A High-Quality Grapevine Downy Mildew Genome Assembly Reveals Rapidly Evolving and Lineage-Specific Putative Host Adaptation Genes

Downy mildews are obligate biotrophic oomycete pathogens that cause devastating plant diseases on economically important crops. Plasmopara viticola is the causal agent of grapevine downy mildew, a major disease in vineyards worldwide. We sequenced the genome of Pl. viticola with PacBio long reads and obtained a new 92.94 Mb assembly with high contiguity (359 scaffolds for a N50 of 706.5 kb) due to a better resolution of repeat regions. This assembly presented a high level of gene completeness, recovering 1,592 genes encoding secreted proteins involved in plant–pathogen interactions. Plasmopara viticola had a two-speed genome architecture, with secreted protein-encoding genes preferentially located in gene-sparse, repeat-rich regions and evolving rapidly, as indicated by pairwise dN/dS values. We also used short reads to assemble the genome of Plasmopara muralis, a closely related species infecting grape ivy (Parthenocissus tricuspidata). The lineage-specific proteins identified by comparative genomics analysis included a large proportion of RxLR cytoplasmic effectors and, more generally, genes with high dN/dS values. We identified 270 candidate genes under positive selection, including several genes encoding transporters and components of the RNA machinery potentially involved in host specialization. Finally, the Pl. viticola genome assembly generated here will allow the development of robust population genomics approaches for investigating the mechanisms involved in adaptation to biotic and abiotic selective pressures in this species

Evolution of cycle length and branching architecture during pearl millet (pennisetum glaucum) domestication

Le mil est une céréale cultivée dans la zone soudano-sahélienne en Afrique. A partir d'un jeu de données comprenant 18 marqueurs microsatellites et 8 séquences nucléotidiques pour 45 populations domestiquées et sauvages, j'ai mis à jour une forte différenciation génétique entre deux pools de mils sauvages. Avec une approche de type Approximate Bayesian Computation, j'ai testé différents scénarios de domestication, et montré que le scénario le plus probable est celui d'une domestication unique à partir des mils sauvages de l'est du Sahel.Deux grands groupes de mil domestiqué existent: des variétés précoces et des variétés tardives. Mes résultats montrent une absence de différenciation génétique neutre entre ces deux groupes, suggérant l'existence de flux de gènes récurrents. J'ai également étudié un gène candidat de la famille Hd3a-like, PgHd3a. Aucune trace de sélection n'a été mise en évidence pour ce gène, et il est vraisemblable qu'il ne participe pas à la différence de phénotype entre mils précoces et tardifs. Son rôle lors de la floraison chez le mil est mis en doute par des résultats d'expression. J'ai également isolé un paralogue de PgHd3a, PgHd3a-2.Finalement, j'ai étudié un cline longitudinal de fréquence à l'échelle continentale pour la présence d'un Miniature Inverted-repeat Transposable Element (MITE) situé dans le gène teosinte-branched1, impliqué dans la détermination du nombre de branches basales chez le maïs. L'étude du taux de différenciation pour le MITE et pour 13 microsatellites neutres et du polymorphisme nucléotidique dans la région entourant le MITE montre que le cline est probablement dû à des processus neutres.Pearl millet is a cereal cultivated in the sudano-sahelian area in Africa. Using a data set consisting of 18 microsatellites and 8 nucleotide sequences for 45 domesticated and wild populations, I showed the existence of a high differentiation between two wild genetic pools. Using the model-based Approximate Bayesian Computation (ABC) approach, I tested different domestication scenarios and showed that a single domestication in eastern Sahel is more likely than other models.There are two main types of domesticated pearl millet: early-flowering and late-flowering varieties. My results show a lack of neutral genetic differentiation between these two types, suggesting the existence of recurrent gene flow. I also studied a candidate gene, PgHd3a, belonging to the Hd3a-like family and which could be implicated in the floral transition. No selection footprint has been detected, and it is likely this gene is not involved in the phenotypic difference between early and late-flowering varieties. Expression results casted some doubt about its role during floral transition in pearl millet. I also isolated a new paralog of PgHd3a, PgHd3a-2.Finally, I studied a longitudinal frequency cline at the continental scale for the presence of a Miniature Inverted-repeat Transposable Element (MITE) located in the teosinte branched1 gene, involved in the regulation of the number of tillers in maize. By comparing differentiation levels between the MITE and 13 neutral microsatellites, and by investigating the nucleotide polymorphism in the genomic region surrounding the MITE, I showed the cline was probably caused by neutral processes

Data from: Inference of domestication history and differentiation between early- and late-flowering varieties in pearl millet

Pearl millet (Pennisetum glaucum) is a staple crop in Sahelian Africa. Farmers usually grow varieties with different cycle lengths and complementary functions in Sahelian agrosystems. Both the level of genetic differentiation of these varieties and the domestication history of pearl millet have been poorly studied. We investigated the neutral genetic diversity and population genetic structure of early- and late-flowering domesticated and wild pearl millet populations using 18 microsatellite loci and 8 nucleotide sequences. Strikingly, early- and late-flowering domesticated varieties were not differentiated over their whole distribution area, despite a clear difference in their isolation-by-distance pattern. Conversely, our data brought evidence for two well-differentiated genetic pools in wild pearl millet, allowing us to test scenarios with different numbers and origins of domestication using approximate Bayesian computation (ABC). The ABC analysis showed the likely existence of asymmetric migration between wild and domesticated populations. The model choice procedure indicated that a single domestication from the eastern wild populations was the more likely scenario to explain the polymorphism patterns observed in cultivated pearl millet

Polymorphism data

Microsatellite and nucleotide sequence data sets, used in the polymorphism study and the ABC analysis. ReadMe files describing the data are included in the directories

Scripts and reference tables of the ABC analysis

Contains the scripts needed to perform the simulations with ABCtoolbox and fastsimcoal for the ABC analysis, and the reference tables for each simulated model. ReadMe files are included in the directories

Europe as a bridgehead in the worldwide invasion history of grapevine downy mildew, Plasmopara viticola

International audienceEurope is the historical cradle of viticulture, but grapevines (Vitis vinifera) have been increasingly threatened by pathogens of American origin. The invasive oomycete Plasmopara viticola causes downy mildew, one of the most devastating grapevine diseases worldwide. Despite major economic consequences, its invasion history remains poorly understood. We analyzed a comprehensive dataset of ∼2,000 samples, collected from the most important wine-producing countries, using nuclear and mitochondrial gene sequences and microsatellite markers. Population genetic analyses revealed very low genetic diversity in invasive downy mildew populations worldwide and little evidence of admixture. All the invasive populations originated from only one of the five native North American lineages, the one parasitizing wild summer grape (V. aestivalis). An approximate Bayesian computation-random forest approach allowed inferring the worldwide invasion scenario of P. viticola. After an initial introduction into Europe, invasive European populations served as a secondary source of introduction into vineyards worldwide, including China, South Africa, and twice independently, Australia. Only the invasion of Argentina probably represents a tertiary introduction, from Australia. Our findings provide a striking example of a global pathogen invasion resulting from secondary dispersal of a successful invasive population. Our study will also help designing quarantine regulations and efficient breeding for resistance against grapevine downy mildew

Quantitative traits of pathogenicity of Plasmopara viticola

Quantitative traits of pathogenicity of Plasmopara viticola when inoculated on two different hosts (susceptible: Vitis vinifera cv. Cabernet sauvignon, CS and partially resistant: Cabernet carbon, CC). 108 isolates from the Bordeaux wine region were used and 8 (on CS) and 6 (on CC) leaf discs were inoculated for each isolate (i.e. repetition). The final sample analysed consisted of 1357 leaf discs corresponding to 214 plant-pathogen interactions. Details for each isolate (geographic locations and genetic data) are described in the « PV_genetic_data » file of this package. Cells left blank indicate missing data. The « Latency_Ndays » column indicates the latency period (number of days between inoculation and the first recorded sporangia). The « Sporangium_prod_per_mm2 » column indicates the number of sporangia per mm2 of leaf disc counted at seven days post inoculation (cumulative, over seven days of infection). The « Sporangium_size_µm » column indicates the weighted average of sporangium size. The « nbzoo_per_sporangium » column indicates the number of zoospores contained by each sporangium (on two leaf discs per isolate). The « T50 » column indicates the time at which the pathogen reached 50% of its maximal sporulation at 7 dpi. The « slope_at_T50 » column indicates the slope of the curve (sporulation vs. time) at T50. The « phenotype_CAA » column indicates whether the isolate is resistant or sensitive to CAA fungicides (see the « PV_genetic_data » file of this package). The « phenotype_QOI » column indicates whether the isolate is resistant or sensitive to QoI fungicides (see the « PV_genetic_data » file of this package). The « Fungicide category » column indicates whether the isolate is resistant and/or sensitive to CAA and QoI fungicides

Data from: Soft selective sweeps in fungicide resistance evolution: recurrent mutations without fitness costs in grapevine downy mildew

Adaptation produces hard or soft selective sweeps depending on the supply of adaptive genetic polymorphism. The evolution of pesticide resistance in parasites is a striking example of rapid adaptation that can shed light on selection processes. Plasmopara viticola, which causes grapevine downy mildew, forms large populations, in which resistance has rapidly evolved due to excessive fungicide use. We investigated the pathways by which fungicide resistance has evolved in this plant pathogen, to determine whether hard or soft selective sweeps were involved. An analysis of nucleotide polymorphism in 108 field isolates from the Bordeaux region revealed recurrent mutations of cytb and CesA3 conferring resistance to quinone outside inhibiting (QoI) and carboxylic acid amide (CAA) fungicides, respectively. Higher levels of genetic differentiation were observed for nucleotide positions involved in resistance than for neutral microsatellites, consistent with local adaptation of the pathogen to fungicide treatments. No hitchhiking was found between selected sites and neighbouring polymorphisms in cytb and CesA3, confirming multiple origins of resistance alleles. We assessed resistance costs, by evaluating the fitness of the 108 isolates through measurements of multiple quantitative pathogenicity traits under controlled conditions. No significant differences were found between sensitive and resistant isolates, suggesting that fitness costs may be absent or negligible. Our results indicate that the rapid evolution of fungicide resistance in P. viticola has involved a soft sweep

Unexpected pattern of pearl millet genetic diversity among ethno-linguistic groups in the Lake Chad Basin

International audienceDespite of a growing interest in considering the role of sociological factors in seed exchanges and their consequences on the evolutionary dynamics of agro-biodiversity, very few studies assessed the link between ethno-linguistic diversity and genetic diversity patterns in small-holder farming systems. This is key for optimal improvement and conservation of crop genetic resources. Here, we investigated genetic diversity at 17 SSR markers of pearl millet landraces (varieties named by farmers) in the Lake Chad Basin. 69 pearl millet populations, representing 27 landraces collected in eight ethno-linguistic farmer groups, were analyzed. We found that the farmers' local taxonomy was not a good proxy for population's genetic differentiation as previously shown at smaller scales. Our results show the existence of a genetic structure of pearl millet mainly associated with ethno-linguistic diversity in the western side of the lake Chad. It suggests there is a limit to gene flow between landraces grown by different ethno-linguistic groups. This result was rather unexpected, because of the highly outcrossing mating system of pearl millet, the high density of pearl millet fields all along the green belt of this Sahelian area and the fact that seed exchanges among ethno-linguistic groups are known to occur. In the eastern side of the Lake, the pattern of genetic diversity suggests a larger efficient circulation of pearl millet genes between ethno-linguistic groups that are less numerous, spatially intermixed and, for some of them, more prone to exogamy. Finally, other historical and environmental factors which may contribute to the observed diversity patterns are discussed