Early Intergenerational Changes in Genetic Diversity During Domestication of Procambarus acanthophorus Using Mitochondrial and Nuclear DNA Markers.

We studied the genetic differentiation, structure, and diversity of Scots pine populations in Lithuania based on nuclear, chloroplast microsatellite, and mitochondrial DNA markers. We focused on revealing evolutionary history, country-wide geneflow patterns, and structuring among the Scots pine populations. We genotyped 439 Scots pine individuals of mature age from 23 natural Scots pine stands in Lithuania and used the AMOVA and a set of genetic-clustering methods. The among-population differentiation was weak for nuclear microsatellite loci (nSSRs) (FST = 0.005) but much stronger for cpSSRs (PhiST = 0.240). The populations were structured into highland and lowland populations based on cpSSRs and eastern highland versus the rest for nSSRs. We detected two mtDNA mitotypes—the universal type A and northeastern type B, and the latter occurred at a markedly higher frequency in eastern Lithuania. Within-population genetic diversity was higher in large pine-dominated forest tracts in the eastern highlands than in fragmented forests in the western highlands. We concluded that phenology-based genetic networks following the temperature climate gradients have a strong effect on shaping the genetic structure of otherwise rather homogeneous gene pools of Scots pine populations in Lithuania. The possible effects of human interference with forests on genetic diversity of Scots pine populations in Lithuania are discussed.

The objective of this study is to inventory the current genetic diversity of the bison quarantine feasibility study (BQFS) herd originating from Yellowstone National Park (YNP) using previously described microsatellite, mitochondrial and nuclear DNA markers with the aim to determine the degree, if any, of cattle DNA introgression in this herd. This work can provide an important tool in monitoring and managing bison genetic diversity as brucellosis‐free reintroduced herds are re‐established throughout the US for conservation purposes. The BQFS composed of 89 Bison bison from YNP that were quarantined and tested to qualify as free of brucellosis in 2006–2007. Understanding genetic diversity of the herd is important to determine if any genetic characteristics such as cattle DNA introgression or low genetic diversity may threaten the herd's protected status. We evaluated genetic diversity at 42 microsatellite loci representing each of the nuclear chromosomes in the bison genome. We found no detectable evidence of cattle DNA introgression in this herd through nuclear markers and mitochondrial DNA analysis. Parentage analysis of the BQFS herd indicated that the majority of mature adults were actively breeding and contributing offspring. Genetic diversity levels in the quarantined herd were high and comparable to the YNP parent herd, suggesting a low risk of genetic loss in the near future. Based on these findings, the genetic diversity currently available within the BQFS herd will provide a strong foundation for bison reintroduced herds and for the preservation of the species.

Genetic variation associated with Picea jezoensis populations of South Korea was investigated using chloroplast (cp), mitochondrial (mt) and nuclear DNA markers. In South Korea, P. jezoensis is distributed across a very restricted area, being found on the summits of three mountains: Mts. Jiri, Dokyu and Gyebang. Examination of five region restriction enzyme combinations for mtDNA and four for cpDNA revealed haplotypes endemic to South Korea. The Gyebang population, the most northerly and most isolated, was genetically distinct from the other populations. Nuclear microsatellite markers indicated, overall, a low level of genetic diversity (H (e) = 0.406) in South Korea; this could be attributed to genetic drift and/or founder effects associated with historical events. The Wilcoxon sign-rank test did not indicate a recent bottleneck in any of the populations irrespective of the model considered (infinite allele model, two-phased model of mutation, and stepwise mutation model). Microsatellite markers also demonstrated that the Gyebang population was distinct from the others. The results of this study could be used as the basis for conservation guidelines for the management of this species in South Korea.

Historical factors, current population size, population connectivity and selective processes at linked loci contribute to shaping contemporary patterns of neutral genetic diversity. It is now widely acknowledged that nuclear and mitochondrial markers react differently to current demography as well as to past history, so the use of both types of markers is often advocated to gain insight on both historical and contemporary processes. We used 12 microsatellite loci genotyped in 13 populations of a mountain lizard (Iberolacerta bonnali) to test whether the historical scenario favoured by a previous mitochondrial study was also supported by nuclear markers and thereby evaluated the consequences of postglacial range movements on nuclear diversity. Congruent signals of recent history were revealed by nuclear and mitochondrial markers using an Approximate Bayesian computation approach, but contemporary patterns of mtDNA and nuclear DNA diversity were radically different. Although dispersal in this species is probably highly restricted at all spatial scales, colonization abilities have been historically good, suggesting capability for reestablishment of locally extinct populations except in fully disconnected habitats.

The study was conducted in 2015 in six spruce stands situated in different forest districts administratively belonging to the Regional Directorate of State Forests in Krosno. Each spruce population was represented by 30 trees and assessed in terms of their current health status. Genetic analyses were performed on shoot samples from each tree using nine nuclear DNA markers and one mitochondrial DNA marker (nad1). The health status of the trees was described according to the classification developed by Szczepkowski and Tarasiuk (2005) and the correlation between health classes and the level of genetic variability was computed with STATISTICA (α = 0.05). Nuclear DNA analyses revealed a low level of genetic variability among spruce populations (only 3% of the total genetic variation (F ST = 0.028) and a high variability within populations (97%). The total heterozygosity in all stands (H T) was calculated as 0.646. Based on UPGMA analysis, the most genetically similar populations are spruce stands in the Bieszczady National Park and the Ustrzyki Dolne Forest District, which have the smallest genetic divergence of all populations (D N = 0.0165). Our analysis of the mitochondrial gene nad1 revealed the presence of six different haplotypes ‘a’, ‘a1’, ‘b’, ‘c’, ‘d’ and ‘d1’. Comprising 56% of all haplotypes, ‘a’ was the most common showing a predominant impact on spruce migration from the Carpathian area. The analysis based on mitochondrial markers (by Nei) revealed a heterozygosity of 0.525. Based on the observations of disease symptoms, 29% of the trees belong to health class 1, 30% to class 2, 28% to class 3 and class 4 contains 13% of trees. The comparison between health status and the level of genetic variation in the analysed stands showed a positive correlation. Spruce stands with better health were also characterised by a greater degree of genetic variability. Since most of the investigated spruce populations shared the mitochondrial haplotype ‘a’, we have ascertained their Hercynian–Carpathian origin. Only one stand (Cisna) had a high frequency (43.3%) of the Nordic haplotype ‘c’ suggesting that this provenance is derived from the Baltic post-glacial refugium of P. abies in europe.

Stocking has had a considerable effect on wild brown trout, Salmo trutta L., populations throughout Europe. To elucidate this impact and to outline further management strategies, the genetic structure of 25 wild populations and five hatchery stocks from Czech Republic and Slovakia were analysed using mitochondrial (control region) and nuclear DNA (microsatellites, LDH‐C1*) markers. Stocking practices have caused massive hybridisation between the Atlantic and Danube brown trout strains in the central Danube basin and have lead to a loss of among‐population divergence in Slovakia and the eastern part of Czech Republic. Comparison with studies from neighbouring countries revealed substantial differences in haplotype, allele frequencies and genetic diversity across Central Europe. Differences in stocking management and origin of breeding stocks appear to be crucial factors for the spatial variability of the genetic structure of brown trout.

Phylogeography of endemic toads and post-Pliocene persistence of the Brazilian Atlantic Forest

Protist diversity is currently a much debated issue in eukaryotic microbiology. Recent evidence suggests that morphological and genetic diversity might be decoupled in some groups of protists, including ciliates, and that these organisms might be much more diverse than their morphology implies. We sought to assess the genetic and morphological diversity of Carchesium polypinum, a widely distributed peritrich ciliate. The mitochondrial marker cytochrome c oxidase subunit I and the nuclear small subunit ribosomal RNA were used to examine genetic diversity. For the morphological assessment, live microscopy and Protargol staining were used. The mitochondrial marker revealed six robust, deeply diverging, and strongly supported clades, while the nuclear gene was congruent for three of these clades. There were no major differences among individuals from the different clades in any of the morphological features examined. Thus, the underlying genetic diversity in C. polypinum is greater than what its morphology suggests, indicating that morphology and genetics are not congruent in this organism. Furthermore, because the clades identified by the mitochondrial marker are so genetically diverse and are confirmed by a conserved nuclear marker in at least three cases, we propose that C. polypinum be designated as a "cryptic species complex." Our results provide another example where species diversity can be underestimated in microbial eukaryotes when using only morphological criteria to estimate species richness.

Sequences of mitochondrial (cytochrome b) and nuclear (recombination activating gene 1–RAG1) DNA markers were obtained for two species of the genus Alburnoides, the Taskent riffle minnow A. oblongus Bulgakov 1923 and the Terek spirlin A. gmelini Bogutskaya and Coad 2009. Phylogenetic analysis revealed that A. oblongus belongs to the genus Alburnus.

BackgroundThe Culex pipiens complex includes the most widespread mosquito species in the world. Cx. pipiens is the primary vector of the West Nile Virus (WNV) in Europe and North America. Cases of WNV have been recorded in Italy since 1998. In particular, wet areas along the Po River are considered some of the most WNV affected areas in Italy. Here, we analyzed the genetic structure of ten Cx. pipiens populations collected in the last part of the Po River including the Delta area.MethodsWe assessed the genetic variability of two mitochondrial markers, cytochrome oxidase 1 (COI) and 2 (COII), for a total of 1200 bp, and one nuclear marker, a fragment of acetylcholinesterase-2 (ace-2), 502 bp long. The effect of the landscape features was evaluated comparing haplotype and nucleotide diversity with the landscape composition.ResultsThe analysis showed a high genetic diversity in both COI and COII gene fragments mainly shared by the populations in the Delta area. The COI-COII network showed that the set of haplotypes found was grouped into three main supported lineages with the higher genetic variability gathered in two of the three lineages. By contrast, ace-2 fragment did not show the same differentiation, displaying alleles grouped in a single clade. Finally, a positive correlation between mitochondrial diversity and natural wetland areas was found.ConclusionsThe high mitochondrial genetic diversity found in Cx. pipiens populations from the Po River Delta contrasts with the low variability of inland populations. The different patterns of genetic diversity found comparing mitochondrial and nuclear markers could be explained by factors such as differences in effective population size between markers, sex biased dispersal or lower fitness of dispersing females. Moreover, the correlation between genetic diversity and wetland areas is consistent with ecosystem stability and lack of insecticide pressure characteristic of this habitat. The mtDNA polymorphism found in the Po River Delta is even more interesting due to possible linkages between the mitochondrial lineages and different biting behaviors of the mosquitoes influencing their vector ability of arboviral infections.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1429-1) contains supplementary material, which is available to authorized users.

The pine processionary moth (Thaumetopoea pityocampa) is an important pest of coniferous forests at the southern edge of its range in Maghreb. Based on mitochondrial markers, a strong genetic differentiation was previously found in this species between western (pityocampa clade) and eastern Maghreb populations (ENA clade), with the contact zone between the clades located in Algeria. We focused on the moth range in Algeria, using both mitochondrial (a 648 bp fragment of the tRNA‐cox2) and nuclear (11 microsatellite loci) markers. A further analysis using a shorter mtDNA fragment and the same microsatellite loci was carried out on a transect in the contact zone between the mitochondrial clades. Mitochondrial diversity showed a strong geographical structure and a well‐defined contact zone between the two clades. In particular, in the pityocampa clade, two inner subclades were found whereas ENA did not show any further structure. Microsatellite analysis outlined a different pattern of differentiation, with two main groups not overlapping with the mitochondrial clades. The inconsistency between mitochondrial and nuclear markers is probably explained by sex‐biased dispersal and recent afforestation efforts that have bridged isolated populations.

We examined genetic variation at 21 polymorphic allozyme loci, 15 nuclear DNA loci, and mitochondrial DNA in four spawning populations of sockeye salmon (Oncorhynchus nerka) from Cook Inlet, Alaska, to test for differences in the patterns of divergence among different types of markers. We were specifically interested in testing the suggestion that natural selection at allozyme loci compromises the effectiveness of these markers for describing the amount and patterns of gene flow among populations. We found concordance among markers in the amount of genetic variation within and among populations, with the striking exception of one allozyme locus (sAH), which exhibited more than three times the amount of among-population differentiation as other loci. A consideration of reports of discordance between allozymes and other loci indicates that these differences usually result from one or two exceptional loci. We conclude that it is important to examine many loci when estimating genetic differentiation to infer historical amounts of gene flow and patterns of genetic exchange among populations. It is less important whether those loci are allozymes or nuclear DNA markers.

Simple SummaryThe main pollinator for the African oil palm, Elaeis guineensis, in Malaysia and Indonesia is Elaeidobius kamerunicus. The weevil species is not native to these countries, but it was introduced from Cameroon, West Africa, in 1981 to improve pollination efficiency, thus improving bunch formation and yield. Forty years after the weevil introduction, recent reductions in bunch yield reported in Malaysia caused by poor bunch formation could be associated with a decrease in pollination efficiency due to the founder effect. Several genetic diversity studies of weevil populations based on morphological and mitochondrial markers have been carried out; however, the studies did not provide sufficient evidence for explaining the genetic variation, particularly at the intra-species level. This study aims to develop a set of robust E. kamerunicus-specific nuclear DNA markers to directly assess the genetic diversity of weevil populations. The marker development lays the foundation for future applications by extending the survey into larger areas where the oil palm is cultivated to obtain a more comprehensive understanding of the genetic diversity and inbreeding occurrence status of E. kamerunicus in these introduced regions. This could facilitate sustainable genetic monitoring and conservation planning of E. kamerunicus, especially in non-native oil palm-growing countries.The oil palm-pollinating weevil (Elaeidobius kamerunicus Faust) was introduced from Cameroon, West Africa, to Malaysia in 1981, and subsequently, to other oil palm-growing countries as well. This study aims to develop a set of robust E. kamerunicus-specific nuclear DNA markers to directly assess the genetic diversity of the weevil populations. A total of 19,148 SNP and 223,200 SSR were discovered from 48 weevils representing three origins (Peninsular Malaysia, Sabah, and Riau) using RAD tag sequencing. Subsequent filtering steps further reduced these to 1000 SNP and 120 SSR. The selected 220 SNP exhibited a polymorphism information content (PIC) of 0.2387 (±0.1280), and 8 SSR had the PIC of 0.5084 (±0.1928). These markers were found to show sufficient polymorphism, making it possible to assign 180 weevils into three major clusters from Ghana, Cameroon, and Southeast Asia (mainly in Malaysia and Indonesia). These DNA markers successfully confirmed the Cameroon origin of the Southeast Asian cluster. However, the presence of null alleles in the SSR markers, due to limited flexibility of the probe design on the short RAD tags, led to an underestimation of heterozygosity within the populations. Hence, the developed SNP markers turned out to be more efficient than the SSR markers in the genetic diversity assessment of the E. kamerunicus populations. The genetic information provides useful insight into developing guidelines for the genetic monitoring and conservation planning of E. kamerunicus.

The black marlin Istiompax indica is a highly migratory species and as a result is expected to show little genetic population structure throughout its broad geographic range. Tissue samples from 183 I. indica were collected from three geographic regions within the central Indo-Pacific and analysed using mitochondrial and nuclear DNA markers. Nuclear genetic heterogeneity was found among populations in the south-western Pacific Ocean, eastern Indian Ocean and South China Sea (significant FST values of 0.013–0.037). Combining information from nuclear markers with published movement and reproductive data suggests that reproductive philopatry plays a role in maintaining contemporary I. indica population structure. Analyses of the mitochondrial control region did not reflect this pattern; however, it identified historical population structure. Differing patterns of genetic population structure revealed by mitochondrial and nuclear markers demonstrated that a transition must have occurred between historical and contemporary population structures. This restructuring presumably reflects a species whose populations have become genetically isolated before experiencing a period of secondary contact. The spatial subdivision evident among populations indicates that I. indica in this central Indo-Pacific region should be managed as three independent stocks, to guide the sustainability of this fisheries resource.

Parsley frogs (Pelodytes) comprise the only genus in the family Pelodytidae, an ancient anuran lineage that split from their closest relatives over 140 million years ago. Pelodytes is a Palearctic group restricted to Western Eurasia including three extant species: the eastern species P. caucasicus, endemic to the Caucasus area, and two closely related species inhabiting Western Europe: the Iberian endemic P. ibericus and the more widespread P. punctatus. Previous studies based on mitochondrial and nuclear DNA markers have revealed the existence of two additional lineages of Parsley frogs in the Iberian Peninsula, which have been flagged as candidate species. Here, we integrate novel molecular, morphological and bioacoustical data to assess the differentiation of the four western Parsley frog lineages. Species trees and Bayesian population assignment analyses based on nuclear markers confirm previous studies and concordantly delineate four parapatric lineages with narrow hybrid zones. Mitochondrial divergence is low (< 2% pairwise distances in the 16S rRNA gene), in line with previously reported low mitochondrial substitution rates in non-neobatrachian frogs. Based on concordance between mitochondrial and nuclear markers, we conclude that four species of Parsley frogs occur in Western Europe: Pelodytes punctatus, distributed from northern Italy to northeastern Spain; Pelodytes ibericus, inhabiting southern Spain and southern Portugal; Pelodytes atlanticus sp. nov., from the Portuguese Atlantic coast; and Pelodytes hespericus sp. nov., occurring in central and eastern Spain. However, bioacoustical and morphological differentiation of these species is low, with no obvious and qualitative diagnostic characters allowing full species discrimination. Differences in the relative size of metacarpal tubercles exist but this character is variable. Pelodytes ibericus and Pelodytes atlanticus are smaller than the other two species, and P. ibericus has shorter limbs and various distinctive osteological characters. Bioacoustically, the pattern by which two different note types are combined in advertisement calls separates P. hespericus from the remaining species. Despite these differences, we emphasize that the taxonomic status of all four western Parsley frogs requires additional investigation, especially the patterns of genetic admixture across contact zones. While a status of separate species best conforms to the currently available data, alternative hypotheses are also discussed.