Restoration genetics of the vernal pool endemic Lasthenia conjugens (Asteraceae)

Restoration of habitat for endangered species often involves translocation of seeds or individuals from source populations to an area targeted for revegetation. Long-term persistence of a species is dependent on the maintenance of sufficient genetic variation within and among populations. Thus, knowledge and maintenance of genetic variability within rare or endangered species is essential for developing effective conservation and restoration strategies. Genetic monitoring of both natural and restored populations can provide an assessment of restoration protocol success in establishing populations that maintain levels of genetic diversity similar to those in natural populations. California’s vernal pools are home to many endangered plants, thus conservation and restoration are large components of their management. Lasthenia conjugens (Asteraceae) is a federally endangered self-incompatible vernal pool annual with gravity- dispersed seeds. Using the molecular technique of intersimple sequence repeats (ISSRs), this study assessed levels and patterns of genetic variability present within natural and restored populations of L. conjugens. At Travis Air Force Base near Fairfield, California, a vernal pool restoration project is underway. Genetic success of the ecologically based seeding protocol was examined through genetic monitoring of natural and restored populations over a three-year period. Genetic diversity remained constant across the three sampled generations. Diversity was also widely distributed across all populations. We conclude that the protocol used to establish restored populations was successful in capturing similar levels and patterns of genetic diversity to those seen within natural pools. This study also demonstrates how genetic markers can be used to inform conservation and restoration decisions.     

Genetic population structure of the Fire-bellied toad <Emphasis Type="Italic">Bombina bombina</Emphasis> in an area of high population density: implications for conservation

In this study, we report the genetic population structure of the Fire-bellied toad Bombina bombina in Brandenburg (East Germany) in the context of conservation. We analysed 298 samples originating from 11 populations in Brandenburg using mitochondrial control region sequences and six polymorphic microsatellite loci. For comparison, we included one population each from Poland and Ukraine into our analysis. Within Brandenburg, we detected a moderate variability in the mitochondrial control region (19 different haplotypes) and at microsatellite loci (9–12 alleles per locus). These polymorphisms revealed a clear population structure among toads in Brandenburg, despite a relatively high overall population density and the moderate size of single populations (100–2000 individuals). The overall genetic population structure is consistent with a postglacial colonization from South East-Europe and a subsequent population expansion. Based on genetic connectivity, we infer Management Units (MUs) as targets for conservation. Our genetic survey identified MUs, within which human infrastructure is currently preventing any genetic exchange. We also detect an unintentional translocation from South East to North West Brandenburg, presumably in the course of fish stocking activities. Provided suitable conservation measures are taken, Brandenburg should continue to harbor large populations of this critically endangered species.     

Ancient Demographics Determine the Effectiveness of Genetic Purging in Endangered Lizards

The purging of deleterious alleles has been hypothesized to mitigate inbreeding depression, but its effectiveness in endangered species remains debatable. To understand how deleterious alleles are purged during population contractions, we analyzed genomes of the endangered Chinese crocodile lizard (Shinisaurus crocodilurus), which is the only surviving species of its family and currently isolated into small populations. Population genomic analyses revealed four genetically distinct conservation units and sharp declines in both effective population size and genetic diversity. By comparing the relative genetic load across populations and conducting genomic simulations, we discovered that seriously deleterious alleles were effectively purged during population contractions in this relict species, although inbreeding generally enhanced the genetic burden. However, despite with the initial purging, our simulations also predicted that seriously deleterious alleles will gradually accumulate under prolonged bottlenecking. Therefore, we emphasize the importance of maintaining a minimum population capacity and increasing the functional genetic diversity in conservation efforts to preserve populations of the crocodile lizard and other endangered species.     

濒危植物居群恢复的遗传学考量

本文针对濒危植物居群的遗传多样性、生殖适合度、基因流、近交和远交衰退等遗传学问题在居群恢复过程中的应用进行了探讨。濒危植物居群的回归重建,既面临遗传多样性的迅速丧失、近交衰退等遗传风险,还因回归引种地存在较多近缘种而带来远交衰退的风险,最终导致遗传适应性降低,生境适应性变窄,繁殖和竞争能力减弱。为提高濒危物种保护的质量和效率,在构建回归居群时,应分批次从同一来源居群的不同母株采集材料,确保种源的遗传纯正性和遗传组成的多样性,还应使回归居群尽可能远离近缘广布种。另外,还需要对回归种群进行持续的监测和管理才能保证回归引种的成功。     

The Irish pollan, Coregonus autumnalis: options for its conservation

The ecology of four relict Irish populations of pollan ( Coregonus autumnalis ) is compared with that of the species elsewhere, and used to advocate conservation. The threats to these populations from introduced/invasive species, habitat degradation, climate warming and commercial exploitation are summarized and the legislation governing conservation of the stocks is reviewed. Conservation options (legislation, habitat restoration, stock translocation and stock augmentation) are outlined and their practicality and efficacy considered. A preliminary search indicates that there are a number of lakes that appear to be suitable for pollan translocation.     

Estimating the Population Size and Genetic Diversity of Amur Tigers in Northeast China

Over the past century, the endangered Amur tiger (Panthera tigris altaica) has experienced a severe contraction in demography and geographic range because of habitat loss, poaching, and prey depletion. In its historical home in Northeast China, there appears to be a single tiger population that includes tigers in Southwest Primorye and Northeast China; however, the current demographic status of this population is uncertain. Information on the abundance, distribution and genetic diversity of this population for assessing the efficacy of conservation interventions are scarce. We used noninvasive genetic detection data from scats, capture-recapture models and an accumulation curve method to estimate the abundance of Amur tigers in Northeast China. We identified 11 individual tigers (6 females and 5 males) using 10 microsatellite loci in three nature reserves between April 2013 and May 2015. These tigers are confined primarily to a Hunchun Nature Reserve along the border with Russia, with an estimated population abundance of 9–11 tigers during the winter of 2014–2015. They showed a low level of genetic diversity. The mean number of alleles per locus was 2.60 and expected and observed heterozygosity were 0.42 and 0.49, respectively. We also documented long-distance dispersal (~270 km) of a male Amur tiger to Huangnihe Nature Reserve from the border, suggesting that the expansion of neighboring Russian populations may eventually help sustain Chinese populations. However, the small and isolated population recorded by this study demonstrate that there is an urgent need for more intensive regional management to create a tiger-permeable landscape and increased genetic connectivity with other populations.     

Variation in the <Emphasis Type="Italic">COI</Emphasis> gene of the freshwater pearl mussel <Emphasis Type="Italic">Margaritifera margaritifera</Emphasis> from River Vuokkijoki

The freshwater pearl mussel Margaritifera margaritifera L. is one of the most endangered freshwater mussels in the world. Effective conservation of threatened species requires not only ecological, but also genetic information from the target species and populations. Since low genetic diversity can reduce the ability of a species to adapt to environmental changes, maintaining genetic diversity has been identified as one of the key elements in successful conservation programs. We examined genetic variation of the freshwater pearl mussel from the River Vuokkijoki, Karelia, Russia. We sequenced a fragment of the cytochrome c oxidase subunit I gene (COI) from 22 individuals and compared the data to 32 previously published COI sequences available in GenBank. We identified 10 different COI haplotypes in the sequenced samples, three of which had not been previously reported. Our results show that the River Vuokkijoki has high genetic diversity and suggest that the colonization of this northern freshwater pearl mussel population might have occurred from multiple and even distant refugia. Therefore, the freshwater pearl mussel population of the River Vuokkijoki is valuable for the conservation of the whole species.     

Assisted Colonization of Foundation Species: Lack of Consideration of the Extended Phenotype Concept—Response to Kreyling et al. (2011)

Assisted colonization (AC), or the intentional translocation of populations to compensate for risks related to climate change, is receiving increasing attention. It has been recently suggested by Kreyling et al. (2011) that rather than relocating endangered species, a focus should be placed on local adaptations of foundation or keystone species, and that these local ecotypes should be moved within their own range. Hence, this type of relocation could be applied with minimal risk in many restoration efforts. We think that caution is needed when considering the translocation of these foundation species, even within their range. Many recent studies have shown that foundation species can influence community structure and ecosystem processes through heritable traits, which suggests a genetic basis for ecosystem services. Thus, the translocation of different genotypes of foundation species might lead to unexpected results of colonization and might not be as “predictable” as Kreyling et al. have argued. Here, in our response, we stress how AC of foundation species can have important evolutionary consequences that might be impossible to reverse. We propose, whenever possible, (1) to favor population mixes of the foundation species to minimize the potential negative effects of specific genotypes; and (2) to collect from adjacent populations along ecological clines of the foundation species to mimic natural processes of migration under climate change.     

Genetic diversity and genetic structure of the endangered Manchurian trout, <Emphasis Type="Italic">Brachymystax lenok tsinlingensis</Emphasis>, at its southern range margin: conservation implications for future restoration

The Manchurian trout, Brachymystax lenok tsinlingensis (family: Salmonidae), is a cold freshwater fish endemic to Northeast Asia. South Korean populations of this species, which comprise its southern range limit, have recently decreased markedly in size and are now becoming critically endangered. We assessed the current population status of this species in South Korea by estimating the levels of genetic diversity and genetic structure of five natural and four restored populations using mitochondrial DNA (mtDNA) control region sequences and eight nuclear microsatellite loci. Levels of within-population genetic diversity were low, suggesting that past effective population sizes (N _e) have been small. Each population had one or a maximum of two mtDNA haplotypes. Microsatellite allelic richness (AR) was significantly higher for natural populations (mean AR?=?3.51; 95% confidence interval, 3.00–4.03) than for restored populations (mean AR?=?2.61; 2.38–2.98). South Korean populations were significantly genetically isolated from one another, with private mtDNA haplotypes and microsatellite alleles, suggesting that limited gene flow has been occurring among populations. A mtDNA phylogeny revealed that South Korean lineages were more closely related to those of China than to those of North Korea and Russia. Overall, we suggest that future restoration efforts aimed at South Korean populations should consider the genetic characteristics reported here, which should help to fulfil effective conservation strategies for this highly cherished species. Our results will inform other conservation efforts, including assisted migration of freshwater fish populations at the equatorial end of the geographical range limit of the species.     

Conservation of rare species with island-like distributions: A case study of Lasthenia conjugens (Asteraceae) using population genetic structure and the distribution of rare markers

Californian vernal pools, a patchy, island-like habitat, are endangered as a result of habitat destruction. Conservation of the remaining vernal pool habitat is essential for the persistence of several endangered species. We present the first study examining DNA-level genetic diversity within and among populations of a vernal pool plant species. We investigated genetic variation across eight populations of the US federally endangered vernal pool endemic Lasthenia conjugens (Asteraceae) using intersimple sequence repeat (ISSR) markers. Genetic diversity within the species was high (Nei's gene diversity estimate was 0.37), with moderate differentiation among populations (Bayesian F _ST analog of 0.124). Using an amova analysis, we found that the majority of the genetic variation (84%) was distributed within populations. There is a significant relationship between geographical distance and pairwise genetic differentiation as measured by the Bayesian estimate θ^B. The alternative hypotheses of historic geological processes within the Central Valley and contemporary gene flow are discussed as explanations of the data. Because of the vulnerability of the populations, we calculated a probability of loss for rare alleles (fragments) in the populations. Calculations show that sampling only one of the eight populations for ex-situ conservation or restoration will capture approximately 54% of the sampled rare fragments. We believe that one of the sampled populations has become extinct since it was sampled. When removing this population from the above-mentioned calculations, sampling one population will capture only 41.3% of the sampled rare fragments. We recommend sampling strategies for future conservation and restoration efforts of L. conjugens.     

Genetic assessment of population restorations of the critically endangered Silene hifacensis in the Iberian Peninsula

In order to preserve endangered plant populations and recover their evolutionary potential and ecological behavior, some restoration measures generally involve the reinforcement of the population size in existing natural populations or the reintroduction of new populations. Genetic monitoring of both natural and restored populations can provide an assessment of restoration protocol success in establishing populations that maintain levels of genetic diversity similar to those in natural populations. The highly threatened Spanish species Silene hifacensis (Caryophyllaceae) has only three natural reduced mainland populations in the Iberian Peninsula, following decline and extinction that occurred during the late 20th century. Preterit restoration strategies were essentially based on the implantation of new populations and reinforcement of certain existing populations using transplants mostly cultivated in greenhouses. In the present contribution, levels and patterns of genetic variability within natural and restored populations of Silene hifacensis were assessed using the molecular technique AFLP. Our results pointed out significant genetic diversity differences across the three existing natural populations though their population fragmentation and progressive loss of individuals have not had an impact on the global genetic diversity of this species. For restored populations, their levels of genetic diversity were similar and even higher than in natural populations. As a result, the past restoration protocols were successful in capturing similar and even higher levels of genetic diversity than those observed within natural pools. However, inbreeding processes have been detected for two restored populations. Finally, the main source of plant material for the long-time restored transplants appears to be the natural population of Cova de les Cendres. This study demonstrates, once again, how genetic markers are useful tools to be taken in consideration for endangered plant species conservation plans.     

Conservation Genetics of Jacquemontia reclinata (Convolvulaceae), an Endangered Species from Southern Florida: Implications for Restoration Management

Guidelines designed to aid in the restoration of rare species have been previously proposed using two primary strategies to select individuals for augmentation and reintroduction: mixing progeny from different populations or separating individuals from different populations. Understanding the genetic structure and diversity of an endangered species can offer insights into conservation management strategies. We used random amplified polymorphic DNA markers to assess the genetic structure and diversity of Jacquemontia reclinata , a federally endangered species endemic to Southeastern Florida. We sampled 20 percent of total number of individuals from eight of the ten known wild populations. Across individuals high levels of polymorphic loci (94.7%) were found and larger populations had greater genetic diversity. Cluster and ordination analyses found that one population was genetically differentiated from all the others; this population grows in a unique habitat. Most genetic variation (77.5%) was found within populations, and genetic distances between populations were not explained by their geographic distances. We recommend the use of two management units in restoration programs for J. reclinata , one consisting of the genetically differentiated population and the second consisting of the other seven populations sampled.     

Conservation genetics of the yellow-bellied toad (Bombina variegata): population structure,genetic diversity and landscape effects in an endangered amphibian

Revealing patterns of genetic diversity and barriers for gene flow are key points for successful conservation in endangered species. Methods based on molecular markers are also often used to delineate conservation units such as evolutionary significant units and management units. Here we combine phylo-geographic analyses (based on mtDNA) with population and landscape genetic analyses (based on microsatellites) for the endangered yellow-bellied toad Bombina variegata over a wide distribution range in Germany. Our analyses show that two genetic clusters are present in the study area, a northern and a southern/central one, but that these clusters are not deeply divergent. The genetic data suggest high fragmentation among toad occurrences and consequently low genetic diversity. Genetic diversity and genetic connectivity showed a negative relationship with road densities and urban areas surrounding toad occurrences, indicating that these landscape features act as barriers to gene flow. To preserve a maximum of genetic diversity, we recommend considering both genetic clusters as management units, and to increase gene flow among toad occurrences with the aim of restoring and protecting functional meta-populations within each of the clusters. Several isolated populations with especially low genetic diversity and signs of inbreeding need particular short-term conservation attention to avoid extinction. We also recommend to allow natural gene flow between both clusters but not to use individuals from one cluster for translocation or reintroduction into the other. Our results underscore the utility of molecular tools for species conservation, highlight outcomes of habitat fragmentation onto the genetic structure of an endangered amphibian and reveal particularly threatened populations in need for urgent conservation efforts.

     

Impacts by feral goats on critically endangered Crepidiastrum grandicollum (Compositae) endemic to the Ogasawara Islands

Oceanic islands are biodiversity hotspots with highly endemic ecosystems that are vulnerable to invasive alien species. Understanding the status of endangered species and identifying threats have the highest priority for insular biodiversity conservation. The two remaining populations of endangered Crepidiastrum grandicollum on Chichi-jima Island were studied for 5 years to describe their status and evaluate the impacts of feral goats (Capra hircus). The main cause of population decline was browsing by goats. The populations protected by the exclosures were stable but declined after an exclosure was removed. Even in the protected population, regeneration was limited outside the exclosures and in 1 year of the survey, a high proportion of feeding damage by moth larvae was observed in one population. These facts indicate that exclosures are not a sufficient conservation measure, and eradication of goats and population restoration in novel habitats are necessary to reduce the extinction risk of C. grandicollum.     

Population genetic and behavioural variation of the two remaining colonies of Providence petrel (<Emphasis Type="Italic">Pterodroma solandri</Emphasis>)

Knowledge of the dispersal capacity of species is crucial to assess their extinction risk, and to establish appropriate monitoring and management strategies. The Providence petrel (Pterodroma solandri) presently breeds only at Lord Howe Island (~32,000 breeding pairs) and Phillip Island-7 km south of Norfolk Island (~20 breeding pairs). A much larger colony previously existed on Norfolk Island (~1,000,000 breeding pairs) but was hunted to extinction in the 18th Century. Differences in time of return to nesting sites are presently observed between the two extant colonies. Information on whether the Phillip Island colony is a relict population from Norfolk Island, or a recent colonization from Lord Howe Island, is essential to assess long-term sustainability and conservation significance of this small colony. Here, we sequenced the mitochondrial cytochrome b gene and 14 nuclear introns, in addition to genotyping 10 microsatellite loci, to investigate connectivity of the two extant P. solandri populations. High gene flow between populations and recent colonization of Phillip Island (95 % HPD 56–200 ya) are inferred, which may delay or prevent the genetic differentiation of these insular populations. These results suggest high plasticity in behaviour in this species and imply limited genetic risks surrounding both the sustainability of the small Phillip Island colony, and a proposal for translocation of Lord Howe Island individuals to re-establish a colony on Norfolk Island.     

Extinction,reintroduction, and restoration of a lizard meta‐population equilibrium in the missouri ozarks

In this issue of Molecular Ecology, Neuwald & Templeton (2013) report on a 22‐year study of natural populations of Collared Lizards (Crotaphytus collaris) that evolved on isolated on rock outcrops (‘glades’) in the Ozark Mountains in eastern Missouri. This ecosystem was originally maintained by frequent fires that kept the forest understory open, but fire‐suppression was adopted as official policy in about 1945, which led to a loss of native biodiversity, including local extinctions of some lizard populations. Policies aimed at restoring biodiversity included controlled burns and re‐introductions of lizards to some glades, which began in 1984. Populations were monitored from 1984–2006, and demographic and genetic data collected from 1 679 lizards were used to documents shifts in meta‐population dynamics over four distinct phases of lizard recovery: 1–an initial translocation of lizards drawn from the same source populations onto three glades that were likely part of one meta‐population; 2–a period of isolation and genetic drift associated with the absence of fires; 3–a period of rapid colonization and population increase following restoration of fire; and 4–stabilization of the meta‐population under regular prescribed burning. This study system thus provides a rare opportunity to characterize the dynamics of a landscape‐scale management strategy on the restoration of the meta‐population of a reintroduced species; long‐term case studies of the extinction, founding, increase, and stabilization of a well‐defined meta‐population, based on both demographic and population genetic data, are rare in the conservation, ecological, and evolutionary literature.     

Effects of forest-dune ecotone management on the endangered heath grasshopper, Chorthippus vagans (Orthoptera: Acrididae)

Dry, oligotrophic ecosystems are highly threatened in Europe due to massive changes in land use and eutrophication. The conservation of these xeric habitats has received much attention, whereas the ecotones between xeric habitats and other habitat types are often disregarded. One species which mainly inhabits the transition zone between pine forests and adjacent xeric habitats is the heath grasshopper, Chorthippus vagans. This species is endangered in large parts of Europe. One of the largest populations in northern Germany is found on a degraded inland dune near Hanover. This population is threatened by dense growth of deciduous trees and litter accumulation. We analyzed changes in the distribution of this population after the implementation of conservation measures (thinning out the forest and removal of leaf litter). Moreover, we examined dispersal distances of the species in order to assess its colonization potential. We also studied the microhabitat preferences of C. vagans to assess key factors influencing its local distribution. Our data show a substantial growth in population size, which might be a consequence of the conservation measures. New patches on the dune were colonized, promoting dispersal between the subpopulations. We propose that restoration of forest-dune ecotones should be considered more often in landscape planning and conservation management.     

Translocation of an endangered insect species,the field cricket (<Emphasis Type="Italic">Gryllus campestris</Emphasis> Linnaeus, 1758) in northern Germany

Relocations of species have become a tool widely used in nature conservation, but insects have rarely been considered as targets. Here, we present a translocation project of the field cricket (Gryllus campestris L. 1758), which is a threatened species at the northern edge of its range. Only ten populations were left in Lower Saxony (Germany), illustrating the need for urgent conservation measures. After 10 years of monitoring and management of an isolated population, 213 nymphs were captured and released at another nature reserve in summer 2001. The size of the new population increased significantly from 27 singing males in spring 2002 to 335 singing males in spring 2005. The occupied area increased from 5.66 ha to 33.14 ha. Altogether, the translocation project was evaluated as successful, but the inland dune proved to be not as suitable for the species as initially expected. Our results indicate that translocations of highly reproductive insect species are promising, as long as the release locality contains sufficiently large areas of suitable habitat and a high number of wild juveniles from a closely located and large source population are released in a climatically favorable period. Management and restoration of habitats, as well as continuous monitoring are of crucial importance for the success of the translocation project. Moreover, the importance of a high quality of cooperation between conservationists, authorities, foresters, farmers, financiers and scientists cannot be overstated.     

Noninvasive genetic sampling allows estimation of capercaillie numbers and population structure in the Bohemian Forest

Wildlife conservation and management of endangered species requires reliable information on the size and structure of populations. One of the flagship species in European wildlife conservation is the forest-dwelling capercaillie (Tetrao urogallus), where several populations are endangered. In the Bohemian Forest, e.g., the population severely declined 30 years ago with only 100 birds remaining in 1985. Subsequently, breeding and release programs were conducted to supplement the local population. The current distribution and population size, however, remained unknown. With recent habitat changes and increasing recreational activities, a reliable population estimate to inform conservation plans was needed. A team of scientists and volunteers collected fresh capercaillie droppings covering an area of about 120,000 ha. We genotyped ten microsatellite loci to estimate the current population size and to determine the population’s spatial and genetic structure. Population size and density estimators revealed a population size of approximately 500 individuals, which is thus one of the two largest relict populations in the low mountain ranges of temperate Europe. The population clustering revealed gene flow across the entire study area. Several genotypes were documented with multiple recaptures at spatial distances between 10 and 30 km additionally corroborating gene flow across the entire landscape of the study area. Males were more closely related than females on small spatial scales up to 3 km, indicating lower dispersal rates in males. We conclude that the population currently appears to have a viable size and shows unrestricted gene flow across state borders and management units of the entire Bohemian Forest. However, long-term viability of this population requires a transboundary strategy to sustainably protect and monitor this isolated capercaillie population in Central Europe.     

Determinants of successful establishment and post‐translocation dispersal of a new population of the critically endangered St. Croix ground lizard (Ameiva polops)

Translocation to areas free of exotic predators, habitat degradation, or disease may be the most viable restoration option for many endangered species. We report on a successful translocation of the critically endangered St. Croix ground lizard, Ameiva polops, extirpated from St. Croix, U.S. Virgin Islands, Caribbean, by predation from introduced mongooses (Herpestes auropunctatus). We translocated 57 adult A. polops from Green Cay to Buck Island in May 2008. We placed 4 females and 3 males each in eight, 100 m², enclosures on Buck Island for 71 days, then the enclosures were opened. During the enclosure period, 20 individuals were identified and 32 others were seen. The average number sighted per survey was only 5.28 (range = 2–10). One hatchling was sighted in an enclosure, indicating a translocated female successfully nested. Body condition of the translocated individuals increased significantly by the end of the enclosure period. Population monitoring surveys at 61 sites across Buck Island showed that 5 years after the initial translocation in June 2013, the new population had grown to an estimated 1,473 individuals and occupied 58.9% of the island. We attribute eradication of mongoose, life history of the species, large propagule size, condition of habitat, soft‐release, use of adults, interagency collaboration, and systematic assessment as primary factors that facilitated this successful translocation. Our findings provide meaningful insights on factors that enhance the potential for successful translocations, and point to new strategies aimed at restoring populations of endangered reptiles in their native ranges.