Genetic Structure of Fragmented Populations of the Endangered Daisy Rutidosis leptorrhynchoides

Abstract: The endangered grassland daisy Rutidosis leptorrhynchoides has been subject to severe habitat destruction and fragmentation over the past century. Using allozyme markers, we examined the genetic diversity and structure of 16 fragmented populations. The species had high genetic variation compared to other plant species, and both polymorphism and allelic richness showed strong positive relationships with log reproductive population size, reflecting a loss of rare alleles (frequency of q < 0.1) in smaller populations. Fixation coefficients were positively related to size, due either to a lack of rare homozygotes in small populations or to Wahlund effects (owing to spatial genetic structure) in large ones. Neither gene diversity nor heterozygosity was related to population size, and other population parameters such as density, spatial contagion, and isolation had no apparent effect on genetic variation. Genetic divergence among populations was low , despite a large north-to-south break in the species' current distribution. To preserve maximum genetic variation, conservation strategies should aim to maintain the five populations larger than 5000 reproductive plants, all of which occur in the north of the range, as well as the largest southern population of 626 plants at Truganina. Only one of these is currently under formal protection. High heterozygosity in smaller populations suggests that they are unlikely to be suffering from inbreeding depression and so are also valuable for conservation. Erosion of allelic richness at self-incompatibility loci, however, may limit the reproductive capacity of populations numbering less than 20 flowering plants.     

Possible Extinction Vortex for a Population of Iberian Lynx on the Verge of Extirpation

Abstract: Theory suggests that demographic and genetic traits deteriorate (i.e., fitness and genetic diversity decrease) when populations become small, and that such deterioration could precipitate positive feedback loops called extinction vortices. We examined whether demographic attributes and genetic traits have changed over time in one of the 2 remaining small populations of the highly endangered Iberian lynx (Lynx pardinus) in Doñana, Spain. From 1983 to 2008, we recorded nontraumatic mortality rates, litter size, offspring survival, age at territory acquisition, and sex ratio. We combined these demographic attributes with measures of inbreeding and genetic diversity at neutral loci (microsatellites) and genes subjected to selection (major histocompatibility complex). Data on demographic traits were obtained through capture and radio tracking, checking dens during breeding, track surveys, and camera trapping. For genetic analyses, we obtained blood or tissue samples from captured or necropsied individuals or from museum specimens. Over time a female‐biased sex ratio developed, age of territory acquisition decreased, mean litter size decreased, and rates of nontraumatic mortality increased, but there were no significant changes in overall mortality rates, standardized individual heterozygosity declined steadily, and allelic diversity of exon 2 of class II major histocompatibility complex DRB genes remained constant (2 allelic variants present in all individuals analyzed). Changes in sex ratio and age of territory acquisition may have resulted from demographic stochasticity, whereas changes in litter size and nontraumatic mortality may be related to observed increases in inbreeding. Concomitant deterioration of both demographic attributes and genetic traits is consistent with an extinction vortex. The co‐occurrence, with or without interaction, of demographic and genetic deterioration may explain the lack of success of conservation efforts with the Doñana population of Iberian lynx.     

Meta-analysis of the differential effects of habitat fragmentation and degradation on plant genetic diversity

Genetic diversity is a key factor for population survival and evolution. However, anthropogenic habitat disturbance can erode it, making populations more prone to extinction. Aiming to assess the global effects of habitat disturbance on plant genetic variation, we conducted a meta-analysis based on 92 case studies obtained from published literature. We compared the effects of habitat fragmentation and degradation on plant allelic richness and gene diversity (equivalent to expected heterozygosity) and tested whether such changes are sensitive to different life-forms, life spans, mating systems, and commonness. Anthropogenic disturbance had a negative effect on allelic richness, but not on gene diversity. Habitat fragmentation had a negative effect on genetic variation, whereas habitat degradation had no effect. When we examined the individual effects in fragmented habitats, allelic richness and gene diversity decreased, but this decrease was strongly dependent on certain plant traits. Specifically, common long-lived trees and self-incompatible species were more susceptible to allelic richness loss. Conversely, gene diversity decreased in common short-lived species (herbs) with self-compatible reproduction. In a wider geographical context, tropical plant communities were more sensitive to allelic richness loss, whereas temperate plant communities were more sensitive to gene diversity loss. Our synthesis showed complex responses to habitat disturbance among plant species. In many cases, the absence of effects could be the result of the time elapsed since the disturbance event or reproductive systems favoring self-pollination, but attention must be paid to those plant species that are more susceptible to losing genetic diversity, and appropriate conservation should be actions taken.     

Predicting the Deleterious Effects of Mutation Load in Fragmented Populations

Abstract:  Human-induced habitat fragmentation constitutes a major threat to biodiversity. Both genetic and demographic factors combine to drive small and isolated populations into extinction vortices. Nevertheless, the deleterious effects of inbreeding and drift load may depend on population structure, migration patterns, and mating systems and are difficult to predict in the absence of crossing experiments. We performed stochastic individual-based simulations aimed at predicting the effects of deleterious mutations on population fitness (offspring viability and median time to extinction) under a variety of settings (landscape configurations, migration models, and mating systems) on the basis of easy-to-collect demographic and genetic information. Pooling all simulations, a large part (70%) of variance in offspring viability was explained by a combination of genetic structure ( F_ST ) and within-deme heterozygosity ( H_S ). A similar part of variance in median time to extinction was explained by a combination of local population size ( N ) and heterozygosity ( H_S ). In both cases the predictive power increased above 80% when information on mating systems was available. These results provide robust predictive models to evaluate the viability prospects of fragmented populations.     

Erosion of Heterozygosity in Fluctuating Populations

Abstract: Demographic, environmental, and genetic stochasticity threaten the persistence of isolated populations. The relative importance of these intertwining factors remains unresolved, but a common view is that random demographic and environmental events will usually drive small populations to the brink of extinction before genetic deterioration poses a serious threat. To evaluate the potential importance of genetic factors, we analyzed a model linking demographic and environmental conditions to the loss of genetic diversity in isolated populations undergoing natural levels of fluctuation. Nongenetic processes—environmental stochasticity and population demography—were modeled according to a bounded diffusion process. Genetic processes were modeled by quantifying the rate of drift according to the effective population size, which was predicted from the same parameters used to describe the nongenetic processes. We combined these models to predict the heterozygosity remaining at the time of extinction, as predicted by the nongenetic portion of the model. Our model predicts that many populations will lose most or all of their neutral genetic diversity before nongenetic random events lead to extinction. Given the abundant evidence for inbreeding depression and recent evidence for elevated extinction rates of inbred populations, our findings suggest that inbreeding may be a greater general threat to population persistence than is generally recognized. Therefore, conservation biologists should not ignore the genetic component of extinction risk when assessing species endangerment and developing recovery plans.     

Temporal Changes in Allele Frequencies and a Population's History of Severe Bottlenecks

Monitoring temporal changes in genetic variation has been suggested as a means of determining if a population has experienced a demographic bottleneck. Simulations have shown that the variance in allele frequencies over time ( F ) can provide reasonable estimates of effective population size ( N_e ). This relationship between F and N_e suggests that changes in allele frequencies may provide a way to determine the severity of recent demographic bottlenecks experienced by a population. We examined allozyme variation in experimental populations of the eastern mosquitofish ( Gambusia holbrooki ) to evaluate the relationship between the severity of demographic bottlenecks and temporal variation in allele frequencies. Estimates of F from both the fish populations and computer simulations were compared to expected rates of drift. We found that different methods for estimating F had little effect on the analysis. The variance in estimates of F was large among both experimental and simulated populations experiencing similar demographic bottlenecks. Temporal changes in allele frequencies suggested that the experimental populations had experienced bottlenecks, but there was no relationship between observed and expected values of F . Furthermore, genetic drift was likely to be underestimated in populations experiencing the most severe bottlenecks. The weak relationship between F and bottleneck severity is probably due to both sampling error associated with the number of polymorphic loci examined and the loss of alleles during the bottlenecks. For populations that may have experienced severe bottlenecks, caution should be used in making evolutionary interpretations or management recommendations based on temporal changes in allele frequencies.     

Conservation Genetics in the Management of Desert Fishes

Abstract: The status and security of fishes in North American deserts have steadily declined in this century due to man's activities in this naturally fragile region. We address genetic aspects of the population structure of desert fishes as applicable to conservation and recovery programs by developing two zoogeographic models of isolation and gene flow. In the Death Valley model populations are isolated, with no chance of natural gene flow among them. Genetic diversity within populations tends to be low, but genetic divergence among populations within a species is high. In the Stream Hierarchy model, a complicated hierarchical genetic structure exists and is a function of geographic proximity and connectivity of habitats. Within-habitat genetic diversity tends to be higher, and among-habitat differentiation lower, than in the Death Valley model. These two systems must be recognized as distinct and managed differently. We also suggest three areas of experimentation needed to better understand and manage genetic stocks of desert fishes: relationships between heterozygosity and fitness, experimental mixing of similar stocks to examine effects of increased heterozygosity, and analysis of the relative roles of genetic adaptation and phenotypic plasticity in local differentiation.     

Preserving Population Allele Frequencies in Ex Situ Conservation Programs

Abstract: Optimization of contributions of parents to progeny by minimizing the average coancestry of the progeny is an effective strategy for maintaining genetic diversity in ex situ conservation programs, but its application on the basis of molecular markers has the negative collateral effect of homogenizing the allelic frequencies at each locus. Because one of the objectives of a conservation program is to preserve the genetic composition of the original endangered population, we devised a method in which markers are used to maintain the allele frequency distribution at each locus as closely as possible to that of the native population. Contributions of parents were obtained so as to minimize changes in allele frequency for a set of molecular markers in a population of reduced size. We used computer simulations, under a range of scenarios, to assess the effectiveness of the method in comparison with methods in which contributions of minimum coancestry are sought, either making use of molecular markers or genealogical information. Our simulations indicated that the proposed method effectively maintained the original distribution of allele frequencies, particularly under strong linkage, and maintained acceptable levels of genetic diversity in the population. Nevertheless, contributions of minimum coancestry determined from pedigree information but ignoring the genealogy previous to the conservation program, was the most effective method for maintaining allelic frequencies in realistic situations.     

Translocations and the Preservation of Allelic Diversity

Translocation is a tool commonly used for the conservation of threatened and endangered fish species. Despite extensive use, the biological implications of translocation remain poorly understood. Of particular interest is the effect of translocation on genetic variability. Maintenance of genetic variability in these "refuge" populations is assumed to be important for both short- and long-term success. We examined allozyme variability at 16 loci for western mosquitofish ( Gambusia affinis ) populations with known histories of introduction. Refuge populations had significantly lower levels of heterozygosity. Refuge populations also had considerably lower levels of allelic diversity than parental populations. All losses were of relatively rare alleles (frequency less than 0.1 in parental population). These losses were probably due to an undocumented bottleneck early in the introduction history. These results were surprising because the initial transplant involved 900 fish and because mosquitofish have numerous reproductive traits that should minimize the effects of bottlenecks on genetic diversity. A literature review revealed that genetic variability is often reduced in refuge populations and that such reductions typically involve the loss of alleles. We suggest that translocated populations be examined periodically for losses of genetic variability.     

Use of Experimental Translocations of Allegheny Woodrat to Decipher Causal Agents of Decline

Translocations are an important tool for wildlife conservation, although progress in the field of reintroduction biology has been hindered by the ad hoc and opportunistic nature of many translocations. We used an experimental translocation to elucidate the role of raccoon roundworm (Baylisascaris procyonis) and inbreeding depression in the decline of the Allegheny woodrat (Neotoma magister), an endangered species. We translocated woodrats from genetically diverse populations in the core of the species range to 4 previously occupied sites (reintroductions) and 2 sites supporting genetically depauperate populations (reinforcements) in Indiana (U.S.A.). In 2 reintroduction sites and 1 reinforcement site, we distributed anthelmintic baits to passively deworm raccoons and reduce the risk of woodrat exposure to roundworms. The remaining sites served as controls. We used raccoon latrine surveys and fecal flotation to monitor temporal variability in roundworm prevalence and effect of treatment. We used live trapping and microsatellite genotyping to monitor the demographic and genetic response of translocated populations over the following 54 months. At the conclusion of the study, 4 of 6 translocations were successfully maintaining abundance through local recruitment. The distribution of anthelmintic baits reduced levels of roundworm contamination, but levels of contamination were also low in 2 of 3 control sites. Reintroductions failed at control sites, one of which was due to high roundworm exposure. The other failed control reintroduction was likely attributable to demographic stochasticity and limited reproductive potential following initial mortality within the first 4 months. In both control and treatment reinforcements, increases in both allelic richness and heterozygosity were accompanied by increases in abundance, which is suggestive of genetic rescue. Our results demonstrate that mitigation of roundworm exposure through the distribution of anthelmintic baits can facilitate woodrat recovery and that diversity within genetically depauperate populations can be restored through the introduction of a limited number of individuals. El Uso de Reubicaciones Experimentales de Neotoma magister para Descifrar Agentes Causales de Disminución     

Patterns of Genetic Diversity and Its Loss in Mammalian Populations

Abstract:  Policy aimed at conserving biodiversity has focused on species diversity. Loss of genetic diversity, however, can affect population persistence, evolutionary potential, and individual fitness. Although mammals are a well-studied taxonomic group, a comprehensive assessment of mammalian genetic diversity based on modern molecular markers is lacking. We examined published microsatellite data from populations of 108 mammalian species to evaluate background patterns of genetic variability across taxa and body masses. We tested for loss of genetic diversity at the population level by asking whether populations that experienced demographic threats exhibited lower levels of genetic diversity. We also evaluated the effect of ascertainment bias (a reduction in variability when microsatellite primers are transferred across species) on our assessment of genetic diversity. Heterozygosity did not vary with body mass across species ranging in size from shrews to whales. Differences across taxonomic groupings were noted at the highest level, between populations of marsupial and placental mammals. We documented consistently lower heterozygosity, however, in populations that had experienced demographic threats across a wide range of mammalian species. We also documented a significant ( p = 0.01) reduction in heterozygosity as a result of ascertainment bias. Our results suggest that populations of both rare and common mammals are currently losing genetic diversity and that conservation efforts focused above the population level may fail to protect the breadth of persisting genetic diversity. Conservation policy makers may need to focus their efforts below the species level to stem further losses of genetic resources.     

Regional Consequences of Local Population Demography and Genetics in Relation to Habitat Management in Gentiana pneumonanthe

Abstract:  A joint demographic and population genetics stage-based model for a subdivided population was applied to Gentiana pneumonanthe , an early successional perennial herb, at a regional (metapopulation) scale. We used numerical simulations to determine the optimal frequency of habitat disturbance (sod cutting) and the intensity of gene flow among populations of G. pneumonanthe to manage both population viability and genetic diversity in this species. The simulations showed that even small populations that initially had near-equal allele frequencies could, if managed properly through sod cutting every 6 to 7 years, sustain their high genetic variation over the long run without gene flow. The more the allele frequencies in the small populations are skewed, however, the higher the probability that in the absence of gene flow, some alleles will be lost and within-population genetic variation will decrease even under proper management. This implies that although local population dynamics should be the major target for management, regional dynamics become important when habitat fragmentation and decreased population size lead to the loss of local genetic diversity. The recommended strategy to improve genetic composition of small populations is the introduction of seeds or seedlings of nonlocal origin.     

Simulating Retention of Rare Alleles in Small Populations to Assess Management Options for Species with Different Life Histories

Preserving allelic diversity is important because it provides the capacity for adaptation and thus enables long‐term population viability. Allele retention is difficult to predict in animals with overlapping generations, so we used a new computer model to simulate retention of rare alleles in small populations of 3 species with contrasting life‐history traits: North Island Brown Kiwi (Apteryx mantelli; monogamous, long‐lived), North Island Robins (Petroica longipes; monogamous, short‐lived), and red deer (Cervus elaphus; polygynous, moderate lifespan). We simulated closed populations under various demographic scenarios and assessed the amounts of artificial immigration needed to achieve a goal of retaining 90% of selectively neutral rare alleles (frequency in the source population = 0.05) after 10 generations. The number of immigrants per generation required to meet the genetic goal ranged from 11 to 30, and there were key similarities and differences among species. None of the species met the genetic goal without immigration, and red deer lost the most allelic diversity due to reproductive skew among polygynous males. However, red deer required only a moderate rate of immigration relative to the other species to meet the genetic goal because nonterritorial breeders had a high turnover. Conversely, North Island Brown Kiwi needed the most immigration because the long lifespan of locally produced territorial breeders prevented a large proportion of immigrants from recruiting. In all species, the amount of immigration needed generally decreased with an increase in carrying capacity, survival, or reproductive output and increased as individual variation in reproductive success increased, indicating the importance of accurately quantifying these parameters to predict the effects of management. Overall, retaining rare alleles in a small, isolated population requires substantial investment of management effort. Use of simulations to explore strategies optimized for the populations in question will help maximize the value of this effort. Simulación de la Retención de Alelos Raros en Poblaciones Pequeñas para Evaluar Opciones de Manejo para Especies con Historias de Vida Diferentes     

An experimental test of alternative population augmentation scenarios

Human land use is fragmenting habitats worldwide and inhibiting dispersal among previously connected populations of organisms, often leading to inbreeding depression and reduced evolutionary potential in the face of rapid environmental change. To combat this augmentation of isolated populations with immigrants is sometimes used to facilitate demographic and genetic rescue. Augmentation with immigrants that are genetically and adaptively similar to the target population effectively increases population fitness, but if immigrants are very genetically or adaptively divergent, augmentation can lead to outbreeding depression. Despite well‐cited guidelines for the best practice selection of immigrant sources, often only highly divergent populations remain, and experimental tests of these riskier augmentation scenarios are essentially nonexistent. We conducted a mesocosm experiment with Trinidadian guppies (Poecilia reticulata) to test the multigenerational demographic and genetic effects of augmenting 2 target populations with 3 types of divergent immigrants. We found no evidence of demographic rescue, but we did observe genetic rescue in one population. Divergent immigrant treatments tended to maintain greater genetic diversity, abundance, and hybrid fitness than controls that received immigrants from the source used to seed the mesocosms. In the second population, divergent immigrants had a slightly negative effect in one treatment, and the benefits of augmentation were less apparent overall, likely because this population started with higher genetic diversity and a lower reproductive rate that limited genetic admixture. Our results add to a growing consensus that gene flow can increase population fitness even when immigrants are more highly divergent and may help reduce uncertainty about the use of augmentation in conservation.     

A Concentric Analysis of the Impact of Urbanization on the Threatened European Tree Frog in an Agricultural Landscape

Abstract:  Pond-breeding amphibians are affected by site-specific factors and regional and landscape-scale patterns of land use. Recent anthropogenic landscape modifications (drainage, agriculture intensification, larger road networks, and increased traffic) affect species by reducing the suitable habitat area and fragmenting remaining populations. Using a robust concentric approach based on permutation tests, we evaluated the impact of recent landscape changes on the presence of the endangered European tree frog ( Hyla arborea ) in wetlands. We analyzed the frequency of 1 traffic and 14 land-use indices at 20 circular ranges (from 100-m up to 2-km radii) around 76 ponds identified in western Switzerland. Urban areas and road surfaces had a strong adverse effect on tree frog presence even at relatively great distances (from 100 m up to 1 km). When traffic measurements were considered instead of road surfaces, the effect increased, suggesting a negative impact due to a vehicle-induced effect. Altogether, our results indicate that urbanization and traffic must be taken into account when pond creation is an option in conservation management plans, as is the case for the European tree frog in western Switzerland. We conclude that our easy-to-use and robust concentric method of analysis can successfully assist managers in identifying potential sites for pond creation, where probability of the presence of tree frogs is maximized.     

Endangered Greater One-horned Rhinoceros Carry High Levels of Genetic Variation

Abstract: The population of Rhinoceros unicornis in the Chitwan Valley, Nepal, was reduced to an estimated effective population size (N_e of 21–28 individuals (60–80 total animals) in 1962. Protein electrophoresis shows that heterozygosity remains very high in this population (H_o= 9.9%) despite its near extinction. We attribute this high heterozygosity to large N_e's prior to the population bottleneck, the recent occurrence of the bottleneck, and long generation time. These results illustrate the importance of considering historical demography and life history parameters when evaluating the possible genetic effects of bottlenecks in wild populations. They also offer support to recent arguments that the erosion of genetic diversity attributed to bottlenecks may be overemphasized.     

The Paradox of Forest Fragmentation Genetics

Abstract:  Theory predicts widespread loss of genetic diversity from drift and inbreeding in trees subjected to habitat fragmentation, yet empirical support of this theory is scarce. We argue that population genetics theory may be misapplied in light of ecological realities that, when recognized, require scrutiny of underlying evolutionary assumptions. One ecological reality is that fragment boundaries often do not represent boundaries for mating populations of trees that benefit from long-distance pollination, sometimes abetted by long-distance seed dispersal. Where fragments do not delineate populations, genetic theory of small populations does not apply. Even in spatially isolated populations, where genetic theory may eventually apply, evolutionary arguments assume that samples from fragmented populations represent trees that have had sufficient time to experience drift, inbreeding, and ultimately inbreeding depression, an unwarranted assumption where stands in fragments are living relicts of largely unrelated predisturbance populations. Genetic degradation may not be as important as ecological degradation for many decades following habitat fragmentation.     

Conservation Genetics of Potentially Endangered Mutualisms: Reduced Levels of Genetic Variation in Specialist versus Generalist Bees

Abstract:  Oligolectic bees collect pollen from one or a few closely related species of plants, whereas polylectic bees visit a variety of flowers for pollen. Because of their more restricted range of host plants, it maybe expected that specialists exist in smaller, more isolated populations, with lower effective population sizes than generalists. Consequently, we hypothesized that oligolectic bees have reduced levels of genetic variation relative to related polylectic species. To test this hypothesis, we used five phylogenetically independent pairs of species in which one member was oligolectic and the other was polylectic. We assayed genetic variation in our species pairs at an average of 32 allozyme loci. Within each species pair, the oligolectic member had fewer polymorphic loci, lower average allelic richness, and lower average expected heterozygosity than its polylectic relative. Averaged over all species pairs, this corresponds to a 21% reduction in allelic richness, a 72% reduction in the proportion of polymorphic loci, and an 83% reduction in expected heterozygosity in specialists compared with generalists. Our data support the hypothesis of reduced effective population size in oligolectic bees and suggest that they may be more prone to extinction as a result. We suggest that in instances in which bee specialists are involved in mutually codependent relationships with their floral hosts, these mutualisms may be endangered for genetic and ecological reasons.     

Effects of Harvesting on Genetic Diversity in Old-Growth Eastern White Pine in Ontario, Canada

Genetic diversity measures at 54 isozyme loci coding for 16 enzymes in megagametophytes were compared between preharvest and postharvest gene pools of two adjacent virgin, old-growth (∼250 years) stands of eastern white pine ( Pinus strobus L.) in the Galloway Lake Old Pine Area of central Ontario. The concurrence of genetic diversity changes between the stands suggests that real and repeatable genetic erosion occurred in these gene pools as a result of harvesting. The total and mean number of alleles detected in each stand were reduced by approximately 25% after tree density reductions of 75%. The percentage of polymorphic loci dropped by about 33% from preharvest levels. About 40% of the low frequency (0.25> p ≥ 0.01) alleles and 80% of the rare ( p < 0.01) alleles were lost from each stand because of harvesting. Hypothetical multilocus gametic diversity was reduced by about 40% in each stand after harvesting. Latent genetic potential of each stand was reduced by about 50%, suggesting that the ability of these gene pools to adapt to changing environmental conditions may have been compromised. Heterozygosity estimates in the postharvest stands did not reflect reductions in allelic richness due to harvesting. Observed heterozygosity increased by 12% in one stand after harvesting, even though other genetic diversity measures decreased. Gene frequency changes due to harvesting imply that gene pools of naturally regenerated progeny stands may be quite different from the original parental stands. Silvicultural practices should ensure that the gene pools of remaining pristine old-growth stands have been reconstituted in the regenerating stands.