Entamoeba moshkovskii, according to recent studies, appears to exert a more significant impact on diarrhoeal infections than previously believed. The efficient identification and genetic characterization of E. moshkovskii isolates from endemic areas worldwide are crucial for understanding the impact of parasite genomes on amoebic infections. In this study, we employed a multilocus sequence typing system to characterize E. moshkovskii isolates, with the aim of assessing the role of genetic variation in the pathogenic potential of E. moshkovskii. We incorporated 3 potential genetic markers: KERP1, a protein rich in lysine and glutamic acid; amoebapore C (apc) and chitinase. Sequencing was attempted for all target loci in 68 positive E. moshkovskii samples, and successfully sequenced a total of 33 samples for all 3 loci. The analysis revealed 17 distinct genotypes, labelled M1–M17, across the tested samples when combining all loci. Notably, genotype M1 demonstrated a statistically significant association with diarrhoeal incidence within E. moshkovskii infection (P = 0.0394). This suggests that M1 may represent a pathogenic strain with the highest potential for causing diarrhoeal symptoms. Additionally, we have identified a few single-nucleotide polymorphisms in the studied loci that can be utilized as genetic markers for recognizing the most potentially pathogenic E. moshkovskii isolates. In our genetic diversity study, the apc locus demonstrated the highest Hd value and π value, indicating its pivotal role in reflecting the evolutionary history and adaptation of the E. moshkovskii population. Furthermore, analyses of linkage disequilibrium and recombination within the E. moshkovskii population suggested that the apc locus could play a crucial role in determining the virulence of E. moshkovskii.

A world-wide coconut germplasm collection (79 genotypes) was analyzed for genetic diversity and population structure based on 48 simple sequence repeat (SSR) loci. The genotypes displayed moderately high amount of genetic diversity, which was strongly structured according to geographical origins. Number of SSR alleles ranged from 2 to 7 with an average of 4.1 per locus. Gene diversity (expected heterozygosity) estimates ranged from 0.162 to 0.811 with a mean of 0.573. Polymorphism information content values ranged from 0.149 to 0.785 with an average of 0.522. Hierarchical clustering analysis grouped the genotypes into two major clusters with two sub-groups in each, which corresponded with the geographic origins. The first cluster comprised of ‘Tall’ genotypes originated from Indo-Atlantic and South Asia regions. The second cluster comprised mostly of ‘Dwarf’ genotypes and some Tall genotypes which originated from Indo-Pacific and South-East Asia regions. Model-based clustering by STRUCTURE analysis also supported the presence of clear genetic structuring in the collection with two major populations (K = 2) and four sub-populations (K = 4). The proportion of SSR locus-pairs in linkage disequilibrium was low (2.4%). Association analysis in a subset of 44 genotypes detected a single SSR locus, CnCir73 (chromosome 1) putatively associated with fruit yield component traits, which corresponded with a previously mapped quantitative trait locus in coconut.

In the present study, we used genomic data, generated with a medium density single nucleotide polymorphisms (SNP) array, to acquire more information on the population structure and evolutionary history of the synthetic Frizarta dairy sheep. First, two typical measures of linkage disequilibrium (LD) were estimated at various physical distances that were then used to make inferences on the effective population size at key past time points. Population structure was also assessed by both multidimensional scaling analysis and k-means clustering on the distance matrix obtained from the animals’ genomic relationships. The Wright’s fixation FST index was also employed to assess herds’ genetic homogeneity and to indirectly estimate past migration rates. The Wright’s fixation FIS index and genomic inbreeding coefficients based on the genomic relationship matrix as well as on runs of homozygosity were also estimated. The Frizarta breed displays relatively low LD levels with r2 and |Dʹ| equal to 0.18 and 0.50, respectively, at an average inter-marker distance of 31 kb. Linkage disequilibrium decayed rapidly by distance and persisted over just a few thousand base pairs. Rate of LD decay (β) varied widely among the 26 autosomes with larger values estimated for shorter chromosomes (e.g. β=0.057, for OAR6) and smaller values for longer ones (e.g. β=0.022, for OAR2). The inferred effective population size at the beginning of the breed’s formation was as high as 549, was then reduced to 463 in 1981 (end of the breed’s formation) and further declined to 187, one generation ago. Multidimensional scaling analysis and k-means clustering suggested a genetically homogenous population, FST estimates indicated relatively low genetic differentiation between herds, whereas a heat map of the animals’ genomic kinship relationships revealed a stratified population, at a herd level. Estimates of genomic inbreeding coefficients suggested that most recent parental relatedness may have been a major determinant of the current effective population size. A denser than the 50k SNP panel may be more beneficial when performing genome wide association studies in the breed.

Analysis of genomic data is becoming increasingly common in the livestock industry and the findings have been an invaluable resource for effective management of breeding programs in small and endangered populations. In this paper, with the goal of highlighting the potential of genomic analysis for small and endangered populations, genome-wide levels of linkage disequilibrium, measured as the squared correlation coefficient of allele frequencies at a pair of loci, effective population size, runs of homozygosity (ROH) and genetic diversity parameters, were estimated in Barbaresca sheep using Illumina OvineSNP50K array data. Moreover, the breed’s genetic structure and its relationship with other breeds were investigated. Levels of pairwise linkage disequilibrium decreased with increasing distance between single nucleotide polymorphisms. An average correlation coefficient <0.25 was found for markers located up to 50 kb apart. Therefore, these results support the need to use denser single nucleotide polymorphism panels for high power association mapping and genomic selection efficiency in future breeding programs. The estimate of past effective population size ranged from 747 animals 250 generations ago to 28 animals five generations ago, whereas the contemporary effective population size was 25 animals. A total of 637 ROH were identified, most of which were short (67%) and ranged from 1 to 10 Mb. The genetic analyses revealed that the Barbaresca breed tended to display lower variability than other Sicilian breeds. Recent inbreeding was evident, according to the ROH analysis. All the investigated parameters showed a comparatively narrow genetic base and indicated an endangered status for Barbaresca. Multidimensional scaling, model-based clustering, measurement of population differentiation, neighbor networks and haplotype sharing distinguished Barbaresca from other breeds, showed a low level of admixture with the other breeds considered in this study, and indicated clear genetic differences compared with other breeds. Attention should be given to the conservation of Barbaresca due to its critical conservation status. In this context, genomic information may have a crucial role in management of small and endangered populations.

Focusing on 116 bread wheat landraces, this study investigated high molecular weight glutenin allele polymorphism, gene diversity, genetic variation and linkage disequilibrium (LD) in Glu-1 loci. To identify gluten alleles, sodium dodesyl sulphate-polyacrylamide, gel electrophoresis was used and for statistical analyses POPGENE software was employed. The results indicated that average genetic variation (h) was the highest in Glu-B1 (0.6421) and the lowest in Glu-A1 locus (0.4548); genetic similarity ratio (I) was the highest in Glu-B1 (1.4170); the highest average genetic diversity (Ht) was observed in Glu-B1 (0.6575) and the lowest diversity was observed in Glu-A1 (0.4558). It was also observed that genetic diversity in Glu-1 locus was largely due to intra-population variations. Inter-population gene flow was also calculated as 4.0051. Marmara and Southeastern Anatolia regions, the results further indicated, had the highest (2.8691) and lowest (0.1694) heterozygosity. Genetic erosion risk for Turkish bread wheat landraces was also seen to be high. Considering the mutual analyses of subunits of nationwide wheat landraces, it is possible to speculate about a limited migration between the landraces. LD of the landraces was largely because of this limited migration and/or epistatic natural selection. Since Turkey is known as the gene centre for major cereals including wheat, barley, rye and oat, where they diversified and spread throughout the world, studying the gluten allele diversity of Turkish bread wheat landraces is important. In addition, this study has revealed the applicability of LD, and neutrality tests to gluten protein diversity for the first time.

Population structure and linkage disequilibrium (LD) were investigated in a set of 395 bread wheat landraces including 154 spring, 193 winter, two facultative wheat and 46 unknown growth type collected from different geographical regions of Iran. A total of 53 microsatellite markers distributed in three genomes of wheat were assayed for polymorphism. The 312 polymorphic alleles were served to estimate population structure and analyse the genome-wide LD. The number of alleles ranged from 2 to 18 with an average of 5.89 alleles/locus. Mean of polymorphic information content was 0.6 with a range of 0.15–0.86 and gene diversity varied from 0.16 to 0.88 with an average of 0.64. The population of landraces was highly structured and based on distance-and model-based cluster analyses the 395 landraces were assigned into eight subpopulations: SG1–SG8. Population structure estimates based on simple sequence repeat (SSR) marker data were quantified in a Q matrix and used in calculation of LD between pair of markers. A low overall LD level found in 12–13% (166) of the SSR marker pairs showed significant pairwise LD in r2 ≥ 0.01 and P ≤ 0.001 and six pair showed r2 ≥ 0.05 with P ≤ 0.001. LD clearly decays within the genetic distance of 40–60 cM with r2 ~ 0.05. The results of this study should provide valuable information for future association mapping using this wheat collection.

Smallholder farmers in Africa and other developing countries raise village chickens under low input production systems characterised by minimal management and veterinary interventions. However, village chickens have a major contribution to village communities as a source of protein, income and cultural needs. The village chickens are described as nondescript birds that have not been developed as a breed and with uncharacterised genetic attributes. Local chickens are found in the most marginalised farming systems, to which they have adapted and survived the harsh production and environmental conditions. Such survival and adaptive attributes need to be characterised, conserved, and utilised. Several studies on Southern African local chickens have revealed high genetic variation within and among the village chicken populations and indicated that village chickens contribute diversity that is unique and different from that of commercial and specialised chicken populations. The availability of whole genome single nucleotide polymorphism (SNP) data facilitates the use of powerful statistical methods for in depth investigations into the evolutionary history and population demographics of village chickens. Linkage disequilibrium (LD), as a function of effective population size, has been used to estimate rate and level of inbreeding as well as selection pressures in the absence of pedigree information. The extent and distribution of LD in the genome has been exploited to shed light on the origin and domestication of animals and facilitate an understanding of breeds’ relatedness. The Illumina iSelect chicken 60K SNP chip has over 54 000 SNPs that have found application in population and quantitative genetics studies and has revealed demographic history, effective population size and level of genetic erosion in commercial and traditional/village chicken populations. The recent launching of the 600 K Affymetrix® Axiom® HD genotyping array for chickens and the ever decreasing cost of sequencing will see improved estimation of LD and the associated population parameters.

Durum wheat (Triticum durum) is predominantly grown as spring type and depending on the production area autumn or spring sowing is used. For the durum production in Austria and Germany, autumn sowing has several advantages, such as yield increase and stability, but this requires the selection for winter hardiness including a good frost tolerance. The aim of this study was to support breeding of winter durum and to facilitate genomic approaches by molecularly characterizing a panel of 170 diverse winter and 14 spring durum lines employing a genotyping-by-sequencing approach. We obtained an unprecedentedly high number of 30,611 polymorphic markers covering the entire genome. The principal coordinate analysis and the cluster analysis revealed the absence of a major population structure but a tendency of lines to group according to their country of origin. Linkage disequilibrium was found to decay within a short distance of approximately 2–5 cM and also showed variable patterns along chromosomes. In summary, our results can assist breeding of durum wheat and pave the way for genomic approaches towards knowledge-based winter durum breeding.

To examine how community-level genetic diversity of the malaria parasite Plasmodium vivax varies across time and space, we investigated the dynamics of parasite polymorphisms during the early phases of occupation of a frontier settlement in the Amazon Basin of Brazil. Microsatellite characterization of 84 isolates of P. vivax sampled over 3 years revealed a moderate-to-high genetic diversity (mean expected heterozygosity, 0·699), with a large proportion (78·5%) of multiple-clone infections (MCI), but also a strong multilocus linkage disequilibrium (LD) consistent with rare outcrossing. Little temporal and no spatial clustering was observed in the distribution of parasite haplotypes. A single microsatellite haplotype was shared by 3 parasites collected during an outbreak; all other 81 haplotypes were recovered only once. The lowest parasite diversity, with the smallest proportion of MCI and the strongest LD, was observed at the time of the outbreak, providing a clear example of epidemic population structure in a human pathogen. Population genetic parameters returned to pre-outbreak values during last 2 years of study, despite the concomitant decline in malaria incidence. We suggest that parasite genotyping can be useful for tracking the spread of new parasite strains associated with outbreaks in areas approaching malaria elimination.

We examined longitudinal associations of vitamin D receptor (VDR) and megalin (LRP2; LDL receptor-related protein-2) gene polymorphisms with central adiposity. We used data from the Baltimore Longitudinal Study of Aging (BLSA), an ongoing prospective open cohort study. Study participants consisted of non-Hispanic white adults residing in Baltimore city, with one or more visits at age ≥50 years, and complete data (n 609–617). Repeated assessments on waist circumference (WC) and waist:hip ratio (WHR) were available. Multiple linear mixed models were used to estimate mid-follow-up age central adiposity level and annual rate of change with cut-points set at the sex-specific 80th percentile. The four binary outcomes were: ‘elevated central adiposity’ (ECA-WC and ECA-WHR) and ‘significant increase in central adiposity’ (SICA-WC and SICA-WHR). SNP for VDR (four SNP: (1) rs11568820 (CdX-2:T/C); (2) rs1544410 (BsmI:G/A); (3) rs7975232 (ApaI:A/C); (4) rs731236 (TaqI:G/A)) and Megalin (three SNP: (1) rs3755166:G/A; (2) rs2075252:C/T; (3) rs4668123:C/T) genes were selected. SNP latent classes (SNPLC) and SNP haplotypes (SNPHAP) were created. Multiple logistic regression analyses indicated that, in men, higher ECA-WHR odds were associated with SNPLC Megalin2:rs3755166[–]/rs2075252[TT]/rs4668123[T–] (v. Megalin1:rs3755166[–]/rs2075252[CC]/rs4668123[–]) (OR 2·87; 95 % CI 1·15, 7·12; P = 0·023) and that SNPLC Megalin3:rs3755166[–]/rs2075252[CT]/rs4668123[–] (v. Megalin1) was linked to lower SICA-WC odds (OR 0·48; 95 % CI 0·26, 0·88; P = 0·019) (P > 0·05 for sex × SNPLC). In women, VDR3 SNPHAP (GAA:bAT) was related to lower odds of ECA-WC (OR 0·37; 95 % CI 0·16, 0·87; P = 0·023) (P < 0·05 for sex × SNPHAP), VDR1 SNPHAP (GCA:baT) was associated with greater odds and VDR3 SNPHAP (GAA:bAT) with lower odds of SICA-WC (P > 0·05 for sex × SNPHAP). Vitamin D-related gene polymorphisms were associated with central adiposity status and change. Future mechanistic studies are needed to confirm those polymorphisms' biological significance to central adiposity.

Single nucleotide polymorphisms (SNPs) are the most widespread source of variation in genomes. While the very large number of SNPs allows for a very precise description of genetic variation, it impedes data processing and significantly increases analysis time. Many of the SNPs located close to each other frequently carry the same or similar information. This problem can be solved by selecting the most informative SNPs (tagSNPs) using linkage disequilibrium information by identifying a set of tagSNPs representative for a chromosome fragment. The goal of this study is to check whether the genetic structure of a population, expressed by relationship and inbreeding coefficients, affects tagSNP selection. Six subsets of 450 bulls are selected out of the 1228 Polish Holstein-Friesian bulls genotyped by the Illumina BovineSNP50 Bead Chip. TagSNPs are selected for each of the subsets, as well as for the whole data set. The average reduction of the SNP number is 77.2% and is very similar in each sub-population. Differences in tagSNP selection between sub-populations are small. On average, 93.88% of the tagSNPs overlap between subsets. The study showed that differences in the genetic structure of the reference population have little influence on tagSNP selection.

The meagre Argyrosomus regius is a large Sciaenid fish known to reproduce in the eastern Atlantic and Mediterranean Sea in just five distinct and restricted geographic areas: along the Mauritanian coast and at estuary openings (Gironde, Tagus, Guadalquivir and Nile). The biological traits of A. regius (high dispersal capabilities, high fecundity, long larval phase, overlapping generations, reproduction until 40 years of age) are, in principle, favourable to high gene flow, which should lead to genetic homogeneity over large geographic scales. Nevertheless, the high geographic distances between the few reproductive areas leads one ask whether there is genetic differentiation in this species. In the present study, the genetic differentiation of the wild A. regius was investigated across most of its natural range from the Atlantic Ocean (France, Portugal, Spain, Mauritania) to the Mediterranean Sea (Egypt, Turkey), using 11 microsatellite markers previously identified in another Sciaenid, the red drum Sciaenops ocellatus. At least two very distinct groups could be identified, separated by the Gibraltar Strait. Genetic divergences (FST values) were intermediate between the Atlantic samples (0.012–0.041), high between Egypt and the Atlantic (0.06–0.107) or Aegean Sea (0.081) and extremely high between the Aegean Sea and the Atlantic (0.098–0.168). A. regius exhibited a very high level of genetic differentiation rarely reported in marine fishes. These results also demonstrate the existence of a sixth independent spawning area in the Menderes delta (Turkey). Factors potentially involved in this very high genetic fragmentation are discussed, including physical barriers, glaciation pulses and biological traits.

Linkage disequilibrium (LD), the association in populations between genes at linked loci, has achieved a high degree of prominence in recent years, primarily because of its use in identifying and cloning genes of medical importance. The field has recently been reviewed by Slatkin (2008). The present article is largely devoted to a review of the theory of LD in populations, including historical aspects.

Recently a paper authored by ourselves and a number of co-authors about the proportion of phenotypic variation in height that is explained by common SNPs was published in Nature Genetics (Yang et al., 2010). Common SNPs explain a large proportion of the heritability for human height (Yang et al.). During the refereeing process (the paper was rejected by two other journals before publication in Nature Genetics) and following the publication of Yang et al. (2010) it became clear to us that the methodology we applied, the interpretation of the results and the consequences of the findings on the genetic architecture of human height and that for other traits such as complex disease are not well understood or appreciated. Here we explain some of these issues in a style that is different from the primary publication, that is, in the form of a number of comments and questions and answers. We also report a number of additional results that show that the estimates of additive genetic variation are not driven by population structure.

The Index of Association (IA) has been proposed by Maynard Smith et al. (1993) as a general method for characterizing the population structures of microorganisms as either: clonal, epidemic, cryptic species or panmictic. With reference to the current debate surrounding the mode of reproduction in parasitic protozoa, this study explores (i) the suitability and limitations of the IA for characterizing populations of Trypanosoma brucei s.l., and (ii) the idea that the significance of genetic differences between populations may be better understood if the evolution, spread and temporal stability of certain parasite genotypes are also considered. Four populations of T. brucei from Côte d'Ivoire, Uganda and Zambia are analysed using the IA and a complementary test for linkage disequilibrium, test f of Tibayrenc, Kjellberg & Ayala (1990). The two populations from Uganda are characterized as epidemic, while the others appear more or less clonal; the merits of the two methods are compared. The implications of the various population classifications are discussed with reference to genotype longevity in each region; the evolutionary and biomedical consequences of the genetic non-homogeneity of T. brucei are reviewed.

In order to estimate the value of population genetics for both the taxonomy of trypanosomes belonging to the species Trypanosoma brucei and a better understanding of Human African Trypanosomiasis (HAT), we undertook a cellulose acetate electrophoresis isoenzyme study involving 55 stocks isolated from man and animals in Congo, Zaire and Cameroun. Out of the 24 loci surveyed, 15 exhibited variability, which made it possible to delimit 23 zymodemes, divided into 2 groups. The first group equated to the classical subspecies Trypanosoma brucei gambiense, while the second corresponded to the classical subspecies Trypanosoma brucei brucei. These results broadly agree with the current taxonomy, and are corroborated by RFLP analysis of kDNA. Statistical analysis indicates a basically clonal reproduction system of the trypanosomes in the area studied; the zymodemes are equivalent to natural clones (or a family of closely related clones), stable in space and time. Epidemiological hypotheses are proposed according to the geographic distribution of the clones in this area.

This study analyses the different populations of Trypanosoma brucei spp. which may coexist within the midgut of wild tsetse flies (Stevens et al. 1994). Cloned trypanosome populations characterized by multilocus enzyme electrophoresis (MLEE) were further analysed by the random amplified polymorphic DNA (RAPD) technique, allowing detection of genetic variation at a finer level than that possible by MLEE. Genetic distance matrices derived from the results of each of the two biochemical methods were calculated and compared using a computer program based on the method of Mantel (1967). The observed correlation was used to investigate the degree of linkage disequilibrium (LD) in the data, association between unrelated polymorphic markers providing a measure of the departure from panmixia. The potential of each biochemical method to detect linkage was evaluated by an extended Mantel test. The MLEE/RAPD correlation test evidenced significant LD within the population, suggesting a predominantly clonal method of reproduction for these West African trypanosomes. Analysis of RAPD data by the extended Mantel test also showed significant LD, while the results with MLEE data were less conclusive, providing an indication of the relative potential of the two techniques to detect fine genetic variation.

The effectiveness of association mapping (AM) based on linkage disequilibrium (LD) is currently being tested in a number of crops. An important prerequisite for the application of AM is the availability of collections of accessions with a suitable level of genetic variation for target traits and with limited spurious LD due to the presence of population structure. Herein, the results of a genome-wide molecular characterization of a collection of elite durum wheat accessions well-adapted to Mediterranean environments are presented. Ninety-seven highly polymorphic simple sequence repeats and 166 amplified fragment length polymorphism markers were used to characterize 189 durum accessions, mainly cultivars and advanced breeding lines. Genome-wide significant and sizeable LD indices at a centimorgan scale were observed, while LD mainly decayed within 10 cM. On the other hand, effects due to spurious LD were notably lower than those previously observed in a durum wheat collection sampling durum gene pools of more diverse origin. These results, coupled with the high level of genetic variability detected for a number of important morpho-physiological traits and their high heritability, indicate the suitability of this collection for AM studies targeting agronomically important traits.