Conservation genetics of the endangered terrestrial orchid Liparis japonica in Northeast China based on AFLP markers

Levels of genetic diversity and population genetic structure of the rare, endangered terrestrial orchid Liparis japonica were examined for eight natural populations (n = 185) in Northeast China using six AFLP primer pairs, where this species has experienced severe habitat loss and fragmentation. Based on 406 DNA bands, a high level of genetic diversity was found at the species level with the PPB of 85.47 %, while the genetic diversity at the population level was low (PPB = 47.48 %). A significantly high degree of population differentiation was found with 42.69 % variation existed among populations as measured by AMOVA, indicating potential restricted gene flow. The genetic distances between populations were independent of the corresponding geographic distances, and the genetic relationship of individuals had no significant correlation with their spatial distribution. The restricted gene flow might be impacted by reduced population size, habitat destruction and fragmentation. The results in this study suggested that habitat protection and keeping a stable environment are critical for the conservation of L. japonica species.     

AFLP and ISSR analysis reveals high genetic variation and inter-population differentiation in fragmented populations of the endangered Litsea szemaois (Lauraceae) from Southwest China

Litsea szemaois (Lauraceae) is an endemic and endangered species from the tropical rain forests of Xishuangbanna, southern Yunnan, SW China, but habitat fragmentation, especially exacerbated by rubber planting, has caused a decline in population size of the species. AFLP and ISSR were used to investigate the genetic diversity and population structure of eight populations from across its known distribution. Three AFLP and ten ISSR primer combinations produced a total of 203 and 77 unambiguous and repeatable bands respectively, of which 164 (80.8%) and 67 (87.0%) were polymorphic for the two markers. These two markers showed that Litsea szemaois exhibits comparatively high genetic diversity at species level (heterozygosity (hs) = 0.2109) relative to some other Lauraceae. Most of the genetic variation was partitioned within populations, but genetic differentiation between populations was significant and relatively high (Φ _st = 0.2420, θ^B = 0.1986) compared with other tropical plants. The genetic characteristics of L. szemaois may be related to its outbreeding system, insect pollination and fragmented distribution. Because L. szemaois is dioecious and slow to mature, ex situ conservation across its genetic diversity is unlikely to succeed, although seedlings grow well under cultivation. Thus, in situ conservation is very important for this endangered species, especially as only 133 adult individuals are known in the wild. In particular, the Nabanhe and Mandian populations should be given a high conservation priority due to their higher genetic diversity, larger population size and better field condition, but wider sampling is required across all populations to determine additional areas with significant genetic conservation value.     

Genetic diversity and structure of Lilium pumilum DC. in southeast of Qinghai–Tibet plateau

Lilium pumilum DC. is a valuable species not only for its showy flowers but also for its edible and medicinal values. As one of the distribution areas of L. pumilum, Qinghai–Tibet plateau has unique environmental features which have high impact on the evolution of the species. No population genetic studies have been done for L. pumilum so far. To provide the first reference data for evolutionary study and understanding the influence of eco-geographic factors on the distribution of genetic variation in L. pumilum, interspecific simple sequence repeat markers were used to investigate genetic diversity and population structure of 28 populations sampled from southeast of Qinghai–Tibet plateau. Fifteen selected primers generated a total of 147 polymorphic bands. The genetic diversity was low within populations (average He = 0.173), but higher at the species level (He = 0.392). A clear population structure and high level of genetic differentiation (F _ST = 0.518) were detected by unweighted pair group method for arithmetic averages, principle coordinate analysis and Bayesian clustering. All clustering approaches supported a division of the 28 populations into 4 major groups for which analysis of molecular variance confirmed a significant variation among groups (34.3 %). These population genetic parameters suggest limited gene flow among populations and evidence for isolation by distance (r = 0.272, P < 0.0001) was found in this study. Altitude, AMT and AMP explained 9.5, 11.5 and 14.0 % of the total variance among populations indicating that eco-geographic factors have a significant effect. Considering the low within-population genetic diversity, high differentiation among populations and the increasing anthropogenic pressure on the species, in situ conservation measures were recommended to preserve L. pumilum in Qinghai–Tibet plateau.     

Phylogeography and conservation of the bull shark (Carcharhinus leucas) inferred from mitochondrial and microsatellite DNA

The bull shark (Carcharhinus leucas) is a widely distributed, large coastal shark species known to travel long distances. These characteristics, coupled with the species?? long life span and late age of maturity, would lead one to predict significant global genetic exchange among bull shark populations. By contrast, data show localized depletion in some areas of large coastal shark fisheries, indicating some geographic isolation may exist. We examined genetic variation in the control region of mitochondrial DNA and at five nuclear microsatellite loci in bull sharks sampled from the western Atlantic to investigate the degree of population subdivision. The average per sample haplotype and nucleotide diversity in the mtDNA (0.51 ± 0.26 and 0.12% ± 0.12, respectively) and expected heterozygosity (0.84) in the microsatellite loci contrast sharply in having lower and higher values (respectively) relative to many other shark species. Significant structure exists between the Brazilian and all northern populations at the mtDNA control region (pairwise ??_ST > 0.8, P < 0.001), but not at the nuclear microsatellite loci. Adjacent northern populations show weak to no genetic differentiation for both markers. These results are congruent with restricted maternal gene flow between populations caused by female site fidelity to nursery areas. We estimate the current effective population size to be around 160,000 and 221,000 individuals for the southern and northern Atlantic populations, respectively. The philopatric habits and the relatively low levels of mtDNA genetic diversity observed in bull sharks must be considered in the conservation of this species. Our results indicate that effective bull shark management strategies will require local, regional, and international attention and cooperation.     

Genetic diversity of the endangered Chinese endemic herb Saruma henryi Oliv. (Aristolochiaceae) and its implications for conservation

Saruma henryi Oliv., the only representative of the monotypic genus Saruma Oliv. (Aristolochiaceae), is an endangered perennial herb endemic to China. It is a phylogenetically, ecologically, and medicinally important species. In the present study, inter-simple sequence repeat (ISSR) markers were employed to investigate the genetic diversity and differentiation of 14 populations. A total of 16 selected primers yielded 175 bright and discernible bands, with an average of 10.94 per primer. POPGENE analysis showed that the genetic diversity was quite low at the population level (h = 0.0447–0.1243; I = 0.0642–0.1853; PPB = 10.29–36.57%), but pretty high at the species level (h = 0.2603; I = 0.3857; PPB = 73.71%). The hierarchical analysis of molecular variance (AMOVA) revealed a high level of genetic differentiation among populations (67.18% of total variance components, P < 0.001), in line with the gene differentiation coefficient (G _ST = 0.6903) and the limited among-population gene flow (N _m = 0.2243). Both Principal Coordinates Analysis (PCoA) and UPGMA cluster analysis supported the grouping of all 14 populations into three geographic groups, among which there occurred a moderate level of genetic differentiation (33.18% of total variance components, P < 0.001) as shown by AMOVA analysis. In addition, Mantel test revealed a significant correlation between genetic and geographic distances among populations (r = 0.7792, P = 0.001), indicating the role of geographic isolation in shaping its present population genetic structure. The present levels and patterns of genetic diversity of S. henryi were assumed to result largely from its breeding system, geographic isolation, clonal growth, its unique biological traits and evolutionary history. The high genetic differentiation among populations implies that the conservation efforts should aim to preserve all the extant populations of this endangered herb.     

Genetic diversity and population structure of the serpentine endemic Ni hyperaccumulator Alyssum lesbiacum

Alyssum lesbiacum is a well-established Ni hyperaccumulator, endemic to serpentine soils of Lesbos Island (Greece). A total of 95 individuals were collected from 17 plots encompassing the only four large populations of this species. ISSR (Inter-simple sequence repeat) markers were used to assess genetic diversity and population structure of A. lesbiacum to provide initial guidance for the development of successful management and conservation measures. A total of ten primers produced 82 bands, 96.34 % being polymorphic. The largest part of total diversity was found within populations, rather than among them. AMOVA analysis partitioned the largest part of diversity within plots (66 %), while 15 % contained among plots within populations and 19 % among populations. Principal coordinates analysis along with dendrogram based on genetic distances among populations showed a high degree of genetic differentiation of the isolated population Loutra. At a smaller scale, distance was not the most significant factor influencing the patterns of genetic diversity, but topography, ecosystem types and connectivity through streams. According to our results, conservation efforts should be organized at the level of watersheds and ecosystem types, considering them as management units. For ex situ conservation and restoration, seed samplings should be representative of the different habitats and watersheds as well as the patches of large and small populations while keeping the east population of Loutra separate from the other three central populations, to avoid any loss of genetic diversity and to preserve the character of the local adapted populations.     

Estimation of genetic variability and population structure in Sapindus trifoliatus L., using DNA fingerprinting methods

Genetic variability and population structure of Sapindus trifoliatus L. (Sapindaceae), collected from Gujarat, Karnataka and Uttar Pradesh states were estimated using three DNA fingerprinting methods viz., random amplified polymorphic DNA (RAPD), directed amplification of minisatellite DNA (DAMD) and inter-simple sequence repeats (ISSR). The cumulative data analysis carried out for all three markers showed 69.42 % polymorphism. The intra-population genetic diversity analysis revealed the highest values of Nei’s genetic diversity (0.16), Shannon information index (0.24) and polymorphic loci (43.99 %) among Bhavnagar (BH) population, whereas lowest values were found in Junagarh (JU) population. The maximum inter-population average genetic distance (0.20) was between Allahabad (AL) and JU populations. Analysis of molecular variance (AMOVA) showed highest percentage of variation among individuals of populations (56 %) followed by 25 % among populations and 19 % among regions. Principal coordinate analysis and UPGMA dendrogram revealed that genetic diversity was in congruence with the geographical diversity. The data strongly suggest that low genetic flow, geographic isolation and to some extent genetic drift are the major factors responsible for high genetic differentiation. Preservation of genetic diversity of S. trifoliatus is important, both to promote adaptability of the populations to changing environment as well as to preserve a large gene pool for future genetic improvement. The present study using RAPD, DAMD and ISSR profiles of S. trifoliatus provide the means of rapid characterization of accessions within the populations, and thus enable the selection of appropriate accessions for further utilization in conservation and prospection programs of this important plant genetic resource.     

Genetic diversity and differentiation in the critically endangered orchid (Amitostigma hemipilioides): implications for conservation

Amitostigma hemipilioides is an endangered terrestrial orchid endemic to China. In the Southwestern China, the species is found restricted growing in the karst limestone or rock in the edge of farmland, moist hillsides or river. In the present study, the genetic diversity and differentiation was estimated within and among habitats, populations and groups of this species by ISSR markers. Using 13 polymorphic primers, an intermediate level of genetic diversity was found at the species level and population level with the percentage of polymorphic bands (P) of 64.7 and 50.9 %, Shannon index of diversity (I) of 0.3873 and 0.2949, respectively. The analysis of molecular variance (AMOVA) showed that the high level of population differentiation was presented with 45.63 % relative to the total genetic variation residing among eight populations. It was noteworthy that as much as 69.17 % of the total diversity was most likely attributed to the difference among the populations in fragmentation habitat conditions, while 16.32 % of the total diversity could be attributed to the difference among the populations in stable and favorable habitat conditions. The in situ conservation is a top strategy, thus the mycorrhizal fungi and pollinators are protected by protecting habitat and avoiding fragmentation which is helpful to the cycle of this endangered orchid species and recovery of its wild populations. In addition, to maintaining the germplasm bank of this species, the ex situ conservation by habitat simulation and reintroduction has been considered in the living collection at Kaili University ecological garden and Yuntai Mountain scenic spot using the plants collected in this study.     

Negligence in the Atlantic forest,northern Brazil: a case study of an endangered orchid

Currently, many Brazilian orchids are threatened with extinction resulting from habitat loss and intense harvesting pressure stemming from their value as ornamental plants. Therefore, the genetic diversity in remaining populations is fundamental to the survival of these species in natural environments. In order to inform conservation strategies, this study evaluated the genetic diversity and structure of Cattleya granulosa populations. The sample consisted of 151 individuals from 12 populations in the Atlantic Forest, northeastern Brazil, evaluated using 91 ISSR markers. Genetic variability was assessed through molecular variance, diversity indexes, clusters of genotypes through Bayesian analysis, and tests for genetic bottlenecks. From all polymorphic loci, genetic diversity (H_E) varied between 0.210 and 0.321 and the Shannon index ranged from 0.323 and 0.472. Significant genetic differentiation between populations (Φ_ST = 0.391; P < 0.0001) resulted in the division of the populations into five groups based on the log-likelihood Bayesian analysis. We found significant positive correlation between geographical and genetic distances between populations (r = 0.794; P = 0.017), indicating isolation by distance. Patterns of allelic diversity within populations suggest the occurrence of bottlenecks in most C. granulosa populations (n = 8). Therefore, in order to maintain the genetic diversity of the species, the conservation of spatially distant groups is necessary.     

A comparison of the population genetic structure of parasitic Viscum album from two landscapes differing in degree of fragmentation

Parasite populations do not necessarily conform to expected patterns of genetic diversity and structure. Parasitic plants may be more vulnerable to the negative consequences of landscape fragmentation because of their specialized life history strategies and dependence on host plants, which are themselves susceptible to genetic erosion and reduced fitness following habitat change. We used AFLP genetic markers to investigate the effects of habitat fragmentation on genetic diversity and structure within and among populations of hemiparasitic Viscum album. Comparing populations from two landscapes differing in the amount of forest fragmentation allowed us to directly quantify habitat fragmentation effects. Populations from both landscapes exhibited significant isolation-by-distance and sex ratios biased towards females. The less severely fragmented landscape had larger and less isolated populations, resulting in lower levels of population genetic structure (F _ST = 0.05 vs. 0.09) and inbreeding (F _IS = 0.13 vs. 0.27). Genetic differentiation between host-tree subpopulations was also higher in the more fragmented landscape. We found no significant differences in within-population gene diversity, percentage of polymorphic loci, or molecular variance between the two regions, nor did we find relationships between genetic diversity measures and germination success. Our results indicate that increasing habitat fragmentation negatively affects population genetic structure and levels of inbreeding in V. album, with the degree of isolation among populations exerting a stronger influence than forest patch size.     

Geographic variation in relict populations: genetics and phenotype of bush-cricket Pholidoptera frivaldskyi (Orthoptera) in Carpathians

A decreasing population size is often causing species extinction, however, relict species persisting in small-sized populations counter this. We analysed spatial genetic variation and past changes in population size at the maternally-inherited mitochondrial DNA level to clarify the origin of all recently known isolated populations of Pholidoptera frivaldskyi occurring in the range of Carpathian Mountains. Along with that we analysed also morphological variation as some phenotypic traits can retain useful information on population genetic structure. We found a relatively low genetic diversity within isolated populations as 778 bp COI gene sequences revealed only 13 unique haplotypes (n = 173 individuals from 10 populations). The spatial analysis of molecular variance identified three geographically homogenous genetic clusters (one in Slovakia and two in Romania) with a high level of differentiation among them, suggesting restricted gene flow, whilst Bayesian skyline simulation reconstructed a negative demographic change through evolutionary time. Inferred genetic pattern clearly coincides with differences in males’ colour phenotype as the extent of pigmentation on the lateral pronotum varied significantly among genetic lineages. We suggest that geographical variation in the species populations has relict-like character and their isolated occurrence is not a result of recent introduction events. Identification of ‘evolutionary units’ may help in the conservation and management of this rare insect species.     

Contrasting coarse and fine scale genetic structure among isolated relic populations of Kmeria septentrionalis

Trees of the Magnoliaceae family are of scientific, cultural and socio-economic importance. Kmeria septentrionalis Dandy (Magnoliaceae) is a dioecious tree, found in small, isolated, relic populations in Southern China, and is subject to extensive protection due to its rarity and high economic values. To improve conservation outcomes and in particular, germplasm collection guidelines, information on spatial genetic structure of the species is required. In this study, we investigated the spatial genetic structure and genetic diversity of 161 individuals of K. septentrionalis collected from five natural populations using AFLP molecular markers. Within-population genetic variation was measured, with percentage of polymorphic bands (PPB) ranged from 63% to 87%, while H _S (genetic diversity within population) varied from 0.185 to 0.244 with a mean of 0.215 ± 0.025. Significant genetic differentiations were revealed between pairwise populations, indicating each population existing as an independent evolutionarily significant unit. Mantel test results showed no pattern of isolation-by-distance among populations separated by large distance. Fine scale spatial patterns of genetic variation suggested significant effects of isolation-by-distance within population at distances of 22 m. The results of contrasting genetic structure at coarse and fine scale in K. septentrionalis may indicate restricted pollen flow and seed dispersal at fine scales, and separated evolution in isolated populations over long period of time at coarser scales. Finally, we make several suggestions for improved management practices that may assist in the conservation of this species.     

Assessing genetic diversity for the USA endemic carnivorous plant <Emphasis Type="Italic">Pinguicula ionantha</Emphasis> R.K. Godfrey (Lentibulariaceae)

Understanding patterns of genetic diversity and population structure for rare, narrowly endemic plant species, such as Pinguicula ionantha (Godfrey’s butterwort; Lentibulariaceae), informs conservation goals and can directly affect management decisions. Pinguicula ionantha is a federally listed species endemic to the Florida Panhandle in the southeastern USA. The main goal of our study was to assess patterns of genetic diversity and structure in 17 P. ionantha populations, and to determine if diversity is associated with geographic location or population characteristics. We scored 240 individuals at a total of 899 AFLP markers (893 polymorphic markers). We found no relationship between the estimated population size with either of two measures of diversity (proportion of loci polymorphic, P = 0.37; Nei’s gene diversity, P = 0.50). We also found low levels of population genetic structure; there was no clear relationship of genetic isolation by distance (P = 0.23) and only a small (but significant) proportion of genetic variation was partitioned amongst regions (2.4 %, P = 0.02) or populations (20.8 %, P < 0.001). STRUCTURE analysis found that the model with two inferred clusters (K = 2) best described the AFLP data; the dominant cluster at each site corresponded to the results from PCoA and Nei’s genetic distance analyses. The observed patterns of genetic diversity suggest that although P. ionantha populations are isolated spatially by distance and both natural and anthropogenic barriers, some gene flow occurs among them or isolation has been too recent to leave a genetic signature. The relatively low level of genetic diversity associated with this species is a concern as it may impair fitness and evolutionary capability in a changing environment. The results of this study provide the foundation for the development of management practices that will assist in the protection of this rare carnivorous plant.     

Patterns of geographic distribution have a considerable influence on population genetic structure in one common and two rare species of Rhododendron (Ericaceae)

Genetic diversity is essential for species to sustain their populations and evolutionary potential. In order to develop effective conservation strategies for rare species, it is necessary to understand differences in patterns of genetic diversity between common and rare species. Data about population genetic structure is important to design effective conservation strategies for rare species. In this study, we compared the genetic diversity and population genetic structure of a common species, Rhododendron weyrichii, to those of two rare species, Rhododendron sanctum and Rhododendron amagianum, with different geographic distributions. We analyzed five microsatellite loci in 16 populations of R. weyrichii, 9 populations of R. sanctum, and 6 populations of R. amagianum. As expected, the level of genetic diversity indicated by allelic richness and gene diversity was lower for the rare species R. sanctum than for the common species R. weyrichii. However, there was no statistically significant difference in genetic diversity between R. weyrichii and the other rare species, R. amagianum. Analyses of the isolation-by-distance pattern, neighbor-joining trees, and Bayesian clustering indicated that R. sanctum had a strong population genetic structure whereas R. amagianum exhibited very weak genetic structure among populations and that there was moderate population genetic structure for R. weyrichii. Therefore, the degree and pattern of population genetic structure in each species was unrelated to its rarity and instead merely reflected its geographic distribution.     

Strong population genetic structure and its implications for the conservation and management of the endangered Itasenpara bitterling

To infer the population genetic structure and genetic diversity of Itasenpara bitterling (Acheilognathus longipinnis), a cyprinid species endemic to Japan and distributed in only three specific regions, we investigated mitochondrial DNA variation. The distribution of the haplotypes among the three regions showed distinct geographic structure, and no common haplotypes were observed among regions. Analysis of molecular variance revealed a significant proportion of the genetic variance was partitioned among regions (93.1 %, P < 0.001), and pairwise estimates of D_A and Φ_ST between regions also revealed strong population structure. Given the strong genetic structure and low genetic diversity within regions, we strongly suggest that each region should be treated as a separate unit in any conservation program and any inter-regional translocations should be avoided.     

Genetic structure and diversity of Coscinium fenestratum: a critically endangered liana of Western Ghats,India

Coscinium fenestratum is a critically endangered medicinal plant, well-known for its bioactive isoquinoline alkaloid berberine. The species has been over harvested from its natural habitats to meet the huge requirement of raw drug market and industrial consumption. This has lead to a rapid decline in the population size and has also led to local population extinction at few locations in the Western Ghats, India. In this study, inter-simple sequence repeat markers were used to investigate the genetic variation and population structure of seven extant populations of C. fenestratum from the central Western Ghats, India. Eight primer combination produced a total of 57 unambiguous bands, of which (47.1 %) were polymorphic. The species exhibited a moderate to low level of intra population genetic diversity (H _s = 0.347 ± 0.008; H _t = 0.378 ± 0.006 (POPGENE) and H _s = 0.262 ± 0.0028; H _t = 0.204 ± 0.020 (HICKORY)). The populations were low to moderately differentiated from one another (G _ST = 0.221) and geographical distance was not significantly correlated with genetic distance, suggesting that these long-lived, geographically distant remnant populations were once connected through gene flow. There was a significant amount of genetic variation among populations (19.85 %). The Bayesian software STRUCTURE and HICKORY were used to further reveal the genetic structure of C. fenestratum. The results revealed weak population structure (K = 2) with one single widespread gene pool, and indicated that gene flow and inbreeding are likely to be the major driving force in shaping current population genetic structure of C. fenestratum. Thus, an understanding of the genetic diversity and population structure of C. fenestratum can provide insight into the conservation and management of this species.     

Genetic assessment of <Emphasis Type="Italic">Abies koreana</Emphasis> (Pinaceae), the endangered Korean fir,and conservation implications

To establish a management plan for endangered and rare species, genetic assessment must first be conducted. The genetic characteristics of plant species are affected by demographic history, reproductive strategy, and distributional range as well as anthropological effects. Abies koreana E. H. Wilson (Pinaceae), Korean fir, is endemic to Korea and found only in sub-alpine areas of the southern Korean Peninsula and Jejudo Island. This species has been designated as critically endangered by the International Union for Conservation of Nature due to a continuous decline in its range and population fragmentation. We genotyped 176 individuals from seven natural populations and two afforested populations on the Korean Peninsula using 19 microsatellite loci. STRUCTURE analysis revealed two genetic clusters in natural populations (F _st  = 0.040 and R _st  = 0.040) despite low differentiation. We did not detect a significant reduction in genetic diversity or the signature of a genetic bottleneck despite population fragmentation and small population size. We deduced that this species exhibits a metapopulation structure, with the population on Jirisan Mountain acting as a source of genetic diversity for other local small populations on the Korean Peninsula, through contemporary asymmetric gene flow. However, the majority of afforested individuals on the Korean Peninsula originated from a different gene cluster. Thus, we recommend a conservation strategy that maintains two genetically unique clusters.     

Genetic Variation in Three Endangered Species of Encholirium (Bromeliaceae) from Cadeia do Espinhaço, Brazil, Setected using RAPD Markers

Encholirium is a Brazilian genus of Bromeliaceae that occurs exclusively in rocky landscapes. This work aimed to generate basic information for the conservation of three Encholirium species that are endemic to the rocky mountains of Cadeia do Espinhaço, employing population genetic analyses. E. pedicellatum and E. biflorum have only one very small population each, both occurring in unprotected, private land sites, being critically endangered. E. subsecundum is more widespread, and some of its populations dwell in protected areas. Five Random Amplified Polymorphic DNA (RAPD) primers generated approximately 60 polymorphic bands for each species. This technique demonstrated the presence of a single RAPD profile for every individual sampled (except for one clone found in E. biflorum). High levels of genetic variability were not expected, due to the clonal habit of the plants and small size of the populations. Populations of E. biflorum and E. pedicellatum presented, respectively, 16.06% (Φ _st  = 0.16, p<0.001) and 8.44% (Φ _st  = 0.08, p<0.001) of the total genetic diversity attributable to genetic differences among groups within the populations. In E. subsecundum, 14.52% (Φ _st  = 0.15, p<0.001) of the total genetic diversity was found among populations. Estimates of the Shannon’s Diversity Index provided similar results. These results are valuable for the development of conservation strategies.     

Genetic diversity of four populations of Qualea grandiflora Mart. in fragments of the Brazilian Cerrado

We analyzed the genetic structure and diversity of Qualea grandiflora Mart., the most abundant woody species in the Brazilian Cerrado. Eight microsatellite loci were used to analyze samples from four populations subjected to different types of anthropic pressure, distributed throughout the state of São Paulo in the regions of Assis, Brotas, Itirapina and Pedregulho. Results indicated a mean number of 12 alleles per locus, but only six effective alleles. Alleles private to particular populations and rare alleles were also detected. An excess of homozygotes and moderate levels of inbreeding were observed. No clones were identified. All populations departed from Hardy–Weinberg equilibrium (p < 0.05). Spatial structure was observed in the distribution of specimens in distance classes ranging from 30 to 40 km and three genetic clusters were identified, with genotypes in the Pedregulho population differing from the others by up to 90 %. The influence of the Wahlund effect on the studied populations lies between 8.5 and 53.3 %. Estimates of effective population size were low (<10), and the minimum viable area for conservation in the short-, medium- and long-term was estimated to be between 4 and 184 ha. Gene flow was high enough to counter the effects of genetic drift. The genetic diversity and divergence between the studied populations indicated that the Pedregulho population should be considered an Evolutionary Significant Unit and a Management Unit.     

Genetic variation and conservation of the endangered endemic Anagyris latifolia Brouss. ex Willd. (Leguminosae) from the Canary Islands

Anagyris latifolia is an endemic and endangered species from the Canary Islands, whose distribution is limited to four islands, with less than 400 individuals in fragmented and isolated localities. RAPD markers have been used to assess the genetic diversity and genetic differentiation of its populations, in order to formulate appropriate management and conservation genetics strategies. Nine polymorphic primers generated 74 polymorphic DNA fragments. Genetic variation levels detected in Anagyris latifolia were significant high (H = 0.200; P% = 97.3), principal coordinates analysis and genetic differentiation coefficient showed a high degree of genetic differentiation between islands, without a define east-to-west stepping stone colonization route. AMOVA analysis showed that of the total genetic variation detected, 32.43% was maintained among islands, 20.73% contained among population within islands, and 46.84% resided within populations. According to these results, management strategies should be focused on each island separately.