Evolution under strong balancing selection: how many codons determine specificity at the female self-incompatibility gene SRK in Brassicaceae?

<p>Abstract</p> <p>Background</p> <p>Molecular lock-and-key systems are common among reproductive proteins, yet their evolution remains a major puzzle in evolutionary biology. In the Brassicaceae, the genes encoding self-incompatibility have been identified, but technical challenges currently prevent detailed analyses of the molecular interaction between the male and female components. In the present study, we investigate sequence polymorphism in the female specificity determinant <it>SRK </it>of <it>Arabidopsis halleri </it>from throughout Europe. Using a comparative approach based on published <it>SRK </it>sequences in <it>A. lyrata </it>and Brassica, we track the signature of frequency-dependent selection acting on these genes at the codon level. Using simulations, we evaluate power and accuracy of our approach and estimate the proportion of codon sites involved in the molecular interaction.</p> <p>Results</p> <p>We identified several members of the S-gene family, together with 22 putative S-haplotypes. Linkage to the S-locus and the presence of a kinase domain were formally demonstrated for four and six of these haplotypes, respectively, and sequence polymorphism was extremely high. Twenty-five codons showed signs of positive selection in at least one species, and clustered significantly (but not exclusively) within hypervariable regions. We checked that this clustering was not an artifact due to variation in evolution rate at synonymous sites. Simulations revealed that the analysis was highly accurate, thus providing a reliable set of candidates for future functional analyses, but with an overall power not higher than 60 %. Assuming similar power, we infer from our results that about 23% of all codons in the S-domain may actually be involved in recognition. Interestingly, while simulations demonstrated that this comparison remained reliable even at very high levels of divergence, codons identified in Brassica had higher posterior rates of non-synonymous to synonymous substitutions than codons identified in <it>A. halleri or A. lyrata</it>, possibly suggesting more intense selection in Brassica.</p> <p>Conclusion</p> <p>The signature of balancing selection can be identified reliably at the codon level even in cases of very high sequence divergence, provided that a sufficiently large set of sequences are analyzed. Altogether, our results indicate that a large proportion of codons may be involved in recognition and confirm the particular importance of hypervariable regions. The more intense signature of positive selection detected in Brassica suggests that allelic diversification in this genus was very recent, possibly following a recent demographic bottleneck.</p

Recent and Ancient Signature of Balancing Selection around the S-Locus in Arabidopsis halleri and A. lyrata

Balancing selection can maintain different alleles over long evolutionary times. Beyond this direct effect on the molecular targets of selection, balancing selection is also expected to increase neutral polymorphism in linked genome regions, in inverse proportion to their genetic map distances from the selected sites. The genes controlling plant self-incompatibility are subject to one of the strongest forms of balancing selection, and they show clear signatures of balancing selection. The genome region containing those genes (the S-locus) is generally described as nonrecombining, and the physical size of the region with low recombination has recently been established in a few species. However, the size of the region showing the indirect footprints of selection due to linkage to the S-locus is only roughly known. Here, we improved estimates of this region by surveying synonymous polymorphism and estimating recombination rates at 12 flanking region loci at known physical distances from the S-locus region boundary, in two closely related self-incompatible plants Arabidopsis halleri and A. lyrata. In addition to studying more loci than previous studies and using known physical distances, we simulated an explicit demographic scenario for the divergence between the two species, to evaluate the extent of the genomic region whose diversity departs significantly from neutral expectations. At the closest flanking loci, we detected signatures of both recent and ancient indirect effects of selection on the S-locus flanking genes, finding ancestral polymorphisms shared by both species, as well as an excess of derived mutations private to either species. However, these effects are detected only in a physically small region, suggesting that recombination in the flanking regions is sufficient to quickly break up linkage disequilibrium with the S-locus. Our approach may be useful for distinguishing cases of ancient versus recently evolved balancing selection in other systems

Controlling for genetic identity of varieties, pollen contamination and stigma receptivity is essential to characterize the self-incompatibility system of Olea europaea L.

open7siBervillé et al. express concern about the existence of the diallelic self-incompatibility (DSI) system in Olea europaea, mainly because our model does not account for results from previous studies from their group that claimed to have documented asymmetry of the incompatibility response in reciprocal crosses. In this answer to their comment, we present original results based on reciprocal stigma tests that contradict conclusions from these studies. We show that, in our hands, not a single case of asymmetry was confirmed, endorsing that symmetry of incompatibility reactions seems to be the rule in Olive. We discuss three important aspects that were not taken into account in the studies cited in their comments and that can explain the discrepancy: (i) the vast uncertainty around the actual genetic identity of vernacular varieties, (ii) the risk of massive contamination associated with the pollination protocols that they used and (iii) the importance of checking for stigma receptivity in controlled crosses. These studies were thus poorly genetically controlled, and we stand by our original conclusion that Olive tree exhibits DSI.openSaumitou-Laprade, Pierre; Vernet, Philippe; Vekemans, Xavier; Castric, Vincent; Barcaccia, Gianni; Khadari, Bouchaib; Baldoni, LucianaSaumitou-Laprade, Pierre; Vernet, Philippe; Vekemans, Xavier; Castric, Vincent; Barcaccia, Gianni; Khadari, Bouchaib; Baldoni, Lucian

Dynamique de l'organisation géographique de la diversité génétique chez l'omble de fontaine Salvelinus fontinalis Mitchill : diversité des habitats et des histoires de vie

Bien que difficiles à identifier, bien que variables, les contraintes du paysage déterminent les possibilités de coalescence entre allèles. La structure de la diversité devrait donc à terme refléter la structure du paysage. Comment alors expliquer que dans certains systèmes, la concordance puisse être si faible ? D’une part, les espèces ont une histoire, souvent liée à celle de leur habitat. D’autre part, les espèces ne sont pas idéales, et leurs paramètres démographiques peuvent varier dans le temps et dans l’espace. Ainsi, les taux de migration peuvent varier d’un point à l’autre de l’aire de répartition sous l’effet des forces qui déterminent l’évolution des forces de dispersion. L’omble de fontaine en milieu côtier se prête particulièrement bien à l’étude empirique des écarts simples aux modèles de génétique des populations, parce que les contraintes de l’habitat sont généralement aisémement identifiables, que l’histoire du nord-est de l’Amérique du Nord est connue, et que la dynamique de la forme anadrome, la forme de dispersion de l’espèce en milieu côtier a été abondamment décrite dans le cadre de l’étude des migrations saisonnières. La faible concordance de la structure de la diversité avec la structure d’un habitat lacustre qui contraint pourtant fortement les patrons de migration a révélé la difficulté associé à cette tâche, particulièrement aux grandes échelles spatiales. Une analyse des variations temporelles des patrons d’isolement par la distance le long d’un gradient de colonisation a ensuite révélé qu’une des causes de la faible concordance pouvait être l’évolution toujours en cours des patrons d’organisation de la diversité. De plus, la forme de dispersion de l’espèce semble avoir évolué au cours de la colonisation en fonction de l’intensité du fardeau de consanguinité, générant ainsi des variations spatiales des paramètres démographiques. La diversité génétique est donc clairement une variable dynamique dont la compréhension nécessite la prise en compte de facteurs biogéographiques. Les résultats de cette thèse mettent en évidence des variations spatiales et temporelles des contraintes de l’habitat qui compromettent l’utilisation de plusieurs méthodes d’inférence démographiques courantes. C’est pourquoi une approche strictement comparative est proposée comme alternative possible.The expected concordance between landscape structure and geographic patterns of genetic structure is due to landscape constraints over potential coalescence among alleles. In the long run, a high level of concordance may thus be expected. Why then is concordance sometimes so weak? First, species have a history, such that landscape disturbances can affect geographic patterns of diversity. Second, species are seldom “ideal”, as defined by population genetic models, such that their demographic properties can vary in space and in time. For instance, migration rates can vary within the species’ range because the form of dispersal can evolve. Brook charr populations in coastal areas is an especially useful model system to empirically investigate the consequences of simple departures from population genetic models because landscape constraints can generally be easily identified, because recent disturbances of north-eastern North America have been extensively described, and because the dynamic of anadromy, brook charr’s form of dispersal in coastal areas, has been ecologically characterized in the frame of seasonal migrations between freshwater and saltwater. Although lakes and rivers strongly constraint migration pathways, geographic patterns of diversity in lacustrine populations were only weakly correlated with landscape structure, especially at large geographic scales. Temporal variation of isolation by distance along a colonization gradient then revealed that spatial patterns of genetic diversity were still evolving toward stable equilibrium. The species’ form of dispersal was also evolving along this gradient, according to variation of the inbreeding load. Our results thus revealed some aspects of the dynamics of genetic diversity in natural populations. Understanding its evolution requires that biogeographic considerations be included into the view of genetic variation in space. Because the accuracy of demographic inference methods would be impaired by spatial and temporal variations in habitat constraints, a strictly comparative approach is proposed as an alternative in empirical population genetics studies

Repeated Adaptive Introgression at a Gene under Multiallelic Balancing Selection

Recently diverged species typically have incomplete reproductive barriers, allowing introgression of genetic material from one species into the genomic background of the other. The role of natural selection in preventing or promoting introgression remains contentious. Because of genomic co-adaptation, some chromosomal fragments are expected to be selected against in the new background and resist introgression. In contrast, natural selection should favor introgression for alleles at genes evolving under multi-allelic balancing selection, such as the MHC in vertebrates, disease resistance, or self-incompatibility genes in plants. Here, we test the prediction that negative, frequency-dependent selection on alleles at the multi-allelic gene controlling pistil self-incompatibility specificity in two closely related species, Arabidopsis halleri and A. lyrata, caused introgression at this locus at a higher rate than the genomic background. Polymorphism at this gene is largely shared, and we have identified 18 pairs of S-alleles that are only slightly divergent between the two species. For these pairs of S-alleles, divergence at four-fold degenerate sites (K = 0.0193) is about four times lower than the genomic background (K = 0.0743). We demonstrate that this difference cannot be explained by differences in effective population size between the two types of loci. Rather, our data are most consistent with a five-fold increase of introgression rates for S-alleles as compared to the genomic background, making this study the first documented example of adaptive introgression facilitated by balancing selection. We suggest that this process plays an important role in the maintenance of high allelic diversity and divergence at the S-locus in flowering plant families. Because genes under balancing selection are expected to be among the last to stop introgressing, their comparison in closely related species provides a lower-bound estimate of the time since the species stopped forming fertile hybrids, thereby complementing the average portrait of divergence between species provided by genomic data

Bulk pollen sequencing reveals rapid evolution of segregation distortion in the male germline of Arabidopsis hybrids

International audienceGenes that do not segregate in heterozygotes at Mendelian ratios are a potentially important evolutionary force in natural populations. Although the impacts of segregation distortion are widely appreciated, we have little quantitative understanding about how often these loci arise and fix within lineages. Here, we develop a statistical approach for detecting segregation distorting genes from the comprehensive comparison of whole genome sequence data obtained from bulk gamete versus somatic tissues. Our approach enables estimation of map positions and confidence intervals, and quantification of effect sizes of segregation distorters. We apply our method to the pollen of two interspecific F1 hybrids of Arabidopsis lyrata and A. halleri and we identify three loci across eight chromosomes showing significant evidence of segregation distortion in both pollen samples. Based on this, we estimate that novel segregation distortion elements evolve and achieve high frequencies within lineages at a rate of approximately one per 244,000 years. Furthermore, we estimate that haploid-acting segregation distortion may contribute between 10% and 30% of reduced pollen viability in F1 individuals. Our results indicate haploid acting factors evolve rapidly and dramatically influence segregation in F1 hybrid individuals

Urea amidolyase of Candida utilis : Characterization of the urea cleavage reactions

Evidence is presented that the enzymes catalyzing the three reactions involved in urea cleavage in Candida utilis, biotin carboxylation, urea carboxylation, and allophanate hydrolysis occur as a complex of enzymes. The allophanatehydrolyzing activity could not be separated from the urea-cleaving activity using common methods of protein purification. Further, urea cleavage and allophanate hydrolysis activities are induced coordinately in cells grown on various nitrogen sources.The reactions involved in urea cleavage can be distinguished from one another on the basis of their sensitivities to (a) heat, (b) pH, and (c) chemical inhibitors. Evidence is presented for the product of the first reaction in urea cleavage, biotin carboxylation. Production of carboxylated enzyme is ATP dependent and avidin sensitive. Carboxylated enzyme is not observed in the presence of 1 mM urea.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/21726/1/0000118.pd

Cryptic MHC Polymorphism Revealed but Not Explained by Selection on the Class IIB Peptide-Binding Region

The immune genes of the major histocompatibility complex (MHC) are characterized by extraordinarily high levels of nucleotide and haplotype diversity. This variation is maintained by pathogen-mediated balancing selection that is operating on the peptide-binding region (PBR). Several recent studies have found, however, that some populations possess large clusters of alleles that are translated into virtually identical proteins. Here, we address the question of how this nucleotide polymorphism is maintained with little or no functional variation for selection to operate on. We investigate circa 750–850 bp of MHC class II DAB genes in four wild populations of the guppy Poecilia reticulata. By sequencing an extended region, we uncovered 40.9% more sequences (alleles), which would have been missed if we had amplified the exon 2 alone. We found evidence of several gene conversion events that may have homogenized sequence variation. This reduces the visible copy number variation (CNV) and can result in a systematic underestimation of the CNV in studies of the MHC and perhaps other multigene families. We then focus on a single cluster, which comprises 27 (of a total of 66) sequences. These sequences are virtually identical and show no signal of selection. We use microsatellites to reconstruct the populations' demography and employ simulations to examine whether so many similar nucleotide sequences can be maintained in the populations. Simulations show that this variation does not behave neutrally. We propose that selection operates outside the PBR, for example, on linked immune genes or on the “sheltered load” that is thought to be associated to the MHC. Future studies on the MHC would benefit from extending the amplicon size to include polymorphisms outside the exon with the PBR. This may capture otherwise cryptic haplotype variation and CNV, and it may help detect other regions in the MHC that are under selection

Coding Gene SNP Mapping Reveals QTL Linked to Growth and Stress Response in Brook Charr (Salvelinus fontinalis)

Growth performance and reduced stress response are traits of major interest in fish production. Growth and stress-related quantitative trait loci (QTL) have been already identified in several salmonid species, but little effort has been devoted to charrs (genus Salvelinus). Moreover, most QTL studies to date focused on one or very few traits, and little investigation has been devoted to QTL identification for gene expression. Here, our objective was to identify QTL for 27 phenotypes related to growth and stress responses in brook charr (Salvelinus fontinalis), which is one of the most economically important freshwater aquaculture species in Canada. Phenotypes included 12 growth parameters, six blood and plasma variables, three hepatic variables, and one plasma hormone level as well as the relative expression measurements of five genes of interest linked to growth regulation. QTL analysis relied on a linkage map recently built from S. fontinalis consisting of both single-nucleotide polymorphism (SNP, n = 266) and microsatellite (n =81) markers in an F2 interstrain hybrid population (n = 171). We identified 63 growth-related QTL and four stress-related QTL across 18 of the 40 linkage groups of the brook charr linkage map. Percent variance explained, confidence interval, and allelic QTL effects also were investigated to provide insight into the genetic architecture of growth- and stress-related QTL. QTL related to growth performance and stress response that were identified could be classified into two groups: (1) a group composed of the numerous, small-effect QTL associated with some traits related to growth (i.e., weight) that may be under the control of a large number of genes or pleiotropic genes, and (2) a group of less numerous QTL associated with growth (i.e., gene expression) and with stress-related QTL that display a larger effect, suggesting that these QTL are under the control of a limited number of genes of major effect. This study represents a first step toward the identification of genes potentially linked to phenotypic variation of growth and stress response in brook charr. The ultimate goal is to provide new tools for developing Molecular Assisted Selection for this species