Nuclear Simple Sequence Repeat Research Articles

Phylogeographic and demographic modeling analyses of the multiple origins of the rheophytic goldenrod Solidago yokusaiana Makino.

Understanding adaptation mechanisms is important in evolutionary biology. Parallel adaptation provides good opportunities to investigate adaptive evolution. To confirm parallel adaptation, it is effective to examine whether the phenotypic similarity has one or multiple origins and to use demographic modeling to consider the gene flow between ecotypes. Solidago yokusaiana is a rheophyte endemic to the Japanese Archipelago that diverged from Solidago virgaurea. This study examined the parallel origins of S. yokusaiana by distinguishing between multiple and single origins and subsequent gene flow. The haplotypes of noncoding chloroplast DNA and genotypes at 14 nuclear simple sequence repeat (nSSR) loci and single-nucleotide polymorphisms (SNPs) revealed by double-digest restriction-associated DNA sequencing (ddRADseq) were used for phylogeographic analysis; the SNPs were also used to model population demographics. Some chloroplast haplotypes were common to S. yokusaiana and its ancestor S. virgaurea. Also, the population genetic structures revealed by nSSR and SNPs did not correspond to the taxonomic species. The demographic modeling supported the multiple origins of S. yokusaiana in at least four districts and rejected a single origin with ongoing gene flow between the two species, implying that S. yokusaiana independently and repeatedly adapted to frequently flooding riversides.

Genetic diversity and population structure analyses in the Alpine plum (Prunus brigantina Vill.) confirm its affiliation to the Armeniaca section

In-depth characterization of the genetic diversity and population structure of wild relatives is of paramount importance for genetic improvement and biodiversity conservation, and is particularly crucial when the wild relatives of crops are endangered. In this study, we sampled the Alpine plum (Briançon apricot) Prunus brigantina Vill. across its natural distribution in the French Alps, where its populations are severely fragmented and its population size strongly impacted by humans. We analysed 71 wild P. brigantina samples with 24 nuclear simple sequence repeat (microsatellite) markers and studied their genetic diversity and population structure, with the aim to inform in situ conservation measures and build a core collection for long-term ex situ conservation. We also examined the genetic relationships of P. brigantina with other species in the Prunophora subgenus, encompassing the Prunus (Eurasian plums), Prunocerasus (North American plums) and Armeniaca (apricots) sections, to check its current taxonomy. We detected a moderate genetic diversity in P. brigantina and a Bayesian model–based clustering approach revealed the existence of three genetically differentiated clusters, endemic to three geographical regions in the Alps, which will be important for in situ conservation measures. Based on genetic diversity and population structure analyses, a subset of 36 accessions were selected for ex situ conservation in a core collection that encompasses the whole detected P. brigantina allelic diversity. Using a dataset of cultivated apricots and wild cherry plums (P. cerasifera) genotyped with the same markers, we detected gene flow neither with European P. armeniaca cultivars nor with diploid plums. Similar to previous studies, dendrograms and networks placed P. brigantina closer to the Armeniaca section than to the Prunus section. Our results thus confirm the classification of P. brigantina within the Armeniaca section; it also illustrates the importance of the sampling size and design in phylogenetic studies.

Genetic structure of coast redwood (Sequoia sempervirens [D. Don] Endl.) populations in and outside of the natural distribution range based on nuclear and chloroplast microsatellite markers.

Coast redwood (Sequoia sempervirens) naturally growing in southern Oregon and northern California is one of the few conifer tree species that are polyploid. Despite its unique ecological and economic importance, its population genetic structure is still insufficiently studied. To obtain additional data on its population genetic structure we genotyped 317 samples collected from populations in California (data set C) and 144 trees growing in a provenance trial in France (data set F) using 12 nuclear (five random nuclear genomic nSSRs and seven expressed sequence tag EST-SSRs) and six chloroplast (cpSSRs) microsatellite or simple sequence repeat (SSR) markers, respectively. These data sets were also used as reference to infer the origin of 147 coast redwood trees growing in Germany (data set G). Coast redwood was introduced to Europe, including Germany as an ornamental species, decades ago. Due to its fast growth and high timber quality, it could be considered as a potential commercial timber species, especially in perspective to climate warming that makes more regions in Germany suitable for its growing. The well performing trees in colder Germany could be potential frost resistant genotypes, but their genetic properties and origin are mostly unknown. Within the natural range in southern Oregon and northern California, only two relatively weak clusters were identified, one northern and one southern, separated by the San Francisco Bay. High genetic diversity, but low differentiation was found based on the 12 nuclear SSR markers for all three data sets F, C and G. We found that investigated 147 German trees represented only 37 different genotypes. They showed genetic diversity at the level less than diversity observed within the natural range in the northern or southern cluster, but more similar to the diversity observed in the southern cluster. It was difficult to assign German trees to the original single native populations using the six cpSSR markers, but rather to either the northern or southern cluster. The high number of haplotypes found in the data sets based on six cpSSR markers and low genetic differentiation based on 12 nuclear SSRs found in this study helps us study and better understand population genetic structure of this complex polyploid tree and supports the selection of potential genotypes for German forestry.

Genetic diversity and genetic structure of Acer monspessulanum L. across Zagros forests of Iran using molecular markers

Acer monspessulanum L. is an important tree species found in the temperate Zagros forests of Iran. Despite its importance, the long-term persistence of its small and fragmented populations is jeopardised by genetic erosion and hence, monitoring its genetic resource and variability is practically required for providing conservation measures of the species germplasm in Zagros woodland ecosystem. The present study aimed to provide the first data on genetic diversity and genetic differentiation pattern of 19 natural populations comprising 240 individuals of A. monspessulanum across its growing area in Zagros forests using three molecular tools including inter-simple sequence repeats (ISSR), start codon targeted (SCoT), and simple sequence repeat (SSR). In total, ISSR and SCoT primers generated a total of 141 and 121 clear and scorable bands for analysis with the polymorphism rate of 90.50 and 90.02% and a mean of 10.85 and 11 fragments per marker, respectively. In addition, 73 alleles were achieved using 10 polymorphic SSR loci from the studied accessions with 100% polymorphism, ranging between 5 and 10 alleles per locus. Average percentage of polymorphic alleles per population for ISSR, SCoT, and SSR data was 84.02%, 83%, and 100%, respectively, and generally, Nei's gene diversity (H) and Shannon's index of diversity (I) values for all populations demonstrated moderate to high levels of genetic diversity (H=0.267-0.707; I=0.38-1.38). The AMOVA results of the three marker systems attributed higher genetic variation to individuals within in each population than among populations. Furthermore, overall GST value for all populations detected the moderate to high levels of genetic differentiation, indicating a limited gene flow occurrence among the populations. STRUCTURE analysis (K=5) clustered the populations into four to five distinct groups, in accordance with geographical distances. These results could represent an important contribution for effective germplasm characterization and could be eventually used in in situ or ex situ conservation of A. monspessulanum genetic resources.

Subspecies divergence and pronounced phylogenetic incongruence in the East-Asia-endemic shrubMagnolia sieboldii

The biogeographic patterns of the East-Asia-endemic shrub Magnolia sieboldii, in which the range of the subsp. sieboldii is interposed with the disjunct distribution of subsp. japonica, implies a complex evolutionary history, involving rapid speciation and hybridization. Here, we aim to reveal the evolutionary and phylogeographic histories of the species with a particular focus on the time of subspecies divergence, the hypothesis of secondary hybridization and the Pleistocene survival of each subspecies, using a combination of genetic analyses and ecological niche modelling. Genetic variation, genetic structures and phylogenetic relationships were elucidated based on nuclear low-copy genes, chloroplast DNA, and nuclear simple sequence repeats (SSRs). A scenario selection analysis and divergence time estimation were performed using coalescent simulation in DIYABC and *BEAST. Ecological niche modelling and a test of niche differentiation were performed using Maxent and ENMTools. All marker types showed deep, but pronouncedly incongruent, west-east genetic divergences, with the subspecies being delineated only by the nuclear low-copy genes. Phylogenetic tree topologies suggested that ancient hybridization and introgression were likely to have occurred; however, this scenario did not receive significant support in the DIYABC analysis. The subspecies differentiated their niches, but both showed a dependence on high humidity and were predicted to have persisted during the last glacial cycle by maintaining a stable latitudinal distribution via migration to lower altitudes. We found a deep genetic divergence and a pronounced phylogenetic incongruence among the two subspecies of M. sieboldii, which may have been driven by major paleogeographic and paleoclimatic events that have occurred since the Neogene in East Asia, including global cooling, climate oscillations and the formation of land bridges. Both subspecies were, however, considered to persist in situ in stable climatic conditions during the late Pleistocene.

Shining a light on species delimitation in the tree genus Engelhardia Leschenault ex Blume (Juglandaceae)

Enhanced efficacy in species delimitation is critically important in biology given the pending biodiversity crisis under global warming and anthropogenic activity. In particular, delineation of traditional classifications in view of the complexity of species requires an integrative approach to effectively define species boundaries, and this is a major focus of systematic biology. Here, we explored species delimitation of Engelhardia in tropical and subtropical Asia. In total, 716 individuals in 71 populations were genotyped using five chloroplast regions, one nuclear DNA region (nrITS), and 11 nuclear simple sequence repeats (nSSR). Phylogenetic trees were constructed and relationships among species were assessed. Molecular analyses were then combined with 14 morphological characteristics of 720 specimens to further explore the species boundaries of Engelhardia. Integrating phylogenetic and morphological clusters provided well-resolved relationships to delineate seven species. The results suggested that: first, that E. fenzelii, E. roxburghiana, E. hainanensis, E. apoensis, and E. serrata are distinct species; second, E. spicata var. spicata, E. spicata var. aceriflora, E. spicata var. colebrookeana, and E. rigida should be combined under E. spicata and treated as a species complex; third, E. serrata var. cambodica should be raised to species level and named E. villosa. We illuminated that bias thresholds determining the cluster number for delimiting species boundaries were substantially reduced when morphological data were incorporated. Our results urge caution when using the concepts of subspecies and varieties in order to prevent confusion, particularly with respect to species delimitation for tropical and subtropical species. In some cases, re-ranking or combining subspecies and/or varieties may enable more accurate species delimitation.

Development of nuclear microsatellite markers to facilitate germplasm conservation and population genetics studies of five groups of tropical perennial plants with edible fruits and shoots: rambutan (Nephelium lappaceum L.), sapodilla (Manilkara zapota (L.) P. Royen), lychee (Litchi chinensis Sonn.), mangosteen (Garcinia mangostana Linn. and Garcinia cochinchinensis (Lour.) Choisy) and bamboo (Bambusa vulgaris Schrad. ex J.C. Wendl and Guadua angustifolia Kunth)

Simple sequence repeat (SSR) enriched libraries for five groups of tropical perennial plants with edible fruits and shoots were prepared and sequenced in a GS-FLX Roche 454: sapodilla (Manilkara zapota (L.) P. Royen), lychee (Litchi chinensis Sonn.), mangosteen (Garcinia mangostana Linn. and G. cochinchinensis (Lour.) Choisy), rambutan (Nephelium lappaceum L.), and bamboo (Bambusa vulgaris Schrad. ex J.C. Wendl and Guadua angustifolia Kunth). For SSR development, these species were organized by their common names in five groups. A total of 3870 SSR primer sets were designed, using capillary electrophoresis 1872 nuclear SSRs were tested on 4 to 10 DNA samples within each plant group, that is 384 loci for each of the four groups of fruit trees and 336 loci for the bamboo group. Only 7.9% of the primers tested did not result in amplification. All 1872 SSRs are provided, we highlight 178 SSRs (between 26 and 47 per group) considered top-quality polymorphic SSRs that amplified all the samples, had strong fluorescence signal, presented no stutters and showed minimum non-specific amplification or background fluorescence. A total of 66,057 contig sequences were submitted to GenBank Database. Markers presented here will be useful not only for conservation efforts in banks of germplasm, but also for in-depth analysis of population genetics which usually requires evaluation of large number of loci.

Phenological shifts and genetic differentiation between sympatric populations of Sargassum horneri (Fucales, Phaeophyceae) in Japan

Marked seasonality, especially in sexual reproduction, is common among seaweed species along temperate coasts and increases the possibility of successful fertilization in outcrossing species. A phenological shift in reproductive seasons, therefore, could be an effective isolation barrier between conspecific seasonal populations, although its power has not been verified in algae.Sargassum horneri, a major component of seaweed beds along the temperate coast of Japan, is known for variability in its reproductive phenology. To understand the significance of phenological shift as an isolation barrier in seaweed species, phenological investigations ofS. horneriseasonal populations on the Sea of Japan coast of central Honshu, Japan, were combined with Bayesian cluster analysis based on a nuclear simple sequence repeat genotype. Results from these analyses concordantly suggest a genetic differentiation between the seasonal populations, although almost 20% of field-collected plants were estimated to be hybrids or have a hybrid origin based on results of Bayesian cluster analyses using experimental hybrids. A collapse of seasonal isolation was also detected at the site of the field investigation, and a high percentage of putative hybrids in the following generation at the site (41%) suggested significant seasonal isolation in the differentiation observed in this study.

Development of nuclear SSR and chloroplast genome markers in diverse Liriodendron chinense germplasm based on low-coverage whole genome sequencing

BackgroundLiriodendron chinense ranges widely in subtropical China and northern Vietnam; however, it inhabits several small, isolated populations and is now an endangered species due to its limited seed production. The objective of this study was to develop a set of nuclear SSR (simple sequence repeats) and multiple chloroplast genome markers for genetic studies in L. chinense and their characterization in diverse germplasm.ResultsWe performed low-coverage whole genome sequencing of the L. chinense from four genotypes, assembled the chloroplast genome and identified nuclear SSR loci by searching in contigs for SSR motifs. Comparative analysis of the four chloroplast genomes of L. chinense revealed 45 SNPs, 17 indels, 49 polymorphic SSR loci, and five small inversions. Most chloroplast intraspecific polymorphisms were located in the interspaces of single-copy regions. In total, 6147 SSR markers were isolated from low-coverage whole genome sequences. The most common SSR motifs were dinucleotide (70.09%), followed by trinucleotide motifs (23.10%). The motif AG/TC (33.51%) was the most abundant, followed by TC/AG (25.53%). A set of 13 SSR primer combinations were tested for amplification and their ability to detect polymorphisms in a set of 109 L. chinense individuals, representing distinct varieties or germplasm. The number of alleles per locus ranged from 8 to 28 with an average of 21 alleles. The expected heterozygosity (He) varied from 0.19 to 0.93 and the observed heterozygosity (Ho) ranged from 0.11 to 0.79.ConclusionsThe genetic resources characterized and tested in this study provide a valuable tool to detect polymorphisms in L. chinense for future genetic studies and breeding programs.

The protected tree Dimorphandra wilsonii (Fabaceae) is a population of inter-specific hybrids: recommendations for conservation in the Brazilian Cerrado/Atlantic Forest ecotone.

Dimorphandra wilsonii Rizzini, a critically endangered and protected tree, has a restricted distribution in the ecotone between the Cerrado and the Atlantic Forest in south-eastern Brazil. In this area, it co-occurs with D. mollis Benth., a common tree from the Cerrado, and D. exaltata Schott., a rare tree from the Atlantic Forest. Previous studies of D. wilsonii indicated heterozygosity excess at the individual level. Field observation of some intermediate phenotypes between D. wilsonii and both congeners suggests hybridization of D. wilsonii with D. mollis and/or D. exaltata. Here, we tested the hypothesis that D. wilsonii may have originated from hybridization between D. exaltata and D. mollis. We also performed cytogenetic analysis to examine if the heterozygosity excess could be explained by polyploidy in D. wilsonii. We evaluated the genetic diversity and population structure of D. wilsonii using 11 nuclear simple sequence repeats (SSRs) genotyped in 152 individuals sampled across the taxon's range. We performed comparative genetic analyses using overlapping SSR markers between D. wilsonii and previously published SSR data in D. mollis and D. exaltata to subsequently perform a series of allelic comparisons, multivariate and Bayesian analysis. Our results suggest that D. wilsonii individuals are most likely to correspond to F1 hybrids between D. exaltata and D. mollis. Cytogenetic analysis indicated that D. wilsonii is diploid with the same chromosome number as D. mollis (2n = 2x = 28). Our study raises questions about the taxonomic status and the evolutionary future of D. wilsonii. We suggest that the conservation and management strategy for D. wilsonii should be revised and that it should take into account both parental Dimorphandra species in the ecotone, with special emphasis on the threatened D. exaltata. Finally, this study highlights the value of genetic information for the design of conservation strategies.

Origins of weedy rice revealed by polymorphisms of chloroplast DNA sequences and nuclear microsatellites

AbstractConspecific weeds that permanently infest worldwide agroecosystems are evolved from their crop species. These weeds cause substantial problems for crop production by competing for resources in agricultural fields. Weedy rice represents such a conspecific weed infesting rice ecosystems, and causing tremendous rice yield losses owing to its strong competitiveness and abundant genetic diversity, likely resulted from its complex origins. Here, we report the use of chloroplast DNA (cpDNA) fingerprints to determine whether weedy rice is evolved from its wild (exo‐feral) or cultivated (endo‐feral) rice progenitor as the maternal donor in recent hybridization events. In addition, we also applied nuclear simple sequence repeat (SSR) markers to confirm the exo‐feral or endo‐feral origins of weedy rice accessions determined by the cpDNA fingerprints. We found that the studied weedy rice accessions evolved either from their wild or cultivated rice progenitor, as the maternal donor, based on the cpDNA network and structure analyses. Combined analyses of cpDNA and nuclear SSR markers indicated that a much greater proportion of weedy rice accessions had the endo‐feral origin. In addition, results from the genetic structure of nuclear SSR markers indicated that weedy rice accessions from the endo‐feral pathway are distinctly associated with either indica or japonica rice cultivars, suggesting their complex origins through crop–weed introgression. The complex pathways of origin and evolution could greatly promote genetic diversity of weedy rice. Therefore, innovative methods should be developed for effective weedy rice control.

Introgression Among Cultivated and Wild Grapevine in Tuscany.

Wild grapevine, Vitis vinifera L. subsp. sylvestris (Gmelin, Hegi) is spontaneous to Europe and common in Tuscany. In this study, wild grapevines were identified in 22 populations from eight locations in Tuscan Maremma (Grosseto and Siena province). The plants were propagated by cuttings, collected in a vineyard, genotyped by nuclear simple sequence repeats (SSRs), chloroplast SSRs and single nucleotide polymorphisms (SNPs), and compared to locally cultivated varieties (Vitis vinifera L. subsp. sativa) and to non-vinifera and non-vitis genotypes. The identity analysis revealed that some individuals were redundant genotypes, suggesting natural vegetative propagation. In addition, four of the supposed V.v. sylvestris were in fact naturalized V.v. sativa. The majority of putative sylvestris genotypes had chlorotype A, while the remainder had chlorotype D, as the majority of Vitis vinifera subsp. sativa cultivated in Italy. Some of the recovered sylvestris genotypes appeared to be natural crosses with cultivated grapevine varieties in Tuscany, and their chlorotype suggests a higher pollen flow from sativa to the sylvestris genotypes than in the opposite direction. In addition, other genotypes appeared to be crosses within sylvestris, sylvestris-sativa or sylvestris-sylvestris siblings, or equivalent relationships. These relationships suggest a noticeably level of sexual reproductive activities among sylvestris and sylvestris-sativa genotypes. A cluster and structure analysis clearly differentiated the true sylvestris from the sativa, and the non-vinifera or non-vitis genotypes, and also highlighted a possible introgression of sylvestris into some Italian and French cultivated varieties. The results therefore suggest that, in addition to the primary ancient center of domestication from the Near East to Central Asia, the introgression among cultivated and wild grapevine occurred in other centers of diversification along the migration routes, contributing to the domestication processes, and suggesting that these processes are still ongoing despite the reduction in populations of sylvestris. The results also highlight that the GrapeReSeq 18K Vitis genotyping chip are suitable for non-vitis genotyping and that the range of SNPs heterozygosity in sylvestris appears to be up to 6% less and does not overlap the heterozygosity range of sativa genotypes.

Development and characterization of 17 microsatellite markers for Sonchus oleraceus.

PremiseThe common sowthistle, Sonchus oleraceus (Asteraceae), is a globally invasive weedy species. In order to investigate its genetic diversity, population genetic structure, and evolutionary history, we developed and characterized nuclear simple sequence repeat markers (SSRs or microsatellites).Methods and ResultsSeventeen microsatellite primer pairs were developed based on the Illumina sequence data. Ten developed SSR loci were polymorphic in four populations sampled from broad geographical regions. The number of alleles per locus ranged from one to 11, and the levels of observed and expected heterozygosity ranged from 0.000 to 1.000 and from 0.000 to 0.801, respectively. Up to 82% of the newly developed primer pairs were successfully amplified in the congeneric taxa S. asper, S. asper subsp. glaucescens, S. canariensis, and S. palmensis.ConclusionsThe SSR markers developed in this study will be useful for future population genetic studies on S. oleraceus and other congeneric species.

Molecular differentiation of Turkish and Common hazels (Corylus colurna L. and Corylus avellana L.) using multiplexed nuclear microsatellite markers

Corylus colurna is considered as important tree species under climate change for dry and warm conditions in Central Europe and was overused because of its valuable wood. Therefore Turkish hazel is now present only in small isolated populations and is protected under IUCN. Genetic conservation of this tree species plays a key role in future sustainable forest development. Turkish hazel co-occurs with Common hazel (C. avellana) in its whole distribution area and may form hybrids. To differentiate between the pure species and their hybrids, cross-species amplifying markers are required. In this study we have evaluated existing simple sequence repeat (SSR) markers using altogether 128 samples of C. avellana and C. colurna. Fifteen nuclear SSRs have generated easy to-score alleles in the two species and 13 of them were highly polymorphic. For all 15 markers the mean allele number, average observed heterozygosity, genetic diversity and polymorphism information index were high. The two most polymorphic SSRs were L1.10 and CaT-B501 with 19 and 16 alleles, respectively. Structure analysis proved the differentiation of the two species C. avellana and C. colurna. No hybridization was detected in the analysed populations. Results also indicated that C. colurna from Balkan Peninsula and Asia Minor belong to separate groups. Our study presents highly polymorphic, easy to score, ready to use SSR-multiplexes, which can be applied in population genetics and gene conservation studies.

Genetic Diversity and Population Structure of Natural Pinus koraiensis Populations

Studying the genetic diversity and population structure of natural forest populations is essential for evaluating their ability to survive under future environmental changes and establishing conservation strategies. Pinus koraiensis is a conifer species with high ecological and economic value in Northeast China. However, its natural forests have been greatly reduced in recent years, mostly due to over exploitation and over utilization. Here, we evaluated the genetic diversity and population structure of seven populations of P. koraiensis located throughout its native distribution. A total of 204 samples were genotyped with nine polymorphic nuclear SSR (simple sequence repeat) markers. The results showed high genetic diversity in all populations, with an average expected heterozygosity of 0.610, and the northern-most populations (Dailin (DL) and Fenglin (FL)) showed slightly higher diversity than the other five populations. The level of genetic differentiation among populations was very low (FST = 0.020). Analysis of molecular variance (AMOVA) showed that only 2.35% of the genetic variation existed among populations. Moreover, STRUCTURE analysis clearly separated the seven populations into two clusters. Populations DL and FL from the Xiaoxinganling Mountains comprised cluster I, while cluster II included the five populations from the Changbai Mountains and adjacent highlands. Our research on the genetic diversity and population structure of P. koraiensis in natural forests of China can provide a basis for the implementation of programs for the conservation and utilization of P. koraiensis genetic resources in the future.

Microsatellite and Morphological Analyses Reveal Unexpected Diversity in Lymantria dispar in China

The gyspy moth Lymantria dispar Linnaeus, a widely distributed leaf-eating pest, is considered geographically isolated in the world, with two Asian gypsy moth subspecies, Lymantria dispar asiatica and Lymantria dispar japonica. In China, only one subspecies, L. d. asiatica, has been observed. In this study, we characterized gypsy moth diversity and divergence using 427 samples covering a wide range of the species distribution, with a focus on sampling along a latitudinal gradient in China. We combine the quantitative analysis of male genitalia and the genetic diversity analysis of nine microsatellite loci of nuclear genes nuclear genes to study the structure of gypsy moth individuals in 23 locations in the world and the male genitalia of gypsy moths in some areas. In mixed ancestry model-based clustering analyses based on nuclear simple sequence repeats, gypsy moths were divided into three well-known subspecies, a unique North American cluster, and a southern Chinese cluster with differentiation between the Asian gypsy moth and European gypsy moth. We also found individuals identified as European gypsy moths in two distant regions in China. The results of a quantitative analysis of male genitalia characteristics were consistent with an analysis of genetic structure and revealed the differentiation of gypsy moths in southern China and of hybrids suspected to be associated with L. d. japonica in the Russian Far East. Admixture in gypsy moths can be explained by many factors such as human transport. In China, we detected European gypsy moths, and found unexpectedly high genetic diversity within populations across a wide range of latitudes.

Diversification and genetic structure of the western-to-eastern progression of European Phaseolus vulgaris L. germplasm

BackgroundCommon bean (Phaseolus vulgaris L.) is the most important food legume for direct human consumption around the world, as it represents a valuable source of components with nutritional and health benefits.ResultsWe conducted a study to define and explain the genetic relatedness and diversification level of common bean (Phaseolus vulgaris L.) germplasm from Portugal to Ukraine, along a western-to-eastern line of southern European countries, including Poland. This was based on the P. vulgaris genetic structure, and was designed to better describe its distribution and domestication pathways in Europe. Using the multi-crop passport descriptors that include geographic origin and different phaseolin types (corresponding to the Mesoamerican and Andean gene pools), 782 accessions were obtained from nine gene banks and 12 geographic origins. We selected 33 genome/ gene-related/ gene-pool-related nuclear simple sequence repeat markers that covered the genetic diversity across the P. vulgaris genome. The overall polymorphic information content was 0.800. Without specifying geographic origin, global structure cluster analysis generated 10 genetic clusters. Among the PvSHP1 markers, the most informative for gene pool assignment of the European P. vulgaris germplasm was PvSHP1-B. Results of AMOVA show that 89% of the molecular variability is shared within the 782 accessions, with 4% molecular variability among the different geographic origins along this western-to-eastern line of southern Europe (including Poland).ConclusionsThis study shows that the diversification line of the European P. vulgaris germplasm followed from the western areas of southern Europe (Portugal, Spain, Italy, Slovenia) to the more eastern areas of southern Europe. This progression defines three geographically separated subgroups, as the northern (Poland, Ukraine, Romania), southern (Albania, Bulgaria), and central (Bosnia and Herzegovina, Serbia, Hungary) areas of eastern Europe.

Spatial Genetic Patterns and Distribution Dynamics of the Rare Oak Quercus chungii: Implications for Biodiversity Conservation in Southeast China

A rapidly changing climate and frequent human activity influences the distribution and community structure of forests. Increasing our knowledge about the genetic diversity and distribution patterns of trees is helpful for forest conservation and management. In this study, nSSRs (nuclear simple sequence repeats) were integrated with a species distribution model (SDM) to investigate the spatial genetic patterns and distribution dynamics of Quercus chungii F.P.Metcalf, a rare oak in the subtropics of southeast China. A total of 188 individuals from 11 populations distributed across the natural range of Q. chungii were genotyped using nine nSSRs. The STRUCTURE analysis indicated that genetic admixture was present in all populations, but the population genetic variation and genetic differentiation were related to their geographical distributions. The SDM result indicated that Q. chungii retreated to the Nanling Mountains and adjacent areas during the Last Glacial Maximum (LGM) period, which corresponds to higher genetic diversity for populations in this region. Landscape genetic analysis showed that the Nanling Mountains served as a corridor for organism dispersal at the glacial and interglacial periods within the Quaternary. Based on these results, we propose that establishing nature reserves to protect the ecological corridor across the Nanling Mountains is necessary for the conservation of regional species genetic diversity, as well as the ecosystem of evergreen broadleaved forests in southern China. The study combines species distribution models and genetic diversity to provide new insight into biodiversity conservation and forest management under future climate change.

Development and characterization of SSR markers for Sanguinaria canadensis based on genome skimming.

PremisePolymorphic nuclear simple sequence repeat (nSSR) markers were developed for Sanguinaria canadensis (Papaveraceae), a spring ephemeral native to eastern North America.Methods and ResultsBased on the genome skimming data of S. canadensis, a total of 240 nSSR primer pairs were designed for 80 loci from the assembled nuclear contigs. Of these primer pairs, 19 were selected for initial validation in four populations (80 individuals). All 19 loci produced heterologous amplification. The numbers of alleles per locus ranged from one to 21; the levels of observed and expected heterozygosity per locus ranged from 0.000 to 1.000 and from 0.000 to 0.847, respectively. Transferability of the loci was tested in the related species Eomecon chionantha.ConclusionsThe developed nSSR markers revealed polymorphism in the four studied populations and may contribute to investigations of the genetic diversity of S. canadensis and E. chionantha.

Regional patterns of genetic structure and environmental differentiation in willow populations (Salix humboldtiana Willd.) from Central Mexico.

AimTo infer the geological and climatic factors that have shaped the genetic diversity and structure of a willow species (Salix humboldtiana) in three basins of Central Mexico.LocationCentral Mexico.MethodsWe collected samples from 11 populations across two hydrological basins (Balsas and Lerma) and one population from another basin (Ameca) within the Mexican Central Plateau (MCP). Individuals were analyzed using sequences of two chloroplast DNA (cpDNA) regions and eight nuclear simple sequence repeats (nSSR). Population genetic diversity and structure were determined from these data. To evaluate whether genetic structure was associated with ecological niche differentiation, we determined whether there is niche equivalence, overlap, or divergence between the Balsas and Lerma basins. Also, we evaluated the relative contributions of geographic distribution and climatic variation on population genetic structuring through redundancy analysis (RDA) and partial RDA.ResultsBoth cpDNA and nSSRs data indicated the presence of three highly differentiated genetic groups, mostly geographically congruent with the three main hydrological basins. According to nSSRs, the three genetic groups can be further subdivided into eight subgroups corresponding to different rivers within the main basins. The niche equivalency test showed that the niches of the species in the Balsas and Lerma basins are significantly nonequivalent. The RDA indicated a significant association of genetic variation among populations with climate variables (particularly those related to the precipitation regime), while controlling for geographic distribution.Main conclusionsThe genetic structure of S. humboldtiana is strongly associated with the historical and current geological configuration of the basins and the rivers within basins. The observed hierarchical genetic differentiation can be due to gene flow limitation resulting from physical barriers to the dispersal of S. humboldtiana, but also to some degree of isolation by environment, as suggested by the significant association between genetic variation among populations and precipitation regime.