Fine mapping of multiple QTL using combined linkage and linkage disequilibrium mapping – A comparison of single QTL and multi QTL methods

Two previously described QTL mapping methods, which combine linkage analysis (LA) and linkage disequilibrium analysis (LD), were compared for their ability to detect and map multiple QTL. The methods were tested on five different simulated data sets in which the exact QTL positions were known. Every simulated data set contained two QTL, but the distances between these QTL were varied from 15 to 150 cM. The results show that the single QTL mapping method (LDLA) gave good results as long as the distance between the QTL was large (> 90 cM). When the distance between the QTL was reduced, the single QTL method had problems positioning the two QTL and tended to position only one QTL, i.e. a "ghost" QTL, in between the two real QTL positions. The multi QTL mapping method (MP-LDLA) gave good results for all evaluated distances between the QTL. For the large distances between the QTL (> 90 cM) the single QTL method more often positioned the QTL in the correct marker bracket, but considering the broader likelihood peaks of the single point method it could be argued that the multi QTL method was more precise. Since the distances were reduced the multi QTL method was clearly more accurate than the single QTL method. The two methods combine well, and together provide a good tool to position single or multiple QTL in practical situations, where the number of QTL and their positions are unknown.     

Mapping the genomic architecture of ecological speciation in the wild: does linkage disequilibrium hold the key?

The hunt for the genes underlying ecological speciation has now closed in on a number of candidates, but making the link from genotype to phenotype continues to pose a significant challenge. This is partly because genetic studies in many systems remain impeded by long generation times or an inability to perform controlled crosses. Now, in this issue of Molecular Ecology, Malek et al. (2012) demonstrate the utility of a novel admixture mapping approach that can be used to identify genomic regions contributing to adaptive trait divergence between natural populations. Remarkably, they validate their approach by mapping traits associated with mate choice in a wild limnetic and benthic threespine stickleback (Gasterosteus aculeatus) species pair, finding several loci associated with male nuptial coloration and shape. While this study benefited from tried‐and‐true microsatellites in a well‐characterized species with a detailed genetic map (and genome sequence), the field is quickly moving towards the use of next‐generation sequencing, especially for nonmodel systems. The ability to characterize molecular polymorphisms for any system suggests that molecular ecologists working on virtually any species may benefit from applying Malek et al.'s approach, if naturally admixed populations are available.     

Do multiple experimenters improve the reproducibility of animal studies?

The credibility of scientific research has been seriously questioned by the widely claimed “reproducibility crisis”. In light of this crisis, there is a growing awareness that the rigorous standardisation of experimental conditions may contribute to poor reproducibility of animal studies. Instead, systematic heterogenisation has been proposed as a tool to enhance reproducibility, but a real-life test across multiple independent laboratories is still pending. The aim of this study was therefore to test whether heterogenisation of experimental conditions by using multiple experimenters improves the reproducibility of research findings compared to standardised conditions with only one experimenter. To this end, we replicated the same animal experiment in 3 independent laboratories, each employing both a heterogenised and a standardised design. Whereas in the standardised design, all animals were tested by a single experimenter; in the heterogenised design, 3 different experimenters were involved in testing the animals. In contrast to our expectation, the inclusion of multiple experimenters in the heterogenised design did not improve the reproducibility of the results across the 3 laboratories. Interestingly, however, a variance component analysis indicated that the variation introduced by the different experimenters was not as high as the variation introduced by the laboratories, probably explaining why this heterogenisation strategy did not bring the anticipated success. Even more interestingly, for the majority of outcome measures, the remaining residual variation was identified as an important source of variance accounting for 41% (CI₉₅ [34%, 49%]) to 72% (CI₉₅ [58%, 88%]) of the observed total variance. Despite some uncertainty surrounding the estimated numbers, these findings argue for systematically including biological variation rather than eliminating it in animal studies and call for future research on effective improvement strategies.

An experimenter heterogenisation was not sufficient to prevent idiosyncratic results in a multi-laboratory setting. Astonishingly, neither the experimenter nor the laboratory accounted for the main portion of the observed variation, but a high amount of residual variation in fact remained unexplained despite strict standardisation regimes.     

Why is there so little intragenic linkage disequilibrium in humans?

The efficient design of association mapping studies relies on a knowledge of the rate of decay of linkage disequilibrium with distance. This rate depends on the population recombination rate, C. An estimate of C for humans is usually obtained from a comparison of physical and genetic maps, assuming an effective population size of approximately 10(4). We demonstrate that under both a constant population size model and a model of long-term exponential growth, there is evidence for more recombination in polymorphism data than is expected from this estimate. An important contribution of gene conversion to meiotic recombination helps to explain our observation, but does not appear to be sufficient. The occurrence of multiple hits at CpG sites and the presence of population structure are not explanations.     

Does fecundity drive the evolution of insect diet?

We investigate whether egg load (a surrogate for fecundity) drives host specificity in a herbivorous insect. In many insects, including our study organism (Edith's checkerspot butterfly), both egg load and tendency to accept low-ranked hosts increase during each search for an oviposition site. Effects on host acceptance of egg load and passage of time are thereby potentially confounded. We conducted two experiments designed to disentangle these effects. In both experiments, we estimated the times of first acceptance of both a high-ranked and a low-ranked host, without allowing the insects to oviposit. In the first experiment, we measured egg load at the time of first acceptance of the low-ranked host. The later the time of first acceptance, the higher was the fecundity. We therefore reject the hypothesis that all insects accepted the low-ranked host at the same predetermined egg load. In the second experiment, we measured egg load 48 h after the high-ranked host was first accepted. We found no relationship between egg load and timing of acceptance of the low-ranked host. Insects with higher rates of egg accumulation did not accept the low-ranked host sooner. Taken together, these results suggest that acceptance of the low-ranked host is not driven directly by egg load. Rather, this acceptance results from some other process that is influenced by time since last oviposition. We conclude that there is no evidence to support the assumption that females with high rates of egg accumulation are more likely to accept low-ranked hosts.     

Does secotioid inertia drive the evolution of false-truffles?

Secotioid inertia is a model implemented to explain the prevalence of highly derived false-truffles with no obvious connection to the Homobasidiomycetes. The model accommodates the apparent lack of epigeous sister taxa for some highly derived hypogeous lineages by assuming that gasteromycetation in some fungi leads to the extinction of their epigeous sister population. The derived state of some hypogeous lineages suggests that they arose early in the evolution of Homobasidiomycetes and that those groups were subject to conditions that favoured hypogeous lineages such that the hypogeous fruit body form became the predominant form for some lineages. The directional selection component of secotioid inertia, termed secotioid drive, led to the extinction of their epigeous sister taxon. Morphological and molecular data from Russulaceae are used to model the evolutionary stages of secotioid inertia. The resulting phylogenetic results are compared with data from the order Leucogastrales, and the genus Destuntzia. The implications of secotioid drive are discussed with reference to gasteromycete phylogenetics, evolution, and conservation. Specifically, secotioid inertia can be used to account for reversals in fruit body morphology and instability in mycorrhizal formation.     

Association mapping, patterns of linkage disequilibrium and selection in the vicinity of the PHYTOCHROME C gene in pearl millet

Key message

Linkage analysis confirmed the association in the region of PHYC in pearl millet. The comparison of genes found in this region suggests that PHYC is the best candidate.

Abstract

Major efforts are currently underway to dissect the phenotype–genotype relationship in plants and animals using existing populations. This method exploits historical recombinations accumulated in these populations. However, linkage disequilibrium sometimes extends over a relatively long distance, particularly in genomic regions containing polymorphisms that have been targets for selection. In this case, many genes in the region could be statistically associated with the trait shaped by the selected polymorphism. Statistical analyses could help in identifying the best candidate genes into such a region where an association is found. In a previous study, we proposed that a fragment of the PHYTOCHROME C gene (PHYC) is associated with flowering time and morphological variations in pearl millet. In the present study, we first performed linkage analyses using three pearl millet F₂ families to confirm the presence of a QTL in the vicinity of PHYC. We then analyzed a wider genomic region of ~100 kb around PHYC to pinpoint the gene that best explains the association with the trait in this region. A panel of 90 pearl millet inbred lines was used to assess the association. We used a Markov chain Monte Carlo approach to compare 75 markers distributed along this 100-kb region. We found the best candidate markers on the PHYC gene. Signatures of selection in this region were assessed in an independent data set and pointed to the same gene. These results foster confidence in the likely role of PHYC in phenotypic variation and encourage the development of functional studies.     

Co-selection of the H63D mutation and the HLA-A29 allele: a new paradigm of linkage disequilibrium?

The major histocompatibility complex (MHC) shows a remarkable conservation of particular HLA antigens and haplotypes in linkage disequilibrium in most human populations, suggesting the existence of a convergent evolution. A recent example of such conservation is the association of particular HLA haplotypes with the HFE mutations. With the objective of exploring the significance of that association, the present paper offers an analysis of the linkage disequilibrium between HLA alleles or haplotypes and the HFE mutations in a Portuguese population. Allele and haplotype associations between HLA and HFE mutations were first reviewed in a population of 43 hemochromatosis families. The results confirmed the linkage disequilibrium of the HLA haplotype HLA-A3-B7 and the HLA-A29 allele, respectively, with the HFE mutations C282Y and H63D. In order to extend the study of the linkage disequilibrium between H63D and the HLA-A29-containing haplotypes in a normal, random population, an additional sample of 398 haplotypes was analyzed. The results reveal significant linkage disequilibrium between the H63D mutation and all HLA-A29-containing haplotypes, favoring the hypothesis of a co-selection of H63D and the HLA-A29 allele itself. An insight into the biological significance of this association is given by the finding of significantly higher CD8(+) T-lymphocyte counts in subjects simultaneously carrying the H63D mutation and the HLA-A29 allele.     

Does domain swapping improve the stability of RNase A?

Self-assembling complexes have potential as novel supramolecular biomaterials but domain swapped complexes have yet to investigated in this capacity. Bovine ribonuclease A (RNase A) is a useful model protein as it is able to form a range of three dimensional domain swapped structures, including dimers, trimers and tetramers that have similar catalytic ability. However, little work has been carried out investigating the physical characteristics of these complexes. In an effort to characterise the strength of these oligomeric interactions, analytical ultracentrifugation was carried out to measure the dissociation of higher order complexes, using fluorescent tags to test for dissociation at very low concentrations. Results of this work suggest that the oligomers form a very tight complex, with no evidence of dissociation down to 250 pM. RNase A oligomers also had similar thermal stability to that of monomeric enzyme, suggesting that the main limiting factor in RNase A stability is the tertiary, rather than quaternary structure. Following thermal unfolding of RNase A, the protein refolded upon cooling, but returned to the monomeric state. This latter result may limit the potential of domain swapping as a means of material assembly.     

Does bacteriorhodopsin energize the membranes of animal mitochondria under light?

A suspension of freeze-thawed mitochondria mixed with purple membranes from Halobacteria was illuminated with visible light. It was found that the light exposure prevented inhibition of succinate oxidation. The illumination also led to a decrease in inhibition of the rate of ferricyanide reduction by rat liver mitochondria in the presence of succinate. Both phenomena are explained by the fact that oxalacetate inhibition of succinate dehydrogenase is prevented by light-induced energization of mitochondrial membranes due to the contact with purple membranes.     

Does the location of enrichment material affect behavior and dirtiness in growing female pigs?

The objective of this study was to investigate if female growing pigs spend similar amounts of time exploring a wooden stick as they do exploring enrichment material regardless of its proximity to the feeder. Forty-eight pigs aged 18 to 26 weeks allocated to 16 pens with three pigs per pen were studied. Fifty percent of the pens had a wooden stick beside the feeder, and the rest had a similar stick opposite to the feeder. Two observers assessed the pigs by means of scan and focal sampling. The pigs spent more time (p < .0001) exploring the wood during the first week than during the rest of the study (10.9% vs 3.6%). The pigs with the wood close to the feeder spent less (p = .0001) time resting (29.9%) and more (p < .0001) time exploring (6.3%) the wood than did pigs with the wood opposite to the feeder (32.4% and 2.5%, respectively). In conclusion, a wooden stick placed close to the feeder was associated with more exploratory behaviorcompared with a similar stick placed opposite to the feeder.     

Does one subgenome become dominant in the formation and evolution of a polyploid?

BackgroundPolyploids are common in flowering plants and they tend to have more expanded ranges of distributions than their diploid progenitors. Possible mechanisms underlying polyploid success have been intensively investigated. Previous studies showed that polyploidy generates novel changes and that subgenomes in allopolyploid species often differ in gene number, gene expression levels and levels of epigenetic alteration. It is widely believed that such differences are the results of conflicts among the subgenomes. These differences have been treated by some as subgenome dominance, and it is claimed that the magnitude of subgenome dominance increases in polyploid evolution.ScopeIn addition to changes which occurred during evolution, differences between subgenomes of a polyploid species may also be affected by differences between the diploid donors and changes which occurred during polyploidization. The variable genome components in many plant species are extensive, which would result in exaggerated differences between a subgenome and its progenitor when a single genotype or a small number of genotypes are used to represent a polyploid or its donors. When artificially resynthesized polyploids are used as surrogates for newly formed genotypes which have not been exposed to evolutionary selection, differences between diploid genotypes available today and those involved in the formation of the natural polyploid genotypes must also be considered.ConclusionsContrary to the now widely held views that subgenome biases in polyploids are the results of conflicts among the subgenomes and that one of the parental subgenomes generally retains more genes which are more highly expressed, available results show that subgenome biases mainly reflect legacy from the progenitors and that they can be detected before the completion of polyploidization events. Further, there is no convincing evidence that the magnitudes of subgenome biases have significantly changed during evolution for any of the allopolyploid species assessed.     

Does the interpolation accuracy of species distribution models come at the expense of transferability?

Model transferability (extrapolative accuracy) is one important feature in species distribution models, required in several ecological and conservation biological applications. This study uses 10 modelling techniques and nationwide data on both (1) species distribution of birds, butterflies, and plants and (2) climate and land cover in Finland to investigate whether good interpolative prediction accuracy for models comes at the expense of transferability – i.e. markedly worse performance in new areas. Models’ interpolation and extrapolation performance was primarily assessed using AUC (the area under the curve of a receiver characteristic plot) and Kappa statistics, with supplementary comparisons examining model sensitivity and specificity values. Our AUC and Kappa results show that extrapolation to new areas is a greater challenge for all included modelling techniques than simple filling of gaps in a well‐sampled area, but there are also differences among the techniques in the degree of transferability. Among the machine‐learning modelling techniques, MAXENT, generalized boosting methods (GBM), and artificial neural networks (ANN) showed good transferability while the performance of GARP and random forest (RF) decreased notably in extrapolation. Among the regression‐based methods, generalized additive models (GAM) and generalized linear models (GLM) showed good transferability. A desirable combination of good prediction accuracy and good transferability was evident for three modelling techniques: MAXENT, GBM, and GAM. However, examination of model sensitivity and specificity revealed that model types may differ in their tendencies to either increased over‐prediction of presences or absences in extrapolation, and some of the methods show contrasting changes in sensitivity vs specificity (e.g. ANN and GARP). Among the three species groups, the best transferability was seen with birds, followed closely by butterflies, whereas reliable extrapolation for plant species distribution models appears to be a major challenge at least at this scale. Overall, detailed knowledge of the behaviour of different techniques in various study settings and with different species groups is of utmost importance in predictive modelling.     

Does the location of a mutation determine the ability to form amyloid fibrils?

We have previously reported studies of fibril formation by a set of protein G B1 domain (beta1) variants, with mutations located around the central parallel beta-strands. In this study, we designed multiple mutations in the edge strands of beta1 to create proteins with a stability range comparable to that of the set of central mutants. All the edge variants are able to form amyloid fibrils when they are incubated at their melting temperatures. This result suggests that overall protein stability is the key determinant for amyloid formation and not the specific location of destabilizing mutations. The edge strand and variants cross-seed with each other and with members of the central variant family. Interesting fibrillar morphology was observed in some cross-seeding cases and its implications for a better understanding of nucleation and elongation events are discussed.     

Does mutual sexual selection explain the evolution of head crests in pterosaurs and dinosaurs?

Hone, D.W.E., Naish, D. & Cuthill, I.C. 2011: Does mutual sexual selection explain the evolution of head crests in pterosaurs and dinosaurs? Lethaia, Vol. 45, pp. 139–156. Cranial ornamentation is widespread throughout the extinct non‐avialian Ornithodira, being present throughout Pterosauria, Ornithischia and Saurischia. Ornaments take many forms, and can be composed of at least a dozen different skull bones, indicating multiple origins. Many of these crests serve no clear survival function and it has been suggested that their primary use was for species recognition or sexual display. The distribution within Ornithodira and the form and position of these crests suggest sexual selection as a key factor, although the role of the latter has often been rejected on the grounds of an apparent lack of sexual dimorphism in many species. Surprisingly, the phenomenon of mutual sexual selection – where both males and females are ornamented and both select mates – has been ignored in research on fossil ornithodirans, despite a rich history of research and frequent expression in modern birds. Here, we review the available evidence for the functions of ornithodiran cranial crests and conclude that mutual sexual selection presents a valid hypothesis for their presence and distribution. The integration of mutual sexual selection into future studies is critical to our understanding of ornithodiran ecology, evolution and particularly questions regarding sexual dimorphism. □Behaviour, Dinosauria, ornaments, Pterosauria, sexual selection.     

Does fish represent an intermediate stage in the evolution of ureotelic cytosolic arginase I?

Arginase catalyses the last step of the urea cycle. At least two isoenzymes of arginase are known; cytosolic ARG I and mitochondrial ARG II. ARG I is predominantly expressed in liver cytosol, as a part of urea cycle in ureotelic animals. The second isoform ARG II is primarily responsible for non-ureogenic functions, expressed in mitochondria of both hepatic and non-hepatic tissues in most vertebrates. Most micro-organisms and invertebrates are known to have only one type of arginase, whose function is unrelated to ornithine-urea cycle (OUC). However, in ureo-osmotic marine elasmobranchs arginase is localized in liver mitochondria as a part of OUC to synthesize urea for osmoregulation. An evolutionary transition occurred in arginase enzyme in terrestrial ureotelic vertebrates, with the evolution of ARG I from a pre-existing ancestral mitochondrial ARG II. This cytosolic ARG I activity is supposed to have first appeared in lung fishes, but the 40% and 60% distribution of arginase I and II activity in liver and kidney tissue of Heteropneustes fossilis indicates reconsideration of the above fact.     

Does constructive neutral evolution play an important role in the origin of cellular complexity?

Recently, constructive neutral evolution has been touted as an important concept for the understanding of the emergence of cellular complexity. It has been invoked to help explain the development and retention of, amongst others, RNA splicing, RNA editing and ribosomal and mitochondrial respiratory chain complexity. The theory originated as a welcome explanation of isolated small scale cellular idiosyncrasies and as a reaction to 'overselectionism'. Here I contend, that in its extended form, it has major conceptual problems, can not explain observed patterns of complex processes, is too easily dismissive of alternative selectionist models, underestimates the creative force of complexity as such, and--if seen as a major evolutionary mechanism for all organisms--could stifle further thought regarding the evolution of highly complex biological processes.