Conservation implications of the distribution of genetic diversity at different scales: a case study using the marsh fritillary butterfly (Euphydryas aurinia)

In the UK, Euphydryas aurinia exists in fragmented habitat patches, and undergoes population fluctuations as a result of a larval parasitoid. Its range is declining in the UK and conservation is thought to require a landscape approach since populations spread over large areas in some years and contract to core breeding patches in others. We examined populations at a range of geographic scales using allozyme electrophoresis to look for evidence of gene flow and differences in genetic diversity among populations. Nationally, our F_ST value was 0.1542 but between population groups within the suspected colonisation range of the butterfly (ca. 20 km), F_ST values were not significantly different from zero. Genetic diversity in terms of number of alleles and heterozygosity was reasonably high in natural populations (H_e=0.267) but low in an introduced, isolated population. We infer that migration between closely spaced subpopulations (in a metapopulation) maintains a high genetic effective population size (large number of individuals in a population that contribute genes to the next generation) which offsets any local reductions in population numbers due to stochastic extinctions or parasitoid effects. We therefore conclude that effective conservation of the species must seek to provide networks of suitable habitat for groups of subpopulations, rather than maintaining habitat for isolated populations.     

Conservation genetics of evolutionary lineages of the endangered mountain yellow-legged frog, Rana muscosa (Amphibia: Ranidae), in southern California

Severe population declines led to the listing of southern California Rana muscosa (Ranidae) as endangered in 2002. Nine small populations inhabit watersheds in three isolated mountain ranges, the San Gabriel, San Bernardino and San Jacinto. One population from the Dark Canyon tributary in the San Jacinto Mountains has been used to establish a captive breeding population at the San Diego Zoo Institute for Conservation Research. Because these populations may still be declining, it is critical to gather information on how genetic variation is structured in these populations and what historical inter-population connectivity existed between populations. Additionally, it is not clear whether these populations are rapidly losing genetic diversity due to population bottlenecks. Using mitochondrial and microsatellite data, we examine patterns of genetic variation in southern California and one of the last remaining populations of R. muscosa in the southern Sierra Nevada. We find low levels of genetic variation within each population and evidence of genetic bottlenecks. Additionally, substantial population structure is evident, suggesting a high degree of historical isolation within and between mountain ranges. Based on estimates from a multi-population isolation with migration analysis, these populations diversified during glacial episodes of the Pleistocene, with little gene flow during population divergence. Our data demonstrate that unique evolutionary lineages of R. muscosa occupy each mountain range in southern California and should be managed separately. The captive breeding program at Dark Canyon is promising, although mitigating the loss of neutral genetic diversity relative to the natural population might require additional breeding frogs.     

Linking the gaps between conservation research and conservation management of rare dipterocarps: A case study of Shorea lumutensis

To conserve a rare plant, conservation programs must be guided by the biological attributes of the species. What constitutes the most critical biological information for plant conservation has been the issue of discussion for the last 30 years. Most scientists promote an approach that is either ecological or genetic in emphasis. Ecological and genetic processes will often interact synergistically to influence the population viability and to determine the persistence of populations in the long run. Consequently, conservation management of plant species, in addition to ecological information, requires a robust understanding of underlying genetic processes as well as the variation within and between populations. Conservation has a cost and the resources available for conservation programs are always limited. Therefore, conservation management strategies should not only be scientifically justified but also practical in terms of resource availability. Shorea lumutensis is a rare and endemic dipterocarp in Peninsular Malaysia. A comprehensive research activity was initiated to assess the population ecology and population genetics of S. lumutensis to elucidate specific ecological and genetic requirements and subsequently to set conservation strategies and priorities. This paper is apparently the first attempt at applying both the ecological and genetic approaches into conservation management of a rare dipterocarp. In addition, this paper also attempts to link the gaps between conservation research and conservation management in a realistic manner. It is our hope that this study will serve as a model for the study of other rare dipterocarps which should be given priority for conservation.     

Extreme population divergence and conservation implications for the rare endangered Atlantic Forest sloth, Bradypus torquatus (Pilosa: Bradypodidae)

The maned sloth (Bradypus torquatus) is an endangered species endemic to the Atlantic Forest of eastern Brazil. This biome has been reduced to 7% of its original extent and the remaining forests are highly fragmented. We analyzed 70 samples from the largest remnant populations in the states of Bahia, Espírito Santo and Rio de Janeiro to characterize their geographic structure and to produce estimates of genetic diversity. The analysis indicated that the remnant populations are reproductively isolated and extremely divergent. The populations present a very discontinuous distribution, with divergent genetic clusters specific to different geographical regions, probably caused by allopatric fragmentation. This pattern is likely related to Pleistocenic climatic and vegetation changes, and indicates the presence of at least two independent evolutionary units. The analyses also indicate that populations separated by more than 100 km should be considered different management units. Thus, devastation of the Atlantic Forest leads to an unrecoverable loss of genetic diversity in this species. These conclusions should direct conservation actions aimed at preserving the distinctiveness of each evolutionary unit, as well as to preserve the demographic isolation of different management units.     

Relationship between population size and genetic diversity in endangered populations of the European bullhead (Cottus gobio): implications for conservation

This study investigated the relationship between the current size of endangered bullhead (Cottus gobio) populations and microsatellite genetic variability. Additionally, the microsatellite data were used to evaluate whether a genetic test for population bottlenecks was able to provide evidence of recent severe population declines. Finally, our results were used to develop conservation priorities and measures. Population size appears to be a crucial parameter in determining the amount of genetic diversity that can be preserved in bullheads, since a significant positive correlation was observed between both variables. Furthermore, in some populations we were able to detect genetic signatures of the documented decline in population size. We suggest that the most immediate goal for bullhead conservation should be to increase the size and the range of the populations, and in doing so minimise or even reverse further genetic erosion. Potential management actions like habitat quality improvement, reduction of river fragmentation and supplementation programmes (translocation, supportive breeding) are discussed.     

Microsatellite analysis of genetic population structure of the endangered cyprinid Anaecypris hispanica in Portugal: implications for conservation

The endangered fish species Anaecypris hispanica is restricted to eight disjunct populations in the Portuguese Guadiana drainage. The genetic structure of these populations was studied in order to determine levels of genetic variation within and among populations and suggest implications for conservation of the species. Based on five microsatellite loci, the null hypothesis of population homogeneity was tested. Tests for genetic differentiation revealed highly significant differences for pairwise comparisons between all populations, and substantial overall population subdivision (F_ST=0.112). All sampled populations contained unique alleles. Our findings indicate marked genetic structuring and emphasise limited dispersal ability. The high levels of genetic diversity detected within and among A. hispanica populations suggest, however, that the observed fragmentation and reduction in population size of some populations during the last two decades, has impacted little on levels of genetic variability. Data imply that most A. hispanica populations should be managed as distinct units and that each has a high conservation value containing unique genetic variation. It is argued that geographic patterns of genetic structuring indicate the existence of eight management units.     

Sperm storage and low incidence of multiple paternity in the European pond turtle, Emys orbicularis: A secure but costly strategy?

The freshwater pond turtle, Emys orbicularis, has recently suffered from population declines throughout its range, mainly due to habitat destruction. The mating strategies of this species were studied using genetic data from successive clutches within and between years. To test for the occurrence and frequency of multiple paternity and sperm storage, genetic paternity at six microsatellite markers was assessed in 114 embryos and hatchlings from single and subsequent clutches of 11 females (including clutches from the same or consecutive years). Multiple paternity was rare and only found in two out of 20 clutches from 11 females. All annual successive clutches and 58% of the clutches in the next year, were fertilized with sperm from the same male. The use of stored sperm is thus a frequent strategy in E. orbicularis. However, hatching rate, hatchling mass, and hatchling length decreased in clutches fertilized by stored sperm, suggesting sperm depletion or deterioration through time. The occurrence of stored sperm despite an associated reduced reproductive output indicated that mating and/or the fertilization process is costly to females. The low incidence of multiple paternity may simply be the residual consequence of the capacity to store viable sperm. These results provide important and innovative insights for the conservation of E. orbicularis. In threatened populations, management strategies may aim to enhance effective copulations in order to increase the reproductive output of females.     

Applying species distribution modelling to identify areas of high conservation value for endangered species: A case study using Margaritifera margaritifera (L.)

The development and implementation of a population supplementation and restoration plan for any endangered species should involve an understanding of the species’ habitat requirements prior to the release of any captive bred individuals. The freshwater pearl mussel, Margaritifera margaritifera, has undergone dramatic declines over the last century and is now globally endangered. In Northern Ireland, the release of captive bred individuals is being used to support wild populations and repatriate the species in areas where it once existed. We employed a combination of maximum entropy modelling (MAXENT) and Generalized Linear Mixed Models (GLMM) to identify ecological parameters necessary to support wild populations using GIS-based landscape scale and ground-truthed habitat scale environmental parameters. The GIS-based landscape scale model suggested that mussel occurrence was associated with altitude and soil characteristics including the carbon, clay, sand, and silt content. Notably, mussels were associated with a relatively narrow band of variance indicating that M. margaritifera has a highly specific landscape niche. The ground-truthed habitat scale model suggested that mussel occurrence was associated with stable consolidated substrates, the extent of bankside trees, presence of indicative macrophyte species and fast flowing water. We propose a three phase conservation strategy for M. margaritifera identifying suitable areas within rivers that (i) have a high conservation value yet needing habitat restoration at a local level, (ii) sites for population supplementation of existing populations and (iii) sites for species reintroduction to rivers where the mussel historically occurred but is now locally extinct. A combined analytical approach including GIS-based landscape scale and ground-truthed habitat scale models provides a robust method by which suitable release sites can be identified for the population supplementation and restoration of an endangered species. Our results will be highly influential in the future management of M. margaritifera in Northern Ireland.     

The use of historical collections to estimate population trends: A case study using Swedish longhorn beetles (Coleoptera: Cerambycidae)

Long term data to estimate population trends among species are generally lacking. However, Natural History Collections (NHCs) can provide such information, but may suffer from biases due to varying sampling effort. To analyze population trends and range-abundance dynamics of Swedish longhorn beetles (Coleoptera: Cerambycidae), we used collections of 108 species stretching over 100 years. We controlled for varying sampling effort by using the total number of database records as a reference for non-red-listed species. Because the general frequency of red-listed species increased over time, a separate estimate of sampling effort was used for that group. We observed large interspecific variation in population changes, from declines of 60% to several hundred percent increases. Most species showed stable or increasing ranges, whereas few seemed to decline in range. Among increasing species, rare species seemed to expand their range more than common species did, but this pattern was not observed in declining species. Historically, rare species did not seem to be at larger risk of local extinction, and population declines were mostly due to lower population density and not loss of sub-populations. We also evaluated the species’ declines under IUCN red-list criterion A, and four currently not red-listed species meet the suggested threshold for Near Threatened (NT). The results also suggested that species’ declines may be overlooked if estimated only from changes in species range.     

Case studies on breeding systems and its consequences for germplasm conservation: 3. Electrophoretic mobility of phaseolins in wild populations of Lima beans (Phaseolus lunatus L.) in the Central Valley of Costa Rica

We studied the variation in the electrophoretic mobility of phaseolins of 202 individual seeds from thirty-eight wild populations of Lima beans (Phaseolus lunatus) in the Central Valley of Costa Rica. We found four main polypeptides or banding regions with molecular weights which ranged from 21.5 to 31 kDaltons. Two of these peptides were polymorphic, and their combination resulted in four distinct genotypes. The frequency of the different genotypes varies among populations, but two of them, namely B¹¹C¹¹, B²²C²², were found in most of the populations studied (37 out of 38 populations examined). Overall, there was a lack of heterozygotes among the individuals examined, which strongly suggests that there is gametic disequilibrium for the encoding regions involved in the synthesis of these polypeptides. When we arranged the populations into eight groups according to their proximity and phenology, we found that the distribution of the peptides was also variable among them. The possible association between the distribution of the patterns of electrophoretic mobility observed and climatic factors, such as mean annual rainfall, mean annual temperature and mean annual relative humidity is discussed.     

AFLP fingerprinting of Anchusa (Boraginaceae) in the Corso-Sardinian system: Genetic diversity, population differentiation and conservation priorities in an insular endemic group threatened with extinction

Although several plants endemic to Corsica and Sardinia are included in various redlists, no attempts have been made to analyse their genetic diversity with molecular techniques. Genus Anchusa occurs with seven taxa in either mountain or coastal habitats of the two islands, but the very restricted range and low population size pose these endemisms in a very precarious conservation status. Highly variable markers (AFLP) were therefore used to analyse the patterns and levels of genetic diversity in a sample of 11 populations from the entire range of the group.Results indicate the separation between a mountain genic pool including Anchusa formosa, Anchusa capellii and Anchusa montelinasana, and four groups of coastal accessions. In spite of small size, mountain taxa show low interpopulation differentiation (F_st = 0.02) and relatively high intrapopulation genetic variation (0.365), while coastal accessions showed on average a stronger differentiation (mean F_st = 0.20) and a lower diversity (0.281), possibly due to higher rates of inbreeding. The particularly low levels of variation found in A. sardoa, A. littorea and A. crispa ssp. maritima from the Coghinas bay are likely due to a historical decrease of populations and to bottleneck events caused by loss of habitat and natural stochastic factors on sand dune ecosystems. While habitat maintenance and regulation of grazing by domestic herbivores should be sufficient to ensure the persistence of the mountain endemics, additional actions of in situ and ex situ conservation are needed for the critically endangered coastal species A. sardoa and A. littorea. A. crispa showed a relatively high variation, especially on Corsica. No correlation between population size and genetic variation was found in the latter species, highlighting the importance of the small patches for its conservation. Also, the genetic separation between subspecies crispa and maritima stresses the need of keeping them distinct in redlists and conservation actions on Sardinia.     

Genetics of a butterfly relocation: large, small and introduced populations of the mountain endemic Erebia epiphron silesiana

Species specific levels of genetic diversity are necessary for the viability of populations. Erebia epiphron (Lepidoptera: Satyrinae) is a relic species in the Sudetan Mountains, Czech Republic. These populations represent a suitable study object to test the population genetic effects of large, small and introduced populations by means of allozyme electrophoresis. A large continuous and viable population (>100,000 individuals) shows a significantly higher genetic diversity than a small isolated and declining population (ca. 5000 individuals), which most probably was founded during the Little Ice Age in the 17th and 18th centuries and became isolated afterwards. In 1932 and 1933, 50 females were transferred from the large population to the Krkonose Mountains, where they established a viable and quickly expanding population. The number of transferred individuals was sufficient to transfer most of the allozyme diversity of the large natural population. Due to the higher elevation of the Krkonose in comparison to the native range, this allocation might safeguard the survival of the Czech race of E. epiphron even under conditions of global warming.     

Potential distribution modelling, niche characterization and conservation status assessment using GIS tools: a case study of Iberian Copris species

Dung beetle populations, in decline, play a critical ecological role in extensive pasture ecosystems by recycling organic matter; thus the importance of their conservation status. Presence data available for Copris hispanus (L.) and Copris lunaris (L.) (Coleoptera, Scarabaeidae) in Comunidad de Madrid (CM), and BIOMAPPER, a GIS-based tool, was used to model their environmental niches. The so derived potential distributions of both species were used to exemplify the utility of this kind of methodologies in conservation assessment, as well as its capacity to describe the potential sympatry between two or more species. Both species, distributed along a Dry-Mediterranean to Wet-Alpine environmental conditions gradient, overlap in areas of moderate temperatures and mean annual precipitations in the north of CM. Copris are poorly conserved in the existing protected sites network, but protection provided by new sites included in the future Natura 2000 Network will improve the general conservation status of these species in CM.     

Spatial covariation of Azotobacter abundance and soil properties: A case study using the wavelet transform

The soil microflora is very heterogeneous in its spatial distribution. The origins of this heterogeneity and its significance for soil function are not well understood. A problem for understanding spatial variation better is the assumption of statistical stationarity that is made in most of the statistical methods used to assess it. These assumptions are made explicit in geostatistical methods that have been increasingly used by soil biologists in recent years. Geostatistical methods are powerful, particularly for local prediction, but they require the assumption that the variability of a property of interest is spatially uniform, which is not always plausible given what is known about the complexity of the soil microflora and the soil environment. We have used the wavelet transform, a relatively new innovation in mathematical analysis, to investigate the spatial variation of abundance of Azotobacter in the soil of a typical agricultural landscape. The wavelet transform entails no assumptions of stationarity and is well suited to the analysis of variables that show intermittent or transient features at different spatial scales.In this study, we computed cross-variograms of Azotobacter abundance with the pH, water content and loss on ignition of the soil. These revealed scale-dependent covariation in all cases. The wavelet transform also showed that the correlation of Azotobacter abundance with all three soil properties depended on spatial scale, the correlation generally increased with spatial scale and was only significantly different from zero at some scales. However, the wavelet analysis also allowed us to show how the correlation changed across the landscape. For example, at one scale Azotobacter abundance was strongly correlated with pH in part of the transect, and not with soil water content, but this was reversed elsewhere on the transect.The results show how scale-dependent variation of potentially limiting environmental factors can induce a complex spatial pattern of abundance in a soil organism. The geostatistical methods that we used here make assumptions that are not consistent with the spatial changes in the covariation of these properties that our wavelet analysis has shown. This suggests that the wavelet transform is a powerful tool for future investigation of the spatial structure and function of soil biota.     

Novel microsatellite loci for the compost earthworm Eisenia fetida: A genetic comparison of three North American vermiculture stocks

Earthworms are important components of soil ecosystems worldwide, and have been used extensively as indicator species in ecotoxicology studies. Our understanding of mating systems, population structure, and genetic diversity in earthworms is limited by the current lack of available genetic tools. To address this gap, we developed 16 novel microsatellite markers for the compost earthworm Eisenia fetida, one of the most widely studied earthworm species. We tested the new markers in 3 putative populations of worms from commercial vermiculture operations in Canada and the U.S.A. All 16 loci were variable in at least one population, with the number of alleles per locus ranging from 3 to 10, observed heterozygosity from 0.000 to 0.783, and polymorphic information content from 0.032 to 0.587. One group had significantly reduced heterozygosity compared to the other 2, but overall there were only minor genetic differences among the tested suppliers, suggesting a possible bottleneck for this species in North America. The microsatellite loci we describe here will be extremely useful tools for future field and laboratory studies of E. fetida. Body size and condition in adult worms varied significantly by supplier, and breeding success and cocoon production were significantly higher in breeding pairs from one supplier compared to the other two. In the absence of evidence for genetic divergence among populations, our results suggest that the early rearing environment may be an important factor affecting adult breeding performance. We caution that the source of worms may affect the outcome of laboratory experiments involving growth and reproduction.     

Conservation of the purple gallinule (Porphyrio porphyrio L.) in Portugal: causes of decline, recovery and expansion

Historical data on the distribution and status of the purple gallinule (Porphyrio porphyrio) in Portugal were collected by bibliographic research, from museum catalogues and from questionnaires to wetland managers, ornithologists and birdwatchers. The population suffered severe decline and range contraction at the end of the 19th century and most of the 20th century, due mainly to habitat loss and degradation and excessive hunting. However, the population has begun to recover in the last decade mainly through direct protection of the species and of the remaining habitat, together with creation of artificial wetlands and a reintroduction programme, and probably also due to immigration from Spain. The population size has increased about 10-fold and the range has expanded to most of its former area. The estimated Portuguese breeding population in 2002 was 49-67 pairs. Densities varied between 0.05 and 7 individuals/ha in recently colonized areas and stable (saturated) areas, respectively. Although the population is recovering, its small size, high fragmentation and several natural and anthropogenic threats lead us to consider it still a species of major concern in Portugal.     

Use of the Biological Flora framework in the United Kingdom Overseas Territories: Euphorbia origanoides L.

The United Kingdom Overseas Territories (UKOTs) are globally important for a high diversity of endemic and threatened plant species but are poorly represented in plant ecological literature. This lack of ecological research is compounded by a lack of funding and skills. Cost effective approaches of compiling conservation relevant information are required. Here we present the first examination of a species from the UKOTs presented within the standard framework of a Biological Flora. This framework allows a convenient way to compile ecological information and assess missing data. The account reviews all available information on Euphorbia origanoides L. (Ascension spurge) from Ascension Island (South Atlantic Ocean) relevant to understanding its ecology and conservation, including soil chemistry, climate and plant community data. E. origanoides is an endemic perennial, found in dry, lava plains of Ascension Island with soils comprised of weathered volcanic scoria. E. origanoides has suffered habitat loss through the introduction of invasive species and survival in the wild is currently under threat. We relate the information gathered for this Biological Flora to the conservation of the species in the wild and propose the framework should be used as one way of compiling information relevant for conservation managers. The framework is beneficial as it allows an evidence-based approach to conservation but also permits the prioritisation of research and can help conservation managers to meet targets for the Convention on Biological Diversity and the Global Strategy for Plant Conservation.     

Population baseline data for monitoring genetic diversity loss for 2010: A case study for Brassica species in the UK

The Convention on Biological Diversity and the subsequent International Treaty on Plant Genetic Resources for Food and Agriculture have proved a watershed in plant genetic resources (PGR) conservation and consequent initiatives have set various PGR conservation targets to be met by 2010. The aim of our paper is to develop ideas and issues concerning the monitoring of natural genetic resources: particularly in terms of developing a baseline from which to measure levels of genetic diversity. Three species of Brassica (B. nigra, B. oleracea, B. rapa) found wild in the UK were assessed for levels of genetic diversity using AFLP. The relationship between genetic distribution and ecogeographic distribution was considered for each species to determine patterns that may be useful in formulating conservation strategies. Genetic distance between populations of B. nigra and B. rapa were correlated to geographic distance. Levels of genetic polymorphism in B. oleracea were correlated to soil pH while in B. rapa they were correlated to soil coarseness. In terms of PGR conservation these findings may suggest an emphasis toward in situ conservation of a selection of disparate populations would be appropriate where possible as such adaptations may be lost in ex situ collections.     

How to calculate nitrogen rhizodeposition: a case study in estimating N rhizodeposition in the pea (Pisum sativum L.) and grasspea (Lathyrus sativus L.) using a continuous N labelling split-root technique

The formulae used in studies with ¹⁵N labelling techniques for estimating the N rhizodeposition (Ndfr) of legumes differ according to the background atom% ¹⁵N values used to determine ¹⁵N excess in the soil and roots grown in soil. Therefore, a continuous ¹⁵N labelling split-root experiment with pea (Pisum sativum L.) and grasspea (Lathyrus sativus L.) was undertaken and the relevant calculations were made to determine a valid method for calculating Ndfr. It is shown that a non-nodulated reference plant or a legume grown on soil without ¹⁵N labelling are required components of experiments which aim to estimate legume-N rhizodeposition, if the ¹⁵N abundance of the total soil N at the start of the experiment and that of the total plant available soil N are different. The standard formula was developed further to calculate Ndfr in a valid way. The impact of using different background atom% ¹⁵N values on the results when estimating Ndfr are demonstrated according to the ¹⁵N abundance of the roots grown in the soil. At physiological maturity, the rhizodeposition of N from roots grown in the soil was 19.8 mg N plant⁻¹ for pea and 14.1 mg N plant⁻¹ for grasspea, which is, respectively, equivalent to 10.5 and 9.2% of their total root and shoot N.     

The extinction of genetic resources of Asian wild rice, Oryza rufipogon Griff.: A case study in Thailand

Natural populations of wild rice, Oryza rufipogon Griff., are now threatened with the disturbance of their natural habitats by various human activities. To obtain basic information on genetic erosion or loss of genetic diversity in wild rice, we investigated how environmental changes of habitat affected the genetic structure of its natural population at a study site in the central plain of Thailand. During 10 years from 1985 to 1994, the wild-rice population at this site was seriously destroyed and fragmented. Using two sets of seed sample collected in 1985 and 1994 from the same population, allozyme variability at 17 loci of 11 enzymes were examined. Isozyme genotypes of mother plants of seed samples were estimated by the segregation in each progeny, and we calculated genetic parameters for the population. Gene diversity severely decreased in the 1994 sample compared with the 1985 sample. It is supposed that declining and fragmentation of the wild rice population, which happened during the 10 years, caused loss of genetic variability and forced the habitually outbreeding plants to inbreed, accelerating a reduction in gene variability. Pgi1-1 allele which was common in Indica rice cultivars of this region was found in the wild rice plants growing at the side of rice fields. Probably, introgression has occurred between wild and cultivated rice plants, and consequently the intrinsic nature of wild rice was gradually blurred by cultivar genes. We must realize that the genetic erosion of wild rice is rapidly proceeding and that an action for their conservation in natural environment, so called in situ conservation, is urgently needed.