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International audience; Hybridization and adaptation to new hosts are important mechanisms of fungal disease emergence. Evaluating the risk of emergence of hybrids with enhanced virulence is then key to develop sustainable crop disease management. We evaluated this risk in Venturia inaequalis, the fungus responsible for the common and serious scab disease on Rosaceae hosts, including apple, pyracantha and loquat. Field isolates from these three hosts and progenies obtained from five crosses between formae speciales isolates collected from pyracantha (f.sp. pyracantha) and apple (f.sp. pomi) were tested for their pathogenicity on the three hosts. We confirmed a strict host specificity between isolates from apple and pyracantha, and showed that most isolates were able to cause disease on loquat. None of the 251 progeny obtained from five crosses between V. inaequalis f.sp. pyracantha and V. inaequalis f. sp. pomi could infect apple. If confirmed on more crosses, the inability of the hybrids to infect apple could lead to a novel biocontrol strategy based on a sexual hijacking of V. inaequalis f.sp. pomi by a massive introduction of V. inaequalis f.sp. pyracantha in apple orchards. This strategy, analogous to the sterile insect approach, could lead to the collapse of the population size of V. inaequalis and dramatically reduce the use of chemicals in orchards.

Hybridization and adaptation to new hosts are important mechanisms of fungal disease emergence. Evaluating the risk of emergence of hybrids with enhanced virulence is then key to develop sustainable crop disease management. We evaluated this risk in

International audience; Today, the genus Scedosporium comprises at least ten species with four of them, Scedosporium apiospermum, Scedosporium boydii, Scedosporium aurantiacum and Scedosporium minutisporum capable of colonizing the lungs of patients with cystic fibrosis. Scedosporium dehoogii, which is also common in the soil, has never been reported as causing human pulmonary infections. Here we report the first genome sequence for S. dehoogii, an invaluable resource to understand the genetic bases of pathogenesis in the genus Scedosporium.

Today, the genus Scedosporium comprises at least ten species with four of them, Scedosporium apiospermum, Scedosporium boydii, Scedosporium aurantiacum and Scedosporium minutisporum capable of colonizing the lungs of patients with cystic fibrosis. Scedosporium dehoogii, which is also common in the soil, has never been reported as causing human pulmonary infections. Here we report the first genome sequence for S. dehoogii, an invaluable resource to understand the genetic bases of pathogenesis in the genus Scedosporium.

The haploid ascomycete Diplocarpon rosae is the causal agent of black spot disease on roses, a widespread and devastating disease in the outdoor landscape. In this study, we established a Eurasian collection of 77 monoconidial strains of D. rosae: 50 strains collected on cultivated roses in Europe and Asia, and 27 strains on wild roses in Kazakhstan. To provide tools to describe its biology and to study its genetic diversity, we sequenced two strains of D. rosae using Illumina paired-end technology. The genome sizes of these two strains were estimated at 31.1 and 35.2 Mb, which are two times smaller than the genome size of the unique strain previously published. A BUSCO analysis confirmed a genome duplication of the strain previously sequenced and partial gene duplication of strains analysed in this study. Using the two genome sequences, 27 polymorphic microsatellite markers were identified. Polymorphism analysis of the 77 strains revealed a strong genetic differentiation between strains from cultivated and wild roses, and a lower diversity within the fungal population from cultivated roses compared to the population from wild roses. Pathogenicity of 10 strains was evaluated on 9 rose cultivars inoculated in the greenhouse. Disease scoring allowed the classification of strains into three groups and the characterization of resistance of rose cultivars. Good correlation observed between resistance scoring in greenhouse conditions and in the field indicates that pathogenicity assays in controlled conditions could be very useful in the near future to rapidly characterize the resistance of new rose varieties to black spot disease., This work was supported by the CASDAR no. C-2014-06 from the French Ministry of Agriculture, Agrifood and Forestry. The authors thank Markus Linde for kindly supplying German infected leaves, Célia Calmels for microsatellite analysis, Sandrine Pierre for the contribution to assays on whole plants, Phenotic and Experimental unit Horticole for plant management in the greenhouse and in the field, respectively, Thibault Leroy for reads assembly of the Kazakh strain, and Biogenouest for read assembly of the German strain. All microsatellite data were obtained on the ANAN platform (M. Bahut). This work was financially supported by SFR 4207 Quasav.

Genetic resistance is a useful strategy to control plant disease, but its effectiveness may be reduced over time due to the emergence of pathogens able to circumvent the defences of the plant. However, the pyramiding of different resistance factors in the same plant can improve the effectiveness and durability of the resistance. To investigate the potential for this approach in apple to control scab disease we surveyed scab incidence in two experimental orchards located more than 300 km apart planted with apple genotypes carrying quantitative resistance and major gene resistance alone or in combination. Our results showed that the effectiveness of pyramiding in controlling scab was dependent on the site and could not be completely explained by the effectiveness level of the resistances alone.

Secondary contact between crops and their wild relatives poses a threat to wild species, not only through gene flow between plants, but also through the dispersal of crop pathogens and genetic exchanges involving these pathogens, particularly those that have become more virulent by indirect selection on resistant crops, a phenomenon known as “pestification”. Joint analyses of wild and domesticated hosts and their pathogens are essential to address this issue, but such analyses remain rare. We used population genetics approaches, demographic inference and pathogenicity tests on host-pathogen pairs of wild or domesticated apple trees from Central Asia and their main fungal pathogen, Venturia inaequalis, which itself has differentiated agricultural and wild-type populations. We confirmed the occurrence of gene flow from cultivated (Malus domestica) to wild (Malus sieversii) apple trees in Asian forests, potentially threatening the persistence of Asian wild apple trees. Pathogenicity tests demonstrated the pestification of V. inaequalis, the agricultural-type population being more virulent on both wild and domesticated trees. SNP markers and the demographic modeling of pathogen populations revealed hybridization following secondary contact between agricultural and wild-type fungal populations, and dispersal of the agricultural-type pathogen population in wild forests, increasing the threat of disease in the wild apple species. We detected a SNP potentially involved in pathogen pestification, generating an early stop codon in a gene encoding a small secreted protein in the agricultural-type fungal population. Our findings, based on joint analyses of paired host and pathogen datasets, highlight the threat posed by cultivating a crop near its center of origin, in terms of pestified pathogen invasions in wild plant populations and introgression in the wild-type pathogen population.

TheVenturiagenus comprises fungal species that are pathogens onRosaceaehost plants, includingV. inaequalisandV. asperataon apple,V. aucupariaeon sorbus andV. pirinaon pear. Although the genetic structure ofV. inaequalispopulations has been investigated in detail, genomic features underlying these subdivisions remain poorly understood. Here, we report whole genome sequencing of 87Venturiastrains that represent each species and each population withinV. inaequalis. We present a PacBio genome assembly for theV. inaequalisEU-B04 reference isolate. The size of selected genomes was determined by flow cytometry, and varied from 45 to 93 Mb. Genome assemblies ofV. inaequalisandV. aucupariaecontain a high content of transposable elements (TEs), most of which belong to the Gypsy or Copia LTR superfamilies and have been inactivated by Repeat-Induced Point mutations. The reference assembly ofV. inaequalispresents a mosaic structure of GC-equilibrated regions that mainly contain predicted genes and AT-rich regions, mainly composed of TEs. Six pairs of strains were identified as clones. Single-Nucleotide Polymorphism (SNP) analysis between these clones revealed a high number of SNPs that are mostly located in AT-rich regions due to misalignments and allowed determining a false discovery rate. The availability of these genome sequences is expected to stimulate genetics and population genomics research ofVenturiapathogens. Especially, it will help understanding the evolutionary history ofVenturiaspecies that are pathogenic on different hosts, a history that has probably been substantially influenced by TEs.

International audience; In pathogens, introgressions through secondary contacts between divergent populations from agricultural and nonagricultural disease reservoirs are expected to have crucial evolutionary and epidemiological implications. Despite the importance of this question for disease management, experimental demonstrations of these implications remain scarce.Recently, we identified a virulent population of the apple scab pathogen Venturia inaequalis that migrated from nonagricultural hosts to European domestic apple orchards. Here, we investigated the occurrence of gene flow between agricultural and nonagricultural populations sampled in two orchards, and thereafter its consequences on the pathogenicity of hybrids.Population genetic structure and demographic inferences based on the genotypes of 104 strains revealed a high amount of gene flow between the two populations in one orchard. In this site, mating between populations was made possible by the presence of a common host. Our results revealed an invasion of the virulent trait in the agricultural population; a main direction of introgression in hybrids from the agricultural to nonagricultural genetic backgrounds; and a population of hybrids with transgressive traits.We demonstrate a secondary contact with gene flow between divergent populations of pathogens. Our findings highlight evolutionary and epidemiological changes in pathogens and have concrete implications for sustainable disease management.

Quantitative plant resistance affects the aggressiveness of pathogens and is usually considered more durable than qualitative resistance. However, the efficiency of a quantitative resistance based on an isolate-specific Quantitative Trait Locus (QTL) is expected to decrease over time due to the selection of isolates with a high level of aggressiveness on resistant plants. To test this hypothesis, we surveyed scab incidence over an eight-year period in an orchard planted with susceptible and quantitatively resistant apple genotypes. We sampled 79 Venturia inaequalis isolates from this orchard at three dates and we tested their level of aggressiveness under controlled conditions. Isolates sampled on resistant genotypes triggered higher lesion density and exhibited a higher sporulation rate on apple carrying the resistance allele of the QTL T1 compared to isolates sampled on susceptible genotypes. Due to this ability to select aggressive isolates, we expected the QTL T1 to be non-durable. However, our results showed that the quantitative resistance based on the QTL T1 remained efficient in orchard over an eight-year period, with only a slow decrease in efficiency and no detectable increase of the aggressiveness of fungal isolates over time. We conclude that knowledge on the specificity of a QTL is not sufficient to evaluate its durability. Deciphering molecular mechanisms associated with resistance QTLs, genetic determinants of aggressiveness and putative trade-offs within pathogen populations is needed to help in understanding the erosion processes.

Understanding how pathogens emerge is essential to bring disease-causing agents under durable human control. Here, we used cross-pathogenicity tests to investigate the changes in life-history traits of the fungal pathogen Venturia inaequalis associated with host-tracking during the domestication of apple and subsequent host-range expansion on the wild European crabapple (Malus sylvestris). Pathogenicity of 40 isolates collected in wild and domesticated ecosystems was assessed on the domesticated apple, its Central Asian main progenitor (M. sieversii) and M. sylvestris. Isolates from wild habitats in the centre of origin of the crop were not pathogenic on the domesticated apple and less aggressive than other isolates on their host of origin. Isolates from the agro-ecosystem in Central Asia infected a higher proportion of plants with higher aggressiveness, on both the domesticated host and its progenitor. Isolates from the European crabapple were still able to cause disease on other species but were less aggressive and less frequently virulent on these hosts than their endemic populations. Our results suggest that the domestication of apple was associated with the acquisition of virulence in the pathogen following host-tracking. The spread of the disease in the agro-ecosystem would also have been accompanied by an increase in overall pathogenicity.

Expanding global trade and the domestication of ecosystems have greatly accelerated the rate of emerging infectious fungal diseases, and host-shift speciation appears to be a major route for disease emergence. There is therefore an increased interest in identifying the factors that drive the evolution of reproductive isolation between populations adapting to different hosts. Here, we used genetic markers and cross-inoculations to assess the level of gene flow and investigate barriers responsible for reproductive isolation between two sympatric populations of Venturia inaequalis, the fungal pathogen causing apple scab disease, one of the fungal populations causing a recent emerging disease on resistant varieties. Our results showed the maintenance over several years of strong and stable differentiation between the two populations in the same orchards, suggesting ongoing ecological divergence following a host shift. We identified strong selection against immigrants (i.e. host specificity) from different host varieties as the strongest and likely most efficient barrier to gene flow between local and emerging populations. Cross-variety disease transmission events were indeed rare in the field and cross-inoculation tests confirmed high host specificity. Because the fungus mates within its host after successful infection and because pathogenicity-related loci prevent infection of nonhost trees, adaptation to specific hosts may alone maintain both genetic differentiation between and adaptive allelic combinations within sympatric populations parasitizing different apple varieties, thus acting as a ‘magic trait’. Additional intrinsic and extrinsic postzygotic barriers might complete reproductive isolation and explain why the rare migrants and F1 hybrids detected do not lead to pervasive gene flow across years.

Evaluating the impact of plant domestication on the population structure of the associated pathogens provides an opportunity to increase our understanding of how and why diseases emerge. Here, we investigated the evolution of the population structure of the apple scab fungus Venturia inaequalis in response to the domestication of its host. Inferences were drawn from multilocus microsatellite data obtained from samples collected on (i) the Central Asian Malus sieversii, the main progenitor of apple, (ii) the European crabapple, Malus sylvestris, a secondary progenitor of apple, and (iii) the cultivated apple, Malus x domestica, in orchards from Europe and Central Asia. Using clustering methods, we identified three distinct populations: (i) a large European population on domesticated and wild apples, (ii) a large Central Asian population on domesticated and wild apples in urban and agricultural areas, and (iii) a more geographically restricted population in M. sieversii forests growing in the eastern mountains of Kazakhstan. Unique allele richness and divergence time estimates supported a host-tracking co-evolutionary scenario in which this latter population represents a relict of the ancestral populations from which current populations found in human-managed habitats were derived. Our analyses indicated that the domestication of apple induced a significant change in the genetic differentiation of populations of V. inaequalis in its centre of origin, but had little impact on its population dynamics and mating system. We discuss how the structure of the apple-based agrosystem may have restricted changes in the population structure of the fungus in response to the domestication of its host.

International audience; We formulate a spatially realistic population-genetic model for ascertaining the synergetic effect between genetic and spatial composition of the host population on the pathogen spread reinforced by evolutionary processes. We show that spatial arrangement of host genotypes is crucial to the efficacy of host genetic diversification. In particular, the reductive effect of multigenic resistance on the pathogen density can be produced by a random patterning of monogenic resistances. Random patterns can reduce both density and genetic diversity of the pathogen population and delay invasion promoted by sexual recombination. By contrast, patchy distributions diversify pathogen population and, hence, reduce the efficacy of resistance genes. The proposed approach provides theoretical support for studying fast emergence and spread of novel pathogen genotypes carrying multiple virulence genes. It has a practical applicability to design innovative strategies for the most appropriate deployment of plant resistance genes.

Publication Inra prise en compte dans l'analyse bibliométrique des publications scientifiques mondiales sur les Fruits, les Légumes et la Pomme de terre. Période 2000-2012. http://prodinra.inra.fr/record/256699; Plant resistance based on a gene-for-gene relationship tends not to be durable because virulent strains can emerge through mutation and colonize formerly resistant cultivars over large areas. Our objective was to determine the origin and colonization history of newly virulent strains of the fungus Venturia inaequalis. Four hundred and eighty-one strains from seven multi-cultivar orchards within a 300 kin wide area were collected from a cultivar carrying the overcome Vf-gene (Vfcv populations) and from non-Vf cultivars (nVfcv populations). Using 9 microsatellite loci, we showed that Vfcv populations were closely related though highly differentiated, indicating they have been recently founded following rare long distance dispersal events from a common origin. Assignment tests revealed that the Vfcv populations had not emerged from any of the nVfcv populations. Vfcv populations had a low diversity and were strongly differentiated from nVfcv populations several years after the first reported breakdown, suggesting reproductive isolation between the two populations

BGPI : équipe 5; International audience; Plant pathogens adapt readily to new crop varieties in agrosystems, and it is crucial to understand the factors underlying the epidemic spread of new virulent strains if we are to develop more efficient strategies to control them. In this study we used multilocus microsatellite typing, molecular epidemiology tools and a large collection of isolates from cultivated, wild and ornamental apples to investigate the origin of new virulent populations of Venturia inaequalis, an ascomycete fungus causing apple scab on varieties carrying the Rvi6 resistance gene. We demonstrated a common origin at the European scale of populations infecting apples (Malus × domestica) carrying the Rvi6 resistance and Malus floribunda, the progenitor of the Rvi6 resistance. Demographic modeling indicated that the Rvi6-virulent lineage separated several thousands of years ago from populations infecting non-Rvi6 hosts, without detectable gene flow between the two lineages. These findings show that 'breakdowns' of plant resistance genes can be caused by the selection and migration of virulent genotypes from standing genetic variation maintained in environmental disease reservoirs, here ornamental crabapples. This work stresses the need to take better account of pathogen diversity in resistance screenings of breeding lines and in resistance deployment strategies, in order to enhance sustainable disease management.

The recent breakdown of Vf, a major resistance gene to apple scab, provided an opportunity to analyze a population genetic process within the matching virulent subpopulation of the fungus Venturia inaequalis. We utilized the amplified fragment length polymorphism technique and allelic variation at four microsatellite loci to assess genetic structure of 133 isolates of V. inaequalis from a single commercial apple orchard sampled from one cultivar carrying the Vf gene (Judeline) and three cultivars devoid of the Vf gene. Both analyses indicated a strong decrease of the genetic diversity among isolates from the Vf cultivar compared with the high level of diversity among isolates from the three other cultivars. This leads to a high genetic differentiation between virVf and avrVf groups (FST > 0.17). Analyses of the genetic distance between AFLP patterns based on the Jaccard index indicate that all virVf isolates could be assigned to a single clonal lineage. These results lead us to conclude that the clonal structure of the population isolated from the Vf cultivar is an example of a founder effect in response to a resistance gene breakdown and it is likely that this event occurred in the orchard during the sampling year.

Theoretical approaches predict that host quantitative resistance selects for pathogens with a high level of pathogenicity, leading to erosion of the resistance. This process of erosion has, however, rarely been experimentally demonstrated. To investigate the erosion of apple quantitative resistance to scab disease, we surveyed scab incidence over time in a network of three orchards planted with susceptible and quantitatively resistant apple genotypes. We sampled Venturia inaequalis isolates from two of these orchards at the beginning of the experiment and we tested their quantitative components of pathogenicity (i.e., global disease severity, lesion density, lesion size, latent period) under controlled conditions. The disease severity produced by the isolates on the quantitatively resistant apple genotypes differed between the sites. Our study showed that quantitative resistance may be subject to erosion and even complete breakdown, depending on the site. We observed this evolution over time for apple genotypes that combine two broad-spectrum scab resistance QTLs, F11 and F17, showing a significant synergic effect of this combination in favour of resistance (i.e., favourable epistatic effect). We showed that isolates sampled in the orchard where the resistance was inefficient presented a similar level of pathogenicity on both apple genotypes with quantitative resistance and susceptible genotypes. As a consequence, our results revealed a case where the use of quantitative resistance may result in the emergence of a generalist pathogen population that has extended its pathogenicity range by performing similarly on susceptible and resistant genotypes. This emphasizes the need to develop quantitative resistances conducive to trade-offs within the pathogen populations concerned. (C) 2014 Elsevier B.V. All rights reserved.

Monogenic plant resistance breakdown is a model for testing evolution in action in pathogens. As a rule, plant pathologists argue that virulence - the allele that allows pathogens to overcome resistance - is due to a new mutation at the avirulence locus within the native/endemic population that infects susceptible crops. In this article, we develop an alternative and neglected scenario where a given virulence pre-exists in a non-agricultural host and might be accidentally released or introduced on the matching resistant cultivar in the field. The main difference between the two scenarios is the divergence time expected between the avirulent and the virulent populations. As a consequence, population genetic approaches such as genome scans and Approximate Bayesian Computation methods allow explicit testing of the two scenarios by timing the divergence. This review then explores the fundamental implications of this alternative scenario for plant breeding, including the invasion of virulence or the evolution of more aggressive hybrids, and proposes concrete solutions to achieve a sustainable resistance. (C) 2014 Elsevier B.V. All rights reserved.

Background Adaptation, which induces differentiation between populations in relation to environmental conditions, can initiate divergence. The balance between gene flow and selection determines the maintenance of such a structure in sympatry. Studying these two antagonistic forces in plant pathogens is made possible because of the high ability of pathogens to disperse and of the strong selective pressures exerted by their hosts. In this article, we analysed the genetic structure of the population of the apple scab fungus, Venturia inaequalis, in a heterogeneous environment composed of various Malus species. Inferences were drawn from microsatellite and AFLP data obtained from 114 strains sampled in a single orchard on nine different Malus species to determine the forces that shape the genetic structure of the pathogen. Results Using clustering methods, we first identified two specialist subpopulations: (i) a virulent subpopulation sampled on Malus trees carrying the Rvi6 resistance gene; and (ii) a subpopulation infecting only Malus trees that did not carry this resistance gene. A genome scan of loci on these two subpopulations did not detect any locus under selection. Additionally, we did not detect any other particular substructure linked to different hosts. However, an isolation-by-distance (IBD) pattern at the orchard scale revealed free gene flow within each subpopulation. Conclusions Our work shows a rare example of a very strong effect of a resistance gene on pathogen populations. Despite the high diversity of Malus hosts, the presence of Rvi6 seems sufficient to explain the observed genetic structure. Moreover, detection of an IBD pattern at the orchard scale revealed a very low average dispersal distance that is particularly significant for epidemiologists and landscape managers for the design of scab control strategies

International audience; Understanding how pathogens evolve according to pressures exerted by their plant hosts is essential for the derivation of strategies aimed at the durable management of resistant cultivars. The spectrum of action of the resistance factors in the partially resistant cultivars is thought to be an important determinant of resistance durability. However, it has not yet been demonstrated whether the pressures exerted by quantitative resistance are different according to their spectrum of action. * To investigate selection pressures exerted by apple genotypes harbouring various resistance quantitative trait loci (QTLs) on a mixed inoculum of the scab disease agent, Venturia inaequalis, we monitored V. inaequalis isolate proportions on diseased apple leaves of an F1 progeny using quantitative pyrosequencing technology and QTL mapping. * Broad-spectrum resistances did not exert any differential selection pressures on the mixed inoculum, whereas narrow-spectrum resistances decreased the frequencies of some isolates in the mixture relative to the susceptible host genotypes. * Our results suggest that the management of resistant cultivars should be different according to the spectrum of action of their resistance factors. The pyramiding of broad-spectrum factors or the use of a mixture of apple genotypes that carry narrow-spectrum resistance factors are two possible strategies for the minimization of resistance erosion.

Twenty-one new polymorphic microsatellite markers were isolated in the phytopathogenic fungus Venturia inaequalis , the causal agent of apple scab. An enrichment protocol was used to isolate microsatellite loci and the level of polymorphism was assessed on 44 European isolates. All loci were polymorphic with an average of 9.1 alleles per locus (range 2–24). Tests of cross-species amplifications suggest that at least some of these microsatellites could be used in different species, mainly Spilocaea pyracanthae and S. eriobotryae .

The apple is the most common and culturally important fruit crop of temperate areas. The elucidation of its origin and domestication history is therefore of great interest. The wild Central Asian species Malus sieversii has previously been identified as the main contributor to the genome of the cultivated apple (Malus domestica), on the basis of morphological, molecular, and historical evidence. The possible contribution of other wild species present along the Silk Route running from Asia to Western Europe remains a matter of debate, particularly with respect to the contribution of the European wild apple. We used microsatellite markers and an unprecedented large sampling of five Malus species throughout Eurasia (839 accessions from China to Spain) to show that multiple species have contributed to the genetic makeup of domesticated apples. The wild European crabapple M. sylvestris, in particular, was a major secondary contributor. Bidirectional gene flow between the domesticated apple and the European crabapple resulted in the current M. domestica being genetically more closely related to this species than to its Central Asian progenitor, M. sieversii. We found no evidence of a domestication bottleneck or clonal population structure in apples, despite the use of vegetative propagation by grafting. We show that the evolution of domesticated apples occurred over a long time period and involved more than one wild species. Our results support the view that self-incompatibility, a long lifespan, and cultural practices such as selection from open-pollinated seeds have facilitated introgression from wild relatives and the maintenance of genetic variation during domestication. This combination of processes may account for the diversification of several long-lived perennial crops, yielding domestication patterns different from those observed for annual species., Author Summary The apple, one of the most ubiquitous and culturally important temperate fruit crops, provides us with a unique opportunity to study the process of domestication in trees. The number and identity of the progenitors of the domesticated apple and the erosion of genetic diversity associated with the domestication process remain debated. The Central Asian wild apple has been identified as the main progenitor, but other closely related species along the Silk Route running from Asia to Western Europe may have contributed to the genome of the domesticated crop. Using rapidly evolving genetic markers to make inferences about the recent evolutionary history of the domesticated apple, we found that the European crabapple has made an unexpectedly large contribution to the genome of the domesticated apple. Bidirectional gene flow between the domesticated apple and the European crabapple resulted in the domesticated apple being currently more similar genetically to this secondary genepool than to the ancestral progenitor, the Central Asian wild apple. We found that domesticated apples have evolved over long time scales, with contributions from at least two wild species in different geographic areas, with no significant erosion of genetic diversity. This process of domestication and diversification may be common to other fruit trees and contrasts with the models documented for annual crops.

International audience

Expanding global trade and the domestication of ecosystems have greatly accelerated the rate of emerging infectious fungal diseases, and host-shift speciation appears to be a major route for disease emergence. There is therefore an increased interest in identifying the factors that drive the evolution of reproductive isolation between populations adapting to different hosts. Here, we used genetic markers and cross-inoculations to assess the level of gene flow and investigate barriers responsible for reproductive isolation between two sympatric populations of Venturia inaequalis, the fungal pathogen causing apple scab disease, one of the fungal populations causing a recent emerging disease on resistant varieties. Our results showed the maintenance over several years of strong and stable differentiation between the two populations in the same orchards, suggesting ongoing ecological divergence following a host shift. We identified strong selection against immigrants (i.e. host specificity) from different host varieties as the strongest and likely most efficient barrier to gene flow between local and emerging populations. Cross-variety disease transmission events were indeed rare in the field and cross-inoculation tests confirmed high host specificity. Because the fungus mates within its host after successful infection and because pathogenicity-related loci prevent infection of nonhost trees, adaptation to specific hosts may alone maintain both genetic differentiation between and adaptive allelic combinations within sympatric populations parasitizing different apple varieties, thus acting as a 'magic trait'. Additional intrinsic and extrinsic postzygotic barriers might complete reproductive isolation and explain why the rare migrants and F1 hybrids detected do not lead to pervasive gene flow across years.

Evaluating the impact of plant domestication on the population structure of the associated pathogens provides an opportunity to increase our understanding of how and why diseases emerge. Here, we investigated the evolution of the population structure of the apple scab fungus Venturia inaequalis in response to the domestication of its host. Inferences were drawn from multilocus microsatellite data obtained from samples collected on (i) the Central Asian Malus sieversii, the main progenitor of apple, (ii) the European crabapple, Malus sylvestris, a secondary progenitor of apple, and (iii) the cultivated apple, Malus x domestica, in orchards from Europe and Central Asia. Using clustering methods, we identified three distinct populations: (i) a large European population on domesticated and wild apples, (ii) a large Central Asian population on domesticated and wild apples in urban and agricultural areas, and (iii) a more geographically restricted population in M. sieversii forests growing in the eastern mountains of Kazakhstan. Unique allele richness and divergence time estimates supported a host-tracking co-evolutionary scenario in which this latter population represents a relict of the ancestral populations from which current populations found in human-managed habitats were derived. Our analyses indicated that the domestication of apple induced a significant change in the genetic differentiation of populations of V. inaequalis in its centre of origin, but had little impact on its population dynamics and mating system. We discuss how the structure of the apple-based agrosystem may have restricted changes in the population structure of the fungus in response to the domestication of its host.

A technique based on the polymerase chain reaction (PCR) was developed for the identification of Venturia nashicola using nucleotide sequence information of the ribosomal DNA region. The complete internal transcribed spacer (ITS) region of V. nashicola strains and phylo-genetically related species was amplified with the two universal ITS1 and ITS4 primers, sequenced, and digested with five restriction enzymes. The alignment of nucleotide sequences and analyses of digestion patterns indicated constant polymorphisms between V. nashicola and related species at nucleotides 126 and 127, which overlapped a TaqI restriction site. An oligonucleotide primer named A126 was designed for identifying this variable region. A primer set (A126 and ITS4) that allowed the amplification of a 391-bp DNA fragment within the ITS region by PCR was specific to V. nashicola when it was checked against fungal genomic DNAs of related fungi. This primer set was a good candidate for a species-specific reagent in a procedure for identification of V. nashicola by PCR.

Genetic relationships, mating crosses, and host specificity of Venturia inaequalis isolates from Malus spp. and of Spilocaea pyracanthae isolates from Pyracantha spp. were evaluated. Sequence analysis of the complete internal transcribed spacer (ITS) region (ITS1-5.8S to ITS2) revealed a total similarity between these two putative species. ITS restriction fragment length polymorphism carried out with five restriction enzymes on a collection of 28 isolates confirmed a lack of diversity in this region between and within these two populations. Additional isolates from three related species (V. pirina, V. nashicola, and S. eriobotryae) were divided into two distinct monophyletic groups in a phylogenetic tree using ITS sequence comparison. These groups were related to their anamorph (i.e., Spilocaea or Fusicladium). When inoculated on their host of origin, fields isolates caused typical symptoms of scab disease, and a host specificity was demonstrated by cross pathogenicity of isolates from Malus × domestica and Pyracantha spp. Mating on dried leaves in vitro between one isolate of each putative species led to production of numerous perithecia. Ninety-six sporulating monoascosporic progenies were isolated from this cross. Based on these genetic and pathogenic data, we proposed that pathogens responsible for scab on Malus spp. and Pyracantha spp. are considered as two formae speciales belonging to V. inaequalis.

International audience

International audience; Les trois phases d’un contournement et les forces évolutives impliquéesDynamique spatiale et temporelle des contournementsLa résistance quantitative pour ralentir les contournements ?Quelle stratégie pour préserver la durabilité des résistances ?Conclusion

Phenotypic and genetic changes occurring during the process of domestication are well described in plants. However, the impact of domestication in life history traits of their pathogens has been poorly studied. In particular, what are the consequences on the life history traits of pathogens that switch from the wild habitat characterized by a high host genetic and spatial heterogeneity to a much more dense and genetically homogeneous agroecosystems? Have pathogen’s genomes particular signatures of demographic changes or of selection related to the domestication of their host? Here we focused on the fungus Venturia inaequalis, causal agent of apple scab, that shares in Central Asia the same center of origin of its wild endemic host Malus sieversii. In the first part of this work we retrace evolutionary history of a population from the wild that was responsible for the rapid breakdown of the Vf resistance gene in apple orchards. We then highlight the threat of wild habitat to scab resistance apple cultivars and thus the necessity to take into account the wild in the management of resistance genes. In the second part, a comparative study based on phenotypic and genomic data was carried out between two populations sampled in Kazakhstan on M. sieversii, either in anthropized areas or in non anthropized area, the latter being identified as a relic of the ancestral population of V. inaequalis. These two populations were genetically differentiated and partially reproductively isolated. The use of two methods of inference (ABC and dadi) based on the alignment of 10 994 genes sampled in the two populations revealed a complex history of a secondary contact event. The fungus would have first followed its host worldwide by "Host-tracking" during the domestication process and then very recently it would re-exchange genes with its ancestral population in Kazakhstan. Secondary contact context is particularly favorable to detect genetic incompatibilities, this particular situation could then allow identification of genes involved in adaptation to habitat (cultivated apple versus M. sieversii) and in post-zygotic reproductive isolation. Analysis of 36 sequenced genomes has identified more than 602 genes with an index of differentiation (Fst) greater than 0.7, what represents numerous candidates genes of potential barriers to homogeneous gene flow between these two populations. Comparative phenotypic analysis between these two populations such spores size and capacity to sporulate showed that apple domestication would have also modified life history traits of V. inaequalis related to its dispersion. This study has allowed identification of candidate loci potentially involved in changes in life history traits, and genetic barriers in pathogen related to the domestication of apple.; Les changements phénotypiques et génétiques consécutifs au processus de domestication et plus largement du passage d'un environnement sauvage aux agro-écosystèmes sont bien décrits chez les espèces végétales. Cependant, l’impact de ces changements de traits d'histoire de vie sur les populations de pathogènes infectant les hôtes domestiqués a jusqu’alors été très peu abordé. En particulier,quelles sont les conséquences sur les traits d’histoire de vie des pathogènes du passage d’un milieu sauvage caractérisé par une grande hétérogénéité d’hôtes à des agro-écosystèmes bien plus denses et homogènes génétiquement? Est-ce que le génome des pathogènes porte des signatures de changements démographiques ou de sélection en relation avec la domestication de leurs hôtes ? Ces questions traitées dans le cadre de la présente thèse ont porté sur le champignon Venturiainaequalis, agent pathogène de la tavelure du pommier, qui partage en Asie Centrale le même centre d’origine que son hôte endémique sauvage Malus sieversii. Les travaux reposent principalement sur 1)l'étude du passage de souches sauvages virulentes sur pommier résistant VF expliquant le contournement rapide de cette résistance et 2)l'étude de génomique des populations et de variations phénotypiques comparant une population de V. inaequalis échantillonnée sur M.sieversii au Kazakhstan, préalablement identifiée comme étant une relique de la population ancestrale, et d'une population Kazakhe de milieu anthropisé. Ces deux populations sont génétiquement différenciées et partiellement isolées reproductivement. L’utilisation des deux méthodes d’inférence l'une l'ABC et l'autre dadi basées sur l’alignement de 10 994 gènes échantillonnés au sein des deux populations révèlent une histoire complexe de contact secondaire, où le champignon aurait dans un premier temps « suivi » la plante hôte à travers le monde au cours de la domestication (modèle de l'hosttracking) puis échangerait à nouveau depuis peu des gènes avec la population ancestrale au Kazakhstan. Cette situation originale de remise en contact secondaire favorisant la mise en évidence d’incompatibilités génétiques, permet à la fois d’envisager de détecter des gènes impliqués dans l’adaptation à l’habitat (pommier cultivé versus Malus sieversii) et dans l’isolement reproductif post-zygotique. L’analyse des 36 génomes séquencés a ainsi permis d’identifier plus de 602 gènes présentant un indice de différentiation (Fst) supérieur à 0,7 autant de gènes verrous faisant potentiellement obstacle aux flux de gènes homogènes entre ces deux populations. Enfin, l’analyse phénotypique de ces deux populations montre que la domestication du pommier a profondément modifié les traits d’histoire de vie de V. inaequalis liés à sa dispersion. L’ensemble de ces analyses a donc permis d’identifier des locus candidats potentiellement impliqués dans des modifications des traits d’histoire de vie, et dans des barrières géniques, en lien avec la domestication du pommier.

Species distributed across a large continental range encounter different socio-economic and ecological situations, which directly impact the distribution of threats and determine the actions for their conservation. At this scale, an integrative approach to the study of populations’ dynamics may be necessary to retrace all rocesses that affect the conservation status of the species. The Corncrake, a grassland bird distributed across the Palearctic during its reproduction season, exhibits a variable conservation status depending on the intensity of agriculture. In western Europe, agricultural intensification lead to a high decrease of populations while numbers remained stable in the central and eastern parts of its range. This work aims at improving knowledge for the conservation of this species using an integrative approach involving species distribution modelling, population genetics, parasitology and theecological analysis of habitat selection.Results confirmed the distribution of the species, which was previously estimated through expert knowledge. We also showed that the variations of agricultural intensity in Europe seem to contribute to a homogenisation of genetic structure as well as a longitudinal gradient of avian malaria prevalence. Finally, we demonstrated that the Corncrake is an umbrella species of poor value for the protection of grassland passerines in Pays-de-Loire. These results should provide new insights to the understanding of Corncrake populations and therefore favour the conservation strategy of the species., Les espèces distribuées sur une large aire de répartition continentale rencontrent des situations socioéconomiqueset des conditions écologiques très variables qui impactent directement la répartition des menaces et déterminent les actions consacrées à leur conservation. A cette échelle, une approche intégrée de l’étude du fonctionnement des populations peut s’avérer nécessaire pour reconstituer l’ensemble des processus affectant la situation de conservation de l’espèce. Le Râle des genêts, oiseau prairial distribué au sein du Paléarctique en période de reproduction, présente ainsi un statut de conservation très variable lié à l’intensité de l’agriculture. En Europe de l’ouest, l’intensification agricole a conduit à un déclin considérable des populations tandis que l’espèce a été épargnée dans les parties centrales et orientales de sa distribution. Cette thèse vise à apporter des éléments de connaissancespour la conservation de l’espèce via une approche intégrée mettant en oeuvre des approches de modélisation de distribution, de génétique des populations, de parasitologie et d’analyses écologiques de sélection d’habitat.Les résultats obtenus ont permis de confirmer la distribution de l’espèce jusqu’alors estimée à dire d’experts. On a également montré que les variations d’intensité agricole au sein de l’Europe semblent favoriser une homogénéisation de la structure génétique ainsi qu’un gradient longitudinal de prévalence en malaria aviaire. Enfin, on a démontré que le Râle des genêts constituait une espèce parapluie d’intérêt modéré pour les passereaux prairiaux en Pays-de-Loire.Ces résultats permettent d’améliorer la compréhension du fonctionnement des populations à différentes échelles, et peuvent ainsi contribuer à la conservation du Râle des genêts.