Phylogeography of the endangered rosewood Dalbergia nigra (Fabaceae): insights into the evolutionary history and conservation of the Brazilian Atlantic Forest

The Brazilian rosewood (Dalbergia nigra) is an endangered tree endemic to the central Brazilian Atlantic Forest, one of the world''s most threatened biomes. The population diversity, phylogeographic structure and demographic history of this species were investigated using the variation in the chloroplast DNA (cpDNA) sequences of 185 individuals from 19 populations along the geographical range of the species. Fifteen haplotypes were detected in the analysis of 1297 bp from two non-coding sequences, trnV-trnM and trnL. We identified a strong genetic structure (F_ST=0.62, P<0.0001), with a latitudinal separation into three phylogeographic groups. The two northernmost groups showed evidence of having maintained historically larger populations than the southernmost group. Estimates of divergence times between these groups pointed to vicariance events in the Middle Pleistocene (ca. 350 000–780 000 years ago). The recurrence of past climatic changes in the central part of the Atlantic forest, with cycles of forest expansion and contraction, may have led to repeated vicariance events, resulting in the genetic differentiation of these groups. Based on comparisons among the populations of large reserves and small, disturbed fragments of the same phylogeographic group, we also found evidence of recent anthropogenic effects on genetic diversity. The results were also analysed with the aim of contributing to the conservation of D. nigra. We suggest that the three phylogeographic groups could be considered as three distinct management units. Based on the genetic diversity and uniqueness of the populations, we also indicate priority areas for conservation.     

Fine-scale genetic structure of the threatened rosewood Dalbergia nigra from the Atlantic Forest: comparing saplings versus adults and small fragment versus continuous forest

Assessing the population genetic structure of threatened species is important for developing successful conservation strategies. In this study, we evaluated the fine-scale spatial genetic structure (SGS) of Dalbergia nigra from a regenerating secondary forest fragment and compared it with previous data from a primary forest of a large reserve. A total of 107 adult and 111 saplings were mapped and genotyped for seven microsatellite loci. The genetic diversity was high and similar in adults (H _e?=?0.682) and saplings (H _e?=?0.680). The spatial extent of SGS was higher in adults than in saplings. Overlapping generations in the potentially reproductive individuals is the likely explanation for the higher SGS in adults (Sp?=?0.016) in relation to the saplings (Sp?=?0.010). The SGS in the adults from the secondary forest fragment was similar to that found in the primary forest. Considering the SGS found in adults, from both the secondary and primary forests, seeds for ex situ conservation should be collected from trees at least 80 m apart to reduce the genetic similarity between samples. These results highlight the importance of preserving small forest fragments to allow successful regeneration and maintenance of the genetic diversity in D. nigra.     

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

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

Short-distance gene flow in <Emphasis Type="Italic">Populus nigra</Emphasis> L. accounts for small-scale spatial genetic structures: implications for in situ conservation measures

The European black poplar (Populus nigra L.) is a major species of riparian softwood forests. Due to human influences, it is one of the most threatened tree species in Europe. For restoration purposes, remaining stands may act as source populations. We analysed a natural population of P. nigra for genetic diversity and spatial genetic structure using seven microsatellite markers. For the first time, paternity analysis of seedlings as well as juveniles from a restricted area of natural regeneration was used to quantify pollen- and seed-mediated gene flow, respectively. In both cases, cultivated P. x canadensis trees in vicinity could act as potential parents. Spatial genetic patterns of the adult tree population suggest small-scale isolation by distance due to short-distance gene flow, the major part of which (i.e. 70%) takes place within distances of less than 1 km. This helps to explain the reduced diversity in the juveniles. It has implications for the spatial management of natural regeneration areas within in situ conservation measures.     

Present,past and future of the European rock fern Asplenium fontanum: combining distribution modelling and population genetics to study the effect of climate change on geographic range and genetic diversity

Background and Aims

Climate change is expected to alter the geographic range of many plant species dramatically. Predicting this response will be critical to managing the conservation of plant resources and the effects of invasive species. The aim of this study was to predict the response of temperate homosporous ferns to climate change.

Methods

Genetic diversity and changes in distribution range were inferred for the diploid rock fern Asplenium fontanum along a South–North transect, extending from its putative last glacial maximum (LGM) refugia in southern France towards southern Germany and eastern-central France. This study reconciles observations from distribution models and phylogeographic analyses derived from plastid and nuclear diversity.

Key Results

Genetic diversity distribution and niche modelling propose that genetic diversity accumulates in the LGM climate refugium in southern France with the formation of a diversity gradient reflecting a slow, post-LGM range expansion towards the current distribution range. Evidence supports the fern''s preference for outcrossing, contradicting the expectation that homosporous ferns would populate new sites by single-spore colonization. Prediction of climate and distribution range change suggests that a dramatic loss of range and genetic diversity in this fern is possible. The observed migration is best described by the phalanx expansion model.

Conclusions

The results suggest that homosporous ferns reproducing preferentially by outcrossing accumulate genetic diversity primarily in LGM climate refugia and may be threatened if these areas disappear due to global climate change.     

Linking beta diversity patterns to protected areas: lessons from the Brazilian Atlantic Rainforest

Understanding the processes that drive patterns of beta diversity is crucial for planning conservation policies and for designing networks of protected area (PAs). Beta diversity can be decomposed into two components: 1—species turnover, the replacement of species by others resulting in a low proportion of shared species; 2—species nestedness—the result of differences in species richness, when a poorer community is a subset of species from a richer community. We aimed to evaluate beta diversity patterns and how they are represented in the network of PAs in southern Brazilian, regarding three forest types: Atlantic Forest s.s., Araucaria Forest, and Seasonal Forest. Beta diversity was partitioned into the turnover and nestedness components. Additionally, we examined spatial patterns of site similarity using distance decay curves. Beta diversity was mainly caused by species turnover (approx. 86%), with only a small contribution of nestedness (approx. 5%) in all three forests types. The patterns of distance decay curves revealed that even at small distances (50–100 km), we found a considerable decrease in similarities, reinforcing turnover patterns. As turnover brought the larger contribution to beta diversity, additional conservation efforts must target an increase in the number of PAs, that should be spread across each one of the regions, to maximize the protection of species diversity. Most of the PAs are currently limited to the eastern region and prioritize the Atlantic Forest s.s. Thus Araucaria Forest and Seasonal Forest should deserve special priority in new conservation actions, as they also contain high levels of species turnover.     

When physical oceanography meets population genetics: The case study of the genetic/evolutionary discontinuity in the endangered goliath grouper (Epinephelus itajara; Perciformes: Epinephelidae) with comments on the conservation of the species

Epinephelus itajara is one of the marine fish species most threatened for extinction and it is considered to be “critically endangered” by the IUCN. The present study evaluated the genetic diversity of the species and the genetic/evolutionary relationships of its populations along the Atlantic coast of South America. The results indicate relatively reduced genetic variation, re-emphasizing the low adaptive potential of the species. One of the populations presented relatively high degrees of genetic diversity and it is evolutionary isolated from the all other populations. The evidences indicate the existence of two Evolutionarily Significant Units comprising E. itajara in the Atlantic coast of South America and the conservation prospects for the species must take these evidences into account.     

Density and habitat use of one of the last jaguar populations of the Brazilian Atlantic Forest: Is there still hope?

The jaguar (Panthera onca) plays an important role in maintaining biodiversity and ecological processes. We evaluated the status of a jaguar population in one of the last stronghold habitats for its conservation in the Atlantic Forest, the Rio Doce State Park (RDSP). We used a random survey design from 2016/17 to estimate jaguar abundance and density as well as its occupancy and detection probabilities in the entire Park''s area. To monitor for temporal fluctuations in density and abundance, we used a systematic survey design in the southern portion of the Park where jaguars were more recorded when using the random approach. We then conducted two surveys in 2017/18 and 2020. Our 2016/17 random survey revealed that jaguar density (0.11 ± SE 0.28 individuals/100 km²) was the lowest obtained for the species across the Atlantic Forest. We noticed that jaguar density increased three times from 2017/18 (0.55 ± SE 0.45 individuals/100 km²) to 2020 (1.61 ± SE 0.6 individuals/100 km²). Jaguar occupancy and detection probability were 0.40 and 0.08, respectively. The low jaguar occupancy probability was positively associated with smaller distances from lakes and records of potential prey. The detection probability was positively associated with prey detection, the rainy season, and smaller distances from lakes. Our work contributes to a growing awareness of the potential conservation value of a protected area in a human‐dominated landscape as one of the last strongholds for jaguars across the Atlantic Forest.     

Biased morph ratios and skewed mating success contribute to loss of genetic diversity in the distylous Pulmonaria officinalis

Background and Aims

In heterostylous plant species, skewed morph ratios are not uncommon and may arise from a range of factors. Despite the recognized importance of skewed morph ratios on overall reproductive success within populations, little is known about the impact of skewed morph ratios on population genetic diversity and differentiation in heterostylous species. This study specifically aimed to clarify the effect of population size and morph bias on population genetic diversity and differentiation in the temperate forest herb Pulmonaria officinalis. This species is characterized by a distylous breeding system and shows morph-specific differences in reproductive success.

Methods

Genetic diversity was determined for 27 P. officinalis populations in northern Belgium by using eight recently developed microsatellite markers. Multiple regressions were used to assess the relationship between genetic diversity, morph bias and population size, and F_ST-values were calculated for short- and long-styled morphs separately to study genetic differentiation as a function of morph type.

Key Results

For all genetic measures used, morph bias was more important in explaining patterns of genetic diversity than population size, and in all cases patterns of population genetic diversity followed a quadratic function, which showed a symmetrical decrease in genetic diversity with increasing morph bias. However, probably due to the reproductive advantage of L-morphs relative to S-morphs, maximum genetic diversity was found in populations showing an excess of L-morphs (60·7 % L-morph). On the other hand, no significant difference in pairwise genetic distances between populations was observed between L- (0·107) and S-morphs (0·106).

Conclusions

Our results indicate that significant deviations from equal morph ratios not only affect plant reproductive success but also population genetic diversity of heterostylous plant species. Hence, when defining conservation measures for populations of heterostylous plant species, morph ratios should be considered as an important trait affecting their long-term population viability.     

Microsatellites reveal substantial among-population genetic differentiation and strong inbreeding in the relict fern Dryopteris aemula

Background and Aims

A previous study detected no allozyme diversity in Iberian populations of the buckler-fern Dryopteris aemula. The use of a more sensitive marker, such as microsatellites, was thus needed to reveal the genetic diversity, breeding system and spatial genetic structure of this species in natural populations.

Methods

Eight microsatellite loci for D. aemula were developed and their cross-amplification with other ferns was tested. Five polymorphic loci were used to characterize the amount and distribution of genetic diversity of D. aemula in three populations from the Iberian Peninsula and one population from the Azores.

Key Results

Most microsatellite markers developed were transferable to taxa close to D. aemula. Overall genetic variation was low (H_T = 0·447), but was higher in the Azorean population than in the Iberian populations of this species. Among-population genetic differentiation was high (F_ST = 0·520). All loci strongly departed from Hardy–Weinberg equilibrium. In the population where genetic structure was studied, no spatial autocorrelation was found in any distance class.

Conclusions

The higher genetic diversity observed in the Azorean population studied suggested a possible refugium in this region from which mainland Europe has been recolonized after the Pleistocene glaciations. High among-population genetic differentiation indicated restricted gene flow (i.e. lack of spore exchange) across the highly fragmented area occupied by D. aemula. The deviations from Hardy–Weinberg equilibrium reflected strong inbreeding in D. aemula, a trait rarely observed in homosporous ferns. The absence of spatial genetic structure indicated effective spore dispersal over short distances. Additionally, the cross-amplification of some D. aemula microsatellites makes them suitable for use in other Dryopteris taxa.     

Low levels of realized seed and pollen gene flow and strong spatial genetic structure in a small,isolated and fragmented population of the tropical tree Copaifera langsdorffii Desf

Over the past century, the Brazilian Atlantic forest has been reduced to small, isolated fragments of forest. Reproductive isolation theories predict a loss of genetic diversity and increases in inbreeding and spatial genetic structure (SGS) in such populations. We analysed eight microsatellite loci to investigate the pollen and seed dispersal patterns, genetic diversity, inbreeding and SGS of the tropical tree Copaifera langsdorffii in a small (4.8 ha), isolated population. All 112 adult trees and 128 seedlings found in the stand were sampled, mapped and genotyped. Seedlings had significantly lower levels of genetic diversity (A=16.5±0.45, mean±95% s.e.; H_e=0.838±0.006) than did adult trees (A=23.2±0.81; H_e=0.893±0.030). Parentage analysis did not indicate any seed immigration (m_seeds=0) and the pollen immigration rate was very low (m_pollen=0.047). The average distance of realized pollen dispersal within the stand was 94 m, with 81% of the pollen travelling <150 m. A significant negative correlation was found between the frequency and distance of pollen dispersal (r=−0.79, P<0.01), indicating that short-distance pollinations were more frequent. A significant SGS for both adults (∼50 m) and seedlings (∼20 m) was also found, indicating that most of the seeds were dispersed over short distances. The results suggested that the spatial isolation of populations by habitat fragmentation can restrict seed and pollen gene flow, increase SGS and affect the genetic diversity of future generations.     

High genetic homogeneity of an intertidal marine invertebrate along 8000 km of the Atlantic coast of the Americas

Phallusia nigra is a cosmopolitan solitary ascidian with a distinct niger-blue tunic that is considered to be an easy diagnostic feature, even in the field. The wide geographic distribution of this species may have been a result of the overconfidence of taxonomists and ecologists in the assignment of unknown specimens to this species, based on this simple diagnostic character. Indeed, there seems to be a correlation between cosmopolitanism and the presence of few, but very conspicuous diagnostic characters in benthic invertebrate species. Almost invariably, genetic studies have demonstrated that the cosmopolitanism of those species is artificial and that, rather, they are made up of groups of morphologically very similar, but genetically distinct, species. Thus, we decided to verify the specific status of Western Atlantic populations of P. nigra, from Miami (USA) to São Paulo (southeastern Brazil) by estimating genetic variation and population structure levels along the entire range of this ascidian in the Western Atlantic. The analysis of 10 allozyme systems shows high levels of mean heterozygosity (H=0.28) so that P. nigra has genetic variation levels more related with other invertebrates than to their, phylogenetically closer, chordate relatives (that show lower levels of heterozygosity). The genetic structure of the population was relatively high (F_ST=0.083) for conspecific populations, but much lower than what would be expected if they belonged to different species. High levels of gene identity also indicate that the Western Atlantic population of P. nigra constitutes a single species. This result suggests an extremely high dispersal capability of the larvae of this ascidian or a relatively recent range expansion of its populations. Since this species is commonly found in harbors, the anthropogenic transport (p.e., in ballast water) may have contributed for the observed genetic homogeneity.     

Morphological variation of the <Emphasis Type="Italic">Caloneis schumanniana</Emphasis> species complex (Bacillariophyceae) from different environmental conditions in North American streams

The aim of the present study is to test by molecular DNA data a hypothesis concerned to speciation by allopatry occurring in Mimagoniates microlepis, associated to the Serra do Mar mountain chain (Atlantic Rain Forest hotspot) in Southern Brazil. Overall genetic diversity and mean genetic distances were high, demonstrating both good conservation status and genetic differentiation. Neighbor-Joining (NJ) and parsimony analyses, together with population genetic parameters (Φ_ST, N_m, G_ST, and AMOVA), identified two main vicariant genetic/evolutionary stocks dividing the upper Iguaçu River samples from those of the coastal plains. Other well-supported intrinsic monophyletic clades were also identified, suggesting fast and remarkable speciation processes. In addition, the genetic, evolutionary, geographic, and phylogeographic evidences reinforced an occurring species complex. Moreover, these evolutionarily significant units (species complex) seem to be inside four natural biogeographic areas. Thus, the genesis and evolution of the Serra do Mar complex might be associated to diversification processes of M. microlepis. Such a consideration suggests that the areas including the upper Iguaçu River and the coastal plains of the states of São Paulo, Paraná, and Santa Catarina require distinct conservation policies involving one of the global biodiversity hotspots, namely, the Brazilian Atlantic Rain Forest.     

Patterns of Vertebrate Diversity and Protection in Brazil

Most conservation decisions take place at national or finer spatial scales. Providing useful information at such decision-making scales is essential for guiding the practice of conservation. Brazil is one of the world’s megadiverse countries, and consequently decisions about conservation in the country have a disproportionate impact on the survival of global biodiversity. For three groups of terrestrial vertebrates (birds, mammals, and amphibians), we examined geographic patterns of diversity and protection in Brazil, including that of endemic, small-ranged, and threatened species. To understand potential limitations of the data, we also explored how spatial bias in collection localities may influence the perceived patterns of diversity. The highest overall species richness is in the Amazon and Atlantic Forests, while the Atlantic Forest dominates in terms of country endemics and small-ranged species. Globally threatened species do not present a consistent pattern. Patterns for birds were similar to overall species richness, with higher concentrations of threatened species in the Atlantic Forest, while mammals show a more generalized pattern across the country and a high concentration in the Amazon. Few amphibians are listed as threatened, mostly in the Atlantic Forest. Data deficient mammals occur across the country, concentrating in the Amazon and southeast Atlantic Forest, and there are no data deficient birds in Brazil. In contrast, nearly a third of amphibians are data deficient, widespread across the country, but with a high concentration in the far southeast. Spatial biases in species locality data, however, possibly influence the perceived patterns of biodiversity. Regions with low sampling density need more biological studies, as do the many data deficient species. All biomes except the Amazon have less than 3% of their area under full protection. Reassuringly though, rates of protection do correlate with higher biodiversity, including higher levels of threatened and small-ranged species. Our results indicate a need for expanded formal protection in Brazil, especially in the Atlantic forest, and with an emphasis on fully protected areas.     

Very low mitochondrial variability in a stingless bee endemic to cerrado

Partamona mulata is a stingless bee species endemic to cerrado, a severely threatened phytogeographical domain. Clearing for pasture without proper soil treatment in the cerrado facilitates the proliferation of termite ground nests, which are the nesting sites for P. mulata. The genetic consequences of these changes in the cerrado environment for bee populations are still understudied. In this work, we analyzed the genetic diversity of 48 colonies of P. mulata collected throughout the species’ distribution range by sequencing two mitochondrial genes, cytochrome oxidase I and cytochrome B. A very low polymorphism rate was observed when compared to another Partamona species from the Atlantic forest. Exclusive haplotypes were observed in two of the five areas sampled. The sharing of two haplotypes between collection sites separated by a distance greater than the flight range of queens indicates an ancient distribution for these haplotypes. The low haplotype and nucleotide diversity observed here suggests that P. mulata is either a young species or one that has been through population bottlenecks. Locally predominant and exclusive haplotypes (H2 and H4) may have been derived from local remnants through cerrado deforestation and the expansion of a few colonies with abundant nesting sites.     

Life in the desert: The impact of geographic and environmental gradients on genetic diversity and population structure of Ivesia webberi

For range‐restricted species with disjunct populations, it is critical to characterize population genetic structure, gene flow, and factors that influence functional connectivity among populations in order to design effective conservation programs. In this study, we genotyped 314 individuals from 16 extant populations of Ivesia webberi, a United States federally threatened Great Basin Desert using six microsatellite loci. We assessed the effects of Euclidean distance, landscape features, and ecological dissimilarity on the pairwise genetic distance of the sampled populations, while also testing for a potential relationship between I. webberi genetic diversity and diversity in the vegetative communities. The results show low levels of genetic diversity overall (H _e = 0.200–0.441; H _o = 0.192–0.605) and high genetic differentiation among populations. Genetic diversity was structured along a geographic gradient, congruent with patterns of isolation by distance. Populations near the species’ range core have relatively high genetic diversity, supporting in part a central‐marginal pattern, while also showing some evidence for a metapopulation dynamic. Peripheral populations have lower genetic diversity, significantly higher genetic distances, and higher relatedness. Genotype cluster admixture results suggest a complex dispersal pattern among populations with dispersal direction and distance varying on the landscape. Pairwise genetic distance strongly correlates with elevation, actual evapotranspiration, and summer seasonal precipitation, indicating a role for isolation by environment, which the observed phenological mismatches among the populations also support. The significant correlation between pairwise genetic distance and floristic dissimilarity in the germinated soil seed bank suggests that annual regeneration in the plant communities contribute to the maintenance of genetic diversity in I. webberi.     

Endophytic and mycorrhizal fungi associated with roots of endangered native orchids from the Atlantic Forest, Brazil

The composition and diversity of fungal communities associated with three endangered orchid species, Hadrolaelia jongheana, Hoffmannseggella caulescens, and Hoffmannseggella cinnabarina, found in different vegetation formations of the Atlantic Forest were determined by constructing clone libraries and by applying diversity and richness indices. Our results demonstrated the presence of Basidiomycetes. Sebacinales (81.61 %) and Cantharellales (12.10 %) were the dominant orders and are potential candidates for orchid mycorrhizal fungi. The Ascomycetes identified included the Helotiales (29.31 %), Capnodiales (18.10 %), and Sordariales (10.34 %), among others. These orders may represent potentially endophytic fungi. A Shannon–Wiener diversity index (H′) analysis showed a relatively high fungal community diversity associated with these tropical orchids. This diversity may offer greater flexibility in terms of the adaptation of the plants to changing environmental conditions and the potential facilitation of reintroduction programs. The Simpson diversity index values showed that all of the libraries included dominant species, and a LIBSHUFF analysis showed that the fungal communities were structurally different from each other, suggesting an influence of local factors on this diversity. This study offers important information for the development of conservation strategies for threatened and endemic species of Brazilian flora in an important and threatened hotspot.     

A geometric morphometric and microsatellite analyses of <Emphasis Type="Italic">Scaptotrigona mexicana</Emphasis> and <Emphasis Type="Italic">S. pectoralis</Emphasis> (Apidae: Meliponini) sheds light on the biodiversity of Mesoamerican stingless bees

Geometric morphometrics and molecular methods are effective tools to study the variability of stingless bee populations and species that merit protection given their worldwide decline. Based on previous evidence of cryptic lineages within the Scaptotrigona genus, we tested the existence of multiple evolutionary lineages within the species S. mexicana and we investigated the status of S. pectoralis. By analyzing their population structure, we found differences between the Pacific and Atlantic populations of each of these species, although geometric morphometrics of the wing only confirmed these results in S. mexicana. There was a tendency towards enhanced genetic differentiation over larger distances in the Atlantic populations of both species but not in the Pacific populations. These results revealed a pattern of differentiation among evolutionary units and a specific distribution of genetic diversity within these Scaptotrigona species in Mesoamerica, suggesting the need for future taxonomic revisions, as well as activities aimed at management and conservation.     

Structure of Pinus nigra Arn. populations in Bulgaria revealed by chloroplast microsatellites and terpenes analysis: Provenance tests

Pinus nigra is a forest and low elevation mountain species found around the Mediterranean Sea that has had its distribution reduced and fragmented by anthropogenic disturbance. Due to commercial interest it is currently being replanted, however, the genetic structure of populations is little known and current planting strategies could threaten its genetic diversity. In the present study we investigated the genetic structure and genetic diversity of P. nigra populations in Bulgaria using chloroplast microsatellite markers and terpene analysis. Nine provenances were chosen throughout the species' range in Bulgaria. Following DNA extraction, chloroplast microsatellite (cpSSR) loci were surveyed using three primer pairs. Between 5 and 9 size variants were identified at each locus. A total of 22 size variants at the 3 loci were identified, that were combined in 68 different haplotypes, of which 7 represent 39.81% of the genetic structure. AMOVA analysis revealed that 6.06% of the variation was found among populations, while 93.94% was expressed within populations. The cpSSR analysis divided European Black pine populations into four groups, the first represented by populations located the eastern Rhodopes, Sr. Gora and St. Planina mountains, while the second group is primarily located in the Phodopes and Slavianca mountains. The populations from Pirin and Osogovo mountains show different genetic patterns. Terpene analysis revealed that most of the monoterpene pool in P. nigra was accounted for by α-pinene followed by β-pinene. The presence of four distinct terpene groups is not consistent with physical distances between populations, and a similar non-significant correlation between genetic distance determined by chloroplast microsatellites analysis and chemotype distance (determined by terpenes) was observed. Our results suggest that the structural pattern of genetic diversity of cpDNA in European Black pine populations is the consequence of historical biogeographic processes.     

Genetic structure of the Atlantic Rainforest tree species Luehea divaricata (Malvaceae)

The Atlantic Rain Forest is one of the most important Brazilian biomes and a hotspot for biodiversity that is characterized by its high level of endemism, where new species are still being described. Luehea divaricata (Malvaceae) is commonly found in riparian forests areas of the Atlantic forest. Because of the importance of this species in reforestation programs, we used nine pairs of microsatellite loci to study the genetic variability of this species along its distribution area and verify if fragmentation is compromising the survival of these populations. A total of 50 alleles were obtained with an average observed and expected heterozygosity of 0.53 and 0.67, respectively. Seven of the nine populations studied showed a heterozygosity deficit. Most of the genetic diversity was found within populations; while the level of genetic differentiation was moderated (6.84) between populations. Different levels of gene flow between the populations were detected. Positive and significant values of Fis were found for seven populations. The signal test for excess of heterozygosity indicated that a recent genetic bottleneck occurred in the fragmented populations. The dendrogram constructed by the UPGMA method revealed the formation of seven clusters, which was confirmed by the Bayesian analysis for number of K clusters. The presence of several pairs of loci in linkage disequilibrium confirms that these populations experienced a loss of genetic diversity caused by genetic drift. The results showed that it is necessary to develop management strategies for the conservation of these populations of L. divaricata as the viability of the next generations are severely compromised.