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.     

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.     

From genetic diversity and structure to conservation: Genetic signature of recent population declines in three mouse lemur species (Microcebus spp.)

The exceptional biodiversity of Madagascar is threatened by anthropogenic landscape changes that took place during the 2000 years of human colonization. This study focuses on the influence of geographic distance and forest fragmentation on genetic diversity and population differentiation of three rare, nocturnal, arboreal lemur species in northwestern Madagascar. Historic declines in population sizes as a consequence of forest fragmentation are quantified and dated. Eighteen sites were visited, and a total of 205 Microcebus ravelobensis, 45 M. bongolavensis and 78 M. danfossi were genotyped with eight microsatellite loci. Genetic differentiation among the sites, as measured by F_ST, ranged from 0.01 to 0.19. These values were significant in almost all cases and indicated genetic structure in the samples. Isolation-by-distance was detected in one species and a STRUCTURE analysis indicated that fragmentation further promoted genetic differentiation. Bayesian methods revealed that populations from all three species underwent a major demographic collapse of around two orders of magnitude. This decrease probably began after the arrival of humans, most likely within the last 500 years. This result suggests that anthropogenic changes may have been limited during the first 1500 years of human colonization in all three ranges. Two of the study species (M. danfossi, M. bongolavensis) lack effectively protected areas in their ranges. Consequently, quick conservation actions are now needed in order to secure the remaining genetic diversity of these species.     

Reproductive biology and conservation implications of three endangered snapdragon species (Antirrhinum, Plantaginaceae)

About 32% of Antirrhinum species are considered to be endangered; however, no field studies have focused on their reproductive biology. In this work, several aspects of the reproductive biology (flowering phenology, floral biology, breeding system) and potential limits on seed quantity and quality (pollen limitation, inbreeding depression) were studied in natural populations of three endangered species of the genus (Antirrhinum charidemi, Antirrhinum subbaeticum, Antirrhinum valentinum). Results disclose that all three species need insect visitors for seed production since fruit set after autonomous self-pollination was lower than under hand cross-pollination. A. charidemi and A. valentinum were mainly self-incompatible, whereas A. subbaeticum was self-compatible but herkogamous. Supplementary pollination in open-pollinated flowers only increased fruit set and seed set relative to controls in a given population of A. valentinum. Preliminary data on inbreeding depression at early life-cycle stages of the self-compatible A. subbaeticum revealed that the cumulative level was low. Despite the three species being closely related and sharing many ecological characteristics, they show different mating systems, and different factors limit seed quantity and quality. Thus, caution should be taken when making a common conservation plan for a group of closely related taxa.     

What genetics tell us about the conservation of the critically endangered Balearic Shearwater?

The Balearic shearwater Puffinus mauretanicus is one of the most critically endangered seabirds in the world. The species is endemic to the Balearic archipelago, and conservation concerns are the low number of breeding pairs, the low adult survival, and the possible hybridization with a sibling species, the morphologically smaller Yelkouan shearwater (P. yelkouan). We sampled almost the entire breeding range of the species and analyzed the genetic variation at two mitochondrial DNA regions. No genetic evidence of population decline was found. Despite the observed philopatry, we detected a weak population structure mainly due to connectivity among colonies higher than expected, but also to a Pleistocene demographic expansion. Some colonies showed a high imbalance between immigration and emigration rates, suggesting spatial heterogeneity in patch quality. Genetic evidence of maternal introgression from the sibling species was reinforced, but almost only in a peripheral colony and not followed, at least to date, by the spread of the introgressed mtDNA lineages. Morphometric differences were not correlated with mtDNA haplotypes and introgression is probably due to a secondary contact between the two species several generations ago. Overall, results suggested that the very recent demographic decline in this critically endangered species has not yet decreased its genetic variability, and connectivity found among most colonies should help to reduce species extinction risk. Spreading of introgression should be monitored, but the species is not jeopardized at the moment by genetic factors and the major conservation actions should concentrate at enhancing adult survival.     

Island differences in population size structure and catch per unit effort and their conservation implications for Komodo dragons

Species inhabiting archipelagos are often characterised by high levels of interpopulation divergence (e.g. size related traits). This divergence may, in turn, influence their life-history. To facilitate better management and conservation of the Komodo dragon (Varanus komodoensis), an island endemic, we identified demographic differences between two island populations in Komodo National Park, Indonesia. Comparison of data collected from dragon populations inhabiting Rinca Island and the much smaller Gili Motang Island indicated that between 1994 and 2004, the Komodo dragon population on Gili Motang significantly decreased its: (1) mean body mass, (2) body condition and (3) relative abundance. These results suggest that the numerically small Gili Motang population was oscillating downwards; in contrast, the Rinca Island population had been relatively stable. More importantly these results emphasize the necessity for managers of this priority conservation species to understand further the inherent functional differences among dragon populations to develop island specific management units. Current management practices (e.g. monitoring) instigated by Komodo National Park management ignore small island dragon populations and thus run the risk of being unable to detect adverse effects for populations that are potentially most prone to decline.     

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.     

Cryptic genetic bottlenecks during restoration of an endangered tropical conifer

Forest restoration programmes aim to use material for re-planting that is genetically diverse and not inbred. However, restricted seed sampling, high variance in reproductive output, and the production of inbred seeds that survive in the nursery but not in the wild can lead to forest restoration stock being genetically compromised. The aim of this study was to evaluate whether the reproductive biology of the New Caledonian endemic conifer Araucaria nemorosa makes it susceptible to these genetic problems and to assess whether there is evidence for genetic bottlenecks and elevated inbreeding in nursery stock compared to seedlings and adults from wild source populations. Reproductive output was low with high variance among trees (only 14% of adult trees surveyed produced mature cones, >50% of examined cones had <10 viable seeds). Evidence for an extreme genetic bottleneck was detected in a nursery population established from cones collected from adult trees. A second nursery population established with seed collected from the forest floor showed no evidence of a genetic bottleneck, but was inbred compared to its wild source population. In light of these results, we do not recommend collecting cones directly from A. nemorosa as an efficient means of establishing genetically diverse stock for restoration programmes. Collecting seed from the forest floor is likely to be more effective, but the planting stock may contain a high proportion of inbred individuals. Collecting established wild seedlings already subjected to natural selection is suggested as an alternative method of maximising the diversity captured, whilst minimising sampling effort and proportion of inbred individuals.     

Population genetic structure and the conservation of isolated populations of Acacia raddiana in the Negev Desert

There is much concern over the high mortality of many populations of Acacia raddiana, a keystone tree species in the Negev desert of Israel. We used random amplified polymorphic DNA (RAPD) to assess patterns of genetic variation within and among 12 populations of A. raddiana from the Arava (Syrian-African Rift) valley and western Negev. A high level of genetic polymorphism was recorded within populations. An analysis of molecular variance (AMOVA) showed that about 59.4% of total genetic variance occurred among populations, which is considerably greater population differentiation than that recorded for other outbreeding species. Cluster and principal coordinates analyses and AMOVA indicate that the western Negev and Arava valley populations are highly differentiated. We suggest that there may have been two invasions of A. raddiana into Israel: one across the northern Sinai/Gaza Strip area into the western Negev, with some plants reaching the Dead Sea and a second invasion across the southern part of the Sinai peninsula, or even from Saudi Arabia, up to the Arava valley. From the conservation point of view, each population should be conserved separately because they are genetically highly differentiated and loss of any one population would lead to a dramatic loss of genetic variation. The mixing of genetically distinct populations may give rise to outbreeding depression (particularly because of GXE interactions). An obvious first step to the maintenance of this species' genetic diversity is the separate management of the western Negev and Arava valley populations because of their different evolutionary histories.     

Seasonal movements in the Sumatran orangutan (Pongo pygmaeus abelii) and consequences for conservation

Preservation of orangutan habitat can only be effective when their range requirements are taken into consideration. Here, the effect of the variation in fruit supply at different altitudes on the seasonal movements of orangutans is investigated. Transects were surveyed every 2 months from 1998 to 2000, at different altitudinal zones in the larger Ketambe area, Indonesia. The density of orangutan and sources of ripe fruit with soft pulp were determined along these transects. Both orangutan density and the availability of fruit varied considerably throughout the survey period at different altitudinal zones. The peaks in fruit production at different altitudes occurred in successive periods. For each altitudinal zone a significant correlation was found between orangutan density and the number of fruit sources, which shows that orangutans follow the production of fruit between these altitudinal zones within the boundaries of their home range. Orangutans with core areas in intermediate and highland areas depend seasonally on fruit production of lowland areas, and vice versa. Thus, conservation of large tracts of primary forest with a range of different altitudes is necessary to preserve the remaining orangutan population.     

Conserving marginal populations of the food plant (Impatiens noli-tangere) of an endangered moth (Eustroma reticulatum) in a changing climate

Impatiens noli-tangere is scarce in the UK and probably only native to the Lake District and Wales. It is the sole food plant for the endangered moth Eustroma reticulatum. Significant annual fluctuations in the size of I. noli-tangere populations endanger the continued presence of E. reticulatum in the UK. In this study, variation in population size was monitored across native populations of I. noli-tangere in the English Lake District and Wales. In 1998, there was a crash in the population size of all metapopulations in the Lake District but not of those found in Wales. A molecular survey of the genetic affinities of samples in 1999 from both regions and a reference population from Switzerland was performed using AFLP and ISSR analyses. The consensus UPGMA dendrogram and a PCO scatter plot revealed clear differentiation between the populations of I. noli-tangere in Wales and those in the Lake District. Most of the genetic variation in the UK (H_T=0.064) was partitioned between (G_ST=0.455) rather than within (H_S=0.034) regions, inferring little gene flow occurs between regions. There was similar bias towards differentiation between metapopulations in Wales, again consistent with low levels of interpopulation gene flow. This contrasts with far lower levels of differentiation in the Lake District which suggests modest rates of gene flow may occur between populations. It is concluded that in the event of local extinction of sites or populations, reintroductions should be restricted to samples collected from the same region. We then surveyed climatic variables to identify those most likely to cause local extinctions. Climatic correlates of population size were sought from two Lake District metapopulations situated close to a meteorological station. A combination of three climatic variables common to both sites explained 81-84% of the variation in plant number between 1990 and 2001. Projected trends for these climatic variables were used in a Monte Carlo simulation which suggested an increased risk of I. noli-tangere population crashes by 2050 at Coniston Water, but not at Derwentwater. Implications of these findings for practical conservation strategies are explored.     

Ecology of the native and introduced crayfishes Austropotamobius pallipes and Procambarus clarkii in southern Spain and implications for conservation of the native species

The former and present distribution of white clawed crayfish (Austropotamobius pallipes) in the province of Granada (southern Spain) is studied. Before 1980 it was widely distributed but at present only 16 populations exist. The decline is related to the presence of the freshwater red-swamp crayfish (Procambarus clarkii), an American species, vector of the aphanomycosis disease, introduced to the Iberian Peninsula in 1974 and now widely distributed in the watercourses and marshes of southern Spain. To establish an appropriate conservation policy for A. pallipes at its southernmost distribution limit, we studied watercourses from two river basins, Genil and Guadiana Menor, (tributaries of the Guadalquivir River). P. clarkii inhabits the medium to lower reaches of these two river basins (with its upper limit at 820 m a.s.l.). The distribution of this species was best explained by the effect of three of the 12 analyzed variables: altitude, water-current and minimum winter temperatures. From our results, the repopulation of the native crayfish is almost impossible in those reaches inhabited by P. clarkii. However, based on the habitat selection study, it is clear that upper reaches are unsuitable for the red-swamp crayfish, where the native white-clawed crayfish may have greater survival possibilities, and these sites can be used for future restocking projects.     

Assessing habitat use of the endangered marine mollusc Patella ferruginea (Gastropoda, Patellidae) in northern Africa: preliminary results and implications for conservation

The limpet Patella ferruginea Gmelin, 1791, endemic to the Mediterranean, is the most endangered marine species on the list of the European Council Directive 92/43/EEC on the Conservation of Natural Habitat of Wild Fauna and Flora (1992). Its Mediterranean range has progressively contracted to a few restricted areas and the species is now threatened with extinction. Seventeen stations were sampled along the littoral zone on the coast of Ceuta, North Africa, for quantifying environmental factors (water movement, siltation and suspended solids), and 70 transects were selected for P. ferruginea sampling. The study revealed the presence of a well-established population of P. ferruginea with values of density and size of 0.67 individuals m⁻¹ ±0.96 and 48.94 mm±11.61 respectively (mean±standard error of mean). The biggest densities of P. ferruginea were found inside the harbour of Ceuta. The limpets were more abundant on artificial harbour stones than on natural rocky shores, and the areas under the highest human pressure were characterised by the lowest densities and the smallest specimens probably due mainly to the predation for food and fishing. Human pressure is probably the main contributing factor to the currently endangered status of P. ferruginea. Programmes of environmental education to avoid its collection for fishing, food or for fun as decorative objects should be conducted, and further experimental studies dealing with the reproductive biology of this species are needed to properly assess the future programmes of conservation.     

Genetic structure, population dynamics, and conservation of Black caiman (Melanosuchus niger)

Microsatellite DNA polymorphisms were screened in seven populations of the largest Neotropical predator, the Black caiman Melanosuchus niger (n = 169), originating from Brazil, French Guiana and Ecuador. Eight loci were used, for a total of 62 alleles. The Ecuadorian population had the lowest number of alleles, heterozygosity and gene diversity; populations of the Guianas region exhibited intermediate diversities; highest values were recorded in the two populations of the Amazon and Rio Negro. During the last century Melanosuchus populations have been reduced to 1-10% of their initial levels because of hunting pressure, but no strong loss of genetic diversity was observed. Both the inter-locus g-test and the Pk distribution suggested no recent important recovery and/or expansion of current populations. On a global scale, the inter-population variation of alleles indicated strong differentiation (F_ST = 0.137).Populations were significantly isolated from each other, with rather limited gene flow; however, these gene flow levels are sufficiently high for recolonization processes to effectively act at regional scales. In French Guiana, genetic structuring is observed between populations of two geographically close but ecologically distinct habitats, an estuary and a swamp. Similar divergence is observed in Brazil between geographically proximate “black water” and “white water” populations. As a consequence, the conservation strategy of the Black caiman should include adequate ecosystem management, with strong attention to preservation of habitat integrity. Distribution of genetic diversity suggests that current populations originated from the central Amazonian region. Dispersal of the species may thus have been deeply influenced by major climatic changes during the Holocene/Pleistocene period, when the Amazonian hydrographic networks were altered. Major ecological changes such as glaciations, marine transgressions and a hypothesized presence of an Amazonian Lake could have resulted in extension of Black caiman habitats followed by isolation.     

Ex situ genetic conservation of endangered Vatica guangxiensis (Dipterocarpaceae) in China

RAPD polymorphisms were applied to check the efficiency of ex situ genetic conservation of endangered Vatica guangxiensis X. L. Mo. endemic to southwestern China. Low level of genetic variation was revealed in three remaining natural populations. Twenty random primers, each with 10 base pairs, generated 231 bands with 53.68% being polymorphic, and with an average of 32.46% being polymorphic in each natural population. Strong population differentiation was revealed by AMOVA (analysis of molecular variance) and Gst value was 0.3764. The population ML ex situ conserved in the Xishuangbanna Tropical Botanical Garden contained an intermediate genetic variation compared with natural populations, with 30.74% bands being polymorphic. Of the total 231 bands generated in V. guangxiensis, 204 bands were also detected in population ML, indicating that 88.31% of the total genetic variations of this species were conserved in ex situ population. If only the alleles with moderate to high frequency (P>0.05) were considered, 204 out of 209 bands (97.61%) occurred in ex situ population ML. RAPD analysis also detected one exclusive band in natural population NS, and five in natural population NP, three of these exclusive bands were generated in every samples of natural population (NP), and other three had moderate to high frequencies. While none of these exclusive bands were detected in ex situ conserved population ML. Our conclusions are that the ex situ conserved population ML contains representative genetic variation to maintain long-term survival and evolutionary process of V. guangxiensis, and that more extensive ex situ sampling in natural population NS and NP is needed to conserve more exclusive alleles in ex situ population. The tropical area in the Botanical Garden would play a more important role in the ex situ conservation of rare and endangered plants.     

Population isolation exacerbates conservation genetic concerns in the endangered Amargosa vole, Microtus californicus scirpensis

The ability of a taxon to maintain adaptive flexibility in a stochastic environment is a function of the genetic diversity within the population. In small, fragmented populations, genetic variation can become depleted more quickly than in larger, more contiguous populations. Characterizing the patterns of genetic variation and differentiation associated with these processes is an important step in establishing conservation priorities. The Amargosa vole, Microtus californicus scirpensis, is an endangered rodent persisting in the small, fragmented marsh complex surrounding the Amargosa River near Death Valley, California. This naturally patchy system has existed since the end of the Pleistocene (approximately 10,000 y.b.p.), however, fragmentation has been exacerbated by recent anthropogenic changes. For this study, I used five nuclear microsatellite loci and the cytochrome-b region of the mitochondrial genome to quantify levels of genetic variation, population substructure, and patterns of gene flow in M.c. scirpensis. These data were compared to a broadly distributed subspecies, Microtus californicus sanctidiegi. Overall levels of nuclear genetic variation were significantly lower in M.c. scirpensis, whether measured in terms of diversity or heterozygosity, compared to more broadly distributed conspecifics. Moreover, only two haplotypes were recovered from the mitochondrial data with over 90% of the observed haplotypes being identical. Despite low genetic diversity, significant genetic subdivision among M.c. scirpensis populations was detected using both pairwise F_ST and Bayesian clustering methods. Furthermore, isolation by distance analyses reveal that an important landscape feature, ephemeral tributaries, is critical for dispersal among population clusters. Recommendations for conservation management are presented.     

Influence of forest fragmentation on an endangered large-bodied lemur in northwestern Madagascar

Madagascar’s diverse and mostly endemic fauna and flora suffer from recent landscape changes that are primarily caused by high levels of human activities. The loss and fragmentation of forest habitats are well known consequences of human activities. In this study, we investigate the effects of forest fragmentation on presence, abundance and genetic diversity in a larger-bodied lemur species, Lepilemur edwardsi, in northwestern Madagascar. In addition, we characterized the genetic differentiation among populations and demographic changes. We found L. edwardsi at only 13 (76.5%) of 17 visited sites, 11 of which were situated in the Ankarafantsika National Park (ANP). We captured between two and 17 individuals per site. We sequenced the mtDNA d-loop of all samples and genotyped 14 microsatellite loci in two exemplary populations for demographic analyses. A negative influence on forest fragmentation could be detected, since the fragments had a lower genetic diversity than sites in the ANP. Genetic differentiation between populations ranged from low to high but was almost always significant. A typical pattern of isolation-by-distance could not be detected and the data could rather be interpreted as results of random genetic drift. The data furthermore revealed signals of a demographic collapse of about two orders of magnitude in the two exemplary sites. This decline probably started during the last few hundred years of intensified human disturbances and population growth. Given the results of this study, urgent conservation actions are needed and should concentrate on an effective protection of the few remaining populations in order to ensure the long-term survival of L. edwardsi.     

Assessing limitations on population growth in two critically endangered Acacia taxa

Flowering phenology, soil seedbank and the impact of fire, weeds and grazing were investigated in two rare Acacia taxa restricted to the agricultural district north of Perth Western Australia. Acacia aprica is known from six extant populations and one extinct population. Five of the extant populations are restricted to linear road reserves with the sixth population located in a small native vegetation remnant. Acacia cochlocarpa ssp. cochlocarpa is known from a single population also situated on a road verge. Size class structure, levels of canopy death and an absence of juveniles indicated that all populations are in decline. Flowering intensity and success varied between populations and years in A. aprica and between years in A. cochlocarpa ssp. cochlocarpa. Seed bank analysis indicated that seeds were patchily distributed while experimental fires demonstrated that such events could break seed dormancy and promote germination. Both taxa have similar germination physiologies and showed increased germination after seeds were exposed to heat. Following emergence, however, competition with annual weeds had a negative impact on seedling growth and survival in both taxa. While vertebrate grazing had some influence, weeds were the major inhibitory influence on recruitment. Reduced fire frequencies since fragmentation may be responsible for population decline in both taxa but other site specific factors such as weeds and grazing may affect the establishment of seedlings following fire.     

Low levels of genetic variation among southern peripheral populations of the threatened herb, Leontice microrhyncha (Berberidaceae) in Korea

Levels of genetic variation and intrapopulation genetic structures of Leontice microrhyncha S. Moore (Berberidaceae) were assessed for six populations in South Korea, representing the southern most range of a species found in Northeast China and the Korean peninsula. Detected genetic diversity (H_es) was very low (0.024) and F_IS values showed large heterozygote deficiencies. The small percentage of polymorphic loci and numbers of alleles per locus suggest that L. microrhyncha has a history of severe or long-lasting population bottlenecks that have eroded genetic diversity. This study suggests that the Korean population appears to consist of two historically isolated and independently evolving populations. It seems likely that these groups have been isolated and unstable for a significant period of time. However, the effects of recent habitat fragmentation on the historically disjunct and fragmented population system found in L. microrhyncha were not those predicted from the lack of significant relationships between population-level patterns of genetic variation and population sizes. Most non-unique genotypes were shared by most individuals and the lower level of diversity, high levels of inbreeding and population differentiation as well as high rate of seed production indicated that this species is autogamous and self-compatible and probably largely selfing. Therefore, to preserve extant genetic variation, all populations must be protected across the small geographic range of the species to retain both allelic and genotypic diversity.     

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.