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Cultivated sunflower (Helianthus annuus L.) exhibits numerous phenotypic and transcriptomic responses to drought. However, the ways in which these responses vary with differences in drought timing and severity are insufficiently understood. We used phenotypic and transcriptomic data to evaluate the response of sunflower to drought scenarios of different timing and severity in a common garden experiment. Using a semi-automated outdoor high-throughput phenotyping platform, we grew six oilseed sunflower lines under control and drought conditions. Our results reveal that similar transcriptomic responses can have disparate phenotypic effects when triggered at different developmental time points. Leaf transcriptomic responses, however, share similarities despite timing and severity differences (e.g., 523 differentially expressed genes (DEGs) were shared across all treatments), though increased severity elicited greater differences in expression, particularly during vegetative growth. Across treatments, DEGs were highly enriched for genes related to photosynthesis and plastid maintenance. A co-expression analysis identified a single module (M8) enriched in all drought stress treatments. Genes related to drought, temperature, proline biosynthesis, and other stress responses were overrepresented in this module. In contrast to transcriptomic responses, phenotypic responses were largely divergent between early and late drought. Early-stressed sunflowers responded to drought with reduced overall growth, but became highly water-acquisitive during recovery irrigation, resulting in overcompensation (higher aboveground biomass and leaf area) and a greater overall shift in phenotypic correlations, whereas late-stressed sunflowers were smaller and more water use-efficient. Taken together, these results suggest that drought stress at an earlier growth stage elicits a change in development that enables greater uptake and transpiration of water during recovery, resulting in higher growth rates despite similar initial transcriptomic responses.

Plant breeding programs design new crop cultivars which, while developed for distinct populations of environments, are nevertheless grown over large areas during their time in the market. Over its cultivation area, the crop is exposed to highly diverse stress patterns caused by climatic uncertainty and multiple management options, which often leads to decreased expected crop performance.In this study, we aim is to assess how finer spatial management of genetic resources could reduce the yield variance explained by genotype × environment interactions in a set of cropping environments and ultimately improve the efficiency and stability of crop production. We used modeling and simulation to predict the crop performance resulting from the interaction between cultivar growth and development, climate and soil conditions, and management practices. We designed a computational experiment that evaluated the performance of a collection of commercial sunflower cultivars in a realistic population of cropping conditions in France, built from extensive agricultural surveys. Distinct farming locations sharing similar simulated abiotic stress patterns were clustered together to specify environment types. We then used optimization methods to search for cultivars × environments combinations leading to increased yield expectations.Results showed that a single cultivar choice adapted to the most frequent environment-type in the population is a robust strategy. However, the relevance of cultivar recommendations to specific locations was gradually increasing with the knowledge of pedo-climatic conditions. We argue that this approach while being operational on current genetic material could act synergistically with plant breeding as more diverse material could enable access to cultivars with distinctive traits, more adapted to specific conditions.Key messageCrop simulation helps to analyze environmental impacts on crops and provides year-independent context information. This information is of major importance when deciding which cultivar to choose at sowing time.

Background Leaf senescence delay impacts positively in grain yield by maintaining the photosynthetic area during the reproductive stage and during grain filling. Therefore a comprehensive understanding of the gene families associated with leaf senescence is essential. NAC transcription factors (TF) form a large plant-specific gene family involved in regulating development, senescence, and responses to biotic and abiotic stresses. The main goal of this work was to identify sunflower NAC TF (HaNAC) and their association with senescence, studying their orthologous to understand possible functional relationships between genes of different species. ResultsTo clarify the orthologous relationships, we used an in-depth comparative study of four divergent taxa, in dicots and monocots, with completely sequenced genomes (Arabidopsis thaliana, Vitis vinifera, Musa acuminata and Oryza sativa). These orthologous groups provide a curated resource for large scale protein sequence annotation of NAC TF. From the 151 HaNAC genes detected in the latest version of the sunflower genome, 50 genes were associated with senescence traits. These genes showed significant differential expression in two contrasting lines according to an RNAseq assay. An assessment of overexpressing the Arabidopsis line for HaNAC001 (a gene of the same orthologous group of Arabidopsis thaliana ORE1) revealed that this line displayed a significantly higher number of senescent leaves and a pronounced change in development rate.ConclutionsThis finding suggests HaNAC001 as an interesting candidate to explore the molecular regulation of senescence in sunflower.

International audience; This article describes how metabolomic data were produced on sunflower plants subjected to water deficit. Twenty-four sunflower ( Helianthus annuus L.) genotypes were selected to represent genetic diversity within cultivated sunflower and included both inbred lines and their hybrids. Drought stress was applied at the vegetative stage to plants cultivated in pots using the high-throughput phenotyping facility Heliaphen. Here, we provide untargeted and targeted metabolomic data of sunflower leaves. These compositional data differentiate both plant water status and different genotype groups. They constitute a valuable resource for the community to study the adaptation of crops to drought and the metabolic bases of heterosis.; Cet article décrit comment les données métabolomiques ont été produites sur des plants de tournesol soumis à un déficit hydrique. Vingt-quatre génotypes de tournesol ( Helianthus annuus L.) ont été sélectionnés pour représenter la diversité génétique du tournesol cultivé et comprennent à la fois des lignées consanguines et leurs hybrides. Une limitation hydrique a été appliquée au stade végétatif aux plantes cultivées en pots à l’aide de la plateforme de phénotypage à haut débit Heliaphen. Ici, nous mettons à disposition des données métabolomiques non ciblées et ciblées de feuilles de tournesol. Ces données de composition permettent de différencier l’état hydrique des plantes et différents groupes de génotypes. Elles constituent une ressource précieuse pour la communauté afin d’étudier l’adaptation des cultures à la sécheresse et les bases métaboliques de l’hétérosis.

SummaryGenes under balancing selection control phenotypes such as immunity, color or sex, but are difficult to identify. Self-incompatibility genes are under negative frequency-dependent selection, a special case of balancing selection, with up to 30 to 50 alleles segregating per population. We developed a method based on pooled Single-Molecule transcriptomics to identify balanced polymorphisms expressed in tissues of interest. We searched for multi-allelic, non-recombining genes causing self-incompatibility in wild sunflower (Helianthus annuus). A diversity scan in pistil identified a gene,Ha7650b,that displayed balanced polymorphism and colocalized with a quantitative trait locus for self-incompatibility. Unexpectedly,Ha7650bdisplayed gigantism (400 kb), which was caused by increase in intron size as a consequence of suppressed recombination.Ha7650bemerged after a whole-genome duplication (29 millions years ago) followed by tandem duplications and neofunctionalisation.Ha7650bshows expression, genetic location, genomic neighbourhood and predicted function that provide strong evidence that it is involved in self-incompatibility. Pooled Single-Molecule transcriptomics is an affordable and powerful new method that makes it possible to identify diversity and structural outliers simultaneously. It will allow a breakthrough in the discovery of self-incompatibility genes and other expressed genes under balancing selection.

International audience; As a rainfed spring-sown crop, sunflower ( Helianthus annuus L.) is increasingly exposed to negative impacts of climate change, especially to high temperatures and drought stress. Incremental, systemic and transformative adaptations have been suggested for reducing the crop vulnerability to these stressful conditions. In addition, innovative cropping systems based on low-input management, organic farming, soil and water conservation practices, intercropping, double-cropping, and/or agroforestry are undergoing marked in agriculture. Because of its plasticity and low-input requirements (nitrogen, water, pesticides), sunflower crop is likely to take part to these new agroecological systems. Aside from current production outputs (yield, oil and cake), ecosystem services ( e.g. bee feeding, soil phytoremediation…), and non-food industrial uses are now expected externalities for the crop. The combination of climatic and societal contexts could deeply modify the characteristics of genotypes to be cultivated in the main production areas (either traditional or adoptive). After reviewing these changes, we identify how innovative cropping systems and new environments could modify the traits classically considered up to now, especially in relation to expected ecosystem services. Finally, we consider how research could provide methods to help identifying traits of interest and design ideotypes.; En tant que culture d’été non irriguée, le tournesol ( Helianthus annuus L.) est davantage exposé aux impacts négatifs du changement climatique, et en particulier aux températures élevées et au stress hydrique. Des adaptations incrémentales, systémiques ou de rupture ont été suggérées pour réduire la vulnérabilité de la culture à ces conditions stressantes. Par ailleurs, des systèmes de culture innovants sont en fort développement qu’il s’agisse de systèmes à bas niveaux d’intrants, d’agriculture biologique, d’agriculture de conservation des sols, d’agroforesterie, ou de l’introduction d’associations de cultures et de doubles cultures. En raison de sa plasticité et de ses faibles besoins en intrants (eau, azote, pesticides), la culture de tournesol est amenée à s’insérer de manière privilégiée dans ces systèmes agro-écologiques. À côté de la production (grains, huile, tourteaux), de nouvelles externalités sont attendues comme la contribution aux autres services écosystémiques (pollinisateurs, phytoremédiation…) et de nouveaux usages non alimentaires. La conjugaison des contextes climatiques et sociétaux pourrait profondément changer les caractéristiques attendues des variétés cultivées dans les principales régions de production qu’elles soient traditionnelles ou en reconquête. Après une revue de ces changements, nous identifions comment systèmes innovants et changement climatique pourraient modifier les caractères phénotypiques considérés classiquement, en particulier dans la perspective de nouveaux services. Enfin, nous verrons comment la recherche pourrait fournir des méthodes pour faciliter l’identification de ces caractères d’intérêt et aider à la conception d’idéotypes.

This article describes how the proteomic data were produced on sunflower plants subjected to water deficit. Twenty-four sunflower genotypes were selected to represent genetic diversity within cultivated sunflower. They included both inbred lines and their hybridsWater deficit was applied to plants in pots at the vegetative stage using the high-throughput phenotyping platform Heliaphen. Here, we provide proteomic data from sunflower leaves corresponding to the identification of 3062 proteins and the quantification of 1211 of them in these 24 genotypes grown in two watering conditions. These data differentiate both treatment and the different genotypes and constitute a valuable resource to the community to study adaptation of crops to drought and the molecular basis of heterosis.

This article presents experimental data describing the physiology and morphology of sunflower plants subjected to water deficit. Twenty-four sunflower genotypes were selected to represent genetic diversity within cultivated sunflower and included both inbred lines and their hybrids.Drought stress was applied to plants in pots at the vegetative stage using the high-throughput phenotyping platform Heliaphen at INRA Toulouse (France). Here, we provide data including specific leaf area, osmotic potential and adjustment, carbon isotope discrimination, leaf transpiration, plant architecture: plant height, leaf number, stem diameter. We also provide leaf areas of individual organs through time and growth rate during the stress period, environmental data such as temperatures, wind and radiation during the experiment. These data differentiate both treatment and the different genotypes and constitute a valuable resource to the community to study adaptation of crops to drought and the physiological basis of heterosis. It is available on the following repository: https://doi.org/10.25794/phenotype/er6lPW7V Keywords: Helianthus, Abiotic stress, Phenotyping platform, Drought stress

Understanding the genetic basis of phenotypic plasticity is crucial for predicting and managing climate change effects on wild plants and crops. Here, we combined crop modeling and quantitative genetics to study the genetic control of oil yield plasticity for multiple abiotic stresses in sunflower. First we developed stress indicators to characterize 14 environments for three abiotic stresses (cold, drought and nitrogen) using the SUNFLO crop model and phenotypic variations of three commercial varieties. The computed plant stress indicators better explain yield variation than descriptors at the climatic or crop levels. In those environments, we observed oil yield of 317 sunflower hybrids and regressed it with three selected stress indicators. The slopes of cold stress norm reaction were used as plasticity phenotypes in the following genome-wide association study. Among the 65,534 tested SNP, we identified nine QTL controlling oil yield plasticity to cold stress. Associated SNP are localized in genes previously shown to be involved in cold stress responses: oligopeptide transporters, LTP, cystatin, alternative oxidase, or root development. This novel approach opens new perspectives to identify genomic regions involved in genotype-by-environment interaction of a complex traits to multiple stresses in realistic natural or agronomical conditions.

Seed germination is regulated by environmental factors, particularly water availability. Water deficits at the time of sowing impair the establishment of crop plants. Transcriptome and proteome profiling was used to document the responses of sunflower (Helianthus annuus) seeds to moderate water stress during germination in two hybrids that are nominally classed as drought sensitive and drought tolerant. Differences in the water stress-dependent accumulation reactive oxygen species and antioxidant enzymes activities were observed between the hybrids. A pathway-based analysis of the hybrid transcriptomes demonstrated that the water stress-dependent responses of seed metabolism were similar to those of the plant, with a decreased abundance of transcripts encoding proteins associated with metabolism and cell expansion. Moreover, germination under water stress conditions was associated with increased levels of transcripts encoding heat shock proteins. Exposure of germinating seeds to water stress specifically affected the abundance of a small number of proteins, including heat shock proteins. Taken together, these data not only identify factors that are likely to play a key role in drought tolerance during seed germination, but they also demonstrate the importance of the female parent in the transmission of water stress tolerance.

This article describes how RNA expression data of 173 genes were produced on 384 sunflowerhybrids grown infield conditions. Sunflower hybrids were selected to represent genetic diversity withincultivated sunflower. The RNA was extracted from mature leaves at one time seven days after anthesis.These data allow to differentiate the different genotype behaviours and constitute a valuable resource to thecommunity to study the adaptation of crops tofield conditions and the molecular basis of heterosis. It isavailable on data.inra.fr repository.; Cet article décrit la production des niveaux d’expression de 173 gènes dans 384 hybrides de tournesol cultivés en conditions de champ. Les hybrides sont issus de parents choisis pour représenter la diversité génétique dans le tournesol cultivé. Les ARN ont été extraits à partir de feuilles matures environ sept jours après la floraison. Ces données permettent de différencier les comportements des différents génotypes et constituent une ressource importante pour les chercheurs intéressés dans l’adaptation des espèces cultivées aux conditions agronomiques et aux bases moléculaires de l’hétérosis. Elles sont disponibles sur le portail Data INRAE : data.inra.fr.

Modern breeding of sunflower (Helianthus annuus L.), which started 100 years ago, increased the number and the diversity of cultivated forms. In addition, for more than 50 years, wild sunflower and other Helianthus species have been collected in North America where they all originated. Collections of both cultivated and wild forms are maintained in gene banks in many countries where sunflower is an important crop, with some specificity according to the availability of germplasm and to local research and breeding programmes. Cultivated material includes land races, open pollinated varieties, synthetics and inbred lines. The majority of wild accessions are ecotypes of wild Helianthus annuus, but also 52 other species of Helianthus and a few related genera. The activities of three gene banks, in USA, France and Serbia, are described in detail, supplemented by data from seven other countries. Past and future uses of the genetic resources for environmental adaptation and breeding are discussed in relation to genomic and improved phenotypic knowledge of the cultivated and wild accessions available in the gene banks. L’amélioration moderne du tournesol (Helianthus annuus L.) a débuté il y a un siècle, diversifiant et augmentant le nombre des formes cultivées du tournesol. De plus, des collectes de tournesols sauvages et d’espèces du genre Helianthus ont lieu depuis 50 ans en Amérique du Nord d’où ils sont tous originaires. Ainsi, des collections de tournesols cultivés et sauvages sont conservées par des centres de ressources génétiques dans de nombreux pays où le tournesol est une culture importante. Chacun d’eux présente des spécificités par rapport aux ressources génétiques maintenues, en fonction des programmes de recherche ou de sélection variétale locales. Le matériel génétique cultivé comprend des écotypes, des populations et des lignées tandis que les accessions sauvages correspondent eux écotypes d’Helianthus annuus sauvages et des 52 autres espèces apparentées du genre Helianthus. Les activités de trois centres de ressources génétiques des États-Unis, de la France et de la Serbie sont décrites en détail, complétées par des données provenant des centres de sept autres pays. L’historique de l’utilisation des ressources génétiques et les perspectives futures pour l’adaptation des variétés à l’environnement sont discutés ainsi que leur caractérisation au niveau génomique et phénotypique. The Supplementary Material is available at [https://www.ocljournal.org/10.1051/ocl/2020004/olm].

Background: Leaf senescence is a complex process, controlled by multiple genetic and environmental variables. In sunflower, leaf senescence is triggered abruptly following anthesis thereby limiting the capacity of plants to keep their green leaf area during grain filling, which subsequently has a strong impact on crop yield. Recently, we performed a selection of contrasting sunflower inbred lines for the progress of leaf senescence through a physiological, cytological and molecular approach. Here we present a large scale transcriptomic analysis using RNA-seq and its integration with metabolic profiles for two contrasting sunflower inbred lines, R453 and B481-6 (early and delayed senescence respectively), with the aim of identifying metabolic pathways associated to leaf senescence. Results: Gene expression profiles revealed a higher number of differentially expressed genes, as well as, higher expression levels in R453, providing evidence for early activation of the senescence program in this line. Metabolic pathways associated with sugars and nutrient recycling were differentially regulated between the lines. Additionally, we identified transcription factors acting as hubs in the co-expression networks; some previously reported as senescence-associated genes in model species but many are novel candidate genes. Conclusions: Understanding the onset and the progress of the senescence process in crops and the identification of these new candidate genes will likely prove highly useful for different management strategies to mitigate the impact of senescence on crop yield. Functional characterization of candidate genes will help to develop molecular tools for biotechnological applications in breeding crop yield. Fil: Moschen, Sebastián Nicolás. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Tucuman-Santiago del Estero. Estación Experimental Agropecuaria Famaillá; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Marino, Johanna Magali. Universidad Nacional de San Martín; Argentina Fil: Nicosia, Salvador. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Higgins, Janet. Earlham Institute; Reino Unido Fil: Alseekh, Saleh. No especifíca; Fil: Astigueta, Francisco Horacio. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Bengoa Luoni, Sofia Ailin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: Rivarola, Maximo Lisandro. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Fernie, Alisdair R.. No especifíca; Fil: Blanchet, Nicolas. Centre National de la Recherche Scientifique; Francia Fil: Langlade, Nicolas B.. Centre National de la Recherche Scientifique; Francia Fil: Paniego, Norma Beatriz. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Fernández, Paula. No especifíca; Fil: Heinz, Ruth Amelia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina

Plant and crop metabolomic analyses may be used to study metabolism across genetic and environmental diversity. Complementary analytical strategies are useful for investigating metabolic changes and searching for biomarkers of response or performance., The experimental material consisted in eight sunflower lines with two line status, four restorers (R, used as males) and four maintainers (B, corresponding to females) routinely used for sunflower hybrid varietal production, respectively to complement or maintain the cytoplasmic male sterility PET1. These lines were either irrigated at full soil capacity (WW) or submitted to drought stress (DS). Our aim was to combine targeted and non-targeted metabolomics to characterize sunflower leaf composition in order to investigate the effect of line status genotypes and environmental conditions and to find the best and smallest set of biomarkers for line status and stress response using a custom-made process of variables selection., H-NMR, MS-based profiles and targeted analyses of major metabolites. Based on statistical analyses, a limited number of markers were able to separate WW and DS samples in a more discriminant manner than previously published physiological data. Another metabolic marker set was able to discriminate line status., This study underlines the potential of metabolic markers for discriminating genotype groups and environmental conditions. Their potential use for prediction is discussed.

International audience; Heliaphen is an outdoor platform designed for high-throughput phenotyping. It allows the automated management of drought scenarios and monitoring of plants throughout their lifecycles. A robot moving between plants growing in 15-L pots monitors the plant water status and phenotypes the leaf or whole-plant morphology. From these measurements, we can compute more complex traits, such as leaf expansion (LE) or transpiration rate (TR) in response to water deficit. Here, we illustrate the capabilities of the platform with two practical cases in sunflower (Helianthus annuus): a genetic and genomic study of the response of yield-related traits to drought, and a modeling study using measured parameters as inputs for a crop simulation. For the genetic study, classical measurements of thousand-kernel weight (TKW) were performed on a biparental population under automatically managed drought stress and control conditions. These data were used for an association study, which identified five genetic markers of the TKW drought response. A complementary transcriptomic analysis identified candidate genes associated with these markers that were differentially expressed in the parental backgrounds in drought conditions. For the simulation study, we used a crop simulation model to predict the impact on crop yield of two traits measured on the platform (LE and TR) for a large number of environments. We conducted simulations in 42 contrasting locations across Europe using 21 years of climate data. We defined the pattern of abiotic stresses occurring at the continental scale and identified ideotypes (i.e., genotypes with specific trait values) that are more adapted to specific environment types. This study exemplifies how phenotyping platforms can assist the identification of the genetic architecture controlling complex response traits and facilitate the estimation of ecophysiological model parameters to define ideotypes adapted to different environmental conditions.

International audience; Domesticated plants and animals often display dramatic responses to selection, but the origins of the genetic diversity underlying these responses remain poorly understood. Despite domestication and improvement bottlenecks, the cultivated sunflower remains highly variable genetically, possibly due to hybridization with wild relatives. To characterize genetic diversity in the sunflower and to quantify contributions from wild relatives, we sequenced 287 cultivated lines, 17 Native American landraces and 189 wild accessions representing 11 compatible wild species. Cultivar sequences failing to map to the sunflower reference were assembled de novo for each genotype to determine the gene repertoire, or 'pan-genome', of the cultivated sunflower. Assembled genes were then compared to the wild species to estimate origins. Results indicate that the cultivated sunflower pan-genome comprises 61,205 genes, of which 27% vary across genotypes. Approximately 10% of the cultivated sunflower pan-genome is derived through introgression from wild sunflower species, and 1.5% of genes originated solely through introgression. Gene ontology functional analyses further indicate that genes associated with biotic resistance are over-represented among introgressed regions, an observation consistent with breeding records. Analyses of allelic variation associated with downy mildew resistance provide an example in which such introgressions have contributed to resistance to a globally challenging disease.

Heliaphen is an outdoor pot platform designed for high-throughput phenotyping. It allows automated management of drought scenarios and plant monitoring during the whole plant cycle. A robot moving between plants growing in 15L pots monitors plant water status and phenotypes plant or leaf morphology, from which we can compute more complex traits such as the response of leaf expansion (LE) or plant transpiration (TR) to water deficit. Here, we illustrate the platform capabilities for sunflower on two practical cases: a genetic and genomics study for the response to drought of yield-related traits and a simulation study, where we use measured parameters as inputs for a crop simulation model. For the genetic study, classical measurements of thousand-kernel weight (TKW) were done on a sunflower bi-parental population under water stress and control conditions managed automatically. The association study using the TKW drought-response highlighted five genetic markers. A complementary transcriptomic experiment identified closeby candidate genes differentially expressed in the parental backgrounds in drought conditions. For the simulation study, we used the SUNFLO crop simulation model to assess the impact of two traits measured on the platform (LE and TR) on crop yield in a large population of environments. We conducted simulations in 42 contrasted locations across Europe and 21 years of climate data. We defined the pattern of abiotic stresses occurring at this continental scale and identified ideotypes (i.e. genotypes with specific traits values) that are more adapted to specific environment types. This study exemplifies how phenotyping platforms can help with the identification of the genetic architecture of complex response traits and the estimation of eco-physiological model parameters in order to define ideotypes adapted to different environmental conditions.

Key message This study compares five models of GWAS, to show the added value of non-additive modeling of allelic effects to identify genomic regions controlling flowering time of sunflower hybrids. Abstract Genome-wide association studies are a powerful and widely used tool to decipher the genetic control of complex traits. One of the main challenges for hybrid crops, such as maize or sunflower, is to model the hybrid vigor in the linear mixed models, considering the relatedness between individuals. Here, we compared two additive and three non-additive association models for their ability to identify genomic regions associated with flowering time in sunflower hybrids. A panel of 452 sunflower hybrids, corresponding to incomplete crossing between 36 male lines and 36 female lines, was phenotyped in five environments and genotyped for 2,204,423 SNPs. Intra-locus effects were estimated in multi-locus models to detect genomic regions associated with flowering time using the different models. Thirteen quantitative trait loci were identified in total, two with both model categories and one with only non-additive models. A quantitative trait loci on LG09, detected by both the additive and non-additive models, is located near a GAI homolog and is presented in detail. Overall, this study shows the added value of non-additive modeling of allelic effects for identifying genomic regions that control traits of interest and that could participate in the heterosis observed in hybrids. Electronic supplementary material The online version of this article (doi:10.1007/s00122-017-3003-4) contains supplementary material, which is available to authorized users.

This article describes how the transcriptomic data were produced on sunflower plants subjected to water deficit. Twenty-four sunflower (Helianthus annuus) genotypes were selected to represent genetic diversity within cultivated sunflower and included both inbred lines and their hybrids. Drought stress was applied to plants in pots at the vegetative stage using the high-throughput phenotyping platform Heliaphen. Here, we provide transcriptomic data from sunflower leaves. These data differentiate both plant water status and the different genotypes. They constitute a valuable resource to the community to study adaptation of crops to drought and the transcriptomic basis of heterosis.; Données transcriptomiques de feuilles de huit lignées de tournesol et de leurs 16 hybrides soumis à un stress hydrique. Cet article décrit la production de données transcriptomiques sur des plantes de tournesol soumises à un stress hydrique. Vingt-quatre génotypes de tournesol (Helianthus annuus) ont été sélectionnés pour représenter la diversité génétique parmi le tournesol cultivé et comprennent à la fois des lignées et leurs hybrides. Le stress hydrique a été appliqué sur des plantes en pots au stade végétatif grâce à la plateforme de phénotypage haut-débit Heliaphen. Ici, nous mettons à disposition les données transcritomiques de feuilles de ces plantes. Les données permettent de différencier à la fois le statut hydrique et les différents génotypes. Elles constituent une ressource importante pour la communauté pour étudier l'adaptation des plantes à la sécheresse et les bases de l'hétérosis au niveau transcriptomique.

International audience; Seed dormancy, which blocks germination in apparently favourable conditions, is a key regulatory control point of plant population establishment. As germination requires de novo translation, its regulation by dormancy is likely to be related to the association of individual transcripts to polysomes. Here, the polysome-associated mRNAs, that is, the translatome, were fractionated and characterized with microarrays in dormant and nondormant sunflower (Helianthus annuus) embryos during their imbibition at 10 degrees C, a temperature preventing germination of dormant embryos. Profiling of mRNAs in polysomal complexes revealed that the translatome differs between germinating and nongerminating embryos. Association of transcripts with polysomes reached a maximum after 15h of imbibition; at this time-point 194 polysome-associated transcripts were specifically found in nondormant embryos and 47 in dormant embryos only. The proteins corresponding to the polysomal mRNAs in nondormant embryos appeared to be very pertinent for germination and were involved mainly in transport, regulation of transcription or cell wall modifications. This work demonstrates that seed germination results from a timely regulated and selective recruitment of mRNAs to polysomes, thus opening novel fields of investigation for the understanding of this developmental process.

Sunflower (Helianthus annuus L.) seed dormancy is regulated by reactive oxygen species (ROS) and can be alleviated by incubating dormant embryos in the presence of methylviologen (MV), a ROS-generating compound. Ethylene alleviates sunflower seed dormancy whereas abscisic acid (ABA) represses germination. The purposes of this study were to identify the molecular basis of ROS effect on seed germination and to investigate their possible relationship with hormone signalling pathways. Ethylene treatment provoked ROS generation in embryonic axis whereas ABA had no effect on their production. The beneficial effect of ethylene on germination was lowered in the presence of antioxidant compounds, and MV suppressed the inhibitory effect of ABA. MV treatment did not alter significantly ethylene nor ABA production during seed imbibition. Microarray analysis showed that MV treatment triggered differential expression of 120 probe sets (59 more abundant and 61 less abundant genes), and most of the identified transcripts were related to cell signalling components. Many transcripts less represented in MV-treated seeds were involved in ABA signalling, thus suggesting an interaction between ROS and ABA signalling pathways at the transcriptional level. Altogether, these results shed new light on the crosstalk between ROS and plant hormones in seed germination.

Plant or soil water status are required in many scientific fields to understand plant responses to drought. Because the transcriptomic response to abiotic conditions, such as water deficit, reflects plant water status, genomic tools could be used to develop a new type of molecular biomarker. Using the sunflower (Helianthus annuus L.) as a model species to study the transcriptomic response to water deficit both in greenhouse and field conditions, we specifically identified three genes that showed an expression pattern highly correlated to plant water status as estimated by the pre-dawn leaf water potential, fraction of transpirable soil water, soil water content or fraction of total soil water in controlled conditions. We developed a generalized linear model to estimate these classical water status indicators from the expression levels of the three selected genes under controlled conditions. This estimation was independent of the four tested genotypes and the stage (pre- or post-flowering) of the plant. We further validated this gene expression biomarker under field conditions for four genotypes in three different trials, over a large range of water status, and we were able to correct their expression values for a large diurnal sampling period.

SNP genotyping of 114 cultivated sunflower populations showed that the multiplication process and the main traits selected during breeding of sunflower cultivars drove molecular diversity of the populations. The molecular diversity in a set of 114 cultivated sunflower populations was studied by single-nucleotide polymorphism genotyping. These populations were chosen as representative of the 400 entries in the INRA collection received or developed between 1962 and 2011 and made up of land races, open-pollinated varieties, and breeding pools. Mean allele number varied from 1.07 to 1.90. Intra-population variability was slightly reduced according to the number of multiplications since entry but some entries were probably largely homozygous when received. A principal component analysis was used to study inter-population variability. The first 3 axes accounted for 17% of total intra-population variability. The first axis was significantly correlated with seed oil content, more closely than just the distinction between oil and confectionary types. The second axis was related to the presence or absence of restorer genes and the third axis to flowering date and possibly to adaptation to different climates. Our results provide arguments highlighting the effect of the maintenance process on the within population genetic variability as well as on the impact of breeding for major agronomic traits on the between population variability of the collection. Propositions are made to improve sunflower population maintenance procedures to keep maximum genetic variability for future breeding.

Climate change is characterized by higher temperatures, elevated atmospheric CO2 concentrations, extreme climatic hazards, and less water available for agriculture. Sunflower, a spring-sown crop often cultivated in southern and eastern regions of Europe, could be more vulnerable to the direct effect of heat stress at anthesis and drought during its growing cycle, both factors resulting in severe yield loss, oil content decrease, and fatty acid alterations. Adaptations through breeding (earliness, stress tolerance), crop management (planting dates), and shifting of growing areas could be developed, assessed and combined to partly cope with these negative impacts. New cultivation opportunities could be expected in northern parts of Europe where sunflower is not grown presently and where it could usefully contribute to diversify cereal-based cropping systems. In addition, sunflower crop could participate to the mitigation solution as a low greenhouse gas emitter compared to cereals and oilseed rape. Sunflower crop models should be revised to account for these emerging environmental factors in order to reduce the uncertainties in yield and oil predictions. The future of sunflower in Europe is probably related to its potential adaptation to climate change but also to its competitiveness and attractiveness for food and energy., Le changement climatique se caractérise par des températures élevées, de plus fortes concentrations atmosphériques en CO2, des risques climatiques extrêmes et moins d'eau disponible pour l'agriculture. Le tournesol, culture semée au printemps dans le sud et l'est de l'Europe, pourrait être plus exposé à l'avenir aux fortes températures et à un déficit hydrique marqué dès la floraison, avec pour conséquences des pertes de rendement, une diminution de la teneur en huile et une altération de la composition en acides gras. Des adaptations sont possibles à court et moyen terme par la sélection (précocité, tolérance aux stress), la conduite de culture (date de semis) et le déplacement des zones de production, permettant de faire face en partie aux impacts négatifs attendus. Ainsi le tournesol pourrait être cultivé plus au nord participant utilement à la nécessaire diversification des bassins céréaliers. En outre, la culture de tournesol étant faiblement émettrice de gaz à effet de serre par rapport aux céréales ou au colza pourrait contribuer davantage à la solution climatique apportée par l'agriculture. Les modèles de culture devraient être revus pour mieux tenir compte de ces facteurs environnementaux émergents si l'on veut réduire les incertitudes dans les prédictions de rendement et de teneur en huile. L'avenir du tournesol en Europe est probablement lié à son potentiel d'adaptation au changement climatique, mais dépendra aussi de sa compétitivité et de son attractivité en tant que fournisseur d'énergie et d'aliments.

Background Phoma macdonaldii has been reported as the causal agent of black stem disease (BS) and premature ripening (PR) on sunflower. PR is considered as the most widespread and detrimental disease on sunflower in France. While genetic variability and QTL mapping for partial resistance of sunflower to stem, collar and roots attacks have been reported on plantlets in controlled conditions, this work aims to describe the genetic variability in a subset of a sunflower lines, and for the first time to map QTL involved in PR resistance evaluated in field conditions using controlled inoculation. Results An efficient and reliable method for inoculation used in field experiments induced stem base necrosis on up to 98% of all plants. A significant genetic variability for PR resistance in the field was detected among the 20 inbred lines of the core collection tested across the two years. For QTL mapping, the PR resistance evaluation was performed on two recombinant inbred lines (RIL) populations derived from the crosses XRQxPSC8 and FUxPAZ2 in two different years. QTL analyses were based on a newly developed consensus genetic map comprising 1007 non-redundant molecular markers. In each of the two RIL populations, different QTL involved in PR partial sunflower resistance were detected. The most significant QTL were detected 49 days post infection (DPI) on LG10 (LOD 7.7) and on LG7 (LOD 12.1) in the XRQxPSC8 and FUxPAZ2 RIL population, respectively. In addition, different QTL were detected on both populations for PR resistance measured between 14 and 35 DPI. In parallel, the incidence of natural attack of P. macdonaldii resulting in BS disease was recorded, showing that in these populations, the genetic of resistance to both diseases is not governed by the same factors. Conclusion This work provides the first insights on the genetic architecture of sunflower PR resistance in the field. Moreover, the separate studies of symptoms on different organs and in time series allowed the identification of a succession of genetic components involved in the sunflower resistance to PR and BS diseases caused by Phoma macdonaldii along the development of the {plant * pathogen} interaction. Electronic supplementary material The online version of this article (10.1186/s12870-017-1116-1) contains supplementary material, which is available to authorized users.

International audience; Background: In the last decade, metabolomics has emerged as a powerful diagnostic and predictive tool in many branches of science. Researchers in microbes, animal, food, medical and plant science have generated a large number of targeted or non-targeted metabolic profiles by using a vast array of analytical methods (GC–MS, LC–MS, 1H-NMR….). Comprehensive analysis of such profiles using adapted statistical methods and modeling has opened up the possibility of using single or combinations of metabolites as markers. Metabolic markers have been proposed as proxy, diagnostic or predictors of key traits in a range of model species and accurate predictions of disease outbreak frequency, developmental stages, food sensory evaluation and crop yield have been obtained. Aim of review : (i) To provide a definition of plant performance and metabolic markers, (ii) to highlight recent key applications involving metabolic markers as tools for monitoring or predicting plant performance, and (iii) to propose a workable and cost-efficient pipeline to generate and use metabolic markers with a special focus on plant breeding. Key message: Using examples in other models and domains, the review proposes that metabolic markers are tending to complement and possibly replace traditional molecular markers in plant science as efficient estimators of performance.

Understanding the genetic basis of phenotypic plasticity is crucial for predicting and managing climate change effects on wild plants and crops. Here, we combined crop modelling and quantitative genetics to study the genetic control of oil yield plasticity for multiple abiotic stresses in sunflower. First, we developed stress indicators to characterize 14 environments for three abiotic stresses (cold, drought and nitrogen) using the SUNFLO crop model and phenotypic variations of three commercial varieties. The computed plant stress indicators better explain yield variation than descriptors at the climatic or crop levels. In those environments, we observed oil yield of 317 sunflower hybrids and regressed it with three selected stress indicators. The slopes of cold stress norm reaction were used as plasticity phenotypes in the following genome-wide association study. Among the 65 534 tested Single Nucleotide Polymorphisms (SNPs), we identified nine quantitative trait loci controlling oil yield plasticity to cold stress. Associated single nucleotide polymorphisms are localized in genes previously shown to be involved in cold stress responses: oligopeptide transporters, lipid transfer protein, cystatin, alternative oxidase or root development. This novel approach opens new perspectives to identify genomic regions involved in genotype-by-environment interaction of a complex traits to multiple stresses in realistic natural or agronomical conditions.

L'article original est publié par The American Society of Plant Biologists; International audience; After-ripening is the mechanism by which dormant seeds become nondormant during their dry storage after harvest. The absence of free water in mature seeds does not allow detectable metabolism; thus, the processes associated with dormancy release under these conditions are largely unknown. We show here that sunflower (Helianthus annuus) seed alleviation of dormancy during after-ripening is associated with mRNA oxidation and that this oxidation is prevented when seeds are maintained dormant. In vitro approaches demonstrate that mRNA oxidation results in artifacts in cDNA-amplified fragment length polymorphim analysis and alters protein translation. The oxidation of transcripts is not random but selective, and, using microarrays, we identified 24 stored mRNAs that became highly oxidized during after-ripening. Oxidized transcripts mainly correspond to genes involved in responses to stress and in cell signaling. Among them, protein phosphatase 2C PPH1, mitogen-activated protein kinase phosphatase 1, and phenyl ammonia lyase 1 were identified. We propose that targeted mRNA oxidation during dry after-ripening of dormant seeds could be a process that governs cell signaling toward germination in the early steps of seed imbibition.

De novo sequencing of complex genomes is one of the main challenges for researchers seeking high-quality reference sequences. Many de novo assemblies are based on short reads, producing fragmented genome sequences. Third-generation sequencing, with read lengths >10 kb, will improve the assembly of complex genomes, but these techniques require high-molecular-weight genomic DNA (gDNA), and gDNA extraction protocols used for obtaining smaller fragments for short-read sequencing are not suitable for this purpose. Methods of preparing gDNA for bacterial artificial chromosome (BAC) libraries could be adapted, but these approaches are time-consuming, and commercial kits for these methods are expensive. Here, we present a protocol for rapid, inexpensive extraction of high-molecular-weight gDNA from bacteria, plants, and animals. Our technique was validated using sunflower leaf samples, producing a mean read length of 12.6 kb and a maximum read length of 80 kb.

International audience; Water de fi cit in fl uences leaf transpiration rate and photosynthetic activity. The genotype-dependent response of the latter has not been assessed in sun fl ower ( Helianthus annuus L.), particularly during the reproductive period when grain fi lling and lipogenesis depend greatly on photosynthate availability. To evaluate genotypic responses to water de fi cit before and after fl owering, two greenhouse experiments were performed. Four genotypes two inbred lines (PSC8, XRQ) and two cultivars (Inedi, Melody) were subjected to progressive water de fi cit. Non-linear regression was used to calculate the soil water de fi cit threshold (FTSWt) at which processes (transpiration and photosynthetic activity) were affected by water de fi cit. In the vegetative growth stage, photosynthetic activity was affected at a lower mean value of FTSWt (0.39) than transpiration (0.55). However, in the reproductive stage, photosynthetic activity was more sensitive to soil water de fi cit (FTSWt = 0.45). We found a signi fi cant ( P = 0.02) effect of plant growth stage on the difference between photosynthesis and transpiration rate thresholds and, a signi fi cant ( P = 0.03) effect of leaf age on transpiration. Such results will improve phenotyping methods and provide paths for integrating genotypic variability into crop models.

A possible contribution of alterations in metabolic sequences involved in citrate catabolism, to intracellular accumulation and subsequent release of citrate was investigated in cluster roots of phosphorus (P)-deficient white lupin (Lupinus albus L.). Citrate accumulation during maturation of root clusters was associated with decreased levels of intracellular soluble Pi and ATP, and with reduced rates of respiration. Inhibitor studies with KCN and salicylhydroxamic acid (SHAM) suggest a reduced capacity of both the cytochrome pathway and of the alternative respiration with a concomitant decrease of immunochemically detectable protein levels of the alternative oxidase. Reduced respiration seems to be related to a general impairment of the respiratory system, rather than to limitation of respiratory substrates such as Pi and adenylates, as indicated by the absence of stimulatory effects of the uncoupler CCCP. The citrate/malate ratio in juvenile root clusters with high rates of respiration and low inherent levels of citrate accumulation was increased by short-term application (4–8 h) of azide and SHAM as respiration inhibitors. During maturation of root clusters, a shift from intracellular malic acid to citric acid accumulation was associated also with down-regulation of ATP citrate lyase (ACL), which catalyzes cleavage of citrate into acetyl-CoA and oxaloacetate with a putative function as anapleurotic source for the production of acetyl-CoA under P-deficient conditions. Inhibition of nitrate uptake and assimilation is a general response to P limitation in many plant species including white lupin. Reduced consumption of the amino acceptor 2-oxoglutaric acid as a product of citrate turnover may therefore contribute to increased citrate accumulation. Accordingly, artificial inhibition of nitrate reduction by localized application of tungstate significantly increased the citrate/malate ratio in juvenile root clusters. Lowering the cytosolic pH by external application of propionate stimulated citrate and malate exudation in non-cluster lateral roots and in developing root clusters. This effect was reverted by preincubation with phosphonate to buffer the cytosol. The results suggest that acidification of the cytosol may be an important factor, triggering the transient release of citrate and protons from mature root clusters in P-deficient white lupin.

In order to cope with phosphate deficiency, white lupin produces bottle-brushed like roots, so-called cluster or proteoid roots which are specialized in malate and citrate excretion. Young, developing cluster roots mainly excrete malate whereas mature cluster roots mainly release citrate. Mature proteoid roots excrete four to six times more carboxylates compared with juvenile proteoid roots. Using a cDNA-amplified restriction fragment length polymorphism (AFLP) approach we identified a gene coding for a putative ATP-citrate lyase (ACL) up-regulated in young cluster roots. Cloning of the lupin ACL revealed that plant ACL is constituted by two polypeptides (ACLA and ACLB) encoded by two different genes. This contrasts with the animal ACL, constituted of one polypeptide which covers ACLA and ACLB. The ACL function of the two lupin gene products has been demonstrated by heterologous expression in yeast. Both subunits are required for ACL activity. In lupin cluster roots, our results suggest that ACL activity could be responsible for the switch between malate and citrate excretion in the different developmental stages of cluster roots. In primary roots of lupin and maize, ACL activity was positively correlated with malate exudation. These results show that ACL is implicated in root exudation of organic acids and hence plays a novel role in addition to lipid synthesis. Our results suggest that in addition to lipid biosynthesis, in plants, ACL is implicated in malate excretion.

International audience; Under phosphorous deficiency, plants of white lupin (Lupinus albus L.) develop root clusters, which are also called proteoid roots due to their preferential presence in the Proteaceae. In their mature stage, these roots acidify the soil and excrete high amounts of carboxylates [up to 1.5 and 7 micromol (g FW)(-1) h(-1) of malate and citrate, respectively] enabling lupins to utilise sparingly available sources of phosphate. Using the amplified fragment length polymorphism (AFLP) technique, we identified genes predominantly expressed in juvenile and mature cluster roots. Transcripts for two enzymes involved in glycolysis, fructokinase and phosphoglucomutase, were identified in juvenile cluster roots and one, sucrose synthase, in mature cluster roots. In order to verify these observations we performed quantitative reverse transcription-polymerase chain reaction (RT-PCR) and could confirm the increased transcript level. Measurements of enzymatic activities showed that fructokinase and phosphoglucomutase activities increased in juvenile cluster roots, whereas sucrose synthase activity was maximal in mature cluster roots. These results indicate that formation of proteoid roots and citrate excretion increase sink strength locally. Production of citrate and inhibition of respiration are likely to result in an increased NADH/NAD+ ratio, which may be toxic for the plant. The fermentation pathway would allow oxidation of NADH by decarboxylation of pyruvate and subsequent reduction of the resulting acetaldehyde. Determination of alcohol dehydrogenase activity showed that this enzyme is strongly induced in mature proteoid roots. However, ethanol production was not increased, indicating that pyruvate is shunted to citrate synthesis and not to ethanol production.

Cluster root formation in white lupin (Lupinus albus L.) is induced mainly by phosphorus (P) starvation, and seems to be regulated by the endogenous P status of the plant. Increased formation of cluster roots, when indole acetic acid is supplied to the growth medium of P sufficient plants, and inhibitory effects of kinetin application suggest the involvement of endogenous phytohormones (auxins and cytokinins), which may act in an antagonistic manner in the P-starvation response. Phosphorus deficiency-induced adaptations of white lupin, involved in P acquisition and mobilization of sparingly available P sources, are predominantly confined to the cluster roots, and moreover to distinct stages during their development. Increased accumulation and exudation of citrate and a concomitant release of protons were found to be mainly restricted to mature root clusters after prolonged culture (3-4 weeks) under P-deficient conditions. Inhibition of citrate exudation by exogenous application of anion channel antagonists such as ethacrynic- and anthracene-9-carboxylic acids may indicate involvement of an anion channel. Phosphorus deficiency-induced accumulation and subsequent exudation of citric acid seems to be a consequence of both enhanced biosynthesis and reduced turnover of citric acid in the cluster root tissue, indicated by enhanced expression of sucrose synthase, fructokinase, phosphoglucomutase, phosphoenol-pyruvate carboxylase, but reduced activity of aconitase and slower root respiration. The release of acid phosphatase and of phenolic compounds (isoflavonoids) as well as the induction of a putative high-affinity P uptake system was more highly expressed in juvenile, mature and even senescent cluster regions than in apical zones of non-proteoid roots. An AFLP-cDNA library for cluster root-specific gene expression was constructed to assist in the identification of further genes involved in cluster root development. Copyright 2000 Annals of Botany Company

International audience; Orthodox seeds are living organisms that survive anhydrobiosis and may display dormancy, an inability to germinate at harvest. Seed germination potential can be acquired during a prolonged period of dry storage called after-ripening. The aim of this work was to determine if gene transcription is an underlying regulatory mechanism for dormancy alleviation during after-ripening. To identify changes in gene transcription strictly associated with the acquisition of germination potential but not with storage, we used seed storage at low relative humidity that maintains dormancy as control. Transcriptome profiling was performed using DNA microarray to compare change in gene transcript abundance between dormant (D), after-ripened non-dormant (ND) and after-ripened dormant seeds (control, C). Quantitative real-time polymerase chain reaction (qPCR) was used to confirm gene expression. Comparison between D and ND showed the differential expression of 115 probesets at cutoff values of twofold change (p,0.05). Comparisons between both D and C with ND in transcript abundance showed that only 13 transcripts, among 115, could be specific to dormancy alleviation. qPCR confirms the expression pattern of these transcripts but without significant variation between conditions. Here we show that sunflower seed dormancy alleviation in the dry state is not related to regulated changes in gene expression.

International audience; High water use efficiency (WUE) can be achieved by coordination of biomass accumulation and water consumption. WUE is physiologically and genetically linked to carbon isotope discrimination (CID) in leaves of plants. A population of 148 recombinant inbred lines (RILs) of sunflower derived from a cross between XRQ and PSC8 lines was studied to identify quantitative trait loci (QTL) controlling WUE and CID, and to compare QTL associated with these traits in different drought scenarios. We conducted greenhouse experiments in 2011 and 2012 by using 100 balances which provided a daily measurement of water transpired, and we determined WUE, CID, biomass and cumulative water transpired by plants. Wide phenotypic variability, significant genotypic effects, and significant negative correlations between WUE and CID were observed in both experiments. A total of nine QTL controlling WUE and eight controlling CID were identified across the two experiments. A QTL for phenotypic response controlling WUE and CID was also significantly identified. The QTL for WUE were specific to the drought scenarios, whereas the QTL for CID were independent of the drought scenarios and could be found in all the experiments. Our results showed that the stable genomic regions controlling CID were located on the linkage groups 06 and 13 (LG06 and LG13). Three QTL for CID were co-localized with the QTL for WUE, biomass and cumulative water transpired. We found that CID and WUE are highly correlated and have common genetic control. Interestingly, the genetic control of these traits showed an interaction with the environment (between the two drought scenarios and control conditions). Our results open a way for breeding higher WUE by using CID and marker-assisted approaches and therefore help to maintain the stability of sunflower crop production.

International audience; Gene regulatory networks (GRNs) govern phenotypic adaptations and reflect the trade-offs between physiological responses and evolutionary adaptation that act at different time-scales. To identify patterns of molecular function and genetic diversity in GRNs, we studied the drought response of the common sunflower, Helianthus annuus, and how the underlying GRN is related to its evolution. We examined the responses of 32 423 expressed sequences to drought and to abscisic acid (ABA) and selected 145 co-expressed transcripts. We characterized their regulatory relationships in nine kinetic studies based on different hormones. From this, we inferred a GRN by meta-analyses of a Gaussian graphical model and a random forest algorithm and studied the genetic differentiation among populations (FST ) at nodes. We identified two main hubs in the network that transport nitrate in guard cells. This suggests that nitrate transport is a critical aspect of the sunflower physiological response to drought. We observed that differentiation of the network genes in elite sunflower cultivars is correlated with their position and connectivity. This systems biology approach combined molecular data at different time-scales and identified important physiological processes. At the evolutionary level, we propose that network topology could influence responses to human selection and possibly adaptation to dry environments.

International audience; Association mapping and linkage mapping were used to identify quantitative trait loci (QTL) and/or causative mutations involved in the control of flowering time in cultivated sunflower Helianthus annuus. A panel of 384 inbred lines was phenotyped through testcrosses with two tester inbred lines across 15 location × year combinations. A recombinant inbred line (RIL) population comprising 273 lines was phenotyped both per se and through testcrosses with one or two testers in 16 location × year combinations. In the association mapping approach, kinship estimation using 5,923 single nucleotide polymorphisms was found to be the best covariate to correct for effects of panel structure. Linkage disequilibrium decay ranged from 0.08 to 0.26 cM for a threshold of 0.20, after correcting for structure effects, depending on the linkage group (LG) and the ancestry of inbred lines. A possible hitchhiking effect is hypothesized for LG10 and LG08. A total of 11 regions across 10 LGs were found to be associated with flowering time, and QTLs were mapped on 11 LGs in the RIL population. Whereas eight regions were demonstrated to be common between the two approaches, the linkage disequilibrium approach did not detect a documented QTL that was confirmed using the linkage mapping approach.

Plant or soil water status is required in many scientific fields to understand plant responses to drought. Because the transcriptomic response to abiotic conditions, such as water deficit, reflects plant water status, genomic tools could be used to develop a new type of molecular biomarker. Using the sunflower (Helianthus annuus L.) as a model species to study the transcriptomic response to water deficit both in greenhouse and field conditions, we specifically identified three genes that showed an expression pattern highly correlated to plant water status as estimated by the pre-dawn leaf water potential, fraction of transpirable soil water, soil water content or fraction of total soil water in controlled conditions. We developed a generalized linear model to estimate these classical water status indicators from the expression levels of the three selected genes under controlled conditions. This estimation was independent of the four tested genotypes and the stage (pre- or post-flowering) of the plant. We further validated this gene expression biomarker under field conditions for four genotypes in three different trials, over a large range of water status, and we were able to correct their expression values for a large diurnal sampling period.

International audience; Major gene resistance to sunflower downy mildew (Plasmopara halstedii) races 304 and 314 was found to segregate independently from the resistance to races 334, 307 and 304 determined by the gene Pl2, already positioned on Linkage Group (LG) 8 of sunflower molecular maps. Using a consensus SSR-SNP map constructed from the INEDI RIL population and a new RIL population FU × PAZ2, the positions of Pl2 and Pl5 were confirmed and the new gene, denoted Pl21, was mapped on LG13, at 8 cM from Pl5. The two RIL populations were observed for their quantitative resistance to downy mildew in the field and both indicated the existence of a QTL on LG8 at 20-40 cM from the major resistance gene cluster. In addition, for the INEDI population, a strong QTL on LG10, reported previously, was confirmed and a third QTL was mapped on LG7. A growth chamber test methodology, significantly correlated with field results, also revealed the major QTL on LG10, explaining 65 % of variability. This QTL mapped in the same area as a gene involved in stomatal opening and root growth, which may be suggested as a possible candidate to explain the control of this character. These results indicate that it should be possible to combine major genes and other resistance mechanisms, a strategy that could help to improve durability of sunflower resistance to downy mildew.

Sunflower (Helianthus annuus L.) contains tocopherol, a non-enzymatic antioxidant known as lipid-soluble vitamin E, and phytosterol, with interesting properties, which can result in decreased risk of chronic diseases in humans and with several beneficial effects in plants. The genetic control of tocopherol and phytosterol content in a population of 123 recombinant inbred lines of sunflower was studied through quantitative trait loci (QTL) analysis using 190 simple sequence repeats and a gene-based linkage map. Seven experiments were conducted in different environments in France and Iran during 2007 and 2008. Each experiment consisted of three replications. Means over all environments were used for QTL mapping. Five QTL for total tocopherol content on linkage groups 1, 8, 10 and 14 accounted for 45% of phenotypic variation, whereas four QTL for total phytosterol content on linkage groups 1, 2, 16 and 17 explained 27% of the phenotypic variation. GST, PAT2, SFH3 and POD genes showed co-localization with QTL for total phytosterol content. SMT2 is also mapped on linkage group 17 near the QTL of total phytosterol content. Four candidate genes, VTE4, HPPD, GST and Droug1, exhibited co-localization with QTL for total tocopherol content. The candidate genes associated with tocopherol and phytosterol, especially HPPD, VTE4 and SMT2, could be used for alternation of the tocopherol and phytosterol content of sunflower seeds through the development of functional markers.