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Plant pathogens are constantly evolving and adapting to their environment, including their host. Virulence alleles emerge, and then increase, and sometimes decrease in frequency within pathogen populations in response to the fluctuating selection pressures imposed by the deployment of resistance genes. In some cases, these strong selection pressures cannot fully explain the evolution observed in pathogen populations. A previous study on the French population ofPuccinia triticina, the causal agent of wheat leaf rust, showed that two major pathotypes — groups of isolates with a particular combination of virulences — predominated but then declined over the 2005-2016 period. The relative dynamics and the domination of these two pathotypes — 166 317 0 and 106 314 0 —, relative to the other pathotypes present in the population at a low frequency although compatible, i.e. virulent on several varieties deployed, could not be explained solely by the frequency ofLrgenes in the landscape. Within these two pathotypes, we identified two main genotypes that emerged in succession. We assessed three components of aggressiveness — infection efficiency, latency period and sporulation capacity — for 44 isolates representative of the fourP. triticinapathotype-genotype combinations. We showed, for both pathotypes, that the more recent genotypes were more aggressive than the older ones. Our findings were highly consistent for the various components of aggressiveness for pathotype 166 317 0 grown on Michigan Amber — a ‘naive’ cultivar never grown in the landscape — or on Apache — a ‘neutral’ cultivar, which does not affect the pathotype frequency in the landscape and therefore was postulated to have no or minor selection effect on the population composition. For pathotype 106 314 0, the most recent genotype had a shorter latency period on several of the cultivars most frequently grown in the landscape, but not on ‘neutral’ and ‘naive’ cultivars. We conclude that the quantitative components of aggressiveness can be significant drivers of evolution in pathogen populations. A gain in aggressiveness stopped the decline in frequency of a pathotype, and subsequently allowed an increase in frequency of this pathotype in the pathogen population, providing evidence that adaptation to a changing varietal landscape not only affects virulence but can also lead to changes in aggressiveness.

Capacity for dispersal is a fundamental fitness component of plant pathogens. Characterization of plant pathogen dispersal is important for understanding how pathogen populations change in time and space. We devised a systematic approach to measure and analyze rain splash-driven dispersal of plant pathogens in field conditions, using the major fungal wheat pathogen Zymoseptoria tritici as a case study. We inoculated field plots of wheat ( Triticum aestivum) with two distinct Z. tritici strains. Next, we measured disease intensity as counts of fruiting bodies (pycnidia) using automated image analysis. These measurements characterized primary disease gradients, which we used to estimate effective dispersal of the pathogen population. Genotyping of reisolated pathogen strains with strain-specific PCR confirmed the conclusions drawn from phenotypic data. Consistently with controlled environment studies, we found that the characteristic scale of dispersal is tens of centimeters. We analyzed the data using a spatially explicit mathematical model that incorporates the spatial extent of the source, rather than assuming a point source, which allows for a more accurate estimation of dispersal kernels. We employed bootstrapping methods for statistical testing and adopted a two-dimensional hypotheses test based on kernel density estimation, enabling more robust inference compared with standard methods. We report the first estimates of dispersal kernels of the pathogen in field conditions. However, repeating the experiment in other environments would lead to more robust conclusions. We put forward advanced methodology that paves the way for further measurements of plant pathogen dispersal in field conditions, which can inform spatially targeted plant disease management. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .

International audience; Cultivar mixtures slow polycyclic epidemics but may also affect the evolution of pathogen populations by diversifying the selection pressures exerted by their plant hosts at field scale. We compared the dynamics of natural populations of the fungal pathogen Zymoseptoria tritici in pure stands and in three binary mixtures of wheat cultivars (one susceptible cultivar and one cultivar carrying the recently broken-down Stb16q gene) over two annual field epidemics. We combined analyses of population "size" based on disease severity, and of population "composition" based on changes in the frequency of virulence against Stb16q in seedling assays with more than 3000 strains. Disease reductions were observed in mixtures late in the epidemic, at the whole-canopy scale and on both cultivars, suggesting the existence of a reciprocal protective effect. The three cultivar proportions in the mixtures (0.25, 0.5, and 0.75) modulated the decrease in (a) the size of the pathogen population relative to the two pure stands, (b) the size of the virulent subpopulation, and (c) the frequency of virulence relative to the pure stand of the cultivar carrying Stb16q. Our findings suggest that optimal proportions may differ slightly between the three indicators considered. We argue potential trade-offs that should be taken into account when deploying a resistance gene in cultivar mixtures: between the dual objectives "efficacy" and "durability," and between the "size" and "frequency" of the virulent subpopulation. Based on current knowledge, it remains unclear whether virulent subpopulation size or frequency has the largest influence on interepidemic virulence transmission.

The fungal pathogen Zymoseptoria tritici is the causal agent of Septoria tritici blotch (STB), a major wheat disease in Western Europe. Microorganisms inhabiting wheat leaves might act as beneficial, biocontrol, or facilitating agents that could limit or stimulate the development of Z. tritici. Improving our understanding of microbial communities in the wheat phyllosphere would lead to new insights into STB management. This article provides fungal and bacterial metabarcoding datasets obtained by sampling wheat leaves with and without symptoms caused by Z. tritici. Tissues were sampled from three commercial wheat varieties on three sampling dates during a cropping season. Weeds around wheat fields were sampled as well. In total, more than 450 leaf samples were collected. The pathogen Z. tritici was quantified using quantitative PCR. We provide the raw metabarcoding datasets, the amplicon sequence variant tables obtained after bioinformatic processing, the metadata associated with each sample (sampling date, wheat variety and tissue health condition), a preliminary descriptive analysis of the data, and the code used for bioinformatic and descriptive statistical analysis. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Human activity impacts the evolutionary trajectories of many species worldwide. Global trade of agricultural goods contributes to the dispersal of pathogens reshaping their genetic makeup and providing opportunities for virulence gains. Understanding how pathogens surmount control strategies and cope with new climates is crucial to predicting the future impact of crop pathogens. Here, we address this by assembling a global thousand-genome panel of Zymoseptoria tritici, a major fungal pathogen of wheat reported in all production areas worldwide. We identify the global invasion routes and ongoing genetic exchange of the pathogen among wheat-growing regions. We find that the global expansion was accompanied by increased activity of transposable elements and weakened genomic defenses. Finally, we find significant standing variation for adaptation to new climates encountered during the global spread. Our work shows how large population genomic panels enable deep insights into the evolutionary trajectory of a major crop pathogen.

The fungal pathogen Zymoseptoria tritici is the causal agent of Septoria tritici blotch (STB), a major wheat disease in Western Europe. Microorganisms inhabiting wheat leaves might act as beneficial, biocontrol or facilitating agents that could limit or stimulate the development of Z. tritici. Improving our understanding of microbial communities in the wheat phyllosphere would lead to new insights into STB management. This resource announcement provides fungal and bacterial metabarcoding datasets obtained by sampling wheat leaves differing in the presence of symptoms caused by Z. tritici. Tissues were sampled from three wheat commercial varieties on three sampling dates during a cropping season. Weeds around wheat fields were sampled as well. In total, more than 450 leaf samples were collected. The pathogen Z. tritici was quantified using qPCR. We provide the raw metabarcoding datasets, the Amplicon Sequence Variant (ASV) tables obtained after bioinformatic processing, the metadata associated to each sample (sampling date, wheat variety and tissue health condition), a preliminary descriptive analysis of the data, and the code used for bioinformatic and descriptive statistical analysis.

This study follows on from a previous study showing that binary mixtures of wheat cultivars affect the evolution of Zymoseptoria tritici populations within a field epidemic from the beginning (t1) to the end (t2) of a growing season. Here, we focused on the impact of interseason sexual reproduction on this evolution. We studied mixtures of susceptible and resistant cultivars (carrying Stb16q, a recently broken-down resistance gene) in proportions of 0.25, 0.5 and 0.75, and their pure stands. We determined the virulence status of 1440 ascospore-derived strains collected from residues of each cultivar by phenotyping on seedlings. Virulence frequencies were lower in mixtures than in pure stands of the resistant cultivar, as at t2, revealing that the impact of mixtures persisted until the beginning of the next epidemic (t3). The frequency of virulence was lower in the offspring population on the susceptible cultivar and, more surprisingly, the frequency of avirulence on the resistant cultivar increased after sexual reproduction. Our findings highlight two epidemiological processes in mixtures: selection within the local pathogen population between t1 and t2 driven by asexual cross-contamination between cultivars (previous study) and sexual crosses between avirulent and virulent strains between t2 and t3 driven by changes in the probabilities of physical encounters (this study). Mixtures therefore appear to be a promising strategy for the deployment of qualitative resistances, not only to limit the intensity of Septoria tritici blotch epidemics, but also to reduce the erosion of resistances by managing evolution of the pathogen population at a pluriannual scale.

International audience; Given the negative environmental effects of conventional agricultural techniques, the need for biodiversity-friendly agriculture systems that rely more on ecosystem services and less on chemical inputs is becoming increasingly urgent. In this paper, we focus on crop protection strategies that are alternatives to the use of pesticides. Diversification of the plant component of agricultural areas at different space and time scales has been presented as a powerful socio-economic and agro-ecological mechanism for the sustainable control of pests. Our interdisciplinary group of scientific experts examined the literature on the ecological effects of plant diversification on pests and their natural enemies, as well as the social science literature on the conditions for farmers to adopt the corresponding practices, to assess the potential offered by plant diversification.We developed a conceptual framework that connects the agro-ecological and socio-economic components of an agricultural landscape in a dynamic loop accounting for interactions among elements at different spatial and temporal scales and their feedback effects. This article presents this framework and illustrates its application to the case of wheat production and protection. By explicitly connecting each level of agro-ecological organization with the potential socio-economic drivers and limitations underpinning the adoption and implementation of plant diversification in landscapes, this framework makes it possible to ana yse the synergies and antagonisms between different modes of diversification and the conditions of their deployment. Exploring this framework is a prerequisite to the identification of opportunities and key feed-back loops for designing diversification strategies that unlock the agro-ecological potential of f uture production systems. We conclude that there is a need for interdisciplinary research in experimental landscapes involving farmers and other local stakeholders to design sustainable future agricultural landscapes that deliver high levels of biological control services..

Plant pathogen populations inhabit patchy environments with contrasting, variable thermal conditions. We investigated the diversity of thermal responses in populations sampled over contrasting spatiotemporal scales, to improve our understanding of their dynamics of adaptation to local conditions. Samples of natural populations of the wheat pathogen Zymoseptoria tritici were collected from sites within the Euro‐Mediterranean region subject to a broad range of climatic conditions. We tested for local adaptation, by accounting for the diversity of responses at the individual and population levels on the basis of key thermal performance curve parameters and “thermotype” (groups of individuals with similar thermal responses) composition. The characterization of phenotypic responses and genotypic structure revealed the following: (i) a high degree of individual plasticity and variation in sensitivity to temperature conditions across spatiotemporal scales and populations; and (ii) geographic variation in thermal response among populations, with major alterations due to seasonal patterns over the wheat‐growing season. The seasonal shifts in functional composition suggest that populations are locally structured by selection, contributing to adaptation patterns. Further studies combining selection experiments and modeling are required to determine how functional group selection drives population dynamics and adaptive potential in response to thermal heterogeneity., We investigated the diversity of thermal responses in fungal plant populations sampled within the Euro‐Mediterranean region. We highlighted (i) a high degree of individual plasticity and variation in sensitivity to temperature conditions across spatiotemporal scales and populations; and (ii) geographic variation in thermal response among populations, with major alterations due to seasonal patterns. The seasonal shifts in functional composition suggest that populations are locally structured by selection and thus reveal two‐tier adaptive dynamics.

Little is known about the impact of host immunity on sexual reproduction in fungal pathogens. In particular, it is unclear whether crossing requires both sexual partners to infect living plant tissues. We addressed this issue in a three-year experiment investigating different scenarios of Zymoseptoria tritici crosses on wheat according to the virulence (‘vir’) or avirulence (‘avr’) of the parents against a qualitative resistance gene. Co-inoculations (‘vir × vir’, ‘avr × vir’, ‘avr × avr’) and single inoculations were performed on a cultivar carrying the resistance gene (Cellule) and a susceptible cultivar (Apache), in the greenhouse. We assessed the intensity of asexual multiplication by scoring disease severity, and the intensity of sexual reproduction by counting the ascospores discharged from wheat residues. As expected, disease severity was more intense on Cellule for ‘vir × vir’ co-inoculations than for ‘avr × vir’ co-inoculations, with no disease for ‘avr × avr’. However, all types of co-inoculation yielded sexual offspring, whether or not the parental strains caused plant symptoms. Parenthood was confirmed by genotyping (SSR markers), and the occurrence of crosses between (co-)inoculated and exogenous strains (other strains from the experiment, or from far away) was determined. We found that symptomatic asexual infection was not required for a strain to participate in sexual reproduction, and that avirulent strains could be maintained asymptomatically “on” or “in” leaf tissues of plants carrying the corresponding resistant gene for long enough to reproduce sexually. In two of the three years, the intensity of sexual reproduction did not differ significantly between the three types of co-inoculation in Cellule, suggesting that crosses involving avirulent strains are not anecdotal. We discuss the possible mechanisms explaining the maintenance of avirulence in Z. tritici populations and supporting the potential efficacy of cultivar mixtures for limiting resistance gene breakdown.HighlightsAvirulent Zymoseptoria tritici strains can reproduce sexually in wheat plants carrying the corresponding resistant gene.Symptomatic infection of plant tissues is not essential for a strain to reproduce sexually.Avirulent strains can be maintained asymptomatically “on” or “in” leaf tissues of plants carrying the corresponding resistant gene for long enough to reproduce sexually.Crosses of virulent strains with virulent and avirulent strains in a plant host carrying the corresponding resistance gene can produce offspring with similar population sizes.Several possible scenarios for sexual crosses can explain the maintenance of avirulence in Zymoseptoria tritici populations evolving in a wheat canopy, particular in cultivar mixtures.

International audience; This study combined culture-dependent (strain isolation plus molecular identification) and culture-independent (metabarcoding) approaches to characterize the diversity of microbiota on wheat and oilseed rape residues. The goal was to develop a methodology to culture microorganisms with the aim of being able to establish synthetic crop residue microbial communities for further study, i.e., testing potential interactions within these communities and characterizing groups of beneficial taxa that could be used as biological control agents against plant pathogens. We generated community-based culture collections. We adapted the isolation strategy to the potential differences in the spatial and temporal distribution of diversity between bacteria and fungi. We performed (i) a high-throughput isolation from few samples with no a priori for bacteria and (ii) a low-throughput isolation from several samples with a priori—i.e., morphotype selection—for fungi. Although isolation using a single medium did not allow us to characterize the microbiome as precisely as metabarcoding, the bacterial diversity (158 ASVs, 36 genera) was relatively higher than the fungal diversity (131 ASVs, 17 genera) known to be limited by competition for growth on non-selective solid media. Isolation and metabarcoding provided consistent and complementary information: they revealed several common but also specific ASVs, leading to close microbial community profiles of the most abundant fungal and bacterial taxa in residues. Finally, by empirically comparing the different profiles, we assessed the cultivability of the most abundant fungal and bacterial taxa obtained in metabarcoding.

The experience presented here relates to 2020, a particularly timely year for plant disease‐related communication (International Year of Plant Health, IYPH2020), but also a unique year because of the COVID‐19 pandemic. Our goal was to illustrate the diversity and beauty of fungal plant pathogens through a naturalist approach that could be followed by any amateur. We achieved this end through “phytopathological strolls”, in which we observed and determined the origin of symptoms on diseased plants found in our garden, in the local streets, and in nearby open spaces, and shared this matter with a broad public. The lockdown imposed in France created an additional motivation to take up the challenge, and to involve our children, even under strong constraints such as movement restrictions. We observed and described fungal pathogens through hundreds of photographs, shared our findings with a large audience on Twitter, and received feedback. The material used was deliberately simple and transportable: a digital reflex camera, an old microscope, a mobile phone, some books, and an internet connection. Between 17 March 2020 and 20 June 2021 we found 196 plant pathogens, including 97 rusts, 27 powdery mildews, and 28 septoria‐like diseases. We discuss here the importance of promoting searches for plant pathogens, their description and conservation, through a combination of classical approaches and digital tools in tune with the times, such as Twitter, by treating pathogen identification like a detective game and, more surprisingly, by making use of the addictive nature of collection approaches, drawing a parallel with Pokémon GO., In the dual context of COVID‐19 lockdown and of the International Year of Plant Health, we used "phytopathological strolls" and communication on the microblogging platform Twitter to promote public education about plant pathogens.

This study is the first to compare culture-dependent (strain isolation plus molecular identification) and culture-independent (whole-community metabarcode sequencing) approaches for characterizing the microbiota of crop residues. We investigated the diversity of fungal and bacterial communities in wheat and oilseed rape residues, using two different culture-dependent strategies to cover the maximum diversity for each kingdom: broad substrate sampling coupled with low-throughput isolation and diversity analysis for fungi, and reduced substrate sampling coupled with high-throughput isolation and diversity analysis for bacteria. The proportion of cultivable microorganisms was neither as low as the ‘1%’ paradigm long associated with the rhizosphere microflora, nor as high as the 50% sometimes reported for the phyllosphere microflora. It was, thus, intermediate between the values for soil and plants. This finding is consistent with residues being considered to constitute an ecotone, at the interface between soil and phyllosphere. Isolation and metabarcoding provided consistent complementary information: they revealed close community profiles, leading to the identification of several common and specific amplicon sequence variants (ASVs). The power of the culture-independent approach was thus confirmed. By contrast the culture-dependent approach was less weak than anticipated. Firstly, it provided complementary information about microbial diversity, with several ASVs not retrieved by metabarcoding being identified after isolation in the community-based culture collection. Secondly, this approach made it possible to preserve and test different taxa either individually or after the creation of synthetic communities, for deciphering the ecological functions of communities beyond merely descriptive aspects.

Contrat : 2018-SA-0248; Commanditaire : Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (France); Type de commande : Commande avec contrat/convention/lettre de saisine; Date de signature : 2020-09-28 Les auteurs mentionnés font partie du groupe de travail (cf. p.15 du rapport) Comité d'experts spécialisé (cf. p. 15-16 du rapport) / Participation ANSES (cf. p. 4 du rapport) / Audition de personnalités extérieures (cf. p.16 du rapport); Un nouveau règlement européen relatif à la lutte contre Xylella fastidiosa (règlement d’exécution UE 2020/1201 de la Commission du 14 août 2020, relatif aux mesures visant à prévenir l’introduction et la dissémination dans l’Union de Xylella fastidiosa) propose aux États membres de l'Union européenne une approche de gestion des foyers de la bactérie plus adaptée à la pluralité des situations phytosanitaires rencontrées sur le terrain que le précédent règlement (décision d’exécution UE 2015/789 de la Commission du 18 mai 2015 modifiée, relative à des mesures visant à éviter l’introduction et la propagation dans l’Union de Xylella fastidiosa). En particulier, ce nouveau règlement assouplit les conditions dérogatoires à l'arrachage des végétaux spécifiés dans les zones infectées, de même que les conditions de replantation de végétaux spécifiés dans les zones infectées ou encore la mise en circulation des végétaux spécifiés en dehors des zones délimitées. Il revient à chaque État membre de décider d'arracher ou de maintenir certaines espèces végétales ou certains individus qui présenteraient un intérêt patrimonial particulier.Il est demandé de « construire un arbre de décision afin d’éclairer et d'apporter un appui technique et scientifique aux services de l'État qui mettent en œuvre les mesures de gestion de foyer en France métropolitaine (y compris en Corse). En particulier, il […] est demandé de lister les différents critères de risque à prendre en considération afin que les services de l’État puissent arbitrer du maintien ou non des végétaux spécifiés dans les zones infectées. Ces critères peuvent être d'ordre biologique, économique ou pédoclimatique. L'évaluation devra porter, par ordre de priorité, sur les espèces suivantes : olivier, agrumes du genre Citrus, laurier-rose, arbres fruitiers d'ornement du genre Prunus (amandier, cerisier), autres espèces spécifiées des sous-espèces multiplex et pauca » de X. fastidiosa.

Durum wheat (Triticum turgidumL. subsp.durum) landraces represent a prominent genetic resource for Mediterranean farming systems and breeding programs. Fourteen landraces sampled in Tunisia were genotyped with 9 microsatellite markers and characterized with 15 morphological descriptors, including resistance to the fungal diseaseSeptoria triticiblotch (STB). The genetic diversity, nearly was as important within landraces populations (45%) than between populations (54%). It was structured in seven genetic groups and was only partly explained by the variety name or the locality of origin. Populations were also greatly diversified phenotypically (Shannon-Weaver H’=0.54) with traits related to spike and awn colours being the most diversified. Resistance to STB was either qualitative in two populations or with varying degrees of quantitative resistance in the others. A Pst-Fstcomparison indicate a local adaptation of the populations. Overall, the genetic structure of Tunisian durum wheat landraces revealed a complex selection trajectory and seed exchanges between farmers.

Mycosphaerella linicola is the causal agent of pasmo, a disease affecting flax (Linum usitatissimum) worldwide. We performed a three-year field survey in France to characterize the dynamics of sexual reproduction in this fungal pathogen on flax straw during the interepidemic period. Cohorts of M. linicola fruiting bodies were sampled from linseed straw during the autumn and winter and carefully observed, focusing on pseudothecia, asci and ascospores. A sequence of experimental steps corresponding to Koch s postulates confirmed, for the first time in France and continental Europe, the widespread presence of the sexual stage of M. linicola in plant host tissues, and its potential role in the disease cycle. The developmental dynamics of pseudothecia on straw, expressed as the change over time in the percentage of mature pseudothecia, was similar in all three years. Pseudothecia appeared in late summer, with peak maturity reached in October. A temporal shift, thought to be due to the climatic conditions in early autumn (rainfall), was highlighted in one of the three years. These observations suggest that sexual reproduction plays a significant role in the epidemiology of pasmo in France. A resurgence of M. linicola infections in spring flax is thought to have occurred in recent years, due to the increase in the area under flax. The presence of the sexual stage of this pathogen probably increased the quantitative impact of residues of winter linseed (used for oil) and flax straw (left on the soil for retting and used for fibers) as an interepidemic "brown bridge". This case study highlights how certain parts of a disease cycle, in this case the sexual phase, can become crucial due to changes in production conditions. Finally, we highlight the importance of further studies of the specific detrimental effects of the sexual stage of this pathogen on flax fiber quality.

Oilseed rape residues are a crucial determinant of stem canker epidemiology as they support the sexual reproduction of the fungal pathogen Leptosphaeria maculans. The aim of this study was to characterize the impact of a resistance gene against L. maculans infection on residue microbial communities and to identify microorganisms interacting with this pathogen during residue degradation. We used near‐isogenic lines to obtain healthy and infected host plants. The microbiome associated with the two types of plant residues was characterized by metabarcoding. A combination of linear discriminant analysis and ecological network analysis was used to compare the microbial communities and to identify microorganisms interacting with L. maculans. Fungal community structure differed between the two lines at harvest, but not subsequently, suggesting that the presence/absence of the resistance gene influences the microbiome at the base of the stem whilst the plant is alive, but that this does not necessarily lead to differential colonization of the residues by fungi. Direct interactions with other members of the community involved many fungal and bacterial amplicon sequence variants (ASVs). L. maculans appeared to play a minor role in networks, whereas one ASV affiliated to Plenodomus biglobosus (synonym Leptosphaeria biglobosa) from the Leptosphaeria species complex may be considered a keystone taxon in the networks at harvest. This approach could be used to identify and promote microorganisms with beneficial effects against residue‐borne pathogens and, more broadly, to decipher the complex interactions between multispecies pathosystems and other microbial components in crop residues., Characterizing the impact of a resistance gene against Leptosphaeria maculans on the microbiome of oilseed rape residues using near‐isogenic lines helps to identify microorganisms interacting with this pathogen.

Deciphering the responses of microbial populations to spatiotemporal changes in their thermal environment is instrumental in improving our understanding of their eco-evolutionary dynamics. Recent studies have shown that current phenotyping protocols do not adequately address all dimensions of phenotype expression. Therefore, these methods can give biased assessments of sensitivity to temperature, leading to misunderstandings concerning the ecological processes underlying thermal plasticity. We describe here a new robust and versatile experimental framework for the accurate investigation of thermal performance and phenotypic diversity in yeasts and yeast-like microorganisms, at the individual and population levels. In addition to proof-of-concept, the application of this framework to the fungal wheat pathogen Zymoseptoria tritici resulted in detailed characterisations for this yeast-like microorganism of (i) the patterns of temperature-dependent changes in performance for four fitness traits; (ii) the consistency in thermal sensitivity rankings of strains between in planta and in vitro growth assessments; (iii) significant interindividual variation in thermal responses, with four principal thermotypes detected in a sample of 66 strains; and (iv) the ecological consequences of this diversity for population-level processes through pairwise competition experiments highlighting temperature-dependent outcomes. These findings extend our knowledge and ability to quantify and categorise the phenotypic heterogeneity of thermal responses. As such, they lay the foundations for further studies elucidating local adaptation patterns and the effects of temperature variations on eco-evolutionary and epidemiological processes.

SummaryPlant pathogen populations inhabit patchy environments with contrasting, variable thermal conditions. We investigated the diversity of thermal responses in populations sampled over contrasting spatiotemporal scales, to improve our understanding of their dynamics of adaptation to local conditions.Samples of natural populations of the wheat pathogenZymoseptoria triticiwere collected from sites within the Euro-Mediterranean region subject to a broad range of environmental conditions. We tested for local adaptation, by accounting for the diversity of responses at the individual and population levels on the basis of key thermal performance curve parameters and ‘thermotype’ (groups of individuals with similar thermal responses) composition.The characterisation of phenotypic responses and genotypic structure revealed: (i) a high degree of individual plasticity and variation in sensitivity to temperature conditions across spatiotemporal scales and populations; (ii) geographic adaptation to local mean temperature conditions, with major alterations due to seasonal patterns over the wheat-growing season.The seasonal shifts in functional composition suggest that populations are locally structured by selection, contributing to shape adaptation patterns. Further studies combining selection experiments and modelling are required to determine how functional group selection drives population dynamics and adaptive potential in response to thermal heterogeneity.

Foliar plant pathogens require liquid or vapour water for at least part of their development, but their response and their adaptive tolerance to moisture conditions have been much less studied than other meteorological factors to date. We examined the impact on the wheat-Zymoseptoria tritici interaction of altering optimal moisture conditions conducive to infection. We assessed the responses in planta of 48 Z. tritici strains collected in two climatologically distinct locations (Ireland and Israel) to four high moisture regimes differing in the timing and the duration of uninterrupted exposure to saturated relative humidity (100% RH) during the first three days of infection. Individual- and population-level moisture reaction norms expressing how the sporulating area of a lesion change with the RH conditions were established based on visual assessments of lesion development at 14, 17 and 20 days post-inoculation (dpi). Our findings highlighted: (i) a critical time-dependent effect on lesion development of uninterrupted periods of exposure to 100% RH during these earliest infection stages; (ii) a marked interindividual variation in the sensitivity to RH conditions both in terms of strain average moisture response and plasticity; (iii) a higher tolerance – expressed at 14 dpi, not later – of the Israeli population to early interruption of optimal moisture conditions. By indicating that sensitivity to sub-optimal moisture conditions may vary greatly between Z. tritici individuals and populations, this study highlights the evidence of moisture adaptation signature in a plant pathogen. This suggests that understanding such variation will be critical to predict their response to changing climatic conditions.

Dispersal is a key ecological process. An individual dispersal event has a source and a destination, both are well localized in space and can be seen as points. A probability to move from a source point to a destination point can be described by a specific probability function, the dispersal kernel. However, when we measure dispersal, the source of dispersing individuals is usually an area, which distorts the shape of the observed dispersal gradient compared to the underlying dispersal kernel. Here, we show with simulations, how different source geometries affect the gradient shape depending on the type of the kernel. We present an explicit mathematical approach for estimating the dispersal kernel from a dispersal gradient data independently of the source dimension. Further, we demonstrate the value of the approach by analysing three experimental dispersal datasets with a conventional method and the proposed method, to show how the estimated dispersal kernels differ between the methods. We use three pre-existing datasets from field experiments measuring dispersal of important plant pathogens. Our results demonstrate how analysis of dispersal data can be improved to achieve more rigorous measures of dispersal. The proposed approach leads to a general measure of dispersal in contrast to results from the conventional method that depend on the design of the dispersal source. This enables a direct comparison between outcomes of different experiments and allows acquiring more knowledge from a large number of previous empirical studies of dispersal.

BackgroundWheat residues are a crucial determinant of the epidemiology of Septoria tritici blotch, as they support the sexual reproduction of the causal agent Zymoseptoria tritici. We aimed to characterize the effect of infection with this fungal pathogen on the microbial communities present on wheat residues, and to identify microorganisms interacting with it. We used metabarcoding to characterize the microbiome associated with wheat residues placed outdoors, with and without preliminary Z. tritici inoculation, comparing a first set of residues in contact with the soil and a second set without contact with the soil, on four sampling dates in two consecutive years.ResultsThe diversity of the tested conditions, leading to the establishment of different microbial communities according to the origins of the constitutive taxa (plant only, or plant and soil), highlighted the effect of Z. tritici on the wheat residue microbiome. Several microorganisms were affected by Z. tritici infection, even after the disappearance of the pathogen. Linear discriminant analyses and ecological network analyses were combined to describe the communities affected by infection. The number of fungi and bacteria promoted or inhibited by inoculation with Z. tritici decreased over time, and was smaller for residues in contact with the soil. The interactions between the pathogen and other microorganisms appeared to be mostly indirect, despite the strong position of the pathogen as a keystone taxon in networks. Direct interactions with other members of the communities mostly involved fungi, including other wheat pathogens. Our results provide essential information about the alterations to the microbial community in wheat residues induced by the mere presence of a fungal pathogen, and vice versa. Species already described as beneficial or biocontrol agents were found to be affected by pathogen inoculation.ConclusionsThe strategy developed here can be viewed as a proof-of-concept focusing on crop residues as a particularly rich ecological compartment, with a high diversity of fungal and bacterial taxa originating from both the plant and soil compartments, and for Z. tritici-wheat as a model pathosystem. By revealing putative antagonistic interactions, this study paves the way for improving the biological control of residue-borne diseases.

First Online: 01 February 2019; International audience; Subpopulations of the wheat pathogen Zymoseptoria tritici (26 sample groups composed of 794 isolates) were collected in two nearby wheat fields in the Paris basin, during both epidemic and inter-epidemic periods of three successive years (2009-2013). In addition to the type of inoculum (ascospores vs. pycnidiospores), the alternative presence of wheat debris allowed taking into account its putative origin (local vs. distant). We used a molecular epidemiology approach, based on population genetic indices derived from SSR marker analysis, to describe putative changes in the structure and genotypic diversity of these subpopulations over 3years, at a spatiotemporal scale consistent with epidemiological observations. Genetic structure was broadly stable over time (within and between years) and between fields, however with weak population differentiation over time. All subpopulations displayed very high diversity and the occurrence of regular sexual reproduction was confirmed in the two fields. A significant increase of the MAT1-1/MAT1-2 ratio was observed over the course of the epidemics. This original finding suggests a competitive advantage of MAT1-1 strains consistently with their greater pathogenicity reported in the literature and may reveal undescribed adaptation. Finally, we found that the period, the type of inoculum and its putative origin had little effect on the short term evolution of the local population of Z. tritici. Fungal population size and diversity are apparently large enough to prevent genetic drift at this fine spatiotemporal scale, and more likely short distance migration contributes strongly to the stabilization of genetic diversity among and within plots.

International audience; This study provides empirical evidence for antagonistic density dependence mechanisms driving sexual reproduction in the wheat fungal pathogen Zymoseptoria tritici. Biparental crosses with 12 increasing inoculum concentrations, in controlled conditions, showed that sexual reproduction in Z. tritici was impacted by an Allee effect due to mate limitation and a competition with asexual multiplication for resource allocation. The highest number of ascospores discharged was reached at intermediate inoculum concentrations (from 5x10(4) conidiamL(-1) to 10(6) conidiamL(-1)). Consistent with these results for controlled co-inoculation, we found that the intensity of sexual reproduction varied with both cropping period and the vertical position of the host tissues in the field, with a maximum between 25 and 35cm above the ground. An optimal lesion density (disease severity of 30 to 45%) maximizing offspring (ascospores) number was established, and its eco-evolutionary consequences are considered here. Two ecological mechanisms may be involved: competition for resources between the two modes of reproduction (decrease in the host resources available for sexual reproduction due to their prior use in asexual multiplication), and competitive disequilibrium between the two parental isolates, due to differential interaction dynamics with the host, for example, leading to an imbalance between mating types. A conceptual model based on these results suggests that sexual reproduction plays a key role in the evolution of pathogenicity traits, including virulence and aggressiveness. Ecological knowledge about the determinants of sexual reproduction in Z. tritici may, therefore, open up new perspectives for the management of other fungal foliar pathogens with dual modes of reproduction.

The negative contribution of crop residues as a source of inoculum for plant diseases is well established. However, microbial ecologists have long reported positive effects of residues on the stability of agrosystems and conservation tillage practices have become increasingly widespread. Most studies have suggested that large microbial communities should be taken into account in plant disease management, but we know little about their ecological interaction with pathogens in the crop residue compartment. This review focuses on microbiomes associated with residues within the context of other microbial habitats in cereal-producing agroecosystems such as phyllosphere or rhizosphere. We connected residue microbiome with the survival of residue-borne fungal plant pathogens, thus combining knowledge in microbial ecology and epidemiology, two disciplines still not sufficiently connected. We provide an overview of the impact of residues on cereal disease epidemics and how dynamic interactions between microbial communities of nonburied residues during their degradation, along with soil and multitude of abiotic factors, can contribute to innovative disease management strategies, including next-generation microbiome-based biocontrol strategies. Starting from the classical but still relevant view of crop residues as a source of pathogen inoculum, we first consider possibilities for limiting the amount of residues on the soil surface to reduce the pathogen pressure. We then describe residues as a transient half-plant/half-soil compartment constituting a key fully fledged microbial ecosystem: in other words, an ecotone which deserves special attention. We focus on microbial communities, the changes in these communities over time and the factors influencing them. Finally, we discuss how the interactions between the microbial communities and the pathogens present on residues could be used: identification of keystone taxa and beneficial assemblages, then preservation of these taxa by adapted agronomic practices or development of synthetic communities, rather than the introduction of a single exogenous biocontrol species designed as a treatment product. [Formula: see text] Copyright © 2019 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

International audience; The wheat pathogen Zymoseptoria tritici is a relevant fungal model organism for investigations of the epidemiological determinants of sexual reproduction. The objective of this experimental study was to determine which intrinsic factors, including parental fitness and timing conditions of infection, affect the numbers of ascospores produced. We first performed 28 crosses on adult wheat plants in semi controlled conditions, with 10 isolates characterized for their fitness traits. We validated the efficiency of the crossing method, opening up new perspectives for epidemiological studies. We found that the ability to reproduce sexually was determined, at least partly, by the parental genotypes. We also found that the number of ascospores released was correlated with the mean size of the sporulating lesions of the parental isolates on the one hand, and the absolute difference in the latent periods of these isolates on the other. No functional trade-off between the two modes of reproduction in Z. tritici was revealed: there was no adaptive compromise between pathogenicity (asexual multiplication on leaves) and transmission (intensity of sexual reproduction on wheat debris). Moreover, a few days' difference in the latent periods of the two parental isolates, such that one progressed more rapidly in the host tissue than the other, seemed to be slightly beneficial to ascosporogenesis. This may be because the first parental isolate breaks down host defenses, thereby facilitating infection for the other parental isolate. However, a larger difference (a few weeks), generated by leaving two to three weeks between the inoculations of the plant with the parental isolates, was clearly detrimental to ascosporogenesis. In this case, the host tissues were likely colonized by the first isolate, leaving less host resources available for the second, consistent with a competition effect during the asexual stage. (C) 2016 Elsevier Inc. All rights reserved.

Crop residues are a crucial ecological niche with a major biological impact on agricultural ecosystems. In this study we used a combined diachronic and synchronic field experiment based on wheat-oilseed rape rotations to test the hypothesis that plant is a structuring factor of microbial communities in crop residues, and that this effect decreases over time with their likely progressive degradation and colonization by other microorganisms. We characterized an entire fungal and bacterial community associated with 150 wheat and oilseed rape residue samples at a plurennial scale by metabarcoding. The impact of plant species on the residue microbiota decreased over time and our data revealed turnover, with the replacement of oligotrophs, often plant-specific genera (such as pathogens) by copiotrophs, belonging to more generalist genera. Within a single cropping season, the plant-specific genera and species were gradually replaced by taxa that are likely to originate from the soil. These changes occurred more rapidly for bacteria than for fungi, known to degrade complex compounds. Overall, our findings suggest that crop residues constitute a key fully-fledged microbial ecosystem. Taking into account this ecosystem, that has been neglected for too long, is essential, not only to improve the quantitative management of residues, the presence of which can be detrimental to crop health, but also to identify groups of beneficial micro-organisms. Our findings are of particular importance, because the wheat-oilseed rape rotation, in which no-till practices are frequent, is particularly widespread in the European arable cropping systems.

The latent period is a crucial life history trait, particularly for polycyclic plant diseases, because it determines how many complete monocycles could theoretically occur during an epidemic. Many empirical studies have focused on the variation of the latent period with pathogen or host genotype, or changes in response to environmental factors. The focus on these aspects is unsurprising, as these factors classically form the three parts of the epidemiological triangle. Experiments in controlled conditions are generally used to assess pathogenicity and host susceptibility, and also provide the opportunity to measure the distribution of latent periods in epidemiological systems. Once estimated for one or several pairs of host-pathogen genotypes, the mean value of this important trait is usually considered to be fixed and is often used "as is" in epidemiological models. We show here that the latent period can display non-negligible variability over the course of a disease epidemic, and that this variability has multiple sources, some of which have complex, antagonistic impacts. We develop arguments for four sources of variation that challenge the implicit assumption that the latent period remains constant: daily fluctuations in leaf temperature, nature of inoculum, host stage or age of host tissues, intra-population competition and selection for aggressiveness traits. We focus on the wheat fungal pathogen Septoria tritici blotch (Zymoseptoria tritici), making use of empirical datasets collected during the first author9s own research projects and a targeted literature review. Such empirical epidemiological knowledge is new and potentially important for modelers. While some studies have demonstrated that the distribution of latent periods around the mean value has consequences for epidemiological dynamics, we show that it might also be important for epidemiological modelers to account for changes in this mean value during an annual epidemic. These results may be of critical importance for improving outbreak forecasting.

This study provides empirical evidence for antagonistic density-dependence mechanisms driving sexual reproduction in the wheat fungal pathogen Zymoseptoria tritici. Biparental crosses with 12 increasing inoculum concentrations, in controlled conditions, showed that sexual reproduction in Z. tritici was impacted by an Allee effect due to mate limitation and a competition with asexual multiplication for resource allocation. We found that asexual multiplication was itself affected by competition for host resources between the two parental isolates, as illustrated by the decrease in asexual fruiting body density and sporulation intensity observed with increasing lesion density. Consistent with these results for controlled co-inoculation, we found that the intensity of sexual reproduction varied with both cropping period and the vertical position of the host tissues in the field. An optimal lesion density maximizing offspring (ascospores) number was established, and its eco-evolutionary consequences are considered here. Two ecological mechanisms may be involved: competition for resources between the two modes of reproduction (decrease in the host resources available for sexual reproduction due to their prior use in asexual multiplication), and competitive disequilibrium between the two parental isolates, due to differential interaction dynamics with the host, for example, leading to an imbalance between mating types. Conceptual models based on these results suggest that sexual reproduction plays a key role in the evolution of pathogenicity traits, including virulence and aggressiveness. Ecological knowledge about the determinants of sexual reproduction in Z. tritici may, therefore, open up new perspectives for the management of other fungal foliar pathogens with dual modes of reproduction.

This preprint has been reviewed and recommended by Peer Community In Evolutionary Biology (http://dx.doi.org/10.24072/pci.evolbiol.100039). The efficiency of plant resistance to fungal pathogen populations is expected to decrease over time, due to its evolution with an increase in the frequency of virulent or highly aggressive strains. This dynamics may differ depending on the scale investigated (annual or pluriannual), particularly for annual crop pathogens with both sexual and asexual reproduction cycles. We assessed this time-scale effect, by comparing aggressiveness changes in a localZymoseptoria triticipopulation over an eight-month cropping season and a six-year period of wheat monoculture. We collected two pairs of subpopulations to represent the annual and pluriannual scales: from leaf lesions at the beginning and end of a single annual epidemic, and from crop debris at the beginning and end of a six-year period. We assessed two aggressiveness traits – latent period and lesion size – on sympatric and allopatric host varieties. A trend toward decreased latent period concomitant with a significant loss of variability was established during the course of the annual epidemic, but not over the six-year period. Furthermore, a significant cultivar effect (sympatric vs. allopatric) on the average aggressiveness of the isolates revealed host adaptation, arguing that the observed patterns could result from selection. We thus provide an experimental body of evidence of an epidemiological trade-off between the intra- and inter-annual scales in the evolution of aggressiveness in a local plant pathogen population. More aggressive isolates were collected from upper leaves, on which disease severity is usually lower than on the lower part of the plants left in the field as crop debris after harvest. We suggest that these isolates play little role in sexual reproduction, due to an Allee effect (difficulty finding mates at low pathogen densities), particularly as the upper parts of the plant are removed from the field, explaining the lack of transmission of increases in aggressiveness between epidemics.

In a cross-infection experiment, we investigated how seasonal changes can affect adaptation patterns in a Zymoseptoria tritici population. The fitness of isolates sampled on wheat leaves at the beginning and at the end of a field epidemic was assessed under environmental conditions (temperature and host stage) to which the local pathogen population was successively exposed. Isolates of the final population were more aggressive, and showed greater sporulation intensity under winter conditions and a shorter latency period (earlier sporulation) under spring conditions, than isolates of the initial population. These differences, complemented by lower between-genotype variability in the final population, exhibited an adaptation pattern with three striking features: (i) the pathogen responded synchronously to temperature and host stage conditions; (ii) the adaptation concerned two key fitness traits; (iii) adaptation to one trait (greater sporulation intensity) was expressed under winter conditions while, subsequently, adaptation to the other trait (shorter latency period) was expressed under spring conditions. This can be interpreted as the result of short-term selection, driven by abiotic and biotic factors. This case study cannot yet be generalized but suggests that seasonality may play an important role in shaping the variability of fitness traits. These results further raise the question of possible counterselection during the interepidemic period. While we did not find any trade-off between clonal multiplication on leaves during the epidemic period and clonal spore production on debris, we suggest that final populations could be counterselected by an Allee effect, mitigating the potential impact of seasonal selection on long-term dynamics.

Subpopulations of the wheat pathogen Zymoseptoria tritici (26 sample groups composed of 794 strains) were collected in two nearby wheat fields in the Paris basin, during both epidemic and inter-epidemic periods of three successive years (2009-2013). In addition to the type of inoculum (ascospores vs. pycnidiospores), the alternative presence of wheat debris allowed taking into account its putative origin (local vs. distant). We used a molecular epidemiology approach, based on population genetic indices derived from SSR marker analysis, to describe putative changes in the structure and genotypic diversity of these subpopulations over three years, at a spatiotemporal scale consistent with epidemiological observations. Genetic structure was stable over time (within and between years) and between fields. All subpopulations displayed very high levels of gene and genotypic diversity. The low levels of linkage disequilibrium and the very low clonal fraction at all stages were consistent with the regular occurrence of sexual reproduction in the two fields. A significant increase of the MAT1-1/MAT1-2 ratio was observed over the course of the epidemics, suggesting a competitive advantage of MAT1-1 strains consistently with their greater pathogenicity reported in the literature. Finally, we found that the period, the type of inoculum and its putative origin had little effect on the short term evolution of the local population of Z. tritici. Fungal population size and diversity are apparently large enough to prevent genetic drift at this fine spatiotemporal scale, and more likely short distance migration contributes strongly to the stabilization of genetic diversity among and within plots.

The latent period is a crucial life history trait, particularly for polycyclic plant diseases, because it determines how many complete monocycles could theoretically occur during an epidemic. Many empirical studies have focused on the variation of the latent period with pathogen or host genotype, or changes in response to environmental factors. The focus on these aspects is unsurprising, as these factors classically form the three parts of the epidemiological triangle. Experiments in controlled conditions are generally used to assess pathogenicity and host susceptibility, and also provide the opportunity to measure the distribution of latent periods in epidemiological systems. Once estimated for one or several pairs of host-pathogen genotypes, the mean value of this important trait is usually considered to be fixed and is often used “as is” in epidemiological models. We show here that the latent period can display non-negligible variability over the course of a disease epidemic, and that this variability has multiple sources, some of which have complex, antagonistic impacts. We develop arguments for four sources of variation that challenge the implicit assumption that the latent period remains constant: daily fluctuations in leaf temperature, nature of inoculum, host stage or age of host tissues, intra-population competition and selection for aggressiveness traits. We focus on the wheat fungal disease Septoria tritici blotch (Zymoseptoria tritici), making use of empirical datasets collected during the first author’s own research projects and a targeted literature review. Such empirical epidemiological knowledge is new and potentially important for modelers. While some studies have demonstrated that the distribution of latent periods around the mean value has consequences for epidemiological dynamics, we show that it might also be important for epidemiological modelers to account for changes in this mean value during an annual epidemic. These results may be of critical importance for improving outbreak forecasting.

BGPI : équipe 5; The structure of pathogen populations is an important driver of epidemics affecting crops and natural plant communities. Comparing the composition of two pathogen populations consisting of assemblages of genotypes or phenotypes is a crucial, recurrent question encountered in many studies in plant disease epidemiology. Determining if there is a significant difference between two sets of proportions is also a generic question for numerous biological fields. When samples are small and data are sparse, it is not straightforward to provide an accurate answer to this simple question because routine statistical tests may not be exactly calibrated. To tackle this issue, we built a computationally-intensive testing procedure, namely the Generalized Monte Carlo Plug-In test with Calibration (GMCPIC test), which is implemented in an R package available at http://dx.doi.org/10.5281/zenodo.53996. A simulation study was carried out to assess the performance of the proposed methodology and to make a comparison with standard statistical tests. This study allows us to give advice on how to apply the proposed method, depending on the sample sizes. The proposed methodology was then applied to real datasets and the results of the analyses were discussed from an epidemiological perspective. The applications to real data sets deal with three topics in plant pathology: the reproduction of Magnaporthe oryzae, the spatial structure of Pseudomonas syringae, and the temporal recurrence of Puccinia triticina.

This chapter provides an analysis specific for Europe of the risk of plant pathogens being used as anti-crop bioweapons, taking into account both the biological and human dimensions of the threat. An historical review of anti-crop bioweapons lays down the starting point of the characterization and contextualizes the threat in Europe. Four types of threat are developed and provide a structure for the analysis: (1) from military state programs to allegations of attacks; (2) from ‘rogue state’ hidden programs to claimed terror attacks; (3) biocrime, sabotage, private allegations and conspiracy theories on social media; (4) from the overzealous application of phytosanitary measures to the deliberate introduction of a regulated pest to justify trade protectionism. A database consisting of 21 important target crops and of 63 potentially dangerous pests (selected from a list of 570 pests) are combined with the development and categorization of ‘scenarios’. This is proposed as a starting point of a prospective approach to quantify the risk of agroterrorism in Europe. Four challenges (‘Convergence Tactics’, ‘Constraints’, ‘Climate’, and ‘Conspiracy’) are suggested to be the most important determinants of the forthcoming evolution of the threat. The prospect for Europe to successfully confront the increasing risk and challenges for the next decade is discussed.