populations), Morocco (two populations), and Tunisia (one population). Unusually, this lark species is the only one among the genus Chersophilus. Our results revealed the early historical divergence of an eastern Dupont's lark lineage (in Tunisia) and a western lineage (in Morocco and Spain), consistent with subspecies taxonomy and distribution. The western lineage subsequently split into two lineages, following the isolation of Iberian and African populations. Such pattern of historical differentiation caused great population genetic structure, with differences among geographic areas explaining more than 80% of total genetic variation. Mismatch distributions and coalescent estimates of divergence time showed that lineage divergence was associated with sudden population expansion events, which apparently took place during the last glaciation, when steppe habitats were widespread across the Mediterranean region. Extant populations from different geographic areas hardly shared any haplotype (only one out of 16 ND2 haplotypes was shared by Tunisian and Moroccan Dupont's larks), and consequently gene flow between geographic areas was found to be virtually absent. Apart from showing great genetic differentiation, Dupont's larks from different geographic areas were morphologically distinct, showing substantial variation in body size and feeding-related traits (length of feet and bill). We conclude that Dupont's lark populations isolated in the Iberian Peninsula, Morocco, and Tunisia are distinct evolutionary entities and should be considered as such in conservation plans. Such circumstance sets a daunting conservation challenge that exemplifies the need of incorporating knowledge of historical processes to our general understanding of the demography of threatened species.     

Population genetics of threatened wild plants in Japan

Approximately one-fourth of Japan's native plant species are threatened with extinction. To conserve these species, it is critical to evaluate genetic diversity at species-level and population-level. Some factors, including population size and geographic distribution, are known to influence the population genetic diversity of wild plant species. This article briefly reviews the population genetic studies that have been conducted on wild threatened plants in Japan. A large population size or wide geographic distribution does not always lead to large genetic diversity, suggesting that historical factors such as speciation processes and population expansion often play more important roles in determining genetic diversity than the number of remnant individuals. The mating system of a species also affects genetic diversity; predominantly selfing species tend to have smaller genetic diversity than outcrossing congeners. Another issue of concern in the conservation genetics of wild plants in Japan is the genetic diversity of insular endemics, because Japan consists of many islands, and the insular flora contains many endemic and threatened species. Previous studies on endemic plants on the Bonin and the Ryukyu Islands are reviewed. Compared to the cases of the Bonin Islands or other oceanic islands, there is much larger genetic diversity in plants endemic to the Ryukyu Islands. This difference is probably the result of the differences in the geological history of these islands. Electronic Publication     

Genetic diversity of Annona crassiflora (Annonaceae) in northern Minas Gerais State

Genetic diversity analyses of tropical tree species are relevant to landscape management, plant genetic resource inventory, and biological conservation of threatened species. Annona crassiflora is an endangered fruit tree native to the Cerrado biome that is threatened by reduction of natural populations and fruit extraction. We examined the intra- and interpopulational genetic diversity of this species in the northern region of Minas Gerais State. Seventy-two individuals from four natural populations were genotyped using RAPD markers. We found moderate genetic diversity among populations, with Shannon's I index varying between 0.31 and 0.44, and Nei's genetic diversity (H(E)) for the population set equal to 0.31. AMOVA indicated a greater genetic variation within (77.38%) rather than among populations (22.62%), tending towards isolation by distance (Mantel's r = 0.914; P = 0.089). Nei's genetic identity estimates among populations revealed a hierarchical pattern of genetic similarity of form [(CA1, CA2), MC], [(GM)], corroborating the high genetic differentiation between spatially isolated populations.     

Population structure and loss of genetic diversity in the endangered white-headed duck, <Emphasis Type="Italic">Oxyura leucocephala</Emphasis>

The white-headed duck is a globally threatened species native to the Palaearctic with a range extending from Spain in the west to the western edge of China in the east. Its populations have become fragmented and undergone major declines in recent decades. To study genetic differences between populations across the range and change in genetic diversity over time, we sequenced a portion of the mitochondrial DNA control region from 67 museum specimens (years 1861–1976) as well as 39 contemporary samples from Spain and seven from Greece (years 1992–2003). In the historical sample, we found a lack of significant genetic structure between populations in different areas. We found evidence that the species experienced a rapid expansion in the past, perhaps from glacial refugia centred around the Mediterranean following the last ice age. In Spain, the population went through a dramatic bottleneck in the 1970s and early 1980s, when only a few dozens individuals remained in the wild. Although population size has since recovered to a few thousand individuals, we found a highly significant loss of mitochondrial haplotype diversity between the historical and contemporary samples. Given ongoing declines in other areas, losses in genetic diversity that may reduce the adaptive potential of white-headed ducks in the future are a continuing concern throughout the geographic range of this species.     

Hierarchical genetic structure of native masu salmon populations in Hokkaido,Japan

Identification of the spatial extent of genetic structuring that may be influenced by evolutionary, ecological and historical factors is critical for effective conservation or management strategies. Masu salmon Oncorhynchus masou is commonly distributed in Far East, however, many local populations have been under threats of decline due to habitat destruction, overexploitation, and genetic introgression. To reveal the spatial genetic structure of native masu salmon populations in Hokkaido, masu salmon samples were collected from 16 rivers in which there was no official record of artificial releases of any masu salmon stock and were analyzed using 15 microsatellite loci. A Bayesian assignment test revealed that masu salmon populations were divided into two genetically distinct groups: the northeastern and southwestern groups. For within-group genetic structure, all populations, except for geographically proximate populations, were significantly different from each other. AMOVA revealed that genetic variation at among-group level based on groups identified assignment test was greater than that of groups based on geographic locations. There was no significant IBD for the 16 populations. However, the Mantel test revealed significant IBD for the northeastern group, but did not for the southwestern group. This study suggested that native masu salmon populations in Hokkaido exhibit a hierarchical genetic structure that is largely a result of their precise homing behavior. The results of this study also highlight the importance of defining populations by using genetic data rather than by using predefined populations based on geographic locations for the correct determination of genetic structure.     

Model-based comparisons of phylogeographic scenarios resolve the intraspecific divergence of cactophilic Drosophila mojavensis

The cactophilic fly Drosophila mojavensis exhibits considerable intraspecific genetic structure across allopatric geographic regions and shows associations with different host cactus species across its range. The divergence between these populations has been studied for more than 60years, yet their exact historical relationships have not been resolved. We analysed sequence data from 15 intronic X-linked loci across populations from Baja California, mainland Sonora-Arizona and Mojave Desert regions under an isolation-with-migration model to assess multiple scenarios of divergence. We also compared the results with a pre-existing sequence data set of eight autosomal loci. We derived a population tree with Baja California placed at its base and link their isolation to Pleistocene climatic oscillations. Our estimates suggest the Baja California population diverged from an ancestral Mojave Desert/mainland Sonora-Arizona group around 230,000-270,000years ago, while the split between the Mojave Desert and mainland Sonora-Arizona populations occurred one glacial cycle later, 117,000-135,000years ago. Although we found these three populations to be effectively allopatric, model ranking could not rule out the possibility of a low level of gene flow between two of them. Finally, the Mojave Desert population showed a small effective population size, consistent with a historical population bottleneck. We show that model-based inference from multiple loci can provide accurate information on the historical relationships of closely related groups allowing us to set into historical context a classic system of incipient ecological speciation.     

Living in two worlds: Evolutionary mechanisms act differently in the native and introduced ranges of an invasive plant

Identifying the factors that influence spatial genetic structure among populations can provide insights into the evolution of invasive plants. In this study, we used the common reed (Phragmites australis), a grass native in Europe and invading North America, to examine the relative importance of geographic, environmental (represented by climate here), and human effects on population genetic structure and its changes during invasion. We collected samples of P. australis from both the invaded North American and native European ranges and used molecular markers to investigate the population genetic structure within and between ranges. We used path analysis to identify the contributions of each of the three factors—geographic, environmental, and human‐related—to the formation of spatial genetic patterns. Genetic differentiation was observed between the introduced and native populations, and their genetic structure in the native and introduced ranges was different. There were strong effects of geography and environment on the genetic structure of populations in the native range, but the human‐related factors manifested through colonization of anthropogenic habitats in the introduced range counteracted the effects of environment. The between‐range genetic differences among populations were mainly explained by the heterogeneous environment between the ranges, with the coefficient 2.6 times higher for the environment than that explained by the geographic distance. Human activities were the primary contributor to the genetic structure of the introduced populations. The significant environmental divergence between ranges and the strong contribution of human activities to the genetic structure in the introduced range suggest that invasive populations of P. australis have evolved to adapt to a different climate and to human‐made habitats in North America.     

Physiological and Behavioral Effects of Zinc and Temperature on Coho Salmon (Oncorhynchus kisutch)

Pacific salmon species including the U.S. federally endangered coho salmon (Oncorhynchus kisutch) and the U.S. federally threatened steelhead trout (Oncorhynchus mykiss) have declined at an alarming rate in the last 40 years. Two of the main causes for the decline in coastal coho populations include increases in temperature and contaminant loads in coastal watersheds. Zinc, in particular, is one of the most common contaminants in aquatic systems. Using an experimental mesocosm design, we examined physiological, biochemical, and behavioral responses of coho salmon to excess dietary zinc and increased temperatures, with the ultimate goal of relating results to wild populations of coho salmon and steelhead in the Navarro River, California. Fish were obtained from a hatchery and divided into four treatments: low water temperature-no dietary zinc, high temperature-no zinc, low temperature-zinc, and high temperature-zinc. Each treatment had four replicate tanks. Zinc concentrations in liver increased during exposure to a high zinc diet. Iron concentrations in liver increased during simultaneous exposure to high zinc diet and increased temperature, and growth was reduced in this experimental treatment. Expression of hsp-70 was not significantly different between treatments, but showed decreasing trends with high dietary zinc and high temperature. Feeding rate increased with exposure to a high zinc diet. Comparison with steelhead trout samples from the Navarro River, California, showed levels of zinc, iron, and hsp-70 greater than those found in the experimental Coho salmon. All comparisons between the hatchery coho salmon and wild steelhead should be viewed with caution due to the differences between species, the laboratory and natural environment, and the genetic differences between wild and hatchery fish.     

Phylogeography of the tidewater goby, Eucyclogobius newberryi (Teleostei, Gobiidae), in coastal California

The tidewater goby, Eucyclogobius newberryi, inhabits discrete, seasonally closed estuaries and lagoons along approximately 1500 km of California coastline. This species is euryhaline but has no explicit marine stage, yet population extirpation and recolonization data suggest tidewater gobies disperse intermittently via the sea. Analyses of mitochondrial control region and cytochrome b sequences demonstrate a deep evolutionary bifurcation in the vicinity of Los Angeles that separates southern California populations from all more northerly populations. Shallower phylogeographic breaks, in the vicinities of Seacliff, Point Buchon, Big Sur, and Point Arena segregate the northerly populations into five groups in three geographic clusters: the Point Conception and Ventura groups between Los Angeles and Point Buchon, a lone Estero Bay group from central California, and San Francisco and Cape Mendocino groups from northern California. The phylogenetic relationships between and patterns of molecular diversity within the six groups are consistent with repeated, and sometimes rapid, northward and southward range expansions out of central California caused by Quaternary climate change. Plio-Pleistocene tectonism, Quaternary coastal geography and hydrography, and historical human activities probably also influenced the modern geographic and genetic structure of E. newberryi. The phylogeography of E. newberryi is concordant with phylogeographic patterns in several other coastal California taxa, suggesting common extrinsic factors have had similar effects on different species. However, there is no evidence of a phylogeographic break coincident with a biogeographic boundary at Point Conception.     

EVOLUTIONARY AND HISTORICAL ANALYSIS OF PROTEIN VARIATION IN THE BLOTCHED FORMS OF SALAMANDERS OF THE ENSATINA COMPLEX (AMPHIBIA: PLETHODONTIDAE)

Geographic variation in 23 to 29 protein-encoding genetic loci was examined in 48 populations of the Ensatina complex, a “ring species” distributed around the Central Valley of California. The samples span two critical links in the chain of morphologically distinct units: the transition from the unblotched to blotched color pattern types in the vicinity of Lassen Peak, northeastern California, and a geographic gap in the range of the complex in the San Gabriel Mountains, southern California. A general pattern of isolation by distance with a regular buildup of genetic distance correlated with increases in geographic distance characterizes the populations studied, with the exception of a little-differentiated group of populations in the northern Sierra Nevada; this region is postulated to be a zone of genetic reticulation characterized by relatively high gene flow. An adaptively significant color pattern is thought to have spread into the northern Sierra Nevada from the south, but protein variants have been introduced both from the north and the south. Genetic distances across the San Gabriel Mountain gap match expectations from the pattern of buildup of genetic distance as a function of geographic distance elsewhere in the complex. A phylogenetic analysis of the protein data supports the reticulation hypothesis; whereas the southernmost populations currently do constitute a monophyletic assemblage, an “extinction experiment” demonstrates that the distinction could be the result of the recent extinction of populations in a present gap in our sampling. The Ensatina complex appears to be a dynamic entity representing several stages in the evolution of species. It is a ring species, and whereas various taxonomic arrangements are possible, no taxonomic changes are proposed.     

Genetic Differentiation of Native Sheep Populations in East and South Asia

Variations of four sheep populations in China were examined by multiloci electrophoresis, and similar data are quoted to analyze the degree of genetic differentiation of native sheep populations in East and South Asia. Among 15 populations, the average heterozygosity is 0.2746, and the effective number of alleles is 1.559. Mongolian sheep possess the highest genetic diversity, and diversity decreases sequentially in the Chinese, Vietnamese, Bangladeshi, and Nepalese populations. Coefficients of genetic differentiation are 0.0126–0.3083, with an average of 0.148, demonstrating that the major genetic variation (85%) exists within populations. Genetic identity and genetic distance all show relatively low genetic differentiation. No relationship was found between geographic distance and genetic distance. Gene flow is common among the mass of populations, which leads to the inconsistency between geographic distance and genetic distance. The 15 native sheep populations in East and South Asia can be divided into two groups, one group including part of the Chinese and Mongolian populations and another including theYunnan population of China and part of the Nepalese and Bangladeshi populations. Other populations did not separate into groups, merging instead into the two main groups.