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Kin recognition and kin selection have long been known to occur in animals where they shape altruistic behavior towards relatives. More recently, studies have found that kin recognition and altered behavior towards kin can also occur in plants. However, inferring the underlying mechanism responsible for variation in plant performance in experimental studies is challenging as often, results can be explained by alternative and non-exclusive mechanisms such as niche differences, kin competition avoidance and genetic variation in growth rate and competitive ability. Plant–plant interactions may change with the life stage of plants, and competition is often most intense towards the end of plants growing season. However, changes in plant–plant interaction intensity across plants life cycle are rarely considered in kin interaction studies. Here, we adapt a model of plant growth over time modified to specifically include effects of kin and non-kin competition. The model decompose competitive interactions at different stages during plant growth from initial growth to the end of the growing season. It estimates genotype specific variation in growth rate, and how sensitive individual genotypes are to competition from neighbors. Furthermore, it estimates size asymmetry among plants accounting for both variation in growth rate, neighbor relatedness and resource variation (here water availability). We use this model to analyze the results from a competition experiment where plants grew in mini-populations with neighbor plants that were either kin or non-kin. We find that when applied to our experiments, this approach can disentangle kin effects from other effects caused by genotypic variation in growth rate and competitive response to neighbors, and thus significantly help to detect whether plants exhibit kin-cooperative behavior.

Background Asian rice Oryza sativa, first domesticated in China, is having considerable success in African fields. While the timing and location of this introduction remain unclear, rice of Asian origin may have evolved specifically during and after migration to Africa, resulting in unique adaptations, particularly in relation to upland cultivation as frequently practiced in Africa. Methods We investigated the genetic differentiation between Asian and African varieties using the 3000 Rice Genomes SNP dataset. African upland cultivars were first characterized using principal component analysis among 292 tropical Japonica accessions of both origins, revealing higher similarity with accessions from Indonesia and Madagascar. Peculiarities of African accessions were then explored using two inference techniques, PCA-KDE for supervised classification and chromosome painting, and ELAI for individual allelic dosage monitoring. Key results Ambiguities of local differentiation between Japonica and other groups pointed at genomic segments that potentially resulted from genetic exchange. Those specific to West African upland accessions were concentrated on chromosome 6 and featured several cAus introgression signals, including a large one between 17.9 and 21.7 Mb. We found iHS statistics in support of positive selection in this region and we provide a list of candidate genes enriched in GO terms for this adaptation to harsh upland cultivation conditions.

Resurrection studies are a useful tool to measure how phenotypic traits have changed in populations through time. If these trait modifications correlate with the environmental changes that occurred during the time period, it suggests that the phenotypic changes could be a response to selection. Selfing, through its reduction of effective size, could challenge the ability of a population to adapt to environmental changes. Here, we used a resurrection study to test for adaptation in a selfing population of Medicago truncatula, by comparing the genetic composition and flowering times across 22 generations. We found evidence for evolution toward earlier flowering times by about two days and a peculiar genetic structure, typical of highly selfing populations, where some multilocus genotypes (MLGs) are persistent through time. We used the change in frequency of the MLGs through time as a multilocus fitness measure and built a selection gradient that suggests evolution toward earlier flowering times. Yet, a simulation model revealed that the observed change in flowering time could be explained by drift alone, provided the effective size of the population is small enough (, Resurrection studies are a useful tool to measure how phenotypic traits have changed in populations and they allow testing whether these traits modifications are a response to selection caused by an environmental change. Selfing, through its reduction of effective size, could challenge the ability of a population to adapt to environmental changes. Here, we used a resurrection study to test for adaptation in a selfing population of Medicago truncatula, by comparing the genetic composition and flowering across 22 generations.

AGAP : Equipe GE2POP; International audience; Most theoretical works predict that selfing should reduce the level of additive genetic variance available for quantitative traits within natural populations. Despite a growing number of quantitative genetic studies undertaken during the last two decades, this prediction is still not well supported empirically. To resolve this issue and confirm or reject theoretical predictions, we reviewed quantitative trait heritability estimates from natural plant populations with different rates of self-fertilization and carried out a meta-analysis. In accordance with models of polygenic traits under stabilizing selection, we found that the fraction of additive genetic variance is negatively correlated with the selfing rate. Although the mating system explains a moderate fraction of the variance, the mean reduction of narrow-sense heritability values between strictly allogamous and predominantly selfing populations is strong, around 60%. Because some nonadditive components of genetic variance become selectable under inbreeding, we determine whether self-fertilization affects the relative contribution of these components to genetic variance by comparing narrow-sense heritability estimates from outcrossing populations with broad-sense heritability estimated in autogamous populations. Results suggest that these nonadditive components of variance may restore some genetic variance in predominantly selfing populations; it remains, however, uncertain how these nonadditive components will contribute to adaptation.

International audience; Studying the consequences of hybridization on plant performance is insightful to understand the adaptive potential of populations, notably at local scales. Due to reduced effective recombination, predominantly selfing species are organized in highly homozygous multi-locus-genotypes (or lines) that accumulate genetic differentiation both among- and within-populations. This high level of homozygosity facilitates the dissection of the genetic basis of hybrid performance in highly selfing species, which gives insights into the mechanisms of reproductive isolation between lines. Here, we explored the fitness consequences of hybridization events between natural inbred lines of the predominantly selfing species Medicago truncatula, at both within- and among-populations scales. We found that hybridization has opposite effects pending on studied fitness proxies, with dry mass showing heterosis, and seed production showing outbreeding depression. Although we found significant patterns of heterosis and outbreeding depression, they did not differ significantly for within- compared to among-population crosses. Family-based analyses allowed us to determine that hybrid differentiation was mostly due to dominance and epistasis. Dominance and/or dominant epistatic interactions increased dry mass, while decreasing seed production, and recessive epistatic interactions mostly had a positive effect on both fitness proxies. Our results illustrate how genetic incompatibilities can accumulate at a very local scale among multi-locus-genotypes, and how non-additive genetic effects contribute to heterosis and outbreeding depression.

SummaryAccounting for intraspecific variation may improve our understanding of species coexistence. However, our knowledge of what factors maintain intraspecific variation is limited. We predicted that 1) a plant grows larger when with non-kin (i.e. different genotypes) than kin (i.e. same genotype) neighbors, 2) abiotic stress alters the outcome of kin vs. non-kin interactions, 3) genetic identity of plants affects composition of soil microbiome.We set up mini-communities ofMedicago truncatula, where focal genotypes were grown together with two kin or two non-kin neighbors from different origins. We analyzed how origin, identity of interacting genotypes and abiotic stress affected growth and fruit production. We also analyzed the composition of soil microbial communities.Focal plants grew larger in non-kin than in kin mini-communities. This pattern was stronger in low level of abiotic stress and when interacting genotypes were from similar origins. However, genotypic variation in growth and response to competition had a stronger effect on growth than mini-community type. Plant genotype identity did not affect soil microbiome.We find that intraspecific variation is affected by genotype-specific traits and abiotic stress. Geographic, rather than genetic, distance among interacting genotypes affects the outcome of intraspecific interactions.

Tracking genetic changes of populations through time allows a more direct study of the evolutionary processes acting on the population than a single contemporary sample. Several statistical methods have been developed to characterize the demography and selection from temporal population genetic data. However, these methods are usually developed under the assumption of outcrossing reproduction and might not be applicable when there is substantial selfing in the population. Here, we focus on a method to detect loci under selection based on a genome scan of temporal differentiation, adapting it to the particularities of selfing populations. Selfing reduces the effective recombination rate and can extend hitch-hiking effects to the whole genome, erasing any local signal of selection on a genome scan. Therefore, selfing is expected to reduce the power of the test. By means of simulations, we evaluate the performance of the method under scenarios of adaptation from new mutations or standing variation at different rates of selfing. We find that the detection of loci under selection in predominantly selfing populations remains challenging even with the adapted method. Still, selective sweeps from standing variation on predominantly selfing populations can leave some signal of selection around the selected site thanks to historical recombination before the sweep. Under this scenario, ancestral advantageous alleles at low frequency leave the strongest local signal, while new advantageous mutations leave no local footprint of the sweep.

AGAP : équipe GE2pop; International audience; Predominantly selfing populations are expected to have reduced effective population sizes due to nonrandom sampling of gametes, demographic stochasticity (bottlenecks or extinction–recolonization), and large scale hitchhiking (reduced effective recombination). Thus, they are expected to display low genetic diversity, which was confirmed by empirical studies. The structure of genetic diversity in predominantly selfing species is dramatically different from outcrossing ones, with populations often dominated by one or a few multilocus genotypes (MLGs) coexisting with several rare genotypes. Therefore, multilocus diversity indices are relevant to describe diversity in selfing populations. Here, we use simulations to provide analytical expectations for multilocus indices and examine whether selfing alone can be responsible for the highfrequency MLGs persistent through time in the absence of selection. We then examine how combining single and multilocus indices of diversity may be insightful to distinguish the effects of selfing, population size, and more complex demographic events (bottlenecks, migration, admixture, or extinction–recolonization). Finally, we examine how temporal changes in MLG frequencies can be insightful to understand the evolutionary trajectory of a given population. We show that combinations of selfing and small demographic sizes can result in high-frequency MLGs, as observed in natural populations. We also show how different demographic scenarios can be distinguished by the parallel analysis of single and multilocus indices of diversity, and we emphasize the importance of temporal data for the study of predominantly selfing populations. Finally, the comparison of our simulations with empirical data on populations of Medicago truncatula confirms the pertinence of our simulation framework.

Standing genetic variation is considered a major contributor to the adaptive potential of species. The low heritable genetic variation observed in self-fertilizing populations has led to the hypothesis that species with this mating system would be less likely to adapt. However, a non-negligible amount of cryptic genetic variation for polygenic traits, accumulated through negative linkage disequilibrium, could prove to be an important source of standing variation in self-fertilizing species. To test this hypothesis, we simulated populations under stabilizing selection subjected to an environmental change. We demonstrate that, when the mutation rate is high (but realistic), selfing populations are better able to store genetic variance than outcrossing populations through genetic associations, notably due to the reduced effective recombination rate associated with predominant selfing. Following an environmental shift, this diversity can be partially remobilized, which increases the additive variance and adaptive potential of predominantly (but not completely) selfing populations. In such conditions, despite initially lower observed genetic variance, selfing populations adapt as readily as outcrossing ones within a few generations. For low mutation rates, purifying selection impedes the storage of diversity through genetic associations, in which case, as previously predicted, the lower genetic variance of selfing populations results in lower adaptability compared to their outcrossing counterparts. The population size and the mutation rate are the main parameters to consider, as they are the best predictors of the amount of stored diversity in selfing populations. Our results and their impact on our knowledge of adaptation under high selfing rates are discussed.

Standing genetic variation is considered a major contributor to the adaptive potential of species. The low heritable genetic variation observed in self-fertilising populations has led to the hypothesis that species with this mating system would be less likely to adapt. However, a non-negligible amount of cryptic genetic variation for polygenic traits, accumulated through negative linkage disequilibrium, could prove to be an important source of standing variation in self-fertilising species. To test this hypothesis we simulated populations under stabilizing selection subjected to an environmental change. We demonstrate that, when the mutation rate is high (but realistic), selfing populations are better able to store genetic variance than outcrossing populations through genetic associations, notably due to the reduced effective recombination rate associated with predominant selfing. Following an environmental shift, this diversity can be partially remobilized, which increases the additive variance and adaptive potential of predominantly (but not completely) selfing populations. In such conditions, despite initially lower observed genetic variance, selfing populations adapt as readily as outcrossing ones within a few generations. For low mutation rates, purifying selection impedes the storage of diversity through genetic associations, in which case, as previously predicted, the lower genetic variance of selfing populations results in lower adaptability compared to their outcrossing counterparts. The population size and the mutation rate are the main parameters to consider, as they are the best predictors of the amount of stored diversity in selfing populations. Our results and their impact on our knowledge of adaptation under high selfing rates are discussed.

Agap : équipe GE2pop et ID; International audience; Sequencing pools of individuals rather than individuals separately reduces the costs of estimating allele frequencies at many loci in many populations. Theoretical and empirical studies show that pool-sequencing a limited number of individuals (typically fewer than 50) provides reliable allele frequency estimates, provided that the DNA pooling and DNA sequencing steps are carefully controlled. Unequal contributions of different individuals to the DNA pool and the mean and variance in sequencing depth both can affect the standard error of allele frequency estimates. To our knowledge, no study separately investigated the effect of these two factors on allele frequency estimates; so that there is currently no method to a priori estimate the relative importance of unequal individual DNA contributions independently of sequencing depth. We develop a new analytical model for allele frequency estimation that explicitly distinguishes these two effects. Our model shows that the DNA pooling variance in a pool-sequencing experiment depends solely on two factors: the number of individuals within the pool and the coefficient of variation of individual DNA contributions to the pool. We present a new method to experimentally estimate this coefficient of variation when planning a pool-sequencing design where samples are either pooled before or after DNA extraction. Using this analytical and experimental framework, we provide guidelines to optimize the design of pool-sequencing experiments. Finally, we sequence replicated pools of inbred lines of the plant Medicago truncatula and show that the predictions from our model generally hold true when estimating the frequency of known multi-locus haplotypes using pool-sequencing.

AGAP : équipe GE2pop; Diversity and structure of populations may differ substantially between morphological traits and molecular markers. Explanations of such discrepancies are crucial for further progress in breeding as well as for the maintenance of genetic resources. Our objective was to compare indices of among-cultivars differentiation for morphological traits (QST) and molecular markers (FST) in alfalfa (Medicago sativa),a legume forage species. Ten cultivars representing the Northern and Southern types commonly grown in Europe were investigated. For each cultivar, 40genotypes were analysed with 16 SSR markers and four morphological traits measured in two locations and several cuts. QST values were in general high(0.02–0.39) compared to the differentiation observed with molecular markers (FST = 0.01), especially for growth habit, indicating that morphological traits were more efficient to structure the diversity than molecular markers. For morphological traits, a clear separation of Northern and Southern cultivars was observed, whereas for molecular markers, no clear structure was detected. Nevertheless, the grazing type cultivar Luzelle was separated from the rest of the cultivars for both morphological traits and molecular markers. Although pairwise differences between cultivars were significant for both morphological traits and molecular markers, the main part of the variation was found within cultivars. This large within-cultivar variation may be explained, besides the outcrossing reproductive mode and auto tetraploid genetics, by the recent history of M. sativa domestication in Europe and the frequent seed exchanges. Selection for morphological traits (QST[FST) was achieved without modification of within-cultivar neutral diversity.

The long-term maintenance of specialized mutualisms remains an evolutionary puzzle. Recent focus has been on factors governing the stability of these mutualisms, including sanctions by the host, partner choice, and coevolutionary constraint, that is, the genetic correlation (rG) between fitness of both partners. So far these studies have been typically carried out in a single environment. Here, we ask if the genetic correlation between fitness of the host plant Medicago truncatula (Fabaceae) and its bacterial symbiont Sinorhizobium meliloti is affected by the presence/absence of a monoterpene (carvacrol) leached into the soil by Thymus vulgaris—a common plant of the Mediterranean vegetation, often co-occuring with Medicago. We show that the presence of carvacrol in the soil dramatically affects fitness of the rhizobial partner and increases the magnitude of rG between plant and rhizobia fitness (rG = 0.02 ± 0.05 vs. rG = 0.57 ± 0.02). This finding emphasizes the importance of heterogeneity in the biotic environment for understanding the evolution of species interactions.

Contact: Joelle.Ronfort@supagro.inra.fr, Fax: 00 33 4 6729 3990; International audience; AMaCAID is an R program designed to analyse multilocus genotypic patterns in large samples. It allows (i) the computation of the number and frequency of the different multilocus patterns available in a molecular data set and (ii) the analysis of discriminatory power of each combination of k markers among n available. It thus enables the identification of the minimum number of markers required to distinguish all the observed genotypes and the subset of markers that maximize the number of distinct genotypes. AMaCAID can be used with any kind of molecular markers, on data sets mixing different kinds of markers, but also on qualitative characters like morphological or taxonomic traits. AMaCAID has been built primarily to select subsets of markers for identifying accessions and monitoring their genetic stability during regeneration cycles in an ex situ genebank. It can, however, also be used to screen any kind of data set that characterizes a set of individuals or species (e.g. taxonomic or phylogenetic studies) for discrimination purposes. The size of the assayed sample has no limitation, but the program only performs computations on all combinations of markers when there are less than 25 markers. For larger number of markers/characters, it is possible to ask AMaCAID to screen a large but limited number of combinations of markers. We apply AMaCAID to three data sets involving either molecular or taxonomic data and give some results on the computing time of the program with respect to the size of the data set

International audience; The occurrence of populations exhibiting high genetic diversity in predominantly selfing species remains a puzzling question, since under regular selfing genetic diversity is expected to be depleted at a faster rate than under outcrossing. Fine-scale population genetics approaches may help to answer this question. Here we study a natural population of the legume Medicago truncatula in which both the fine-scale spatial structure and the selfing rate are characterized using three different methods. Selfing rate estimates were very high (B99%) irrespective of the method used. A clear pattern of isolation by distance reflecting small seed dispersal distances was detected. Combining genotypic data over loci, we could define 34 multilocus genotypes. Among those, six highly inbred genotypes (lines) represented more than 75% of the individuals studied and harboured all the allelic variation present in the population. We also detected a large set of multilocus genotypes resembling recombinant inbred lines between the most frequent lines occurring in the population. This finding illustrates the importance of rare recombination in redistributing available allelic diversity into new genotypic combinations. This study shows how multilocus and fine-scale spatial analyses may help to understand the population history of self-fertilizing species, especially to make inferences about the relative role of foundation/migration and recombination events in such populations

AGAP : équipe GEpop; International audience; Local climatic conditions likely constitute an important selective pressure on genes underlying important fitness-related traits such as flowering time and in many species flowering phenology and climatic gradients strongly covary. To test whether climate shapes genetic variation on flowering time genes and to identify candidate flowering genes involved in the adaptation to environmental heterogeneity, we used a large M. truncatula core collection to examine the association between nucleotide polymorphisms at 224 candidate genes and both climate variables and flowering phenotypes. Unlike genome-wide studies, candidate gene approaches are expected to enrich for the number of meaningful trait associations because they specifically target genes that are known to affect the trait of interest. We found that flowering time mediates adaptation to climatic conditions mainly by variation at genes located upstream in the flowering pathways, close to the environmental stimuli. Variables related to the annual precipitation regime reflected selective constraints on flowering time genes better than the other variables tested (temperature, altitude, latitude or longitude). By comparing phenotype and climate associations, we identified 12 flowering genes as the most promising candidates responsible for phenological adaptation to climate. Four of these genes were located in the known flowering time QTL region on chromosome 7. However, climate and flowering associations also highlighted largely distinct gene sets, suggesting different genetic architectures for adaptation to climate and flowering onset.

Correspondance: glemin@univ-montp2.fr; International audience; Various methods have been proposed to estimate inbreeding depression and to assess its consequences for natural populations. As an alternative to controlled crosses, the use of molecular markers has allowed direct investigation of inbreeding depression in natural populations, but usually suffers from low statistical power. Here, we investigated the effect of inbreeding depression on survival in two populations of the rare species Brassica insularis, using both controlled crosses and a marker-based approach. We compare the respective merits of the two approaches for studying inbreeding depression. We also use information from the molecular markers to dissect in detail patterns of inbreeding depression in this species. A posteriori, we find that combining the approaches was not necessary to obtain simple point estimates of inbreeding depression. However, using molecular markers may give insight into the genetic basis of inbreeding depression, such as the occurrence of epistatic interactions among deleterious alleles or purging.

DNA sequence polymorphism carries genealogical information and allows for testing hypotheses on selection and population history, especially through coalescent-based analysis. Understanding the evolutionary forces at work in plant domestication and subsequent selection is of critical importance for the management of genetic resources. In this study, we surveyed DNA sequence diversity at two assumed neutral nuclear loci in the wild-domesticated species complex of alfalfa (Medicago sativa L.). A high level of polymorphism was detected. The domesticated pool contains on average 31% less diversity than the wild pool, but with a high heterogeneity among loci. Coalescent simulations of the domestication process showed that this result cannot be explained by assuming a constant population size but is rather consistent with a demographic bottleneck during domestication. A very low level of divergence was detected between the wild and the domesticated forms as well as between the related subspecies of the M. sativa species complex. However, the originality of the Spanish wild populations, already observed based on mitochondrial DNA polymorphism, was confirmed. These results, together with patterns of intrapopulation polymorphism, suggest that nuclear sequence polymorphism could be a promising tool, complementary to mitochondrial DNA and phenotypic evaluations, to investigate historical demographic and evolutionary processes.

International audience; In habitats where colonization and extinction are recurrent, the distribution of gene frequencies among patches of suitable habitat may reflect the age structure of different populations. In this study, we quantify population genetic structure for a pioneer tree species, Antirhea borbonica, in a chrono-sequence of primary succession on the lava flows of the Piton de La Fournaise volcano (La Réunion). Using microsatellite loci and amplified fragment length polymorphism (AFLP) markers, we quantified genetic variation within and among populations for early- and late-succession populations in a landscape where extinction and recolonization are recurrent (the 'Grand Brûlé') and for late-succession populations in a more stable landscape. This study produced three main results. First, we detected no evidence that founder events increase genetic differentiation among colonizing populations; FST values among early- and among late-succession populations were similar. Second, we found no evidence for isolation by distance; genetic distance was not correlated with spatial distance within and among populations. Third, FIS values are consistently high in all populations, despite the fact that A. borbonica populations are functionally close to dioecy and thus expected to have an outcrossing mating system. Multiple colonization events from different sources may limit differentiation among young populations and spatial isolation may enhance differentiation among late-succession populations. Ecological processes acting during colonization may create the conditions for spatial aggregation within pioneer populations, and thus contribute to the high FIS values.

In order to clarify the relationships (genetic exchange and shared ancestry) between natural and cultivated populations of alfalfa (Medicago sativa L.) in Spain, we investigated the patterns of mitochondrial DNA variation (characterized through restriction fragment length polymorphism) for 248 individuals in seven natural and six cultivated populations of this species. Mitochondrial variation was evidenced in both natural and cultivated populations of M. sativa. Among the seven mitotypes idengified in the species, two were specific of the natural populations, a result attesting the fact that the Spanish wild form of M. sativa is an original genetic pool compared to the cultivated one. Other mitotypes were observed in both natural and cultivated populations, suggesting the occurrence of gene flow through seeds from cultivated towards natural populations. Comparisons with previously gathered nuclear and phenotypic data give insights into the different evolutionary forces acting on the different kinds of Spanish natural populations examined so far.

Summary � The use of quantitative disease resistance (QDR) is a promising strategy for promoting durable resistance to plant pathogens, but genes involved in QDR are largely unknown. To identify genetic components and accelerate improvement of QDR in legumes to the root pathogen Aphanomyces euteiches, we took advantage of both the recently generated massive genomic data for Medicago truncatula and natural variation of this model legume. � A high-density (� 5.1 million single nucleotide polymorphisms (SNPs)) genome-wide association study (GWAS) was performed with both in vitro and glasshouse phenotyping data collected for 179 lines. � GWAS identified several candidate genes and pinpointed two independent major loci on the top of chromosome 3 that were detected in both phenotyping methods. Candidate SNPs in the most significant locus (r 2 = 23%) were in the promoter and coding regions of an F-box protein coding gene. Subsequent qRT-PCR and bioinformatic analyses performed on 20 lines demonstrated that resistance is associated with mutations directly affecting the interaction domain of the F-box protein rather than gene expression. � These results refine the position of previously identified QTL to specific candidate genes, suggest potential molecular mechanisms, and identify new loci explaining QDR against A. euteiches.

The outcome of intrafamily selection on variation in the load from deleterious mutations and on the magnitude of inbreeding depression has been investigated through an analytical one-locus model of mutation-selection balance, in partially selfing and in partially parthenogenetic populations. It is shown that, in contrast with an ordinary selection model, when intrafamily selection is assumed, increasing selfing rates are associated with increasing frequencies of a recessive deleterious allele, as well as with increasing magnitudes of inbreeding depression. With the same selective regime, a similar behaviour could be observed for the frequency of a deleterious allele in the case of parthenogenesis. On the basis of these results, intrafamily selection could have important consequences on the evolution of reproductive systems.

Extensive genomic resources are available in the model legume Medicago truncatula. Here, we present the discovery and design of the first array of single-nucleotide polymorphism (SNP) markers in M. truncatula through large-scale Sanger resequencing of genomic fragments spanning the genome, in a diverse panel of 16 M. truncatula accessions. Both anonymous fragments and fragments targeting candidate genes for flowering phenology and symbiosis were surveyed for nucleotide variation in almost 230 kb of unique genomic regions. A set of 384 SNP markers was designed for an Illumina's GoldenGate assay, genotyped on a collection of 192 inbred lines (CC192) representing the geographical range of the species and used to survey the diversity of two natural populations. Finally, 86% of the tested SNPs were of high quality and exhibited polymorphism in the CC192 collection. Even at the population level, we detected polymorphism for more than 50% of the selected SNPs. Analysis of the allele frequency spectrum in the CC192 showed a reduced ascertainment bias, mostly limited to very rare alleles (frequency

AGAP : équipe Génomique évolutive et gestion des populations (GE²pop); Mating systems are expected to have a strong influence on both the dynamic of adaptation and the genetic architecture of adaptive traits. In particular, the bias toward the fixation of dominant or partially dominant beneficialmutations predicted under outcrossing (Haldane’s sieve) is expected to be reduced under self-fertilization. To test this prediction in plants, we considered domestication as an example of adaptation. We compiled data from studies reporting the degree of dominance of quantitative trait loci (QTL) involved in the domestication syndrome. We found that adaptation to cultivation mostly proceeded through the selection of recessive and partially recessive genes in predominantly selfing species whereas a much larger fraction of domestication-related QTL were dominant or partially dominant in outcrossers, as expected under Haldane’s sieve. Our study also showed that levels of dominance in mixed mating crop species resemble those observed in selfers, suggesting that recessive alleles can contribute to adaptation even under moderate selfing rates. Although these results rely on a particular example of adaptation, they constitute one of the first attempts to test theoretical expectations on the level of dominance of genes involved in plant adaptation.

Thanks to genome-scale diversity data, present-day studies can provide a detailed view of how natural and cultivated species adapt to their environment and particularly to environmental gradients. However, due to their sensitivity, up-to-date studies might be more sensitive to undocumented demographic effects such as the pattern of migration and the reproduction regime. In this study, we provide guidelines for the use of popular or recently developed statistical methods to detect footprints of selection. We simulated 100 populations along a selective gradient and explored different migration models, sampling schemes and rates of self-fertilization. We investigated the power and robustness of eight methods to detect loci potentially under selection: three designed to detect genotype-environment correlations and five designed to detect adaptive differentiation (based on F(ST) or similar measures). We show that genotype-environment correlation methods have substantially more power to detect selection than differentiation-based methods but that they generally suffer from high rates of false positives. This effect is exacerbated whenever allele frequencies are correlated, either between populations or within populations. Our results suggest that, when the underlying genetic structure of the data is unknown, a number of robust methods are preferable. Moreover, in the simulated scenario we used, sampling many populations led to better results than sampling many individuals per population. Finally, care should be taken when using methods to identify genotype-environment correlations without correcting for allele frequency autocorrelation because of the risk of spurious signals due to allele frequency correlations between populations.

Background Gene duplications are a molecular mechanism potentially mediating generation of functional novelty. However, the probabilities of maintenance and functional divergence of duplicated genes are shaped by selective pressures acting on gene copies immediately after the duplication event. The ratio of non-synonymous to synonymous substitution rates in protein-coding sequences provides a means to investigate selective pressures based on genic sequences. Three molecular signatures can reveal early stages of functional divergence between gene copies: change in the level of purifying selection between paralogous genes, occurrence of positive selection, and transient relaxed purifying selection following gene duplication. We studied three pairs of genes that are known to be involved in an interaction with symbiotic bacteria and were recently duplicated in the history of the Medicago genus (Fabaceae). We sequenced two pairs of polygalacturonase genes (Pg11-Pg3 and Pg11a-Pg11c) and one pair of auxine transporter-like genes (Lax2-Lax4) in 17 species belonging to the Medicago genus, and sought for molecular signatures of differentiation between copies. Results Selective histories revealed by these three signatures of molecular differentiation were found to be markedly different between each pair of paralogs. We found sites under positive selection in the Pg11 paralogs while Pg3 has mainly evolved under purifying selection. The most recent paralogs examined Pg11a and Pg11c, are both undergoing positive selection and might be acquiring new functions. Lax2 and Lax4 paralogs are both under strong purifying selection, but still underwent a temporary relaxation of purifying selection immediately after duplication. Conclusions This study illustrates the variety of selective pressures undergone by duplicated genes and the effect of age of the duplication. We found that relaxation of selective constraints immediately after duplication might promote adaptive divergence.

E-mail Addresses: siol@supagro.inra.fr; International audience; Despite its significance in evolutionary and conservation biology, few estimates of effective population size (N-e) are available in plant species. Self-fertilization is expected to affect N-e, through both its effect on homozygosity and population dynamics. Here, we estimated N-e using temporal variation in allele frequencies for two contrasted populations of the selfing annual Medicago truncatula: a large and continuous population and a subdivided population. Estimated N-e values were around 5-10% of the population census size suggesting that other factors than selfing must contribute to variation in allele frequencies. Further comparisons between monolocus allelic variation and changes in the multilocus genotypic composition of the populations show that the local dynamics of inbred lines can play an important role in the fluctuations of allele frequencies. Finally, comparing N-e estimates and levels of genetic variation suggest that H-e is a poor estimator of the contemporaneous variance effective population size.

Symbiotic nitrogen-fixing rhizobia are able to trigger root deformation in their Fabaceae host plants, allowing their intracellular accommodation. They do so by delivering molecules called Nod factors. We analyzed the patterns of nucleotide polymorphism of five genes controlling early Nod factor perception and signaling in the Fabaceae Medicago truncatula to understand the selective forces shaping the evolution of these genes. We used 30 M. truncatula genotypes sampled in a genetically homogeneous region of the species distribution range. We first sequenced 24 independent loci and detected a genomewide departure from the hypothesis of neutrality and demographic equilibrium that suggests a population expansion. These data were used to estimate parameters of a simple demographic model incorporating population expansion. The selective neutrality of genes controlling Nod factor perception was then examined using a combination of two complementary neutrality tests, Tajima's D and Fay and Wu's standardized H. The joint distribution of D and H expected under neutrality was obtained under the fitted population expansion model. Only the gene DMI1, which is expected to regulate the downstream signal, shows a pattern consistent with a putative selective event. In contrast, the receptor-encoding genes NFP and NORK show no significant signatures of selection. Among the genes that we analyzed, only DMI1 should be viewed as a candidate for adaptation in the recent history of M. truncatula.

Email: jean-marie.prosperi@supagro.inra.fr.; International audience; One hundred and three natural populations of Medicago L. were collected in Spain, mainly from roadsides, non-irrigated or grazed lands. This germplasm was evaluated at Montpellier (France) with control cultivars and Spanish landraces under completely random block design, replicated four times, and observed for 64 qualitative or quantitative characters. Differences between natural populations and cultivated controls are highly significant, but gene flow occurs between wild and cultivated compartment and hybrid populations were identified. On the basis of multivariable analysis, the accessions were grouped into four clusters depending on their proximity with the cultivated pool to facilitate their management and ex situ conservation. The relationship between environment of the site of collection and phenotypic characteristics of the natural populations was also discussed. Different policies of conservation of these genetic resources are suggested to avoid their disappearance even when they disappeared from the other regions of the western Mediterranean. Spanish wild pool of alfalfa, also called ‘Mielga’, appears of great interest for the breeding of alfalfa because it contains a large diversity of characteristics (prostrate habit, rhizomes) linked to tolerance to grazing or drought environment. With the need for more sustainable systems in agriculture, the erosion of natural habitats and the necessity for rehabilitation of degraded ecosystems, the importance of this wild pool is really inestimable

International audience; Understanding the selective constraints of partner specificity in mutually beneficial symbiosis is a significant, yet largely unexplored, prospect of evolutionary biology. These selective constraints can be explored through the study of nucleotide polymorphism at loci controlling specificity. The membrane-anchored receptor NORK (nodulation receptor kinase) of the legume Medicago truncatula controls early steps of root infection by two symbiotic microorganisms: nitrogen-fixing bacteria (rhizobia) and endomycorrhizal fungi (Glomales). We analyzed the diversity of the gene NORK by sequencing 4 kilobases in 28 inbred lines sampled from natural populations. We detected 33 polymorphic sites with only one nonsynonymous change. Analysis based on Tajima’s D and Fay and Wu’s H summary statistics revealed no departure from the neutral model. We analyzed divergence using sequences from the closely related species M. coerulea. The McDonald-Kreitman test indicated a significant excess of nonsynonymous changes contributing to this divergence. Furthermore, maximum-likelihood analysis of a molecular phylogeny of a few legume species indicated that a number of amino acid sites, likely located in the receptor domain of the protein, evolved under the regime of positive selection. Further research should focus on the rate and direction of molecular coevolution between microorganisms’ signaling molecules and legumes’ receptors

Alfalfa (Medicago sativa) is an autotetraploid, allogamous and heterozygous species whose cultivars are synthetic populations. The breeders apply selection pressure for some agronomic traits within a breeding pool to increase the frequency of favorable individuals. The objective of this study was to investigate the differentiation level among seven cultivars originating from one breeding program, and between these cultivars and the breeding pool, with eight SSR markers. These highly polymorphic and codominant markers, together with recent population genetic statistics extended to autotetraploids, offer tools to analyse genetic diversity in alfalfa. The number of alleles per locus varied between 3 and 24. All loci were at a panmictic equilibrium in the cultivars, except one, probably because of null alleles. With seven SSR loci, each cultivar was at panmictic equilibrium. The mean gene diversity was high, ranging from 0.665 to 0.717 in the cultivars. The parameter F ST indicated a low but significant diversity among cultivars. Among 21 pairs of cultivars, 15 were significantly different. The breeding pool also had a high diversity, and was significantly different from each cultivar except the most recent one. Considering the characteristics of the breeding program and the mode of cultivar elaboration, we found that they were unable to generate a large variety differentiation. Estimation of population genetics parameters at SSR loci can be applied for assessing the differences between cultivars or populations, either for variety distinction or the management of genetic resources.

Inbreeding depression is a general phenomenon that is due mainly to recessive deleterious mutations, the so-called mutation load. It has been much studied theoretically. However, until very recently, population structure has not been taken into account, even though it can be an important factor in the evolution of populations. Population subdivision modifies the dynamics of deleterious mutations because the outcome of selection depends on processes both within populations (selection and drift) and between populations (migration). Here, we present a general model that permits us to gain insight into patterns of inbreeding depression, heterosis, and the load in subdivided populations. We show that they can be interpreted with reference to single-population theory, using an appropriate local effective population size that integrates the effects of drift, selection, and migration. We term this the “effective population size of selection” (NeS). For the infinite island model, for example, it is equal to NeS=N(1+m∕hs), where N is the local population size, m the migration rate, and h and s the dominance and selection coefficients of deleterious mutation. Our results have implications for the estimation and interpretation of inbreeding depression in subdivided populations, especially regarding conservation issues. We also discuss the possible effects of migration and subdivision on the evolution of mating systems.

Crop-to-wild gene flow has received close attention over the past ten years in connection with the development and cultivation of transgenic crops. In this paper, we review key examples of crop/wild sympatry and overlapping flowering phenology, pollen and seed dispersal, the barriers to hybridisation and introgression, the evolution and fate of interspecific hybrids, their fitness, and the potential cost of transgenes. We pay particular attention to ways in which the evolution and divergence between crops and their wild relatives may interfere with these successive steps. Our review suggests that crop-to-weed gene flow is highly idiosyncratic and that crop gene dispersion will certainly be very difficult to preclude totally. Future directions for research should thus focus on the long-term establishment and effects of transgenes on natural communities.

The evolutionary history of a wild-cultivated complex is the outcome of different factors, among which are the domestication of the cultivated form, its geographical and demographic expansion, as well as gene flow between natural and cultivated populations. To clarify this history for alfalfa (Medicago sativa), we adopted a phylogeographical approach based on a sample of natural and cultivated populations of the M. sativa species complex. This sample was characterized for mtDNA variation through restriction fragment length polymorphisms. Twenty-two mitotypes were identified in the whole data set (155 individuals). In the wild pool from the presumed area of origin (Near East to Central Asia), the diversity was high but the absence of geographical differentiation hinders a more precise location of the centre of domestication. Within the cultivated alfalfa, the geographical structure suggests strongly the existence of at least two independent routes of dissemination of alfalfa from its centre of origin. In particular, original mitotypes detected in some regions suggest that wild populations not sampled in this study, and located outside the presumed area of domestication, may have contributed to the diversity of the cultivated pool. Finally, analysis of the mtDNA variation shows that natural populations collected in Spain constitute an endemic wild pool now introgressed partially by cultivated alfalfa.

In order to understand the dynamics of contact zones between natural and cultivated populations of alfalfa (Medicago sativa L.) in Spain, we investigated the patterns of mitochondrial variation (characterized through restriction fragment length polymorphisms) for 173 individuals in four natural and four cultivated populations of this species. We also examined the relationship between mtDNA and phenotypic variation. This study shows that cytoplasmic variation occurs in both natural and cultivated populations of Medicago sativa. Among the five mitotypes identified in the species, two are specific from the natural populations. The distribution of shared mitotypes between wild and cultivated forms attested to the occurrence of gene flow through seed from cultivated towards natural populations. Comparisons between cytoplasmic and phenotypic patterns of variation give information on the patterns of gene exchange that occur in contact zones over successive generations.

Self-incompatibility (SI) is a widespread mechanism that prevents inbreeding in flowering plants. In many species, SI is controlled by a single locus (the S locus) where numerous alleles are maintained by negative frequency-dependent selection. Inbreeding depression, the decline in fitness of selfed individuals compared to outcrossed ones, is an essential factor in the evolution of SI systems. Conversely, breeding systems influence levels of inbreeding depression. Little is known about the joint effect of SI and drift on inbreeding depression. Here we studied, using a two-locus model, the effect of SI (frequency-dependent selection) on a locus subject to recurrent deleterious mutations causing inbreeding depression. Simulations were performed to assess the effect of population size and linkage between the two loci on the level of inbreeding depression and genetic load. We show that the sheltering of deleterious alleles linked to the S locus strengthens inbreeding depression in small populations. We discuss the implications of our results for the evolution of SI systems.

International audience; In order to elucidate the mechanisms underlying the large amount of RAPD polymorphism found in 1990 in a population of the selfing annual Medicago truncatula GAERTN. (Fabaceae), we have analysed most of the individuals (n = 363) from the same population 6 years later using microsatellite loci. We confirm the result of the earlier study, namely that this population is very polymorphic and highly subdivided, with approximately 37% of the variance distributed among subpopulations, only 50 m apart one from another. We use standard F-statistics analyses, linkage disequilibria, minimum spanning network, multilocus assignment tests and spatial autocorrelation analyses to test the hypotheses that spatial structure and outcrossing events are involved in maintaining the large amount of genetic diversity at the level of each subpopulation. Interestingly, fine-scale spatial structure could be observed in only one subpopulation suggesting that other mechanisms are acting elsewhere. To the best of our knowledge, this is the first study of fine spatial genetic structure in a predominantly selfing species.

The conservation of a crop's wild relatives as genetic resources requires an understanding of the way genetic diversity is maintained in their populations, notably the effect of crop-to-wild gene flow. In this study, the amount of differentiation between natural and cultivated populations of Medicago sativa was analysed using random amplified polymorphic DNA (RAPD) markers and an extension of the AMOVA procedure adapted to autotetraploid organisms. Simulations of structured populations were performed to test whether AMOVA provides estimates of population structure in autotetraploids that can be directly compared to those obtained for allozyme data. Simulations showed that straight phi-statistics allow a good estimation of population differentiation when unbiased allelic frequencies are used to correct the conditional expectations of squared genetic distances. But such unbiased estimates can not be practically guaranteed, and population structure is notably overestimated when some populations are fixed for the presence of amplified fragments. However, removing fixed loci from the data set improves the statistical power of the test for population structure. The genetic variation of 15 natural and six cultivated populations of M. sativa was analysed at 25 RAPD loci and compared to estimates computed with allozymes on the same set of populations. Although RAPD markers revealed less within-population genetic diversity than allozymes, the quantitative and qualitative patterns of population structure were in full agreement with allozymes. This confirmed the conclusions drawn from the allozymic survey: crop-to-wild gene flow occurred in many locations, but some other mechanisms opposed cultivated traits to be maintained into natural populations.

Amounts of genetic variability, genetic differentiation among taxa and populations, and population sizes were studied in five populations of Centaurea maculosa ssp. maculosa (a widespread taxon), all six populations of C. corymbosa (a narrowly endemic species), and the single population of C. maculosa ssp. albida. Seventeen isozyme loci were studied, of which nine were polymorphic. Results suggest that C. corymbosa and C. maculosa ssp. albida are likely derived from C. maculosa ssp. maculosa because the former represent a sample of the diversity of the latter. The percentage of polymorphic loci and Nei’s genetic diversity were positively and significantly correlated with population size over all populations, but not within each taxon. Populations of both the widespread C. maculosa ssp. maculosa and the rare C. corymbosa were strongly differentiated: overall, FST values were 0.26 and 0.34, respectively. Differentiation among populations of different taxa was of the same order of magnitude as that observed among populations within taxa. Nevertheless, significant differentiation among the three taxa was found by a hierarchical analysis of variance on allele frequencies. We suggest that bottlenecks or founder effects associated with colonization events and ecological specialization in some populations of C. maculosa ssp. maculosa have led to new taxa such as C. corymbosa and C. maculosa ssp. albida. This may be a direct consequence of the particularly strong differentiation among populations of the widespread C. maculosa ssp. maculosa. Our study highlights the utility of considering closely related widespread taxa in order to understand the population biology and evolution of rare species, as well as to design proper management programs. Predicciones de Endemismo a Partir de la Estructura Poblacional de una Especie con Amplia Distribucion: Estudio del Caso de Centaurea maculosa Lam. (Asteraceae) So determino la variabilidad genetica, la diferenciacion genetica entre taxones y entre poblaciones, y el tamano de poblacion en 5 poblaciones de Centaurea maculosa ssp. maculosa (un taxon de amplia distribucion), las 6 unicas poblaciones de C. corymbosa (especie endemica), y la unica poblacion de C. maculosa ssp. albida existente. Se estudiaron diecisiete loci enzimaticos, de los cuales 9 fueron polimorficos. Los resultados sugieren que los dos taxones endemicos probablemente derivaron del taxon de amplia distribucion, ya que los primeros representan una muestra de la diversidad de este ultimo. Tanto el porcentaje de loci polimorficos como la diversidad genetica (indice de Nei) mostraron una correlacion positiva con el tamano de la poblacion. Se encontro una fuerte diferenciacion entre las poblaciones tanto en el taxon endemico como en el de amplia distribucion: los valores de FST globales fueron 0.34 para C. corymbosa y 0.26 para C. maculosa ssp. maculosa. La diferenciacion entre poblaciones de taxones diferentes fue del mismo orden de magnitud que la observada entre poblaciones dentro de un mismo taxon. Sin embargo, un analisis de la varianza jerarquico de las frecuencias genicas revelo una diferenciacion significativa entre los tres taxones. Se sugiere que la aparicion de taxones endemicos como C. corymbosa y C. maculosa ssp. albida se debe a efectos fundacionales, como consecuencia de “cuellos de botella” o procesos de colonizacion, y posterior especializacion ecologica en algunas poblaciones de C. maculosa ssp. maculosa. Esto seria una consecuencia directa de la fuerte diferenciacion entre poblaciones de C. maculosa ssp. maculosa. Este estudio remarca la utilidad del uso de especies de amplia distribucion estrechamente emparentadas con especies endemicas para comprender la biologia y evolucion de especies raras, asi como para disenar programas de manejo adecuados.

Crop/weed gene flow is often of great concern for a better understanding of germplasm diversity. This paper provides an overview of the levels and partition of genetic diversity assessed between wild and cultivated populations of Medicago sativa L. in Spain for both molecular markers and agromorphological traits. We report that most wild populations preserve a great agronomic and morphological originality despite their occasional parapatry with cultivated forms and the occurrence of crop/weed gene flow in many locations. However, populations that display either a continuous range of intermediate plants between the typical wild and cultivated forms, or a mosaic of individuals that can be assigned to each gene pool respectively, were also identified. Providing that reproductive isolation is not strict, we hypothesise that contrasted selective pressure between wild and cultivated populations mainly accounted for the maintenance of their phenotypic integrity., L’existence d’un flux génique entre des populations cultivées et sauvages joue souvent un rôle prépondérant dans l’évolution de la diversité génétique de ces dernières. Nous présentons ici une vue générale des niveaux, et de l’organisation de la diversité génétique mesurée sur la base de marqueurs neutres et de caractères agromorphologiques dans des populations sauvages et cultivées de luzerne (M. sativa L.)en Espagne. La plupart des populations sauvages maintiennent une claire originalité agronomique et morphologique bien qu’elles soient parapatriques de populations cultivées et qu’elles fassent parfois l’objet d’introgressions régulières de leur part. Néanmoins notre étude a permis de mettre en évidence un ensemble de populations sauvages moins typiques, présentant un continuum de formes intermédiaires ou une mosaïque d’individus caractéristiques de chaque forme. Notre étude suggère que l’isolement reproducteur entre populations sauvages et cultivées n’est pas total et qu’il est vraisemblable que de fortes pressions de sélection engendrées par le milieu jouent un rôle déterminant dans le maintien de l’intégrité phénotypique des formes sauvages.

The factor “2” is lacking in the initial version. This is only a typo error and the different figures were drawn with the correct equation. Note that there is the same typo error in equation (16) in Glemin (2007). However, Aneil Agrawal pointed us to a potential problem with this equation. Our aim was to use a not too arbitrary function for the α term, so we used a rather crude approximation. This equation can be obtained for instance from equation (8) in Hudson and Kaplan (1995) using H + F − HF instead of H and C(1 − F) instead of C for dominance and recombination coefficient, respectively. Equation (8) of Hudson and Kaplan (1995) then becomes

International audience; Mitochondrial variability was investigated in natural populations of wild carrot (Daucus carota ssp carota) in different regions: South of France, Greece, and various sites in the Mediterranean Basin and Asia. Total DNA was digested with two restriction endonucleases (EcoRV and HindIII) and probed with three mitochon-drial DNA-specific genes (coxI, atp6, and coxlI). Twenty-five different mitochondrial types were found in 80 analyzed individuals. Thirteen mitotypes were found among the 7 French populations studied. On average, 4.4 different mitotypes were observed per population, and these mitotypes were well-distributed among the populations. All of the mitochondrial types were specific to a single region. However, the proportion of shared restriction fragments between 2 mitotypes from different regions was not particularly lower than that which occurred among mitotypes from a single region. On the basis of the sexual phenotype [male-sterile (MS) or hermaphrodite] of the plants studied in situ and that of their progeny, 2 mitotypes were found to be highly associated with male sterility. Eighty percent of the plants bearing these mitotypes were MS in situ, and all of these plants produced more than 30% MS plants in Communicated by R. Hagemann J. Ronfort ([]) D6partement de biologie des populations, Centre d'Ecologie Fon-ctionnelle et Evolutive (CEFE), CNRS, 1919, Route de Mende, B.P. 5051-34033 Montpellier Cedex France P. Saumitou-Laprade Laboratoire de G6netique et Evolution des populations v6g6tales, 237, Universitb Paris vi, 75252 Paris Cedex France their progeny. This association with male sterility was consistent in several populations, suggesting an association with a cytoplasmic male-sterility system. Moreover , these two mitotypes had very similar mitochon-drial DNA restriction patterns and were well-differentiated from the other mitotypes observed in wild plants and also from those observed in the two CMS types already known in the cultivated carrot. This suggests that they correspond to a third cytoplasmic sterility. Key words RFLP 9 Mitochondrial DNA diversity 9 Natural populations 9 Cytoplasmic male sterility 9 Daucus carota L.

SummaryPrevious theoretical studies of the evolution of the selfing rate have shown that mixed mating systems are not evolutionary stable states. Such models have, however, not included the effects of population structure and thus biparental inbreeding together with the evolution of selfing rates and inbreeding depression. In order to examine selection on selfing rates in structured populations, a stochastic model simulating a finite population with partial selfing and restricted pollen and seed dispersal has been developed. Selection on the mating system was followed by introducing modifiers affecting the selfing rate. The major result was that, with density dependent recruitment, a process which maintains the population structure necessary for biparental inbreeding to occur, a mixed mating system could be maintained. This result was associated with an increase of the mutation load with high selfing rates, and the selected selfing rate depended on the degree of population structure rather than on the initial selfing rate. With low dominance of deleterious alleles, complete allogamy can be selected for. Further studies showed that the more general condition of spatial heterogeneity of recruitment can lead to similar results, the most important condition being the maintenance of genetic structure within populations. A brief survey of the empirical literature shows that a positive relationship between the magnitude of inbreeding depression and the inbreeding coefficient within populations has been observed, in support of the present model.

Equalization of reproductive success of individuals, although it results in an increase of effective population size, leads also to an increase of the mutation load. The magnitude of this increase depends highly on the mode of fitness interactions between deleterious mutations, and is higher in the case of inbreeding. Recommended practices in conservation genetics must be evaluated in regards to these differing consequences of an increase of effective population size. To keep a balance between retaining genetic variability and minimizing the increase of the mutation load, equalization of reproductive success of a set of individuals rather than of every individual might be more advantageous.