This finding is further supported by the pairwise FST of Malay sub-populations, of which the north and south regions showed the highest differentiation (FST [North–south] = 0.0011). The collective findings therefore suggest that population sub-structure of Malays are more heterogenous than previously expected even within a small geographical region, possibly due to factors like different genetic origins, geographical isolation, could result in spurious association as demonstrated in our analysis. We suggest that cautions should be taken during the stage of study design or interpreting the association signals in disease mapping studies which are expected to be conducted in Malay population in the near future.Fine scale population structure of Malays - the major population in Malaysia, has not been well studied. This may have important implications for both evolutionary and medical studies. Here, we investigated the population sub-structure of Malay involving 431 samples collected from all states from peninsular Malaysia and Singapore. We identified two major clusters of individuals corresponding to the north and south peninsular Malaysia. On an even finer scale, the genetic coordinates of the geographical Malay populations are in correlation with the latitudes (RThe online version of this article (doi:10.1186/s40246-015-0039-x) contains supplementary material, which is available to authorized users. Malaysia, a multi-ethnic, multi-lingual, multi-cultural and multi-religious country, is located at the crossroads of Southeast Asia. It is separated by the South China Sea into two land masses namely, the Peninsular Malaysia and East Malaysia . Malaysia has a total population of about 30 million people, of which approximately 26 million populate the Peninsular Malaysia. Among the major populations in Peninsular Malaysia, the Malays are the largest ethnic group and make up to 63% of the total population follow by Chinese, Indians and other minority ethnic groups.Many Malays are of Malayo-Polynesian (Austronesian) origin that are culturally and historically heterogeneous . The MalEarlier studies had indicated potential genetic sub-structure among the different groups of Malays from Peninsular Malaysia , 4, 5, wIn this study, we showed that the genetic diversity and population sub-structure of the Malays from Peninsular Malaysia are correlated to the geographical latitude. Notably, we observed the main differentiations between populations corresponding to the north and south Peninsular Malaysia. In addition, simulation analyses carried out also revealed that the genetic association is greatly affected by population sub-structure, suggesting that consideration of population stratification of samples at the stage of study design and careful interpretation of the association signals are necessary when mapping complex diseases in Malay populations.smartPCA. Two clusters were observed representing the north and south regions, respectively  showed closer affinity to the South Asia populations (GIH) Fig. . We thenely Fig. .Fig. 1PrIn ADMIXTURE analysis, a significant difference was observed between the Malays from the north and south in the major component, with 57% and 65% in the north and south, respectively   synthesis. Another highly differentiated SNP, rs4148475, is located at the candidate gene ABCC4. This gene is a member of the superfamily of ATP-binding Cassette (ABC) transporters, which may play a role in cellular detoxification , Simulations on genome-wide association study (GWAS) were performed using PLINK 1.07, following the procedure of Xu et al. (2009) .


Nothofagus  from Chile. To estimate population genetic parameters and infer the influence of the last ice age on the spatial and genetic distribution of these species, we examined and analyzed genetic variability at seven polymorphic microsatellite DNA loci in 640 individuals from 40 populations covering most of the ranges of these species in Chile. Populations showed no significant inbreeding and exhibited relatively high levels of genetic diversity (HE = 0.502–0.662) and slight, but significant, genetic structure (RST = 8.7–16.0%). However, in N. obliqua, the small amount of genetic structure was spatially organized into three well-defined latitudinal groups. Our data may also suggest some introgression of N. alpina genes into N. obliqua in the northern populations. These results allowed us to reconstruct the influence of the last ice age on the genetic structure of these species, suggesting several centers of genetic diversity for N. obliqua and N. alpina, in agreement with the multiple refugia hypothesis.The effect of glaciation on the levels and patterns of genetic variation has been well studied in the Northern Hemisphere. However, although glaciation has undoubtedly shaped the genetic structure of plants in the Southern Hemisphere, fewer studies have characterized the effect, and almost none of them using microsatellites. Particularly, complex patterns of genetic structure might be expected in areas such as the Andes, where both latitudinal and altitudinal glacial advance and retreat have molded modern plant communities. We therefore studied the population genetics of three closely related, hybridizing species of  The current genetic structure and diversity of natural plant populations can be seen as the resulting product of the interaction between biology, geography, and climatic change Hewitt . ImportaThe west coast of southern South America in Chile and western Argentina between 33°00′S and 41°30′S is crossed longitudinally by two mountain ranges separated by the Central Valley. The Coastal Range has altitudes between 500 and 2000 m above sea level (m a.s.l.), and the Andes have altitudes between 3000 and 6000 m a.s.l., both becoming generally lower from north to south. The Mediterranean forests are found between 33°00′S and 36°30′S, and the temperate rainforests south of 37°30′S, with an ecotonal zone – the transitional forests – in between Donoso .Nothofagus obliqua (Mirb.) Oerst., N. alpina (Poepp. et Endl.) Oerst. (= N. nervosa), and N. glauca (Phil.) Krasser. are sympatric South American endemics. Together with two species from Australia (N. cunninghamii (Hook.) Oerst. and N. moorei (Muell.) Krasser.) and one from New Zealand (N. menziessii (Hook.) Oerst.), these species constitute subgenus Lophozonia , glaciers covered a large proportion of the current forest, and additionally, palynological research has identified periglacial effects  gradually brought drier and warmer conditions to the area, pushing Nothofagus forests to their current distributions in the mountains.The Central Valley of central Chile (33°00′–36°00′S) is characterized by warm and dry summers and supports sclerophyllous forests. However, with more precipitation at higher elevations, it is possible to find populations of S Donoso . During  Heusser  and probN. alpina, which has its main distribution in the Andes, occur on the top of the Nahuelbuta Mountains in the Coastal Range. These forest islands have been interpreted as the remnants of glacial populations that were once growing in the Central Valley during the ice age  to live in the Central Valley. According to pollen records, recolonization of Nahuelbuta by Nothofagus spp. started about 6000 years ago in the Holocene (Villagran Similar events occurred in the middle portion of the ranges of the three species (36°00′–39°00′S). Today, isolated populations of N. obliqua, N. alpina) might have expanded slowly to this area in the Holocene  when climatic conditions started to be warm and moist enough to support that vegetation   Donoso ). Thus, es Table .Lophozonia (Nothofagus) to infer how these genetic parameters are influenced by climatic change, geography, and biological factors. Thus, in this study we will compare results obtained here with studies using other biparentally inherited markers in N. alpina: allozymes , consisting of 31 and 14 N. obliqua and N. alpina populations, respectively. We sampled populations where there were typically 10 mother trees  represented by five planted individuals per family; therefore, at each population, we sampled one offspring for each mother tree, and the remaining six sampled individuals are half-siblings to one of the originally sampled individuals. For N. glauca, and some populations of N. obliqua and N. alpina, we collected dormant buds in July 2003 from randomly selected trees separated, when possible, by at least 20 meters in natural populations , diluted samples to the concentrations of 50–200 ng/μL, and stored samples at −20°C until use.After pulverizing buds and leaf tissue in a bead mill, we extracted total genomic DNA using a modified CTAB protocol for silica-dried tissue  at the ICBR facility, University of Florida. We used four fluorescent dyes to label the M13 primer to combine four loci in each run by pooling them together in each well of a 96-well plate. We performed fragment analysis and scoring of alleles using GeneMapper® 4.0 , repeating unsuccessful amplifications once and treating second-round failures as missing data.icro-Checker 2.2.3  and a Markov chain algorithm with 20 batches and 5000 iterations per batch on each population to detect linkage between pairs of loci. For comparison, we used the likelihood ratio LD procedure (with 1000 permutations) in Arlequin 3.11 , observed (HO) and expected (HE) heterozygosity, inbreeding coefficient (FIS), and allele size-based inbreeding coefficient (ρIS). The parameter A is equivalent to allelic richness when, as in this study, sample sizes among populations are similar, and it is the most important parameter when evaluating glacial refugia  across latitude and elevation obtaining Pearson correlation coefficients (r), and among species, locations , latitudinal groups , glacial refugia hypothesis , and sources of plant material , using nonparametric Kruskal–Wallis tests. Additionally, we tested for differences in inbreeding coefficients (FIS and ρIS) between sources of plant material. We applied the standard Bonferroni technique  using the standard maximum K as the number of populations plus two: for N. obliqua, K = 1 to 22; N. alpina, K = 1 to 14; and N. glauca, K = 1 to 10. For each species, we performed 10 separate runs at each K. We used a burn-in period of 100,000 and 200,000 Markov chain Monte Carlo (MCMC) iterations for each run, obtaining posterior probabilities (LnP[D]) to detect the most likely K. We chose K at the highest LnP[D] and using Evanno's ΔK  in Arlequin 3.11  following the infinite island population model  analysis, following the recommendation by Frantz et al.  trees were constructed using pairwise 6.6 Page . Before tructure 2.3.2  over 487 possible combinations (3.3%), and none was consistent across populations. The most consistent event was the LD between ncutas06 and ncutas13, but it was only significant in three of 40 populations, indicating that it was probably a relationship due to chance alone. All other cases of LD were significant in only one or two populations.Combining ncutas04) in all populations of N. obliqua and N. alpina, but not in N. glauca (Table A and HE) were always lower in N. glauca than in N. alpina, and lower in N. alpina than in N. obliqua (Table N. obliqua and N. glauca for both parameters (P < 0.001), significant results between N. obliqua and N. alpina for A (P < 0.05), and between N. alpina and N. glauca for HE (P < 0.05). None of the populations from any of the species had significant signs of recent genetic bottlenecks, with P-values always greater than 0.5 for the bottleneck analysis.While we did not measure polymorphism in our study because it is usually close to 100% when using microsatellite loci selected specifically for their polymorphism, there was one monomorphic locus  across all loci, there is a general tendency in N. obliqua and N. glauca to have positive inbreeding coefficients, FIS and ρIS, across populations, in clear contrast to N. alpina (Table Even though only populations La Campana (1) and Hueyusca (32) of na Table .In general, there was no significant correlation or consistent trend between diversity measures and latitude or elevation for the species. Only in comparisons between populations from the Coast and the Andes was there a general trend of higher levels of genetic variation in the Coast for all species, but this result was not significant.N. obliqua or N. alpina .We hypothesized that samples obtained from provenance trials would show inbreeding and lower levels of genetic diversity in comparison with samples from natural populations because in the trials, six of the16 sampled individuals were half-siblings. However, nonparametric Kruskal–Wallis tests did not show any significance or trends showing higher inbreeding or lower genetic diversity in samples obtained from the trials in either N. obliqua and N. alpina to contrast the levels of genetic diversity between these two latitudinal groups: north, including all populations from the Mediterranean forests north of the parallel 36°30′S, and south, including all others  for each K (number of assumed clusters). In all three species, the number of optimal clusters was much lower than the number of sampled populations, indicating extensive gene flow, either historical or ongoing. The maximum LnP[D] was at K = 3 for N. obliqua, K = 7 for N. alpina, and K = 2 for N. glauca  and N. glauca , ΔK agrees with the optimal K found by LnP[D], but in N. alpina , ΔK does not agree with the K found by LnP[D] and supports an optimal value of K = 2 instead of 7 , corresponding to the Mediterranean forests, are clearly differentiated from all other populations. Among the latter, a transition group (populations 7–12) is distinct from the southern populations (13–20), which correspond to the temperate rainforests. Only Ninhue (6), which belongs geographically to the northern group, clusters with populations from the south. Significant gene flow among groups is inferred from the levels of admixture seen in the Structure diagram  and Bullileo Alto (23). Both populations are located in the Andes, in the northern tip of the distribution of N. alpina, separated by 100 km and relatively isolated from other populations  and the southernmost population Quilleco (40) are clearly distinct and also very isolated from other populations of the species  levels of genetic differentiation among populations  as 2.0, 1.3, and 2.6 individuals per generation, respectively. The RST values were similar to those obtained using the FST statistic, with FST = 0.12, 0.14, and 0.074 for N. obliqua, N. alpina, and N. glauca, respectively. In N. obliqua, Structure also found that the species could be divided into three spatially defined and homogeneous groups of populations. As a result, we ran an AMOVA including that extra group level. In this analysis, the variation among groups (RCT) was 0.114 (P < 0.00001) and the variation among populations within groups (RSC) was only 0.037 (P < 0.00001), also indicating homogeneity within groups with substantially more intragroup (Nem = 6.5) than intergroup (Nem = 1.9) migration. Consequently, the total variation among populations (RST) was 0.146 (P < 0.00001), higher than the original values for N. obliqua (RST = 0.11), because we excluded Ninhue (6), which did not have a well-defined membership to any group.The within-species AMOVAs indicated that the genetic variation within populations was always greater than 80%. The ns Table . Using tRST  with the morphological identity of the species for all N. alpina and N. glauca populations, and for populations (6) to (20) in N. obliqua. Northern populations (1) to (5) in N. obliqua had higher admixture, but their identity was still greater than 50% (data not shown). Under this scenario, in N. obliqua there was an average admixture proportion of 10.3% coming from N. alpina and 1.9% coming from N. glauca, far higher than the admixture going to N. alpina or N. glauca (Table By running Sulations  to (20) N. obliqua is not distributed evenly among all populations; most of it is in the northern populations. Due to this uneven distribution in admixture proportions, we also inspected data using K = 4, which we knew placed these northern populations in a different cluster, allowing us to have a separate estimation of admixture. Most of the interspecies admixture in N. obliqua disappeared in this analysis, reducing the overall admixture proportion in N. obliqua to 3.4% coming from N. alpina and 1.4% coming from N. glauca, with only minor changes to the values going to N. alpina and N. glauca  and Loncha (2) with 86% each.It is clear, however, that the admixture in ca Table . In addiNothofagus are consistent with results for other related, large-sized, long-lived, outcrossing, wind-pollinated trees (Table HE = 0.452–0.496) and number of alleles per locus (A = 3.7–4.4) for Nothofagus in Argentina  from the Northern Hemisphere are similar to those found in our study . However, our values are higher than those reported for Nothofagus in Argentina and fall in the lower end of the ranges for Fagus and Quercus. We attribute the latter to the span of the geographical distributions of the species in our study, which are more narrowly distributed than those species from the Northern Hemisphere.The high levels of nuclear microsatellite genetic variation in these species of es Table . For nucNothofagus alpina from Chile has lower RAPD diversity than other long-lived forest species, a result attributed to intense exploitation and substitution during the last century  in relation to geographic range are consistent with general expectations. Using the studies with nuclear microsatellite markers for Fagus and Quercus listed in Table HE = 0.68–0.87) for widespread species such as F. sylvatica and intermediate values (HE = 0.60–0.66) for regional species such as Q. garryana, similar to values for N. alpina (HE = 0.62) and N. obliqua (HE = 0.66). Accordingly, narrowly distributed N. glauca has lower diversity (HE = 0.50).Although it may be misleading to compare this range-influenced trend of genetic variation obtained for the Chilean species of subgenus Nothofagus have a largely outcrossing breeding system  in N. alpina. However, despite the lack of significant inbreeding for most of the populations, there is a tendency for inbreeding coefficients  to be greater than zero in N. obliqua and N. glauca , the FIS and ρIS values are probably due to chance alone. The general correspondence of populations of N. alpina with HWE could mean that this species has more efficient ways of avoiding inbreeding such as larger effective population sizes or its regeneration strategy, but so far all the evidence in the literature states the opposite : La Campana (1) in ca Table . This coe Pineda .N. obliqua and Hueyusca (32) at the southern limit of N. alpina  at the northern limit of  at the nIBD would have an effect on the clustering algorithm in Structure, mainly by making the posterior probability of the data become asymptotic, thus overestimating the number of “real” clusters. Fortunately, even though we have strong evidence of IBD in N. obliqua and some evidence in N. glauca, the posterior probabilities in our Structure analyses were not asymptotic and had a peak that was also coincident with the optimal number of clusters given by ΔK. In addition, Frantz et al.  is moderate in comparison with values for N. alpina and N. glauca in our study, and with other Nothofagaceae and Fagaceae (Table Nem = 2.0) maintains the genetic homogeneity of the species quite well. Additionally, IBD plays an important role , but it is more restricted than the gene flow among populations within groups (Nem = 6.5). These groups are distributed latitudinally, each with members from very different environments  to an oceanic climate (Cfb) based on the Köppen–Geiger classification , and it is among the highest values reported for Nothofagus and the closely related Fagaceae  is lower than those values for N. obliqua and N. alpina but still moderate in comparison with other Nothofagus and Fagaceae (Table Nem = 2.6). There is a weak tendency for the populations to be isolated by distance  and Andean Alico (39) and the genetic dissimilarity between Bullileo (38) and Alico (39), two Andean populations geographically close but on different sides of a group of mountains  and Quilleco (40), with no apparent geographical groups  , many cases occur in populations that never were in sympatry, and there are no reports of hybridization between these two species.A potential source of genetic variation and structure in the studied populations is hybridization within the subgenus. Natural F) Donoso , and popre Table  agree witructure analysis to interpret admixture among the three species. When using the optimal number of clusters (K = 3), with each species belonging to its own cluster, we found high levels of gene flow going from N. alpina toward N. obliqua in the highlands of the northern part of the distribution in the Mediterranean forests between 33°00′S and 36°30′S. This result was very surprising because currently in this area the species do not seem to interbreed and the populations with more admixture, that is, La Campana (1), Loncha (2), and Bellavista (3), are isolated from any N. alpina populations. Interestingly, according to Vazquez and Rodriguez  on the distribution patterns and genetic variation of Our results agree with the multiple refugia hypothesis and, therefore, with the existence of several centers of genetic diversity across the entire geographical ranges of these species. Also, the lack of signs of bottlenecks or repeated founder events in our microsatellite data indicates that these refugia harbored considerable variation and that pollen flow has been an important agent in keeping populations genetically diverse and genetically connected from the time of the first recolonization events after the LGM.Nothofagus obliqua , in agreement with palynological records  in the Coast Range near 41°00′S seems to also be a glacial refugium for N. obliqua that, like most of the Coastal Range refugia, did not participate in postglacial colonization  was probably only mildly influenced by the climatic changes since the last glaciation. Our data indicate that the limited genetic structure of this species is due to two very isolated populations  is the result of a 100-km dispersal event. More likely, there were small N. glauca stands in the Andean foothills and Central Valley connecting all the populations of the species during the LGM about 20,000 years ago when moist conditions at low elevations were ideal for maintaining Nothofagus forest. Drier conditions could have caused most of the populations to gradually disappear in the area, except Quilleco (40). Moreover, this population has been isolated from other putative N. glauca populations in the area by the great alluvial cone of Lake Laja deposited in the Laja valley about 10,000 years ago (Thiele et al. The population genetics of ons Fig. : the norons Fig. . The larN. obliqua, N. alpina, and N. glauca are high and correspond to the values of other tree species with similar natural histories and ranges of distribution. There is limited, but significant, genetic structure explained by isolation by distance within population continua, geographic isolation of certain populations, and some hypothesized geographic barriers to gene flow. Also, the tendency of these species to form stands with different levels of kinship did not result in significant inbreeding. On the other hand, the evidence of ancient and current hybridization as an important contributor to the intraspecific genetic diversity within the subgenus indicates that it may be a major source of variability and differentiation for the northern populations of N. obliqua. Finally, the patterns of genetic variation that we observed agree with the multiple refugia hypothesis for N. obliqua and N. alpina and suggest several centers of genetic diversity across the areas of distribution for all three species.Our results indicate that the levels of neutral genetic variation exhibited by


Sequoiadendron giganteum (giant sequoia) that is restricted to a narrow belt of groves in the Sierra Nevada Mountains. We ask whether the current distribution is a result of northward colonization since the last glacial maximum (LGM), restriction of a broader range in the recent past (LGM) or independent colonizations in the deeper past. Genetic diversity at eleven microsatellite loci decreased with increasing latitude, but partial regressions suggested this was a function of smaller population sizes in the north. Disjunct populations north of the Kings River were divergent from those south of the Kings River that formed a single cluster in Bayesian assignment tests. Demographic inferences supported a demographic contraction just prior to the LGM as the most likely scenario for the current disjunct range of the species. This contraction appeared to be superimposed upon a long‐term decline in giant sequoia over the last 2 million years, associated with increasing aridity due to the Mediterranean climate. Overall, low genetic diversity, together with competition in an environment to which giant sequoia is likely already poorly adapted, will pose major constraints on its success in the face of increasing aridity.Mediterranean ecosystems comprise a high proportion of endemic taxa whose response to climate change will depend on their evolutionary origins. In the California flora, relatively little attention has been given to the evolutionary history of paleoendemics from a molecular perspective, yet they number among some of the world's most iconic plant species. Here, we address questions of demographic change in  The relationship between genetic diversity and species' range is a fundamental question in conservation genetics. Although meta‐analyses have suggested that restricted species on average have less genetic diversity than their widespread congeners Karron , many siin situ, and paleoendemics that have dispersed into the new Mediterranean habitat , a paleoendemic species that occupies the western slope of the Sierra Nevada Mountains of California  is endemic to the western slope of the Sierra Nevada range in Central California. It occurs in about 75 groves stretching in a narrow belt (not more than 24 km wide and about 420 km long) from the American River in Placer County (Placer Grove) to southern Tulare County (Deer Creek Grove). The northern two‐thirds of the range comprise eight small disjunct groves. The remaining groves, including all the large ones, occur from the Kings River drainage south to Deer Creek , 0.5 units of TAQ DNA polymerase , 1 μL −10x PCR buffer with 15 mmol/L MgCl2, 0.4 μmol/L reverse, 0.16 μmol/L FAM‐labeled M13 primer, and approximately 20 ng of extracted DNA. The solution was run through an initial denaturation at 95°C for 15 min, followed by 30 cycles of 30 sec at 95°C, 45 sec at 56°C, and 45 sec at 72°C. This was followed by 8 cycles of 95°C for 30 sec, 53°C for 45 sec, and 72°C for 45 sec. Final extension consisted of 30 min at 72 C. Then, 1.5 μL of the PCR product was mixed with a solution of 8 μL formamide and 0.5 μL Genescan LIZ500 size standard . The resulting solution was run through an ABI 3730 automated sequencer. Microsatellite fragments were analyzed with Genescan 3.7 and Genotyper 3.7 software (Applied Biosystems).The DNA was extracted using the cetyltrimethyl ammonium bromide (CTAB) method, following Cullings . Individand Dodd . All DNAand Dodd . PCRs coWe estimated pairwise relatedness coefficients (Lynch and Ritland HE) and allelic richness (AR) ).Microsatellite loci were tested for deviation from Hardy–Weinberg expectations using unbiased Markov chain exact tests in GENEPOP v. 3.4 . Because geographic distance could be confounded by the strong geographic pattern of northern disjunct populations and the southern more continuous range, we created a binary indicator matrix, in which pairwise population comparisons were coded as 0 if both populations in the comparison came from a like group and 1 if they came from a unlike group. Four groups were formed on the basis of their approximate contiguity: (1) two Calaveras groves, (2) Mariposa and Nelder groves, (3) McKinley grove, and (4) remaining groves south of the King River. Tests of IBD were performed with 30,000 randomizations of Rousset's distance [FST/(1 − FST)] and geographic distances based on population centroids of latitude and longitude coordinates. Rousset  may confound discrete population genetic structure. Pritchard et al.  caution  Rousset  recommenWe used two Bayesian model‐based approaches to identify population genetic structure: a spatially explicit model in BAPS v 6.0  to vary from 2 to 23, to obtain an estimate of the number of partitions. For final runs, we set K to vary from 2 to 15, and we ran 10 simulations at each value of K. Subsequently, we ran fixed K group partition comparisons to test for the most likely evolutionary‐based grouping of sampled populations. We tested four scenarios: (1) two groups: northern disjunct stands (including McKinley grove and groves further north) and the remaining southern groves; (2) six groups: the same north–south divide as in (1), but northern groves were separated into spatially distinct groups ; (3) nine groups as in (2), but all northern groves were treated as separate; and (4) nine groups, divided by major drainage systems . We set uniform priors of 0.25 to each of the scenarios.Because we did not have the original coordinates for each sampled individual, we randomly selected their latitude and longitude coordinates from Google Earth maps of each grove. For the purposes of this study, we were not interested in within‐grove fine‐scale spatial structure, so the random coordinates were intended only to provide unique coordinates for each sample. We inferred the optimal partition in BAPS using the spatial clustering of groups' option. In the initial runs, we set K) present in the dataset and assigned individuals to these clusters, using the admixture model with correlated allele frequencies, with no a priori information about collection sites. For all final simulations performed in STRUCTURE, we used a “burn‐in” period of 5 × 104 followed by a run length of 5 × 105 MCMC iterations, which were sufficient to give a stable α and estimate of the log probability of the data. For preliminary runs, we included the whole dataset with K ranging from 1 to 23 (the number of sampled populations). These preliminary runs indicated minor changes in log probability of the data for K greater than 8; therefore, for final MCMC runs, we selected the upper K value as 8 and ran 10 simulations for each value of K. To estimate the most likely K value, we used the ΔK statistic . Permuting in this way erases the effect of the stepwise mutational model of microsatellite alleles, but retains the divergence among populations due to allele frequency differences and approximates FST  framework using DIYABC v. 2.0  Colonization of northern groves from the south during the late Pleistocene (recent colonization). Here, it was assumed that migration northward was the most likely direction, given the larger populations and greater genetic diversity see  in the sNorthward stepping‐stone colonization model: McKinley colonized from the southern group and provided seed for colonization of the Nelder and Mariposa groves. In turn, Nelder and Mariposa provided seed for colonization of Merced and Tuolumne groves. Merced and Tuolumne provided seed for the Calaveras groves. Under these scenarios, it was assumed that the founder populations were small, and so we simulated initial bottlenecks.Scenario 1a: Northward mass colonization model: All northern groves were colonized at the same time from the southern group. Again, the founder populations were assumed to be small and were simulated with bottlenecks.Scenario 1b: Scenario 2: Divergence among northern groves and the southern group of groves following restriction in size of an ancestral population during the late Pleistocene. This scenario would suggest a broader and more continuous range prior to the last glacial.Scenario 3: Independent origins from an ancestral population in the deep past. This scenario would be consistent with early Pleistocene migrations across the Sierra Nevada from populations occupying the eastern slope.FST and δμ2 among population pairwise parameters. For each scenario, we ran 4 x 106 simulated datasets to estimate posterior probabilities using logistic regression. We selected the scenario with the highest posterior probability and estimated the values of the associated demographic parameters. To evaluate confidence in the choice of scenario, we created 500 pseudo‐observed datasets . The microsatellite loci were divided into two groups: (1) all dinucleotide repeat loci and (2) the single trinucleotide repeat locus. Default values were used for the microsatellite mutation models. We estimated the following summary statistics: mean number of alleles, mean genic diversity, and mean allele size variance for each population and Splitting times were converted to absolute scales assuming a range of generation times for this long‐lived woody plant. Generation time in giant sequoia was estimated following Lande :α) as ranging from 60 to 150 years  was rejected for only eight of the 231 population–primer combinations (see Table S1). Of the eight combinations not in HWE, five were for primers 4 and 6 in the northern disjunct group of populations. We considered the number of rejections of HWE to be low and not to justify the removal of populations or primers in further analyses. Among all populations, we detected 45 primer pair combinations not in linkage equilibrium, out of a total of 1232 tests (see Table S2). These were scattered across populations and primer combinations suggesting that the instances of LD would not seriously affect further analyses. No evidence for null alleles, or scoring errors, was detected.AR) ranged from 2.08 at Placer to 4.07 at Windy Gulch and measured as expected heterozygosity (HE) from 0.29 at Placer to 0.70 at Nelder (Tables N), and (3) sample size that might bias diversity estimates (n). Partial correlations showed no significant effect of latitude on allelic diversity  after accounting for the effects of grove size (N) and sample size (n), but significant effects of grove size  and sample size . No significant partial correlations were found for expected heterozygosity .We detected a total of 118 alleles across the 11 loci . Across populations, genetic diversity measured as allelic richness  and with the indicator matrix . However, after controlling for the indicator matrix, the partial correlation of genetic similarity and geographic distance was not significant , whereas genetic distance was significantly correlated with the indicator matrix after controlling for geographic distance . Because of the lack of a significant signal of IBD, we continued with tests to detect population genetic structure.Rousset's distance was significantly correlated with the log of geographic distance . Of these clusters, two comprised only two individuals each, so that the bulk of individuals was assigned to one of 9 clusters. Individuals from each of the northern groves were either exclusively  or mostly assigned  to single clusters. All southern groves were almost exclusively assigned to a single cluster. Among the four partition comparisons, posterior probabilities were estimated as Scenario 1 Prob 0, Scenario 2 Prob 0, Scenario 3 Prob 1, and Scenario 4 Prob 0, indicating that the genetic data best fit the third scenario of nine groups including single‐grove clusters for all northern groves and a single cluster for groves south of McKinley.K in STRUCTURE increased more slowly from K = 6–8 . The ΔK parameter [Evanno et al. K = 5 . Assignments were mostly to one of four single clusters within northern populations, with the exception of Tuolumne and Merced groves, whereas in the south, there was no clear correspondence between populations of origin and cluster assignment . The permuted RST is equivalent to an FST estimate, so that a significantly observed RST indicates a phylogeographic signal as a result of mutational divergence. Also, we tested for phylogeographic signal for northern and southern populations separately. Observed RST was greater than permuted RST for the northern populations, but the difference was not significant . For southern populations, permuted RST was slightly greater than observed RST .Global Ndiv = 1870; 95% CI; 589–6640), but about one‐third of the ancestral population size of 23,000 . RMAEs for parameter estimates of contemporary populations were low, with higher values for the past demographic estimates: Nanc = 0.37 and tanc = 0.35  was the most likely scenario  forest: End of a dynasty. The demise of giant sequoia was seen as the consequence of fire suppression that encourages competing vegetation and inhibits its germination and establishment. Although today, management of the groves is aimed at providing the conditions suitable for regeneration, the new threat comes from climatic change. It is worrisome that genetic diversity was generally moderate to low and particularly low in small northern groves. Under natural colonization, the northern groves might be expected to serve as source populations for colonization of new suitable habitat at higher latitudes, or at higher elevations, although the latter would likely result in smaller populations with competition among species for limited space. Our inferences of past demographic changes suggest that the decrease in summer rainfall has led to a general decline in giant sequoia that has been exacerbated by cold, dry glacial cycles. If, as predicted, future climate in the Sierra Nevada will entail reduced winter snow pack and reduced summer rainfall, giant sequoia will face increasing pressure to compete in its present range. As suggested by the spatial shifts that likely occurred during the late Pleistocene and Holocene, local displacements may be possible if the rapidity of climate change is slow enough that dispersal and establishment can keep pace. However, if conditions in the current range of the species become drier and warmer, the long‐term decline that we have detected is likely to continue, and the future of this iconic California paleoendemic may be a signal of what can be expected for other Mediterranean ecosystem paleoendemics. In the absence of human intervention to plant in suitable habitat, the dynasty may indeed be at an end.Axelrod titled his 1986 manuscript The Sierra redwood (None declared.Raw microsatellite genotype data for the full dataset of 357 individuals will be submitted to datadryad.org if the manuscript is accepted for publication. The individuals removed based on the criterion of 0.25 pairwise relatedness coefficient will be indicated in the microsatellite dataset. Conditions and priors for the DIYABC analyses are in the accompanying Supporting information for online publication.Table S1. Summary of microsatellite data by population.Table S2. Linkage equilibrium.Table S3. Priors for all demographic parameters used in DIYABC.Table S4. Partition from BAPS spatial clustering of individuals.Fig. S1. Lynch and Ritlands pairwise relatedness coefficients for the full dataset of 357 individuals.Fig. S2. Log probability of the data for increasing values of K in STRUCTURE.Fig. S3. Evanno et al. (K parameter for increasing values of K.o et al.  ΔK paramFig. S4. Scenario evaluation in DIYABC.Click here for additional data file.


A pattern of distribution of segments identical by descent between groups of East-West and South Slavs suggests shared ancestry or a modest gene flow between those two groups, which might derive from the historic spread of Slavic people.The Slavic branch of the Balto-Slavic sub-family of Indo-European languages underwent rapid divergence as a result of the spatial expansion of its speakers from Central-East Europe, in early medieval times. This expansion–mainly to East Europe and the northern Balkans–resulted in the incorporation of genetic components from numerous autochthonous populations into the Slavic gene pools. Here, we characterize genetic variation in all extant ethnic groups speaking Balto-Slavic languages by analyzing mitochondrial DNA , Y-chromosomes  and genome-wide SNP profiles (n = 296), within the context of other European populations. We also reassess the phylogeny of Slavic languages within the Balto-Slavic branch of Indo-European. We find that genetic distances among Balto-Slavic populations, based on autosomal and Y-chromosomal loci, show a high correlation (0.9) both with each other and with geography, but a slightly lower correlation (0.7) with mitochondrial DNA and linguistic affiliation. The data suggest that genetic diversity of the present-day Slavs was predominantly shaped  Balto-Slavic speakers comprise around one-third of present-day Europeans and occupy nearly a half of the European subcontinent. There is a near consensus among linguists that the Baltic and Slavic languages stem from a common root, Proto-Balto-Slavic, which separated from other Indo-European languages around 4,500–7,000 years before present (YBP) –8 and whAlthough there is no single archaeological signature for their spread, historical records suggest that a major Slavic expansion across Europe took place approximately 1,400–1,000 YBP –19; reviThe question of to what extent this recent cultural transformation within Europe affected its genetic landscape has been the subject of numerous studies. Uniparental genetic markers, mitochondrial DNA (mtDNA) and the non-recombining part of the Y-chromosome (NRY), indicate that the genetic composition of Slavs does not differ significantly from that of their neighboring non-Slavic populations –34. In aHere, our aim is to contribute to a comprehensive understanding of patrilineal, matrilineal and autosomal genetic variation in the Balto-Slavic-speaking peoples. The Balto-Slavic “case” allows us to test correlations across these three genetic systems in well-established linguistic and geographic space, and to address questions about the genetic history of the carriers of this large linguistic subfamily within the neighboring non-Balto-Slavic Indo-European, Finno-Ugric, North Caucasian and Turkic speakers. To do so, we analyze 6,876 mtDNAs, 6,079 NRYs and 296 whole genome SNP profiles representing all extant Balto-Slavic populations, of which 917, 2,392 and 70, respectively, are reported here for the first time. We complement our genetic study with linguistic evidence, in particular by refining the phylogeny of the extant Slavic languages.vsPC3, NRY and mtDNA MDS analyses, respectively    , indicating thereby an extended geographic area with low genetic differentiation among the majority of Slavic speakers across Central-East Europe.The genetic structuring of Balto-Slavic populations  in a Eurectively . In the ectively . In theiectively . This grNei = 0.239 for NRY; FST = 0.0009 for autosomal data)  and eastern (Macedonians and Bulgarians) regions of the Balkan Peninsula with Serbians placed in-between . The meaBoth extant Baltic-speaking populations, Latvians and Lithuanians, lie in the vicinity of Finno-Ugric-speaking Estonians according to their Y-chromosome diversity , whilst The patterns of genetic structure of the Balto-Slavic populations agree particularly between autosomal and NRY data. However, the maternal gene pool of the Balto-Slavic populations, although less structured possibly due to somewhat lower phylogenetic resolution of the dataset , bears sdark blue (k3) and the light blue (k2), albeit in different proportions   . It is n yellow) , S2 Fig.fIBD algorithm [To analyze further the patterns of gene flow among the Balto-Slavic populations and their non-Slavic neighbors as well as to explore the genetic heritage of the suggested Slavic migration from Central-East to the Balkan region of Europe, we focused on the pairwise sharing of IBD segments ,52 and algorithm . We creaIBD analysis  reveals South Slavs in their turn share a similar number of IBD segments with East-West Slavs and with the ‘inter-Slavic’ Romanian, Hungarian and Gagauz populations . NotablyAltogether, the analysis of IBD segment distributions revealed even patterns of IBD sharing among East-West Slavs–‘inter-Slavic’ populations –and South Slavs, i.e. across an area of assumed historic movements of people including Slavs.We applied a lexicostatistical approach to refine the phylogeny of the extant Balto-Slavic languages ,7,54, foAnalysis of molecular variance (AMOVA) partitions the overall genetic diversity in a group of populations into fractions according to hierarchical levels of population structure. We analyzed the distribution of the NRY diversity among three levels of the linguistic tree of Balto-Slavic languages see , S5 Fig.A Mantel test was applied to compare the roles which geography and language have played in shaping the genetic variation of the Balto-Slavic populations . The tesThe results of our study have shown the close genetic proximity of the majority of West and East Slavic populations inhabiting the geographic area from Poland in the west, to the Volga River in the East . Some mtIn contrast to this apparent genetic homogeneity of the majority of West and East Slavs, the gene pool of South Slavs, who are confined to the geographically smaller Balkan Peninsula, differs substantially and shows internal differentiation, as testified by their NRY and autosomal variation . ConsequThe importance of these substrata in shaping the genetic diversity of the present-day Slavs is evident from the observed lower IBD relatedness between the combined group of East-West Slavs and South Slavs than with north-east Europeans, including Baltic speakers . The latMost West and East Slavs of Central-East Europe form genetically a compact group of populations that, as a general rule, differ from their western (Germanic-speaking) and eastern (Finno-Ugric-speaking) neighbors . Howeverin situ gene pool shaping, are needed to explain the genetic patterns observed on the eastern, north-eastern and western margins of the current ‘Slavic area’ within Central-East Europe.Taken together, several mechanisms including cultural assimilation of the autochthonous populations by expanding Slavs while maintaining the pre-Slavic genetic boundaries, and The presence of two distinct genetic substrata in the genomes of East-West and South Slavs would imply cultural assimilation of indigenous populations by bearers of Slavic languages as a major mechanism of the spread of Slavic languages to the Balkan Peninsula. Yet, it is worthwhile to add here evidence from the analysis of IBD segments: the majority of Slavs from Central-East Europe (West and East) share as many IBD segments with the South Slavs in the Balkan Peninsula as they share with non-Slavic populations residing nowadays between Slavs . This evExpansion of Slavic languages took place in an area already occupied by speakers of the Baltic languages ,50. DespComparing genetic and linguistic reconstructions with geography has a long tradition in human population genetics . Here, wCombining all lines of evidence, we suggest that the major part of the within-Balto-Slavic genetic variation can be primarily attributed to the assimilation of the pre-existing regional genetic components, which differed for West, East and South Slavic-speaking peoples as we know them today.The DNA samples analysed in the study were collected after having obtained written informed consent. The procedure has been approved by Ethics Committees of the appropriate Institutions, including the Research Ethics Committee of the University of Tartu (UT REC) (no 225/T-9) and the Ethics Committee of the Research Centre for Medical Genetics, Russian Academy of Sciences.The NRY data comprises 6,079 samples, including 1,254 reported here for the first time and 1,138 samples updated from previous work  technology as described . MtDNA asmartpca program of the EIGENSOFT package [MDS analysis based on genetic distances  was perf package ; sets of package ; sets ofvs South Slavs (group2) (i.e. IBD relatedness within Slavs) to the IBD relatedness between each group of Slavs vs their respective neighboring groups of mostly non-Slavic populaitons (Table F in fIBD) algorithm [http://faculty.washington.edu/browning/beagle/beagle.html) was used to detect pairwise IBD segments. Sets of Illumina-only cross-platform SNPs  were used in the analysis. See We aimed to compare the level of IBD relatedness between the combined group of East-West Slavs (group1) lgorithm  implemenAMOVA (implemented in the Arlequin 3.11) was applied to estimate genetic differentiation when Balto-Slavic populations were grouped according to the three hierarchical levels of the tree of Balto-Slavic languages , S5 Fig.20 wordlists of extant Balto-Slavic languages were used to reconstruct their phylogeny. The consensus tree  was drawS1 Datasetbslav.dbf, bslav.var, bslav.inf, lexical dataset in STARLING format . This dataset exported in MS EXCEL format is available as Table A in bslav.nex, the same dataset as a binary matrix in NEXUS format.*.tre, some of the discussed trees in NEWICK format;NEXUS files for NeighborNet networks.(zip-archive).(ZIP)Click here for additional data file.S1 Fig(PDF)Click here for additional data file.S2 Fig(PDF)Click here for additional data file.S3 Fighttp://www.conceptdraw.com/How-To-Guide/geo-map-europe.Populations within each group are listed in Table F in (PDF)Click here for additional data file.S4 FigRussians from Northern region of European part of Russia are considered separately from the group of north-east Europeans. The x-axis indicates ten classes of IBD segment length (in cM); the y-axis indicates the average number of shared IBD segments per pair of individuals within each length class.(PDF)Click here for additional data file.S5 Fig(PDF)Click here for additional data file.S1 File(XLSX)Click here for additional data file.S2 FileFig A in S2 File. Geographical distribution of extant Slavic and East Baltic languages and dialects used in the study. Map was prepared by Yuri Koryakov. Fig B in S2 File. Dated phylogenetic tree of the Balto-Slavic lects produced by the StarlingNJ method from the multistate matrix (binary nodes only). Bootstrap values are shown near the nodes . Fig C in S2 File. Phylogenetic tree of the Balto-Slavic lects produced by the NJ method from the binary matrix in the SplitsTree4 software. Bootstrap values are shown near the nodes . Branch length reflects the relative rate of cognate replacement as suggested by SplitsTree4. The BioNJ method yields the same topology. Fig D in S2 File. Phylogenetic tree of the Balto-Slavic lects produced by the UPGMA method from the binary matrix in the SplitsTree4 software. Bootstrap values are shown near the nodes . Branch length reflects the relative rate of cognate replacement as suggested by SplitsTree4. Fig E in S2 File. Consensus phylogenetic tree of the Balto-Slavic lects produced by the Bayesian MCMC method from the binary matrix in the MrBayes software. Bayesian posterior probabilities are shown near the nodes (not shown for stable nodes with P ≥ 0.95). Branch length reflects the relative rate of cognate replacement as suggested by MrBayes. Fig F in S2 File. Optimal phylogenetic tree of the Balto-Slavic lects produced by the UMP method from the binary matrix in the TNT software. Bootstrap values are shown near the nodes . Branch length reflects the relative rate of cognate replacement as suggested by TNT. Fig G in (PDF)Click here for additional data file.S3 File(XLSX)Click here for additional data file.S1 Text(DOCX)Click here for additional data file.


Plasmodiumfalciparum malaria in China. However, few studies have investigated the characteristics of this parasite. Therefore, this study aimed to evaluate the genetic diversity and population structure of P. falciparum to predict the geographic origin of falciparum malaria.Yunnan and Hainan provinces are the two major endemic regions for P. falciparum isolates obtained from blood samples collected from Yunnan and Hainan provinces of South China. The isolates were analysed for genetic diversity, linkage disequilibrium, and population structure. The parasite populations were clustered into two subgroups  and a classification algorithm was used to identify molecular markers for classifying the P. falciparum populations.Thirteen highly polymorphic microsatellite loci were studied to estimate the genetic diversity and population structure of 425 He) in Yunnan and Hainan was 0.766 ± 0.036 and 0.677 ± 0.039, respectively, revealing a moderate high level of genetic diversity. Significant linkage disequilibrium was found for some regions of Yunnan (Lazan county and Xishuangbanna region) and Hainan (Dongfang city and Sanya city) province. According to the classification algorithm, a combination of three microsatellites could be used as a discriminatory marker to identify the origin of P. falciparum isolates.All 13 microsatellite loci were highly polymorphic, with the number of alleles per locus varying from 5 to 20. The mean expected heterozygosity  contains supplementary material, which is available to authorized users. Plasmodium falciparum  is the most severe form and can be fatal n, [T]n, and [TAA]n repeats  (1 − ∑Pi2), where n is the number of sampled infections and Pi is the frequency of the ith allele.Population genetics were preferably assessed only in samples with single infections, because the use of samples with multiple infections could result in bias. To measure the genetic diversity, the number of haplotypes (h), number of different alleles  was used to assess multilocus LD in each P. falciparum population from Yunnan and Hainan provinces. The IAS values were calculated using the formula (VD/Ve − 1)/(r − 1) with permutation testing of the null hypothesis of IAS considered as 0 (complete LD), where VD represents the observed variance in mismatch values, Ve represents the expected variance in mismatch values, and r represents the number of analysed loci. The significance of the IAS values was tested using the Monte Carlo method.Multilocus LD, defined as the nonrandom association among alleles from all 13 screened loci, was analysed using the program LIAN version 3.5 . The staThe pairwise Fst indices was employed to quantify the genetic distance between pairs of populations, and was calculated using Arlequin 3.1 . The genThe program STRUCTURE version 2.3.4 , 36 was P. falciparum isolates between Yunnan and Hainan provinces, data were analysed using Weka software (version 3.7.1) [To identify the most reliable discriminatory microsatellite marker sets that would best differentiate n 3.7.1) . The clan 3.7.1) , with stOf the total of 425 blood samples, 358 samples were successfully genotyped at all 13 microsatellites , Np (30), and mean He (0.766 ± 0.036) compared with the isolates from Hainan, who showed corresponding values of 6.923 ± 0.923, 4, and 0.677 ± 0.039, respectively.Next, comparison of the overall genetic diversity of Na (9.000 ± 1.092) for the isolates from TC, and the highest mean He for the isolates from LZ (0.726 ± 0.047) and DH (0.726 ± 0.053).Comparison of the diversity of the 13 microsatellites for the isolates from the seven regions showed the highest mean LD was measured first for all complete haplotypes, including single and multiple infections n = 358), and subsequently tested only for single infections by removing alleles from multiple infections (n = 306). Because all haplotypes in the dataset were unique, it was not necessary to measure LD among unique haplotypes, as observed in a previous study . A stron58, and sP. falciparum populations studied were shown in Table Pairwise comporations of genetic distance between all P. falciparum populations according to the allele frequencies at each locus. According to the 13 microsatellite data set, cluster analysis showed that the primary peak of ΔK was observed at K = 2 . In Pop1, 93.8% members from the Yunnan–Myanmar border regions and 6.2% members from Hainan province were clustered together. In contrast, 72.1% members from Hainan province clustered with 27.9% members from the Yunnan province in Pop2. Hence, the parasite populations were clustered based on their geographic origins. When the number of subgroups was increased from K = 2 to K = 5  was launched in 2010 with the goal to eliminate malaria from all regions by 2015, except the border regions in Yunnan province, and to completely eliminate malaria all over China by 2020 . To guidP. falciparum in South China using highly polymorphic neutral markers and samples of field isolates collected from 2004 to 2008. All 13 trinucleotide microsatellite markers were extremely polymorphic for both Yunnan and Hainan provinces. The genetic diversity observed in the two P. falciparum populations was slightly different. For Yunnan Province, the microsatellite markers revealed high variation, the mean Na was 11.692 ± 1.216, and the mean He was 0.766 ± 0.036. While, for Hainan province, the microsatellite markers showed intermediate variation, the mean Na was 6.923 ± 0.923, and the mean He was 0.677 ± 0.039. However, the genetic diversity of plasmodium falciparum in South China (0.642–0.726) was slightly higher than that reported in several studies in southeast Asian countries [Currently, falciparum malaria has been eliminated in most provinces because of the large-scale control activities. However, Yunnan and Hainan provinces of Southern China remain to be major endemic regions , 3. Thisountries ,such as ountries , 20.P. falciparum populations from some regions of Yunnan and Hainan provinces. Deviate from the general pattern established by Anderson et al.: microsatellite data revealed a spectrum of population structures in a single parasite species, that is, a strong LD was observed for regions of low transmission, while a weak LD was observed for regions of high transmission [P. falciparum due to some epidemiological or ecological reason.Despite the high levels of genetic diversity, a significant linkage disequilibrium was found for the smission . The LD smission . The samP. falciparum populations, the proportion of multiple infections are related to the transmission intensity, which is an important parameter affecting P. falciparum mating because cross fertilization and recombination may occur between parasite gametes of either the same or different genotypes of the mosquito vector [Generally, with regard to local o vector . High leo vector , 44, 45,The level of genetic differentiation were moderate (Fst 0.039–0.214) between the seven populations in South China, and comparable with that of Philippines (Fst 0.096–0.144), and lower than that of Malaysian Borneo (0.038–0.376). High level of genetic differentiation of the populations in Malaysian Borneo are considered to be a result of fragmentation of the population structure owing to an effective malaria control and low migration rate of people in the endemic areas. Nowadays, due to the intense population movements within the country, there were high migration rate of people in the endemic areas, which may causing low level of genetic differentiation.P. falciparum populations from South China were primarily structured into two major clusters by STRUCTURE software. The populations were found to be clearly clustered according to their geographic origin, except some isolates collected from LeD and SY, probably because of their limited sample size. Furthermore, the median-joining network diagram tree was used to clarify these findings. A further analysis using the STRUCTURE program at K = 3–5 revealed that, although microsatellite analysis can still discriminate P. falciaprum populations from Yunnan and Hainan, it can have limitations in distinguishing P. falciparum cases from each site. Among the Yunnan populations, we observed a large amount of mixing among the LZ, TC, and DH populations. This was probably due to the high level of transmission in China-Myanmar border [P. falciparum cases using only microsatellite markers. However, the P. falciparum population from BN, which is also located in Yunnan province but far away from TC and DH, was quiet different from that from the other three regions and was relatively simple. This could be due to the limited human movements. For the Hainan populations, especially in DF population, the P. falciparum isolates collected from 2004 to 2007 seems have a common ancestor with LeD and SY. While, the isolates collected in 2008 may have a different ancestor. The analysis results of isolates collected from different years, suggesting another outbreak of falciparum malaria for unknown reasons . Nevertheless, despite the limitations in distinguishing P. falciparum cases from each site, microsatellite-based analysis with the STRUCTURE and Network programs confirmed the presence of two subgroups in Southern China, namely Yunnan and Hainan.Population assignment is an important aspect of epidemiological studies. As demonstrated by the dendrogram based on the pairwise Fst matrix, the r border , and it P. falciparum isolates from South China was performed. Two marker sets containing three microsatellite loci were selected and used to establish a stepwise classification, which correctly classified more than 92% isolates. The results clearly demonstrate the feasibility of accurate classification of individual isolates from a geographic region and the effectiveness of the selected marker sets as applicable tools for tracing the origin of falciparum malaria outbreaks in the future.Further statistical analysis to identify effective marker sets for discriminating the origin of P. falciparum parasites in Southern China using 13 highly polymorphic microsatellite loci. The results showed a high level of genetic diversity co-occurring with significant multilocus LD. In addition, the parasite populations were clustered into two subgroups according to their geographic locations, and the intensity of human movements could affect population assignment. Finally, two marker sets were selected to be used for tracing the origin of falciparum malaria outbreaks in the future. These findings provide important information for understanding the population structure of P. falciparum in Southern China and will allow investigators to focus their attention on very small geographic regions and rapidly formulate appropriate therapeutic strategies, which will eventually control and eliminate this disease completely.This is the first study to report the genetic diversity, transmission patterns, and population structure of


Ficus insipida subsp. insipida in order to investigate patterns of spatial genetic structure across the Neotropics and within Amazonia.To examine the phylogeography of Neotropics.trnH–psbA; 410 individuals from 54 populations) and nuclear ribosomal internal transcribed spacer  sequences were sampled from Mexico to Bolivia, representing the full extent of the taxon's distribution. Divergence of plastid lineages was dated using a Bayesian coalescent approach. Genetic diversity was assessed with indices of haplotype and nucleotide diversities, and genetic structure was examined using spatial analysis of molecular variance (SAMOVA) and haplotype networks. Population expansion within Amazonia was tested using neutrality and mismatch distribution tests.Plastid DNA (trnH–psbA sequences yielded 19 haplotypes restricted to either Mesoamerica or Amazonia; six haplotypes were found among ITS sequences. Diversification of the plastid DNA haplotypes began c. 14.6 Ma. Haplotype diversity for trnH–psbA was higher in Amazonia. Seven genetically differentiated SAMOVA groups were described for trnH–psbA, of which two were also supported by the presence of unique ITS sequences. Population expansion was suggested for both markers for the SAMOVA group that contains most Amazonian populations.F. insipida between Mesoamerica and Amazonia, implying that the Andes and seasonally dry areas of northern South America are eco-climatic barriers to its migration. This pattern is shared with other widespread pioneer species affiliated to wet habitats, indicating that the ecological characteristics of species may impact upon large-scale phylogeography. Ficus insipida also shows genetic structure in north-western Amazonia potentially related to pre-Pleistocene historical events. In contrast, evident population expansion elsewhere in Amazonia, in particular the presence of genetically uniform populations across the south-west, indicate recent colonization. Our findings are consistent with palaeoecological data that suggest recent post-glacial expansion of Amazonian forests in the south.Our results show marked population genetic structure in  Phylogeographical studies can give insights into past changes in species distributions that can be related to environmental change and the history of landscapes Avise, . Phylogeet al., et al., et al., et al., At a broad-scale, low genetic structure has been inferred from nuclear markers of pioneer species across the Neotropics indicating high gene flow for pollen and/or seeds and/or recent colonization over large areas  in south-eastern Peru suggests a zone of secondary contact between two historically isolated populations  at a broad-scale across the Neotropics and at a regional-scale within Amazonia. This subspecies is a key element of early successional Neotropical rain forest communities. It is a good exemplar taxon for widespread Neotropical rain forest trees because it is distributed across the Andes and into Mesoamerica, is a long-lived pioneer tree confined to humid environments, and, like the majority of rain forest trees, has animal-dispersed seeds. We used an intensive range-wide geographical sampling scheme and worked with both plastid and nuclear DNA markers. Our specific objectives were: (1) to estimate divergence time among lineages; (2) to compare genetic diversity within and among populations and between Mesoamerica and Amazonia; (3) to test population genetic structure and the spatial location of genetic breaks; (4) to establish where populations of F. insipida subsp. insipida show genetic uniformity or distinctiveness in Amazonia; and (5) to establish if the regional patterns we find might reflect Quaternary climate changes or the legacies of more ancient geological events.Here, we examine the phylogeography of Ficus L. comprises c. 750 species  and Paraguay have similar leaf morphology to F. insipida subsp. insipida but have smaller stipules (1–4 cm long), different coloration in the mature fruits, and non-overlapping geographical ranges  and Amazonia (n = 23 sites). At least eight individuals were collected at each site, although fewer samples were sourced from several sites where the species was rare. In some cases our sampling was supplemented using herbarium specimens. Leaf samples were dried and stored in silica gel and the locations of individuals were recorded using a handheld GPS. For Amazonian populations, at least one herbarium voucher was collected from each population (see Appendix S1 in Supporting Information).Our study is based principally on leaf samples collected from the field for 410 individual trees of rpl32–trnL, trnQ–5′-rps16, 3′trnV–ndHC, atpI–atpH, trnD–trnT, trnH–psbA and trnL–trnF  of the nuclear ribosomal DNA was amplified and sequenced for 77 samples from Amazonia using the ITS1 and ITS4 primers , 0.2 μm of each primer and 1 U JumpStartTM Taq polymerase  , 0.2 μL of Taq polymerase  and 6.8 μL of distilled H2O. However, identical cycling programs were used for samples of both geographical regions. The thermal cycle for trnH–psbA (and ITS) was 94 °C for 5 min (3 min), followed by 35 (30) cycles at 94 °C for 30 s (1 min), 55 °C (56 °C) for 30 s (1 min) and 72 °C for 1 min (90 s), and a final extension at 72 °C for 10 min (5 min). Mesoamerican PCR products were sequenced as in Poelchau & Hamrick  and nucleotide substitutions, indels (i.e. insertions or deletions) and inversions were visually checked against the original electropherograms. The alignment was created manually in Mesquite 2.74 , Americana , Sycomorus (EU213825) and Galoglychia (EU213821) and Poulsenia armata (tribe Castilleae) were used as multiple outgroups. Inversions and indels were excluded from the analysis.Bayesian inference was used to estimate divergence time among plastid DNA haplotypes. The ingroup comprised all plastid haplotypes of beast 1.6.2 .The phylogenetic analysis suggested that three outgroup taxa, pida Fig.. These strnH–psbA and ITS sequences using statistical parsimony in tcs 1.21  using the packages ape  and by using haplotype frequencies (GST). The presence of a phylogenetic component to the phylogeographical structure was assessed by testing whether NST is significantly higher than GST based on 10,000 permutations in PermutCpSSR 2.0  and 100 annealing simulations for each K. In each run, populations were clustered into genetically and geographically homogenous groups . Genetic structure among groups of populations defined by geographical region and by SAMOVA was further examined by analysis of molecular variance computing a distance matrix in Arlequin 3.5. Significance of genetic structure indices was tested using a nonparametric randomization procedure.Genetic differentiation among populations was estimated by computing a distance matrix based on the number of mutational steps between haplotypes , time since the expansion (t) can be calculated as τ =2μt, where μ is the mutation rate for the total length of each DNA marker. A mean mutation rate of 1.64 × 10−9 substitutions site−1 year−1 (s s−1 yr−1) for ITS was taken from Dick et al. . The single accessions representing each of these three species come from Barro Colorado Island, Panama (Mesoamerica), but F. tonduzii and F. maxima have sequences related to Amazonian haplotypes of F. insipida and therefore do not cluster geographically with Mesoamerican F. insipida haplotypes. Finally, a GenBank accession of F. insipida subsp. insipida (GQ982221) also collected in Barro Colorado Island is nested within Mesoamerican F. insipida haplotypes .Diversification of plastid DNA haplotypes in trnH–psbA marker, a total of 410 individuals collected in 54 populations were successfully sequenced (Tablen = 31 sites) and 182 from Amazonia (n = 23 sites). After the exclusion of two variable mononucleotide repeats, 340 bp of aligned sequences remained. A total of 19 polymorphic sites were detected including 14 substitutions, four indels and one inversion (see Appendix S3); these mutations defined a total of 19 haplotypes , with higher mean genetic distance between populations in different regions (0.18) than within each region (Amazonia = 0.06 and Mesoamerica = 0.05). Genetic differentiation among populations was significantly higher when computed using a distance matrix (NST = 0.81) than when using haplotype frequencies , indicating some significant phylogeographical structuring. The SAMOVA analysis showed increasing values of differentiation among groups up to a K value of 7 (ΦCT = 0.8). Four groups were defined in Mesoamerica, primarily segregating populations containing haplotypes H8 (group I), H9 (group II) and H7 (group III) from the rest of the Mesoamerican populations  for ITS  were found in the 635 bp of aligned sequences. These mutations defined six haplotypes of which haplotype H1 is widespread across Amazonia and extends to Panama in Central America. Haplotype H3 occurs in three localities in Amazonia and the remaining four haplotypes each occur in single populations in Amazonia and Mesoamerica  is the result of hybridization with F. insipida subsp. insipida. However, we can at least conclude that hybridization is not frequent given that the species are morphologically distinct from other Ficus species, and intermediates have not been observed in the field . However, the accessions of F. tonduzii and F. maxima from Barro Colorado Island, Panama, share haplotypes with Amazonian accessions of F. insipida subsp. insipida (see Appendix S2b), and if the phylogeographical patterns we observe are due to hybridization we would expect to see geographical sharing of haplotypes. More fundamentally, the key patterns of the population genetic analyses of F. insipida subsp. insipida we observe and discuss below are robust regardless of concerns about species monophyly or the origin of haplotypes. These are a clear divide in chloroplast lineages between geographical areas, which can only be explained via lack of seed flow, and the presence of widespread haplotypes, which suggests recent population expansion.Our phylogenetic analysis showed that three of the outgroup taxa, pida Fig.. The lacF. insipida subsp. insipida, with differentiation between Mesoamerican and Amazonian haplotypes demonstrated by the haplotype network  with subsequent dispersal from Amazonia to Mesoamerica only in the case of haplotype H9 . Therefore, the Andes are a significant potential dispersal barrier and must have been since 3 Ma when the most recent uplift of the Eastern Cordillera caused it to reach 2500 m a.s.l.  greatly exceeding the current elevational limits of lowland rain forest trees. ht Berg, . Thus, tSchizolobium parahyba , Ecuador (32 and 33) and Bolivia (53), are separated from SAMOVA group VII containing the remaining populations. The plastid haplotype found at Sacta (population 53 in Bolivia) may represent an extreme case of ancestral polymorphism, or alternatively, it may reflect a recent long-distance dispersal event from Ecuador, the only other area where we observed its presence. Despite the extensive sampling carried out in this study, it is also possible that this haplotype is present in the intervening area in unsampled trees. While the three SAMOVA groups have widespread haplotypes (e.g. H1 and H5), they additionally contain a haplotype set which is restricted to this area and belongs to a phylogenetically distinct clade that diversified approximately 4.8 Ma  and early Pliocene (c. 4.5 Ma)  has experienced recent demographic expansion, most likely during the Pleistocene  may have experienced range contraction at the southern end of its range and a reduction in effective population size. By contrast, the presence of populations containing high haplotype and nucleotide diversity (e.g. population 33 in Ecuador) for the plastid marker suggest that F. insipida subsp. insipida may have persisted in ‘refugia’ in wetter forest of north-western Amazonia.The population differentiation of ene Table. MoreoveF. insipida subsp. insipida is consistent with episodes of effective seed dispersal. Fig fruits are important for frugivores in the Neotropics, and fish and bats are the primary seed dispersers of F. insipida (Banack et al., et al., F. insipida, which is a light-demanding species that grows in riparian and disturbed areas (Berg, et al., The occurrence of widespread plastid haplotypes in some areas of the range of F. insipida subsp. insipida showed marked genetic differentiation between Amazonian and Mesoamerican populations in the plastid DNA marker trnH–psbA. This contrasts with previous studies of other Neotropical trees – for example Cordia alliodora, Ceiba pentandra, Jacaranda copaia, Trema micrantha – that have little differentiation between these areas. Although all these species are pioneers, only F. insipida subsp. insipida is confined to ever-wet habitats. This suggests that the ecological characteristics of species, in this case a requirement of moist conditions for regeneration and survival, seem to be important drivers of phylogeographical patterns in Neotropical trees. The tolerance of seasonally dry climates in the other species suggests that seasonally dry tropical forest and savannas in northern South America did not represent long-term barriers to their migration. In contrast, areas with seasonally dry climates may have significantly restricted seed dispersal of F. insipida subsp. insipida. Further phylogeographical studies of widespread species will be needed to determine the extent to which ecological characteristics have determined historical migration patterns of other Neotropical tree species.Our phylogeographical study of F. insipida subsp. insipida also shows genetic differentiation among populations of the northern basin. Pre-Pleistocene events related to the changes of the landscape caused by the uplift of the Andes may be responsible for the initiation of population differentiation there. In contrast, the tentative evidence for demographic expansion in the rest of the basin, in particular the presence of genetically uniform populations across southern Amazonia, may indicate more recent colonization events, a scenario consistent with the palaeoecological data that indicates post-glacial rain forest expansion close to the southern margins of the forest.Within Amazonia, the plastid marker of


Background: Many amphibian species are negatively affected by habitat change due to anthropogenic activities. Populations distributed over modified landscapes may be subject to local extinction or may be relegated to the remaining—likely isolated and possibly degraded—patches of available habitat. Isolation without gene flow could lead to variability in phenotypic traits owing to differences in local selective pressures such as environmental structure, microclimate, or site-specific species assemblages.Methods: Here, we tested the microevolution hypothesis by evaluating the acoustic parameters of 349 advertisement calls from 15 males from six populations of the endangered amphibian species Proceratophrys moratoi. In addition, we analyzed the genetic distances among populations and the genetic diversity with a haplotype network analysis. We performed cluster analysis on acoustic data based on the Bray-Curtis index of similarity, using the UPGMA method. We correlated acoustic dissimilarities  with geographical and genetic distances among populations.Results: Spectral traits of the advertisement call of P. moratoi presented lower coefficients of variation than did temporal traits, both within and among males. Cluster analyses placed individuals without congruence in population or geographical distance, but recovered the species topology in relation to sister species. The genetic distance among populations was low; it did not exceed 0.4% for the most distant populations, and was not correlated with acoustic distance.Discussion: Both acoustic features and genetic sequences are highly conserved, suggesting that populations could be connected by recent migrations, and that they are subject to stabilizing selective forces. Although further studies are required, these findings add to a growing body of literature suggesting that this species would be a good candidate for a reintroduction program without negative effects on communication or genetic impact. The greatest threat to endangered amphibians is habitat change caused by anthropogenic activities, which alters resource availability, environmental quality, and ecological processes . Such neSurprisingly, several species thrive in modified sites even after profound anthropogenic transformation. Because these landscapes often exhibit physical, climatic, and biological  shifts, the ability of the remaining species to persist is likely a consequence of phenotypic plasticity in traits such as behavior, morphology, and reproduction .One of the most important phenotypic traits in evolutionary studies of anurans is the male advertisement call, because components of these calls are fundamental to species recognition and mate choice, and are thus under sexual selection . MoreoveThe anuran advertisement call is a multidimensional signal that can be viewed as a collection of spectral and temporal acoustic traits that are influenced, for example, by body size, air temperature, and social context . TherefoDifferent traits of advertisement calls may have distinct roles in anuran communication and, therefore, may evolve by distinct selective pressures . As a reHowever, acoustic variation is not always related to genetic variation at the population level . In thesProceratophrys moratoi. We tested the hypothesis that unique selective pressures among these remaining populations have led to divergence in acoustic traits and increased genetic structure.Herein, we speculated that historical modification of landscapes by agricultural crops has created barriers among persistent populations, and affected phenotypic and genetic traits in an endangered  NeotropiProceratophrys includes 40 South American frog species  an Audiotechnica AT 835b microphone and a Marantz PMD-222 recorder, (2) a Dynamic microphone and an Uher 4000 recorder, (3) a Sennheiser ME67/K6 microphone and a Boss 864 recorder, (4) a Sennheiser ME67/K6 microphone and a Marantz PMD671 recorder, or (5) a Sennheiser ME66/K6 microphone and an M–audio Microtrack II recorder. All recordings were made with sample rate of 44.1 or 48 kHz, and at 16-bit resolution. Acoustic recordings used in the present study are available in the Fonoteca Neotropical Jacques Vielliard, with collection numbers FNJV 10498, 10577, 12222–12224, 12228, the Smithsonian Institution website (http://vertebrates.si.edu/herps/frogs_boraceia/list.htm), and the personal collection of Ariovaldo A. Giaretta, which are detailed in the appendix of We analyzed 349 calls from 15 males of Acoustic analyses were performed in Raven Pro 64 1.4 for Windows (Cornell Lab of Ornithology), with the following settings: FFT (Fast Fourier Transformation) = 1,024; Overlap = 50 for spectral evaluations; and FFT = 256 and Overlap = 50 for temporal variables. Both temporal and spectral values were extracted from the spectrogram. We analyzed the following quantitative traits: frequency range, maximum frequency, minimum frequency, peak of dominant frequency, call duration, number of pulses per note, and pulse rate (pulses per second). Spectral measurements were obtained by selecting four variables in the source “choose measurements” in Raven: (1) Frequency 5% (Hz); (2) Frequency 95% (Hz)—these two measures include maximum frequency and minimum frequency, ignoring 5% below and above the total energy in the selected call; (3) Bandwidth 90% (Hz)—frequency range that included 90% of the energy distribution, i.e., the difference between Frequency 95% and Frequency 5%; (4) Max Frequency (Hz)—peak of dominant frequency . For temporal properties, we made precise selections on calls in the spectrogram, and visually counted the pulses.We calculated the variation in quantitative acoustic variables through the coefficient of variation  for both the among-males and within-males level. As defined by P. moratoi individuals from six populations were collected: four populations in the state of São Paulo: Itirapina (n = 5), São Carlos (n = 5), Bauru (n = 5), and Avaré (n = 2); and two populations in the state of Minas Gerais: Ituiutaba (n = 4) and Uberlândia (n = 5). This small sample size is in part attributable to the rarity of the species, and collection restrictions, as it is endangered and apparently extinct in at least two populations. Tissues were preserved in 95% ethanol. Voucher specimens were deposited in the Coleção Científica Jorge Jim indexed in Museu Nacional, Rio de Janeiro, Brazil . Tissue collection can be found in the Collection of tissue and chromosome preparation Shirlei Maria Recco Pimentel, Universidade Estadual de Campinas (UNICAMP), Campinas, São Paulo, Brazil . Total genomic DNA was extracted according to p distances) were computed from mitochondrial loci using MEGA 5.1 .Cluster analysis placed ividuals . IndividP. moratoi averaged 0.2% (0.0–0.4) between populations .We found 7 haplotypes in the 26 partial sequences of the mitochondrial 16S gene . Most haulations  and 0.25P. moratoi . Taken together, these results indicate minimal pressure for signal divergence, which could be explained by the following, non-exclusive hypotheses: (1) the reproductive environment in the sampled localities is similar with respect to habitat structure and background noise; (2) the female choice drives stable selection, which equalizes the general acoustic features of males from different localities; (3) the populations were recently connected, presenting traces of recent genetic flow; and (4) random evolutionary processes act on the calls  and elsewhere. Thus, the findings of the present study add to the growing body of literature supporting P. moratoi as a potential candidate for conservation actions, although additional work is necessary before an action plan could be initiated.Although our dataset is limited, we observed that acoustic and genetic variation appears to be conserved among individuals distributed across human-altered landscapes. Our preliminary results showed similar call types and genotypes  among different populations. Such reduced acoustic and possible genetic structure could be considered in future conservation actions; for example, these findings suggest that communication barriers (prezygotic) would pose no obstacle to reproduction  if a rei


Tuta absoluta, is native to South America and is invasive in the Mediterranean basin. The species' routes of invasion were investigated. The genetic variability of samples collected in South America, Europe, Africa and Middle East was analyzed using microsatellite markers to infer precisely the source of the invasive populations and to test the hypothesis of a single versus multiple introductions into the old world continents. This analysis provides strong evidence that the origin of the invading populations was unique and was close to or in Chile, and probably in Central Chile near the town of Talca in the district of Maule.The Lepidopteran pest of tomato,  Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), is a serious pest species native to South America that recently became a major threat to tomato production in the Mediterranean Basin and may become a problem in most countries of Africa, and Eurasia1Tuta absoluta was first observed outside its native range in Eastern Spain in 2006, and can now be found throughout Southern Europe, North Africa and the Middle East1T. absoluta invading the Mediterranean basin is required to help develop strategies to control this speciesT. absoluta. Here, we analyzed the genetic variability of samples collected in South America, Europe, Africa and the Middle East to infer precisely the source of the T. absoluta populations invading the Mediterranean Basin and to test the hypothesis of a single versus multiple introductions into the old world continents.The tomato borer, STF values and genotypic differentiation tests  to 0.31 (Argentina vs Venezuela), and p < 10−5). Samples from the same country in South America were not significantly differentiated with the notable exception of Southern and Northern Chile that were differentiated from Central Chile . Most samples from the invaded areas in Africa, Europe and Asia were moderately differentiated with mean STF values of 0.024, except the samples from South-Eastern Spain which displayed a high level of differentiation from all the other samples from the Mediterranean basin  tree grouped the samples of invasive  cluster . This stor K = 4 . In addiesponded . Moreoveesponded . These rT. absoluta invading the Mediterranean Basin with posterior probabilities >0.9 and non overlapping confidence intervals. Low type I and mean type II errors were obtained for both prior sets  analyses are shown in ior sets . When Cherformed . They clulations , with poulations .T. absoluta displayed very weak genetic structure so that it was not possible to infer the origin of invasive populations at a finer geographical scale. The absence of genetic structure in this region, as determined by the use of neutral genetic markers, indicates that the region surroundings Talca contains a single population of T. absoluta. The choice of new natural enemies (e.g. parasitoids) for possible use in biological control should take into account this new finding concerning the origin of the invasive population T. absoluta in the Mediterranean, as suggested by Roderick & NavajasThe main result of this study is that the origin of the invading populations around the Mediterranean was identified as unique and was in or close to Chile, and probably Central Chile in the district of Maule. Within this region, T. absoluta in South America is far from genetically homogeneous. Substantial genetic differentiation was found between northern and southern regions of South America, with more than 20% of the allele frequency variance found between southern Chile and the group of Venezuela and Colombia. Such a high level of differentiation is also found over smaller distances, for instance between Argentina and Chile or between Venezuela and Colombia. Such strong genetic structure in the native area does not usually facilitate the precise inference of the source of invasive populations because it requires an extensive sampling scheme.The second important conclusion of this study is that the native population of Harmonia axyridis expansion in FranceDiabrotica virgifera virgifera expansion in North America and Central EuropeThe third main result was that there was an almost complete absence of genetic structuring in the invaded areas, from southern Spain to Israel and from Israel to Morocco. This genetic homogeneity over space was measured using hyper variable microsatellite markers and therefore was probably not the consequence of a low power of analysis. Instead, it very probably corresponds to a single introduction in Africa or Spain followed by an expansion without noticeable a demographic bottleneck, which would have led to genetic differentiation through space. The same situation has already been found in other recent insect invasions, such as the Asian ladybeetle T. absoluta larvae were collected on tomato plants in greenhouses or in open field and stored in ethanol (>90%) prior to DNA extraction. Total genomic DNA of each sampled individual was extracted using the DNeasy Tissue Kit (Qiagen) following manufacturer's instructions. The genotypes of 966 individuals were obtained at 12 microsatellite markers FST valuesWe collected samples from various regions in South America, and from the invaded area in North Africa, Europe and Asia . T. absoethanol >0% prior FST values between each invasive population sample and each South American sample. 2) We then computed the mean individual assignment likelihoodLi→s) of each invading population sample i, to each possible South American source population using GENECLASS2i was determined as the South American population whose sample displays the minimum corrected FST values with i and the maximum mean individual assignment likelihood of i. 3) We also plotted a neighbor joining (NJ) treehttp://bioinformatics.org/~tryphon/populations/). It is expected that the source of a target invasive population's sample i is located in close proximity to i in the tree. 4) Finally, a Bayesian clustering analysis was performed using the STUCTURE software6 iterations for the MCMC and 2 × 105 iterations for the burn-in period were used. The most probable source of each target invasive population's sample i was determined as the population whose samples were the last ones to still cluster with i with increasing values of K. The most likely value of K was determined using the method of Evanno et al.The most probable source population of the European and African invasive populations was investigated as followsst observation dates of the samples) and on historical and genetic parameters priors . The analyses were conducted twice, with different parameter prior distributions and with 2 different sets of samples representative of the STRUCTURE clusters. Spa_cas and Mar_lar (see An Approximate Bayesian Computation Analysiss priors . We conts priors . The puts priors  plus a n_lar see  samples  (ti i = , 2, 3, 4T.G., A.T. and N.D. conceived the sampling scheme and the experiments, T.G., E.L. and A.B. analyzed the data, I.L. and A.B. performed the experiments. N.D., M.R., E.T., A.T. and L.N. organized the sampling. All authors participated to the writing of the manuscript.Supplementary Information