Effects of Pleistocene climate change on genetic structure and diversity of <Emphasis Type="Italic">Shorea macrophylla</Emphasis> in Kalimantan Rainforest

The island of Borneo is the diversity center of the Dipterocarpaceae, the most important family of tropical rainforest trees in Southeast Asia. However, changes in land use and climate have affected dipterocarp distributions on the island, raising concerns about the vulnerability (inter alia) of the endemic riparian species Shorea macrophylla. Thus, to aid conservation efforts, we have investigated the genetic diversity, structure, and demographic history of S. macrophylla. The species’ genetic diversity and structure in Kalimantan (part of Indonesia, covering 75% of the island) were explored by examining genotypes of 377 individuals representing 13 populations in three regions (Northeast, Central, and West Kalimantan) using 14 newly developed microsatellite loci. Higher genetic diversity was found, at all loci, in samples from Northeast Kalimantan than in samples from the other regions. Moderate genetic differentiation between populations was detected (F_ST 0.093). Bayesian clustering, principal coordinate, and neighbor joining tree analyses of the population structure consistently identified two genetically distinct groups, one in the Northeast and the other in the Central and West regions. The higher diversity of the diverged populations in Northeast Kalimantan indicates that the region may have hosted rainforest refugia during the ice age. Accordingly, analysis using DIY ABC software indicated that the Northeast and Central-West Kalimantan groups diverged 194,000 years ago. We conclude that global climate change during the Pleistocene strongly influenced the genetic diversity and structure of S. macrophylla populations in Kalimantan.     

Invasion history of <Emphasis Type="Italic">Cardamine hirsuta</Emphasis> in Japan inferred from genetic analyses of herbarium specimens and current populations

Multiple introductions of a species are thought to enhance its invasion success by increasing genotypic diversity; this involves frequent crossing among different lineages. However, genetic diversity through crossing is less likely in autogamous species. To understand the impact of multiple introductions on the colonization success of autogamous species, we studied hairy bittercress, Cardamine hirsuta, which invaded Japan several decades ago. We detected temporal changes in its population structure using nine microsatellite markers amplified from leaf samples collected from 87 sites between 2009 and 2010, and herbarium specimens collected between 1988 and 2007. To examine whether the phenotypic variation corresponded with the genetic population structure, we also investigated the geographic variation in the lateral stamen number of this species across 49 sites. The present populations can be divided into three genetic groups, which are distributed in northern, eastern, and western Japan. This finding suggests that there are three invasive lineages (North, East, and West) in Japan. The geographic variation in lateral stamen number corresponded to the distributions of these lineages. The former distributions of the North and West lineages mostly corresponded to those found at present, but they were also historically found in eastern Japan. However, the East lineage has apparently expanded into eastern Japan, resulting in a change in dominant lineages over only a few decades. For the autogamous C. hirsuta, multiple introductions contributed toward colonization success over a wider range, which was associated with a local change in the dominant lineages.     

Co-occurring patterns of endophyte infection and genetic structure in the alpine grass, <Emphasis Type="Italic">Festuca eskia</Emphasis>: implications for seed sourcing in ecological restoration

Choosing the provenance of seed used in ecological restoration could entail its success. An alternative approach to examine local adaptation in seed sourcing is the assessment of genetic structure and diversity based on molecular markers. These types of analyses focus on the genetics of the target plant itself and eliminate the genetic influence of associated organisms, such as Epichloë/Neotyphodium endophytes in grasses. By impacting the fitness of their host, such symbionts may influence population genetic structure and diversity. Therefore, seed sourcing for grasses must consider the influence of their endophytes to increase seed translocation success and minimize the risks associated with this practice. To delineate seed zones for restoration of the alpine fescue Festuca eskia Ramond ex. DC. (Poaceae), we assessed population genetic differentiation and diversity patterns in the species including endophyte occurrence along altitudinal and longitudinal gradients in the Pyrenees Mountains. Twenty-three populations were analysed for endophyte status, and three STS and one SSR marker were used to examine genetic differentiation and diversity patterns. Results showed that F. eskia hosts an asexual form of Epichloë and infection frequency within populations decreased from East to West (100 vs. 8–25%). Molecular markers separated F. eskia into two East and West groups, and endophyte infection and genetic patterns were congruent with molecular data. Little evidence for genetic differentiation or difference in endophyte occurrence associated with altitude was detected. Little variation was found in within population diversity, regardless of provenance altitude and site, and/or endophyte infection frequency. The results of this study suggested the establishment of two distinct management units for F. eskia seed sourcing restoration.     

CONCORDANCE BETWEEN GENETIC AND SPECIES DIVERSITY IN CORAL REEF FISHES ACROSS THE PACIFIC OCEAN BIODIVERSITY GRADIENT

The relationship between genetic diversity and species diversity provides insights into biogeography and historic patterns of evolution and is critical for developing contemporary strategies for biodiversity conservation. Although concordant large‐scale clines in genetic and species diversity have been described for terrestrial organisms, whether these parameters co‐vary in marine species remains largely unknown. We examined patterns of genetic diversity for 11 coral reef fish species sampled at three locations across the Pacific Ocean species diversity gradient (Australia: ～1600 species; New Caledonia: ～1400 species; French Polynesia: ～800 species). Combined genetic diversity for all 11 species paralleled the decline in species diversity from West to East, with French Polynesia exhibiting lowest total haplotype and nucleotide diversities. Haplotype diversity consistently declined toward French Polynesia in all and nucleotide diversity in the majority of species. The French Polynesian population of most species also exhibited significant genetic differentiation from populations in the West Pacific. A number of factors may have contributed to the general positive correlation between genetic and species diversity, including location and time of species origin, vicariance events, reduced gene flow with increasing isolation, and decreasing habitat area from West to East. However, isolation and habitat area, resulting in reduced population size, are likely to be the most influential.     

No evident spatial genetic structuring in the rapidly declining Black-tailed Godwit <Emphasis Type="Italic">Limosa limosa</Emphasis><Emphasis Type="Italic">limosa</Emphasis> in The Netherlands

With 40% of the European Black-tailed Godwit population breeding in The Netherlands, this country harbours internationally significant numbers of this species. However, ongoing agricultural intensification has resulted in the fragmentation of the population and drastic population declines since 1967. By establishing genetic diversity, genetic differentiation and gene flow on the basis of 12 microsatellites, we investigated whether the population genetic structure of the Dutch Black-tailed Godwit bears the marks of these changes. Genetic diversity appeared to be moderate, and Bayesian model-based analysis of individual genotypes revealed no clustering in the Dutch populations. This was supported by pairwise F_ST values and AMOVA, which indicated no differentiation among the nine breeding areas. Gene flow estimates were larger than “one migrant per generation” between sample locations, and no isolation by distance was demonstrated. Our results indicate the maintenance of moderate levels of genetic diversity and genetic connectivity between breeding sites throughout the Dutch Black-tailed Godwit breeding population. We suggest that the Dutch Black-tailed Godwit breeding areas should be managed as a single panmictic unit, much as it is presently done.     

Genetic diversity and population differentiation of traditional fonio millet (<Emphasis Type="Italic">Digitaria </Emphasis>spp.) landraces from different agro-ecological zones of West Africa

Fonio millets (Digitaria exilis Stapf, D. iburua Stapf) are valuable indigenous staple food crops in West Africa. In order to investigate the genetic diversity and population differentiation in these millets, a total of 122 accessions from five countries (Benin, Burkina Faso, Guinea, Mali and Togo) were analysed by Amplified Fragment Length Polymorphisms (AFLPs). Genetic distance-based UPGMA clustering and principal coordinate analysis revealed a clear-cut differentiation between the two species and a clustering of D. exilis accessions in three major genetic groups fitting to their geographical origins. Shannon’s diversity index detected in D. iburua was low (H = 0.02). In D. exilis, the most widespread cultivated species, moderate levels of genetic diversity (Shannon’s diversity H = 0.267; Nei’s gene diversity H′ = 0.355) were detected. This genetic diversity is unequally distributed with the essential part observed in the Upper Niger River basin while a very low diversity is present in the Atacora mountain zone. Analysis of molecular variance (AMOVA) revealed that a large part of the genetic variation resides among the genetic groups (70%) and the country of origin (56%), indicating a clear genetic differentiation within D. exilis. Influence of mating system (inbreeding or apomixis), agricultural selection and ecological adaptations as well as founding effects in the genetic make-up of the landraces were visible and seemed to jointly contribute to the genetic structure detected in this species. The genetic variability found between the analysed accessions was weakly correlated with their phenotypic attributes. However, the genetic groups identified differed significantly in their mean performance for some agro-morphologic traits. The results obtained are relevant for fonio millets breeding, conservation and management of their genetic resources in West Africa.     

Genetic diversity and population structure of two important medicinal plant species <Emphasis Type="Italic">Schisandra chinensis</Emphasis> and <Emphasis Type="Italic">Schisandra sphenanthera</Emphasis> revealed by nuclear microsatellites

Seven polymorphic and transferable nuclear microsatellites were used to investigate the population structure of genetic diversity of Schisandra chinensis and Schisandra sphenanthera for facilitating their conservation and sustainable utilization. High levels of gene diversity were revealed in these two medicinal species, the majority of genetic diversity was harbored within populations, and population structure was might due to restricted gene flow among populations. Isolation by distance was close to significance in S. chinensis but not in S. sphenanthera. In S. chinensis, null alleles were identified as a cause for excess of homozygotes at loci G24 and WGA60, but inbreeding might also be partly responsible for the positive F _IS values in this species. In contrast, null allele frequencies were high at all the seven loci in S. sphenanthera and resulted in overestimation of fixation index. The strategy for ex situ conservation of these two medicinal species is discussed based on the genetic results.     

Isolation and characterization of species-specific microsatellite markers for blue and black wildebeest (<Emphasis Type="BoldItalic">Connochaetes taurinus</Emphasis> and <Emphasis Type="BoldItalic">C. gnou</Emphasis>)

The blue wildebeest (Connochaetes taurinus) is distributed throughout southern and east Africa while the black wildebeest (Connochaetes gnou) is endemic to South Africa and was driven to near extinction in the early 1900s due to hunting pressure and disease outbreaks. Extensive translocation of both species throughout South Africa is threatening the genetic integrity of blue and black wildebeest. To effectively manage these species, genetic tools that can be used to detect hybrid individuals, identify genetically unique subpopulations and determine the levels of genetic diversity are required. In this study, 11 microsatellite markers were developed for wildebeest through next-generation sequencing. The microsatellite loci displayed 2.00–4.14 alleles, unbiased heterozygosity values ranged from 0.32 to 0.60 and observed heterozygosity values ranged from 0.26 to 0.52. The comparatively high level of polymorphism observed in the microsatellite markers indicates that these markers can contribute significantly to our knowledge of population genetic structure, relatedness, genetic diversity and hybridization in these species.     

Habitat predictors of genetic diversity for two sympatric wetland‐breeding amphibian species

Population genetic diversity is widely accepted as important to the conservation and management of wildlife. However, habitat features may differentially affect evolutionary processes that facilitate population genetic diversity among sympatric species. We measured genetic diversity for two pond‐breeding amphibian species (Dwarf salamanders, Eurycea quadridigitata; and Southern Leopard frogs, Lithobates sphenocephalus) to understand how habitat characteristics and spatial scale affect genetic diversity across a landscape. Samples were collected from wetlands on a longleaf pine reserve in Georgia. We genotyped microsatellite loci for both species to assess population structures and determine which habitat features were most closely associated with observed heterozygosity and rarefied allelic richness. Both species exhibited significant population genetic structure; however, structure in Southern Leopard frogs was driven primarily by one outlier site. Dwarf salamander allelic richness was greater at sites with less surrounding road area within 0.5 km and more wetland area within 1.0 and 2.5 km, and heterozygosity was greater at sites with more wetland area within 0.5 km. In contrast, neither measure of Southern Leopard frog genetic diversity was associated with any habitat features at any scale we evaluated. Genetic diversity in the Dwarf salamander was strongly associated with land cover variables up to 2.5 km away from breeding wetlands, and/or results suggest that minimizing roads in wetland buffers may be beneficial to the maintenance of population genetic diversity. This study suggests that patterns of genetic differentiation and genetic diversity have associations with different habitat features across different spatial scales for two syntopic pond‐breeding amphibian species.     

The genetic population structure of lacustrine sockeye salmon, <Emphasis Type="Italic">Oncorhynchus nerka,</Emphasis> in Japan as the endangered species

Lacustrine sockeye salmon (Oncorhynchus nerka) are listed as an endangered species in Japan despite little genetic information on their population structure. In order to clarify the genetic diversity and structure of Japanese populations for evaluating on the bottleneck effect and an endangered species, we analyzed the ND5 region of mitochondrial DNA (mtDNA) and 45 single nucleotide polymorphisms (SNPs) in 640 lacustrine sockeye salmon in Japan and 80 anadromous sockeye salmon in Iliamna Lake of Alaska. The genetic diversity of the Japanese population in both mtDNA and SNPs was significantly less than that of the Iliamna Lake population. Moreover, all Japanese populations had SNP loci deviating from the HWE. In spite of low genetic diversity, the SNP analyses resulted that the Japanese population was significantly divided into three groups. These suggest that Japanese sockeye salmon populations should be protected as an endangered species and genetically disturbed by the hatchery program and transplantations.     

Genetic diversity and structure analyses on the natural populations of diploids and triploids of tiger lily, <Emphasis Type="Italic">Lilium lancifolium</Emphasis> Thunb., from Korea,China, and Japan

Tiger lily, Lilium lancifolium Thumb. is an endemic species in Korea. Genetic structure and diversity of diploid and triploid tiger lily accessions were analyzed using SSRP and IRAP molecular markers. All genetic indices analyzed showed that the diploids hold higher genetic variations compared to the triploids which were collected from Korea, China, and Japan. All variations found in triploids were also found in diploids to support the autotriploidy of L. lancifolium. It was also revealed that the triploid populations have experienced population regrowth from small population after population bottleneck. The population and phylogenetic analyses showed that analyzed tiger lilies were partitioned into three populations, Pop 1 from West Sea of Korean Peninsula, Pop 2 from South Sea of Korean Peninsula and Pop 3 of triploids in which many accessions of West Sea and South Sea were admixed in each partitioned group. Although the level of population differentiation is not extensive, F_ST analysis revealed some level of differentiation among the three populations. The analyses of genetic structure and phylogenetic dendrogram revealed admixture in some accessions among populations. All the results obtained in our analyses imply that autotriploid tiger lilies were originated from a small number of accessions of diploid L. lancifolium.     

Estimation of the Genetic Diversity of Willow Warbler Populations of the Subspecies <Emphasis Type="Italic">Phylloscopus trochilus acredula</Emphasis> (L.) in Different Parts of Its Nesting Area in the European Part of Russia

The genetic diversity (GD) of the Phylloscopus trochilus acredula subspecies populations was studied in different parts of the breeding area in the European part of Russia using microsatellite analysis. It was detected that the level of GD sufficient for the preservation of population genetic structure of the species with optimal ratio of intra- and interpopulation components of the gene diversity and minimal deviations from panmixia is maintained in the studied populations. The maximal values of GD indices were registered in an ecologically suboptimum zone on the periphery of the breeding range.     

Genetic evidence of recent migration among isolated-by-sea populations of the freshwater blenny (<Emphasis Type="Italic">Salaria fluviatilis</Emphasis>)

Genetic diversity is the raw material for evolutionary change, so a species’ capacity to maintain its genetic diversity is a major concern in conservation genetics. Although genetic diversity within a population is reduced through time by genetic drift, gene flow among populations can act to recover or add new genetic variants. The goal of this study was to infer potential connectivity among isolated-by-sea populations of the vulnerable freshwater blenny (Salaria fluviatilis) and to determine if gene flow could contribute to maintaining genetic diversity in connected populations. Four genetic clusters (one small at the North, one large at the South for both East and West coasts) were detected with different clustering methods (FLOCK, STUCTURE, UPGMA, AMOVA). The two larger genetic clusters with higher migration-rate estimates among localities had higher genetic diversity and allelic richness and lower relatedness between individuals, compared to isolated localities found in smaller clusters. Our results also suggest that sea currents may facilitate fish movements among neighbouring rivers. Overall, gene flow among isolated-by-sea but close rivers could maintain the evolutionary potential of freshwater blenny populations. This finding should be considered when elaborating a conservation program for this species.     

Polymorphic microsatellite loci for population studies of the razor clam, <Emphasis Type="Italic">Sinonovacula constricta</Emphasis>

The razor clam (Sinonovacula constricta) is an important aquacultured bivalve in China. The natural populations of this species are decreasing quickly. To facilitate studies on genetic diversity and population structure of wild populations, microsatellites were isolated from a CA enriched genomic library. Eight microsatellite loci were polymorphic in 30 individuals from Chongming in Shanghai, China. The number of alleles per polymorphic locus varied from 6 to 13 and the values of observed heterozygosity and expected heterozygosity ranged from 0.350 to 1.000 and from 0.602 to 0.902, respectively. These microsatellites are being used in studying population differentiation and genetic diversity for effective conservation and management genetic resources of S. constricta.     

Population and Genetic Consequences of Hurricanes for Three Species of West Indian Phyllostomid Bats

Strong hurricanes can cause population reductions in West Indian birds and bats, but the genetic consequences of such reductions have not been documented. For three species of phyllostomid bats, we report on the genetic effects of three strong hurricanes that struck the northern West Indies in 2004. Hurricane Ivan devastated Grand Cayman and severely depressed populations of several bat species. Despite being smaller than pre-hurricane levels, the population of Artibeus jamaicensis (the only species we could resample) on Grand Cayman contained greater mitochondrial haplotype diversity but similar microsatellite allelic diversity compared to pre-Ivan levels. We suggest that hurricane-aided dispersal from Cayman Brac introduced two new haplotypes into the Grand Cayman population. In the Bahamas, two other phyllostomids ( Erophylla sezekorni and Macrotus waterhousii ) did not suffer population losses or changes in genetic diversity as a result of Hurricanes Frances and Jeanne. Our results suggest that strong hurricanes usually have greater demographic than genetic effects but that hurricane-aided dispersal can occasionally introduce new genotypes or haplotypes into island bat populations.     

Species diversity and population density affect genetic structure and gene dispersal in a subtropical understory shrub

Aims The dispersal of pollen and seeds is spatially restricted and may vary among plant populations because of varying biotic interactions, population histories or abiotic conditions. Because gene dispersal is spatially restricted, it will eventually result in the development of spatial genetic structure (SGS), which in turn can allow insights into gene dispersal processes. Here, we assessed the effect of habitat characteristics like population density and community structure on small-scale SGS and estimate historical gene dispersal at different spatial scales.Methods In a set of 12 populations of the subtropical understory shrub Ardisia crenata, we assessed genetic variation at 7 microsatellite loci within and among populations. We investigated small-scale genetic structure with spatial genetic autocorrelation statistics and heterogeneity tests and estimated gene dispersal distances based on population differentiation and on within-population SGS. SGS was related to habitat characteristics by multiple regression.Important findings The populations showed high genetic diversity (H e = 0.64) within populations and rather strong genetic differentiation (F ′ ST = 0.208) among populations, following an isolation-by-distance pattern, which suggests that populations are in gene flow–drift equilibrium. Significant SGS was present within populations (mean Sp = 0.027). Population density and species diversity had a joint effect on SGS with low population density and high species diversity leading to stronger small-scale SGS. Estimates of historical gene dispersal from between-population differentiation and from within-population SGS resulted in similar values between 4.8 and 22.9 m. The results indicate that local-ranged pollen dispersal and inefficient long-distance seed dispersal, both affected by population density and species diversity, contributed to the genetic population structure of the species. We suggest that SGS in shrubs is more similar to that of herbs than to trees and that in communities with high species diversity gene flow is more restricted than at low species diversity. This may represent a process that retards the development of a positive species diversity–genetic diversity relationship.     

Genetic population structure and demography of an apex predator,the tiger shark Galeocerdo cuvier

Population genetics has been increasingly applied to study large sharks over the last decade. Whilst large shark species are often difficult to study with direct methods, improved knowledge is needed for both population management and conservation, especially for species vulnerable to anthropogenic and climatic impacts. The tiger shark, Galeocerdo cuvier, is an apex predator known to play important direct and indirect roles in tropical and subtropical marine ecosystems. While the global and Indo‐West Pacific population genetic structure of this species has recently been investigated, questions remain over population structure and demographic history within the western Indian (WIO) and within the western Pacific Oceans (WPO). To address the knowledge gap in tiger shark regional population structures, the genetic diversity of 286 individuals sampled in seven localities was investigated using 27 microsatellite loci and three mitochondrial genes (CR, COI, and cytb). A weak genetic differentiation was observed between the WIO and the WPO, suggesting high genetic connectivity. This result agrees with previous studies and highlights the importance of the pelagic behavior of this species to ensure gene flow. Using approximate Bayesian computation to couple information from both nuclear and mitochondrial markers, evidence of a recent bottleneck in the Holocene (2,000–3,000 years ago) was found, which is the most probable cause for the low genetic diversity observed. A contemporary effective population size as low as 111 [43,369] was estimated during the bottleneck. Together, these results indicate low genetic diversity that may reflect a vulnerable population sensitive to regional pressures. Conservation measures are thus needed to protect a species that is classified as Near Threatened.     

Population size,center–periphery,and seed dispersers’ effects on the genetic diversity and population structure of the Mediterranean relict shrub Cneorum tricoccon

The effect of population size on population genetic diversity and structure has rarely been studied jointly with other factors such as the position of a population within the species’ distribution range or the presence of mutualistic partners influencing dispersal. Understanding these determining factors for genetic variation is critical for conservation of relict plants that are generally suffering from genetic deterioration. Working with 16 populations of the vulnerable relict shrub Cneorum tricoccon throughout the majority of its western Mediterranean distribution range, and using nine polymorphic microsatellite markers, we examined the effects of periphery (peripheral vs. central), population size (large vs. small), and seed disperser (introduced carnivores vs. endemic lizards) on the genetic diversity and population structure of the species. Contrasting genetic variation (H_E: 0.04–0.476) was found across populations. Peripheral populations showed lower genetic diversity, but this was dependent on population size. Large peripheral populations showed high levels of genetic diversity, whereas small central populations were less diverse. Significant isolation by distance was detected, indicating that the effect of long‐distance gene flow is limited relative to that of genetic drift, probably due to high selfing rates (F_IS = 0.155–0.887), restricted pollen flow, and ineffective seed dispersal. Bayesian clustering also supported the strong population differentiation and highly fragmented structure. Contrary to expectations, the type of disperser showed no significant effect on either population genetic diversity or structure. Our results challenge the idea of an effect of periphery per se that can be mainly explained by population size, drawing attention to the need of integrative approaches considering different determinants of genetic variation. Furthermore, the very low genetic diversity observed in several small populations and the strong among‐population differentiation highlight the conservation value of large populations throughout the species’ range, particularly in light of climate change and direct human threats.     

Population genetics of the bigeye thresher shark <Emphasis Type="Italic">Alopias superciliosus</Emphasis> in the Atlantic and Indian Oceans: implications for conservation

Population structure and genetic connectivity are pivotal contributions to the establishment of conservation strategies for fisheries management, in particular for highly migratory species that are affected by commercial fisheries. This study used partial sequences of mitochondrial DNA control region to determine the genetic structure of the bigeye thresher shark Alopias superciliosus in the Atlantic and Indian Oceans. A total of 858 base pairs of mtDNA CR from 228 individuals were analyzed. The resulting nucleotide diversity (π) was 0.0011?±?0.0008 and the haplotype diversity (h) was 0.127?±?0.030. These are the lowest diversities registered in elasmobranchs with this genetic marker. Two genetically distinct lineages were identified, one of them represented by 3.9% of the analyzed individuals and none restricted to any particular area. Simulated scenarios of population structure, tested with AMOVA and pairwise Φ_ST did not result in significant values indicating high connectivity among all sampled groups. The absence of population structure, even between Atlantic and Indian Oceans, corroborates the high dispersal ability of this species. The low genetic diversity detected in this species and the identification of two historical lineages occurring in sympatry, one represented by a very small number of individuals, should be considered in the conservation efforts and management plans of A. superciliosus.     

Genetic diversity of insular natural populations of <Emphasis Type="Italic">Festuca pratensis</Emphasis> Huds.: RAPD analysis

In natural populations of Festuca pratensis Huds. from the islands of Onega Lake, the level of genetic diversity was evaluated. In three populations variability of 64 RAPD loci was tested. The level of genetic diversity (P _95% = 30.2; H _exp = 0.093) was low for a cross-pollinating plant species. Furthermore, genetic similarity between the plants from insular populations was found to be high (I _N = 0.887). It was demonstrated that genetic variation among the population accounted for at most 5.3% of total genetic diversity, which, however, was higher than the F _ST values for continental populations (F _ST = 0.022). It was suggested that specific features of the genetic structure of insular population, i.e., low gene diversity within the populations along with high differentiation among the populations, were caused by the gene flow attenuation, as a result of isolation, and intensification of inbreeding. These features had negative effect on total population adaptation.