By combining high-throughput sequencing (HTS) with experimental evolution, we can observe the within-host dynamics of pathogen variants of biomedical or ecological interest. We studied the evolutionary dynamics of five variants of Potato virus Y (PVY) in 15 doubled-haploid lines of pepper. All plants were inoculated with the same mixture of virus variants and variant frequencies were determined by HTS in eight plants of each pepper line at each of six sampling dates. We developed a method for estimating the intensities of selection and genetic drift in a multi-allelic Wright-Fisher model, applicable whether these forces are strong or weak, and in the absence of neutral markers. This method requires variant frequency determination at several time points, in independent hosts. The parameters are the selection coefficients for each PVY variant and four effective population sizes Ne at different time-points of the experiment. Numerical simulations of asexual haploid Wright-Fisher populations subjected to contrasting genetic drift (Ne ∈ [10,2000]) and selection (|s| ∈ [0,0. 15]) regimes were used to validate the method proposed. The experiment in closely related pepper host genotypes revealed that viruses experienced a considerable diversity of selection and genetic drift regimes. The resulting variant dynamics were accurately described by Wright-Fisher models. The fitness ranks of the variants were almost identical between host genotypes. By contrast, the dynamics of Ne were highly variable, although a bottleneck was often identified during the systemic movement of the virus. We demonstrated that, for a fixed initial PVY population, virus effective population size is a heritable trait in plants. These findings pave the way for the breeding of plant varieties exposing viruses to stronger genetic drift, thereby slowing virus adaptation.
Evolution in isolated populations results from the interplay between several forces, including mutation, selection, and genetic drift. Mutation creates genetic diversity within a population. Subsequent selection and genetic drift drive the evolution of diversity within the population. Selection is a deterministic force that increases the frequency of the fittest variants at the expense of the weakest ones. It can be characterized by the selection coefficient s, commonly calculated, at a specific locus, as the relative difference in fitness conferred by two alleles. Genetic drift, unlike selection, acts equally on all variants. It is the outcome of random sampling effects between generations, resulting in stochastic fluctuations in variant frequencies [1]. The strength of genetic drift is frequently evaluated by determining the effective population size Ne [1]. Ne is defined as the size of an ideal panmictic population of constant size with non-overlapping generations that would display the same degree of randomness in allele frequencies as the population studied [2]. Ne is often much lower than the census population size [3,4], but it can be seen as its evolutionary analog [5]. When Ne is small, sampling effects are magnified between generations, and allele frequencies therefore fluctuate strongly. For populations varying in size over time, the effective population size over a given number of generations can be approximated by the harmonic mean N ¯ e of effective population sizes at each generation. This approximation holds provided that the number of generations is much smaller than N ¯ e [6–8] and that mutation can be neglected [9]. Population size may vary over time due to bottlenecks, which are common in natural populations. As they greatly decrease population size, they have a disproportionate effect on the overall value of N ¯ e [1]. When selection and genetic drift act simultaneously, the probability of fixation of a new mutation (with a selection coefficient s), and, more generally, its evolutionary dynamics, is controlled by the product Ne × |s| [1,10]. If Ne × |s| ≪ 1, then genetic drift predominates over selection and evolution is mostly stochastic. If Ne × |s| ≫ 1, then selection becomes effective and evolution is mostly deterministic [10]. This rule of thumb can be applied to the evolutionary dynamics of pathogen variants of biomedical or ecological interest, during the course of infection of a single host, for microbe variants escaping the immune response of their host, or becoming resistant to drug therapy (e. g. [11]) or, in the case of plant pathogens, for variants adapting to host resistance genes (e. g. [12]). In this study, we combined high-throughput sequencing (HTS) with experimental evolution to measure the within-host dynamics of five variants of Potato virus Y (PVY, genus Potyvirus, family Potyviridae) in closely related plant genotypes [13]. It remains challenging to unravel the effects of genetic drift and selection in the absence of neutral markers, in studies of the adaptation dynamics of pathogens. This situation is frequently encountered for pathogens with small genomes, especially viruses [14,15]. Various approaches based on moment [16,17] or likelihood [18–20] methods have been proposed for estimating Ne, but all require the genetic markers studied to be neutral. Various methods have also been proposed for detecting selection and estimating selection coefficients. These methods require at least some prior information about Ne (e. g. [21]) or assume that genetic drift is negligible (e. g. [22]). However, in the absence of neutral markers and without prior estimates of Ne, both selection and genetic drift must be taken into account, as these two forces act simultaneously on evolution. This greatly complicates the estimation of Ne and s. Only a few methods have been proposed for the joint estimation of Ne and s from time-sampled data (see [11] and [23] for a review). For large effective population sizes (typically Ne > 5000) and small selection coefficients (typically |s| < 0. 01), several likelihood methods based on diffusion approximations of the Wright-Fisher model [1,11] are available [24–27]. In the situations in which these methods are valid, the ranges of Ne and s values obtained are rather restrictive for many microorganisms, particularly viruses [28–31]. Foll et al. [32] recently proposed the use of approximate Bayesian computation (ABC) for the joint estimation of Ne and s in a Wright-Fisher model. Their method can deal with both weak and strong selection regimes, but still requires multilocus genome-wide data with mostly neutral loci to estimate Ne accurately. In this study, we investigated the evolutionary dynamics of five variants of PVY in 15 closely related pepper genotypes. All plants were inoculated with the same mixture of virus variants and variant frequencies were determined with HTS in eight plants of each genotype at each of six sampling dates after inoculation. A diverse range of evolutionary patterns was observed. We developed a method for estimating the parameters of a multi-allelic Wright-Fisher model with selection and genetic drift, to investigate the underlying evolutionary processes. This method has two main advantages: it applies to a large range of selection and genetic drift intensities and it works efficiently in the absence of neutral markers. The parameters of the Wright-Fisher model (i. e. selection coefficients for each virus variant and effective population sizes at given time points) can be estimated by coupling maximum likelihood and ABC methods and applying them to a set of variant frequencies determined at several time points in independent hosts. We tested the method with numerical simulations mimicking the datasets obtained with HTS in evolve-and-resequence experiments [33]. The simulations covered an extensive range of Ne and s values. We were then able to estimate the selection coefficient of each PVY variant in each pepper genotype and the changes in effective population size over time during the colonization of the plant by the virus. Finally, by varying pepper genotypes and fixing the initial PVY population, we provided evidence that the effective population size of PVY is a heritable plant trait. This finding paves the way for the breeding of plant cultivars exposing viruses to greater genetic drift and/or smaller selection effects.
We developed a method for estimating the parameters of a multi-allelic Wright-Fisher model with selection and genetic drift for a haploid population. The parameters and state variables of the model and the observed variables are summarized in Table 1. Before using the estimation method on the datasets corresponding to the biological experiment, we performed several batches of simulations to assess its ability to infer effective population sizes and selection coefficients accurately (see S2 Text for details). Briefly, in experiment 1, we first simulated the changes in frequencies of five virus variants under 750 selection and genetic drift regimes with a Wright-Fisher model for haploid individuals. The simulations were designed to fit the experimental setup of our datasets (48 independent host plants regularly analyzed at 6 sampling dates). For each of the 750 datasets obtained, the true parameters θtrue were known and could be compared with the estimated parameters θ ^. In experiment 2, we assessed the sensitivity of the estimation method to the presence of a sixth undetected virus variant. This sixth variant was selectively neutral (its selection coefficient is zero), present in the inoculum at a frequency of 3% and still present at the last sampling date (34 dpi) in all plants analyzed, at frequencies ranging from 1% to 6%. It affected the dynamics of the five variants of interest in all plants but was not detected, so the variant frequencies measured by HTS (and used to estimate θ ^) are noisy with respect to their true values. In all, 350 simulated datasets were analyzed in this second test.
The frequencies of the five virus variants were assessed in completely isolated populations during the course of infection, in 15 different plant genotypes. For each of these 15 pepper genotypes, 48 plants were inoculated with the same equimolar mixture of the five variants, and the frequencies of the virus variants were determined in eight plants at each of six sampling dates, from 6 to 34 days post-inoculation (Fig 2). In a few cases, no viruses were detected in plant samples (lacking bars in Fig 2). These negative samples may reflect the presence of an extreme bottleneck at inoculation, leading to virus population extinction, or a long time lag to systemic infection of the plant (for measurements from 10 to 34 dpi), resulting in the sampling of leaves not yet infected (e. g. DH line 2321). Negative samples were most frequent for the first two dates on which systemically infected leaves were analyzed, i. e. at 10 and 14 dpi, probably indicating a time lag to systemic infection in some DH lines. Negative samples were observed in only four DH lines (e. g. DH lines 219 and 2321). No infection was observed in a mean of 3. 5 (resp. 2. 0) plant samples 10 (resp. 14) dpi for the four DH lines concerned. The virus populations present in all infected plants and in the common inoculum were analyzed by HTS, to determine the frequencies of the five PVY variants. Inoculum analysis confirmed that all variants were present in roughly equimolar proportions, with 22. 6% of variant G, 17. 5% of N, 20. 6% of K, 17. 1% of GK and 22. 2% of KN. The raw data for variant frequency dynamics provided considerably different patterns between the 15 pepper genotypes (Fig 2, S2 and S3 Figs). Variant frequencies were similar between virus populations sampled on the same date in some plant genotypes, consistent with weak genetic drift (e. g. DH lines 240 and 2430, Fig 2A and 2B), whereas they differed in other plant genotypes (e. g. DH lines 2321 and 219, Fig 2D and 2E). Furthermore, the heterogeneity of variant frequencies between the eight plants analyzed fluctuated between dates, probably due to changes in effective population size during the course of infection (e. g. DH line 2344, Fig 2C). The four pepper genotypes for which some samples were virus-negative were also characterized by the highest heterogeneity in variant frequencies, consistent with an extreme bottleneck at inoculation and/or during systemic movement of the virus (see DH lines 2321,219,2256 and 2400 in Fig 2D and 2E, S2D and S3I Figs). Selection regimes also differed between lines. In some DH lines, all variants remained present at all dates (e. g. DH line 240, Fig 2A), whereas one variant (e. g. DH line 219, Fig 2E), or up to two variants (e. g. DH lines 2430,2344,2321, Fig 2B–2D) became extinct in others. Before its application to the experimental dataset, we validated the estimation method proposed by numerical simulations of a Wright-Fisher model with selection and genetic drift for haploid individuals. We estimated the Ne (t) and ri of the PVY populations in each DH line with a Wright-Fisher model including selection and genetic drift. By contrast to the numerical experiments, the evolutionary parameters underlying the true dynamics of virus populations in their hosts were unknown. The Wright-Fisher model fitted the data very satisfactorily (Fig 5). The best-fit line between observed and fitted mean variant frequencies (averaged over all virus populations and sampling times) was very close to the first bisector (Fig 5A; slope = 0. 92, intercept = 0. 01, R2 = 0. 92). This was also the case for the variability of variant frequencies between virus populations at each sampling date td (Fig 5B; slope = 0. 92, intercept = -0. 09, R2 = 0. 84). A Wright-Fisher model including selection and genetic drift accurately described the mean evolutionary dynamics of a virus population and the variability of these dynamics between hosts. Due to an identifiability issue (we observed the relative frequencies of variants rather than variant densities), we had to fix the number of generations per day γ. We set this number to 1, a value close to that reported by Khelifa et al. [40]. Different γ values would change ri and Ne (t) estimates to r i 1 / γ and γNe (t), but would have no effect on their ranking. Relative fitness values (ri) ranged from 0. 43 to 1. 25 (corresponding to |s|: 5% quantile = 0. 004, mean = 0. 12,95% quantile = 0. 27) and were associated with narrow 90% confidence intervals (S3 Table). The fitness ranks of the PVY variants were very similar in most DH lines (Fig 6A and 6C). Variant G was the weakest in all DH lines, followed by variant N in 13 DH lines. Variant GK was the fittest variant in 13 DH lines, with variant K the fittest variant in the remaining two lines (DH lines 2256 and 2430). Overall, variants K and GK were the two fittest variants in 12 DH lines; variants GK and KN were the two fittest in DH lines 2349 and 2321, and variants N and GK the two fittest in DH line 219. The fitness difference between the weakest and the fittest variants ranged from 0. 14 for DH line 219 to 0. 81 for DH line 2349. We further estimated the dynamics of effective population size over the time course of the experiment, as modeled by a piecewise function Ne (t), using a model selection procedure. Four models with one to four parameters were considered. The most general model M 4 distinguished four successive effective population sizes (one in the inoculated organ and three during systemic infection). M 4 was the model best supported by the data for five DH lines (2173,2321,2328,2344 and 2367). Model M 3 distinguished three successive effective population sizes (one in the inoculated organ and two during systemic infection). It was best supported by the data for five DH lines (219,221,2256,240 and 2430). Model M 2, which distinguished two successive effective population sizes (one in the inoculated organ and one during systemic infection), was selected for a single DH line (2426). Finally, with M 1, the effective population size of the virus population remained constant. This model was selected in the four remaining DH lines (2123,2264,2349 and 2400). The corresponding posterior probabilities of each model are shown in S4 Table, together with effective population size estimates and 90% credibility intervals. At the first sampling date, considerable variability was observed (Fig 6B and 6D), with effective population sizes ranging from 13 for DH lines 219 and 2256 to 1515 for DH line 240. This was not surprising, given that we chose the DH lines on the basis of the density of primary infection foci in inoculated organs [34] (S1 Fig). A much narrower range of effective population sizes, from 18 to 462, was observed across all plant genotypes at 10 dpi, the first date on which systemic infection was observed. From 6 to 10 dpi, effective population sizes decreased in eight DH lines (Fig 6B), remained approximately constant in six DH lines (Fig 6D) and increased slightly in a single plant genotype (DH line 2173, Fig 6D). Later on, from 10 to 34 dpi, effective population size increased in eight DH lines (mostly DH lines displaying a bottleneck from 6 to 10 dpi, Fig 6B) and remained approximately constant in the others (mostly in DH lines with lower, i. e. < 500, effective population sizes in the inoculated organ, Fig 6D). By creating two dataset replicates of 24 randomly chosen plants for each DH line, we estimated the heritability of two plant traits corresponding to the evolutionary forces exerted by the plant on virus populations: selection and genetic drift. These forces were estimated by (i) intrinsic rates of increase in viral variants and (ii) effective population sizes for PVY. With 24 plants in each dataset, we used the function Ne (t) of model M 2 with two parameters. In this approach, we used the contrasting behavior of PVY populations, which were fixed and identical at the time of inoculation in all plants, on different pepper genotypes to characterize the phenotype of each host. Very high heritability estimates were obtained for the intrinsic rates of increase (mean heritability over the five variant estimates: h2 = 0. 94). Somewhat lower, but nevertheless substantial heritability estimates were obtained for effective population size in the inoculated organ (mean heritability, h2 = 0. 64) and for effective population size during systemic infection (mean heritability, h2 = 0. 63). The details of the calculation are provided in S3 Text.
We present here a method for the estimation of selection and genetic drift in a haploid and asexual organism, as modeled by a Wright-Fisher process. As for any model-based approach, the population of interest must not be too far from an ideal Wright-Fisher population with suitable parameters [10]. The estimation method did not require neutral markers. It was validated for small effective population sizes (Ne ≪ 100) and a wide range of both positive and negative selection coefficients (weak (|s| ≃ 0. 01) or strong (|s| ≃ 0. 15) selection), using simulated datasets. Recent reviews [23,32] have highlighted the small number of methods available for the inference of selection and genetic drift over the whole range of variation, particularly in the case of small effective population sizes (Ne ≪ 1000) and strong selection coefficients (|s| ≃ 0. 1). Indeed, these conditions do not fulfill the hypotheses underlying most approximations of the Wright-Fisher model. The classical approximation, with a standard diffusion process, requires both selection and genetic drift to be weak [23]. Approximations based on Gaussian diffusion require the stochastic effects of genetic drift to decrease more rapidly than the effects of selection [23]. The work of Foll et al. [11,32] constituted a major step forward, but their method requires a large proportion of the genetic markers studied to be neutral. This assumption is not valid for many pathogens with small genomes, such as viruses. For example, only 22. 7% of 66 randomly chosen mutations in the genome of Tobacco etch virus (TEV, genus Potyvirus), a plant RNA virus, were found to be consistent with neutrality [45]. As the statistical power to detect departure from neutrality is limited, the true proportion of neutral mutations is probably much lower. Similar results have been obtained for bacteria (e. g. [46]). The estimation method proposed does not require neutral markers, an appealing feature for studying pathogens with small genomes. Lacerda and Seoighe [47] recently developed another method that does not require neutral markers. Their method provided satisfactory estimates of both Ne and s (estimated at a single locus) for a relatively small effective population size of 1000 individuals and values of s up to 0. 5. They did not test the performance of their method for Ne ≪ 1000. By comparison, the method developed here was effective for much lower Ne values, in the range of a few tens of individuals, and for inferring the time course of Ne over a few tens of generations. However, although the range of selection coefficients s included cases of strong selection (|s| ≃ 0. 1, as defined by Malaspinas [23]), none of the simulation experiments included values as high as 0. 5. It may be possible to infer such high selection coefficients with the estimation method proposed, provided that the first generations are sampled more densely, typically every day after inoculation in our set-up. Lacerda and Seoighe [47], for example, used samples taken at each generation, for 20 generations. This makes it possible to record the trajectories of variant frequencies before variant loss or fixation. The use of the proposed estimation method requires observation of the evolution of isolated populations derived from the same parental population, each population being sampled only once. This design is particularly suitable for studying within-host microbial evolution when several genetically-identical hosts (48 plants for each pepper genotype in our case study) can easily be included in the experiment. With this experimental design, we observed a set of variant frequencies at several time points, in independent hosts. This set contained footprints of selection and genetic drift. In the method developed, selection is evaluated from the mean trajectories of variant frequencies. Genetic drift is evaluated at several time points, by assessing differences in variant frequencies between the replicated populations during the time-course of the experiment. Even for populations with small effective sizes, for which genetic drift and selection have confounding effects on the fate of variants (Fig 2), a moderate number of replicates contains sufficient information to disentangle the two mechanisms. Here, we estimated four selection coefficients and four effective population sizes (i. e. 8 parameters) with 48 samples (6 sampling dates × 8 replicates). The proposed estimation method could be improved further. It explicitly accounts for the technical sampling noise resulting from the assessment of variant frequencies from finite counts of virus sequences. However, HTS also introduces sequencing errors, albeit at a low rate of about 1 substitution per 400 bases for MiSeq technology [48], which were not explicitly accounted for in our framework. Several models have been proposed for separating true genetic variation from technical artifacts [48], and these models could be integrated into the method through a hierarchical Bayesian modeling framework [49], for example. Finally, the method could be extended to take mutation and recombination into account, particularly for experiments over longer periods, in which new variants might appear and displace those currently most abundant. In our short-term experiment, we have already observed de novo substitutions in a few plants (removed plant samples, see S1 Text). The inclusion of recombination is not relevant for our case study, as the nucleotide positions differentiating the variants are located only a few codons apart. Recombination can thus be ignored in this study [50], particularly given the small number of generations considered [32]. On the host side, our experiment involved 15 DH lines of pepper, all carrying the major resistance gene pvr23, but differing in terms of their genetic backgrounds [12]. These DH lines were derived from the F1 hybrid between two pepper lines, Perennial and Yolo Wonder. Consequently, on average, any pair of DH lines have 50 percent of alleles in common for markers differentiating between Perennial and Yolo Wonder. This is the first study, to our knowledge, to show such a high level of diversity in selection and genetic drift regimes experienced by virus populations from the same viral inoculum in closely related host genotypes (Fig 2, S2 and S3 Figs). On the pathogen side, we used five virus variants: the G and N variants displayed weaker adaptation to pvr23 than the K, GK and KN variants. The ranking of the selection coefficients of the five variants was mostly identical in the 15 plant genotypes. We were therefore unable to identify any host genotype, among those tested, able to counterselect against the virus variants best adapted to pvr23. This may be due to (i) the strong selective effect exerted by the major-effect resistance gene pvr23, which is present in all the DH lines studied here and probably exceeds the additional selective effect of the plant genetic background and/or (ii) the close genetic relatedness of the DH lines analyzed. Other genetic resources for pepper should be explored, to identify genotypes capable of counterselecting against the K, GK and KN variants, which were the fittest in our study. The best candidates for this would be pepper genotypes carrying pvr2 resistance alleles other than pvr23, with a different specificity in the face of PVY diversity [51], or pepper genotypes devoid of resistance alleles at the pvr2 locus, as shown by Quenouille et al. [12]. Combinations of plant genotypes exerting opposite selective pressures on pathogen populations are particularly interesting for the sustainable management of plant resistance at landscape level, and can be implemented in cultivar rotations, mixtures or mosaics [52]. However, in our study, the difference in fitness between the weakest and fittest variants differed between host genotypes. The dynamics of selection for the fittest variants were under plant genetic control and could therefore be modulated by the choice of plant genotypes grown. For example, growing the pepper DH lines with the smallest differential selection between the five PVY variants would be particularly useful for delaying PVY adaptation in pvr23-carrying plants, in which a two-step mutational trajectory may be required [12]. Indeed, the G and N variants are most likely to appear initially, because they require transitions, whereas the K variant requires a transversion, and transitions are more frequent than transversions [53]. However, an additional substitution, in a second step, is required to confer a sufficient level of fitness for the emergence of GK and KN variants. These mutational trajectories were observed in PVY adaptation to the Perennial pepper genotype, the resistant parent of all the DH lines studied here [12]. We also inferred the time course of the genetic drift experienced by the viruses in the 15 host environments during the experiment. Genetic drift intensities were highly variable with time and between plant genotypes, revealing an unprecedented level of variability between closely related host genotypes. Our estimates of Ne (t) ranged from 18 to 462 just after the colonization of apical leaves at 10 dpi, and from 13 to 1515 in the inoculated leaves four days previously (at 6 dpi). Eight of the 15 DH lines displayed a high Ne in the inoculated leaves at 6 dpi (from 421 to 1515), a decrease at 10 dpi (Ne (10 dpi) values of 1. 5 to 83. 5% of the value at 6 dpi) and a subsequent increase (Fig 6B). This pattern suggests a founder effect, in which a new PVY population in apical leaves is set up by a few members of the original population in the inoculated leaf. In the remaining seven DH lines, the Ne of the inoculated leaves at 6 dpi was much lower (from 13 to 462), and Ne values often remained low in the apical leaves (Fig 6D). However, an increase in Ne was observed in DH lines 219 and 2173, after 14 dpi. This result sheds new light on the importance of the within-host bottlenecks experienced by virus populations, as discussed in a recent article by Zwart et al. [54], who reported that the Ne of TEV in the first systemically infected leaf of tobacco plants was determined largely by inoculum viral load. They then hypothesized that genetic drift occurred mostly during the inoculation process. Previous estimations of Ne for viruses did not focus on Ne dynamics at the whole-plant level as in this study. Instead, they considered the multiplicity of infection (MOI) during cell-to-cell movement or Ne during the colonization of apical leaves (for a comprehensive review, see Gutiérrez et al. [4]). Direct comparisons with these studies are, therefore, not appropriate. Gutiérrez et al. [55] recently showed that Turnip mosaic virus (genus Potyvirus) infections are characterized by a very low MOI (≃ 1) when cells are infected with virus particles moving in the plant vasculature, and a much higher MOI (≃ 30) during subsequent cell-to-cell movement in the mesophyll. The general picture that emerges when we consider both these MOI patterns and plant growth dynamics is consistent with our observations. Indeed, the lowest Ne values were observed at 10 dpi, corresponding to the onset of systemic infection, when plants were small and consisted essentially of a few infected leaves. Ne tends often to increase with time, because (i) increasing numbers of leaves are infected and behave as virus sources as the plant grows and (ii) leaf areas increase, probably increasing the relative proportion of cell-to-cell, as opposed to long-distance, virus movement. One of the key results of this study is the finding that the effective population size of PVY is a heritable plant trait. The high heritability estimated for Ne (partially due to the use of a DH progeny of pepper genotypes) indicates that plant resistance could potentially be improved through breeding programs. Indeed, our findings pave the way for the breeding of plant cultivars exposing viruses to greater genetic drift. This would provide a twofold benefit against viruses. First, in asexual populations, genetic drift favors the accumulation of deleterious mutations, decreasing viral fitness (Muller’s ratchet) [56]. Second, genetic drift decreases the fixation probability of beneficial mutations, such as those responsible for overcoming plant resistance genes [57]. Breeding for greater genetic drift in virus populations would thus constitute a novel approach to increasing the durability of resistance to plant viruses in agricultural landscapes [52,58,59]. Another key result is the finding that the Wright-Fisher model accurately captures the major processes driving the within-host dynamics of a set of virus variants (Fig 5), despite being much simpler than the underlying mechanisms involved in the infection of highly structured hosts. Over longer periods, mutation and recombination increase in importance and this can easily be encompassed in the Wright-Fisher model [60]. This model can thus serve as a valuable cornerstone for linking the within- and between-host scales of disease dynamics and studying, for example, how breeding for greater genetic drift can delay the emergence of a new pathogen variant.
A growing number of experimental evolution studies are using an “evolve-and-resequence” approach to observe the within-host dynamics of pathogen variants of biomedical or ecological interest. The resulting data are particularly appropriate for studying the effects of evolutionary forces, such as selection and genetic drift, on the emergence of new pathogen variants. However, it remains challenging to unravel the effects of selection and genetic drift in the absence of neutral markers, a situation frequently encountered for microbes, such as viruses, due to their small constrained genomes. Using such an approach on a plant virus, we observed that the same set of virus variants displayed highly diverse dynamics in closely related plant genotypes. We developed and validated a method that does not require neutral markers, for estimating selection coefficients and effective population sizes from these experimental evolution data. We found that the viruses experienced considerable diversity in genetic drift regimes, depending on host genotype. Importantly, genetic drift experienced by virus populations was shown to be a heritable plant trait. These findings pave the way for the breeding of plant varieties exposing viruses to strong genetic drift, thereby slowing virus adaptation.
organismal evolution
plant anatomy
population genetics
variant genotypes
microbiology
genetic mapping
plant science
effective population size
microbial evolution
population biology
leaves
evolutionary genetics
viral evolution
genetic drift
population metrics
population size
virology
heredity
genetics
biology and life sciences
evolutionary biology
evolutionary processes
Estimating virus effective population size and selection without neutral markers