Spatially explicit ecological modeling improves empirical characterization of plant pathogen dispersal.

Dispersal is a key ecological process, but it remains difficult to measure. By recording numbers of dispersed individuals at different distances from the source, one acquires a dispersal gradient. Dispersal gradients contain information on dispersal, but they are influenced by the spatial extent of the source. How can we separate the two contributions to extract knowledge about dispersal? One could use a small, point-like source for which a dispersal gradient represents a dispersal kernel, which quantifies the probability of an individual dispersal event from a source to a destination. However, the validity of this approximation cannot be established before conducting measurements. This represents a key challenge hindering progress in characterization of dispersal. To overcome it, we formulated a theory that incorporates the spatial extent of sources to estimate dispersal kernels from dispersal gradients. Using this theory, we re-analyzed published dispersal gradients for three major plant pathogens. We demonstrated that the three pathogens disperse over substantially shorter distances compared to conventional estimates. This method will allow the researchers to re-analyze a vast number of existing dispersal gradients to improve our knowledge about dispersal. The improved knowledge has potential to advance our understanding of species' range expansions and shifts, and inform management of weeds and diseases in crops.

Population-level deep sequencing reveals the interplay of clonal and sexual reproduction in the fungal wheat pathogen Zymoseptoria tritici.

Pathogens cause significant challenges to global food security. On annual crops, pathogens must re-infect from environmental sources in every growing season. Fungal pathogens have evolved mixed reproductive strategies to cope with the distinct challenges of colonizing growing plants. However, how pathogen diversity evolves during growing seasons remains largely unknown. Here, we performed a deep hierarchical sampling in a single experimental wheat field infected by the major fungal pathogen Zymoseptoria tritici. We analysed whole genome sequences of 177 isolates collected from 12 distinct cultivars replicated in space at three time points of the growing season to maximize capture of genetic diversity. The field population was highly diverse with 37 SNPs per kilobase, a linkage disequilibrium decay within 200-700 bp and a high effective population size. Using experimental infections, we tested a subset of the collected isolates on the dominant cultivar planted in the field. However, we found no significant difference in virulence of isolates collected from the same cultivar compared to isolates collected on other cultivars. About 20 % of the isolate genotypes were grouped into 15 clonal groups. Pairs of clones were disproportionally found at short distances (<5 m), consistent with experimental estimates for per-generation dispersal distances performed in the same field. This confirms predominant leaf-to-leaf transmission during the growing season. Surprisingly, levels of clonality did not increase over time in the field although reproduction is thought to be exclusively asexual during the growing season. Our study shows that the pathogen establishes vast and stable gene pools in single fields. Monitoring short-term evolutionary changes in crop pathogens will inform more durable strategies to contain diseases.

A population-level invasion by transposable elements triggers genome expansion in a fungal pathogen.

Genome evolution is driven by the activity of transposable elements (TEs). The spread of TEs can have deleterious effects including the destabilization of genome integrity and expansions. However, the precise triggers of genome expansions remain poorly understood because genome size evolution is typically investigated only among deeply divergent lineages. Here, we use a large population genomics dataset of 284 individuals from populations across the globe of Zymoseptoria tritici, a major fungal wheat pathogen. We built a robust map of genome-wide TE insertions and deletions to track a total of 2456 polymorphic loci within the species. We show that purifying selection substantially depressed TE frequencies in most populations, but some rare TEs have recently risen in frequency and likely confer benefits. We found that specific TE families have undergone a substantial genome-wide expansion from the pathogen's center of origin to more recently founded populations. The most dramatic increase in TE insertions occurred between a pair of North American populations collected in the same field at an interval of 25 years. We find that both genome-wide counts of TE insertions and genome size have increased with colonization bottlenecks. Hence, the demographic history likely played a major role in shaping genome evolution within the species. We show that both the activation of specific TEs and relaxed purifying selection underpin this incipient expansion of the genome. Our study establishes a model to recapitulate TE-driven genome evolution over deeper evolutionary timescales.

In-Field Detection and Quantification of Septoria Tritici Blotch in Diverse Wheat Germplasm Using Spectral-Temporal Features.

Hyperspectral remote sensing holds the potential to detect and quantify crop diseases in a rapid and non-invasive manner. Such tools could greatly benefit resistance breeding, but their adoption is hampered by i) a lack of specificity to disease-related effects and ii) insufficient robustness to variation in reflectance caused by genotypic diversity and varying environmental conditions, which are fundamental elements of resistance breeding. We hypothesized that relying exclusively on temporal changes in canopy reflectance during pathogenesis may allow to specifically detect and quantify crop diseases while minimizing the confounding effects of genotype and environment. To test this hypothesis, we collected time-resolved canopy hyperspectral reflectance data for 18 diverse genotypes on infected and disease-free plots and engineered spectral-temporal features representing this hypothesis. Our results confirm the lack of specificity and robustness of disease assessments based on reflectance spectra at individual time points. We show that changes in spectral reflectance over time are indicative of the presence and severity of Septoria tritici blotch (STB) infections. Furthermore, the proposed time-integrated approach facilitated the delineation of disease from physiological senescence, which is pivotal for efficient selection of STB-resistant material under field conditions. A validation of models based on spectral-temporal features on a diverse panel of 330 wheat genotypes offered evidence for the robustness of the proposed method. This study demonstrates the potential of time-resolved canopy reflectance measurements for robust assessments of foliar diseases in the context of resistance breeding.

Joint evolution of dispersal and connectivity.

Functional connectivity, the realized flow of individuals between the suitable sites of a heterogeneous landscape, is a prime determinant of the maintenance and evolution of populations in fragmented habitats. While a large body of literature examines the evolution of dispersal propensity, it is less known how evolution shapes functional connectivity via traits that influence the distribution of the dispersers. Here, we use a simple model to demonstrate that, in a heterogeneous environment with clustered and solitary sites (i.e., with variable structural connectivity), the evolutionarily stable population contains strains that are strongly differentiated in their pattern of connectivity (local vs. global dispersal), but not necessarily in the fraction of dispersed individuals. Also during evolutionary branching, selection is disruptive predominantly on the pattern of connectivity rather than on dispersal propensity itself. Our model predicts diversification along a hitherto neglected axis of dispersal strategies and highlights the role of the solitary sites-the more isolated and therefore seemingly less important patches of habitat-in maintaining global dispersal that keeps all sites connected.

Precision Phenotyping Reveals Novel Loci for Quantitative Resistance to Septoria Tritici Blotch.

Accurate, high-throughput phenotyping for quantitative traits is a limiting factor for progress in plant breeding. We developed an automated image analysis to measure quantitative resistance to septoria tritici blotch (STB), a globally important wheat disease, enabling identification of small chromosome intervals containing plausible candidate genes for STB resistance. 335 winter wheat cultivars were included in a replicated field experiment that experienced natural epidemic development by a highly diverse but fungicide-resistant pathogen population. More than 5.4 million automatically generated phenotypes were associated with 13,648 SNP markers to perform the GWAS. We identified 26 chromosome intervals explaining 1.9-10.6% of the variance associated with four independent resistance traits. Sixteen of the intervals overlapped with known STB resistance intervals, suggesting that our phenotyping approach can identify simultaneously (i.e., in a single experiment) many previously defined STB resistance intervals. Seventeen of the intervals were less than 5 Mbp in size and encoded only 173 genes, including many genes associated with disease resistance. Five intervals contained four or fewer genes, providing high priority targets for functional validation. Ten chromosome intervals were not previously associated with STB resistance, perhaps representing resistance to pathogen strains that had not been tested in earlier experiments. The SNP markers associated with these chromosome intervals can be used to recombine different forms of quantitative STB resistance that are likely to be more durable than pyramids of major resistance genes. Our experiment illustrates how high-throughput automated phenotyping can accelerate breeding for quantitative disease resistance.

Hyperspectral Canopy Sensing of Wheat Septoria Tritici Blotch Disease.

Producing quantitative and reliable measures of crop disease is essential for resistance breeding, but is challenging and time consuming using traditional phenotyping methods. Hyperspectral remote sensing has shown potential for the detection of plant diseases, but its utility for phenotyping large and diverse populations of plants under field conditions requires further evaluation. In this study, we collected canopy hyperspectral data from 335 wheat varieties using a spectroradiometer, and we investigated the use of canopy reflectance for detecting the Septoria tritici blotch (STB) disease and for quantifying the severity of infection. Canopy- and leaf-level infection metrics of STB based on traditional visual assessments and automated analyses of leaf images were used as ground truth data. Results showed (i) that canopy reflectance and the selected spectral indices show promise for quantifying STB infections, and (ii) that the normalized difference water index (NDWI) showed the best performance in detecting STB compared to other spectral indices. Moreover, partial least squares (PLS) regression models allowed for an improvement in the prediction of STB metrics. The PLS discriminant analysis (PLSDA) model calibrated based on the spectral data of four reference varieties was able to discriminate between the diseased and healthy canopies among the 335 varieties with an accuracy of 93% (Kappa = 0.60). Finally, the PLSDA model predictions allowed for the identification of wheat genotypes that are potentially more susceptible to STB, which was confirmed by the STB visual assessment. This study demonstrates the great potential of using canopy hyperspectral remote sensing to improve foliar disease assessment and to facilitate plant breeding for disease resistance.

Ranking Quantitative Resistance to Septoria tritici Blotch in Elite Wheat Cultivars Using Automated Image Analysis.

Quantitative resistance is likely to be more durable than major gene resistance for controlling Septoria tritici blotch (STB) on wheat. Earlier studies hypothesized that resistance affecting the degree of host damage, as measured by the percentage of leaf area covered by STB lesions, is distinct from resistance that affects pathogen reproduction, as measured by the density of pycnidia produced within lesions. We tested this hypothesis using a collection of 335 elite European winter wheat cultivars that was naturally infected by a diverse population of Zymoseptoria tritici in a replicated field experiment. We used automated image analysis of 21,420 scanned wheat leaves to obtain quantitative measures of conditional STB intensity that were precise, objective, and reproducible. These measures allowed us to explicitly separate resistance affecting host damage from resistance affecting pathogen reproduction, enabling us to confirm that these resistance traits are largely independent. The cultivar rankings based on host damage were different from the rankings based on pathogen reproduction, indicating that the two forms of resistance should be considered separately in breeding programs aiming to increase STB resistance. We hypothesize that these different forms of resistance are under separate genetic control, enabling them to be recombined to form new cultivars that are highly resistant to STB. We found a significant correlation between rankings based on automated image analysis and rankings based on traditional visual scoring, suggesting that image analysis can complement conventional measurements of STB resistance, based largely on host damage, while enabling a much more precise measure of pathogen reproduction. We showed that measures of pathogen reproduction early in the growing season were the best predictors of host damage late in the growing season, illustrating the importance of breeding for resistance that reduces pathogen reproduction in order to minimize yield losses caused by STB. These data can already be used by breeding programs to choose wheat cultivars that are broadly resistant to naturally diverse Z. tritici populations according to the different classes of resistance.

Evolution of dispersal under variable connectivity.

The pattern of connectivity between local populations or between microsites supporting individuals within a population is a poorly understood factor affecting the evolution of dispersal. We modify the well-known Hamilton-May model of dispersal evolution to allow for variable connectivity between microsites. For simplicity, we assume that the microsites are either solitary, i.e., weakly connected through costly dispersal, or part of a well-connected cluster of sites with low-cost dispersal within the cluster. We use adaptive dynamics to investigate the evolution of dispersal, obtaining analytic results for monomorphic evolution and numerical results for the co-evolution of two dispersal strategies. A monomorphic population always evolves to a unique singular dispersal strategy, which may be an evolutionarily stable strategy or an evolutionary branching point. Evolutionary branching happens if the contrast between connectivities is sufficiently high and the solitary microsites are common. The dimorphic evolutionary singularity, when it exists, is always evolutionarily and convergence stable. The model exhibits both protected and unprotected dimorphisms of dispersal strategies, but the dimorphic singularity is always protected. Contrasting connectivities can thus maintain dispersal polymorphisms in temporally stable environments.