Search

Managing Salmonella enterica Enteritidis (SE) carriage in chicken is necessary to ensure human food safety and enhance chicken breeding viability. Salmonella can contaminate poultry products, causing human foodborne disease and economic losses for farmers. Both genetic selection for a decreased carriage and gut microbiota modulation strategies could reduce Salmonella propagation in farms.Two-hundred and twenty animals from the White Leghorn inbred lines N and 61 were raised together on floor, infected by SE at 7 days of age, transferred into isolators to prevent oro-fecal recontamination and euthanized at 19 days. Caecal content DNA was used to measure individual Salmonella counts (ISC) by droplet digital PCR. A RNA sequencing approach was used to measure gene expression levels in caecal tonsils after infection of 48 chicks with low or high ISC.The analysis between lines identified 7516 differentially expressed genes (DEGs) corresponding to 62 enriched Gene Ontology (GO) Biological Processes (BP) terms. A comparison between low and high carriers allowed us to identify 97 DEGs and 23 enriched GO BP terms within line 61, and 1034 DEGs and 288 enriched GO BP terms within line N. Among these genes, we identified several candidate genes based on their putative functions, including FUT2 or MUC4, which could be involved in the control of SE infection, maybe through interactions with commensal bacteria. Altogether, we were able to identify several genes and pathways associated with differences in SE carriage level. These results are discussed in relation to individual caecal microbiota compositions, obtained for the same animals in a previous study, which may interact with host gene expression levels for the control of the caecal SE load.

BackgroundInteractions between the gut microbiota and the immune system may be involved in the vaccine response. In the present study, we studied the correlations between caecal microbiota composition and the immune response in six experimental laying hen lines harboring different haplotypes at the Major Histocompatibility complex (MHC), 7 weeks after their first vaccination against the infectious bronchitis virus (IBV). Two lines were previously considered as high responders (HR) to IBV vaccination and two other ones as low responders (LR). We explored to what extent the gut microbiota could be related to this variability through the characterization of caecal bacterial communities with a 16S rRNA gene amplicon sequencing approach, one week after an IBV infectious challenge. ResultsWe observed significant effects of both the vaccination and the genetic line on the microbiota, with lower bacterial richness in vaccinated chickens, especially in the Ruminococcaceae family. We also observed dissimilar caecal communities among the different MHC lines, and among the vaccinated and non-vaccinated animals. The effect of vaccination was similar in all the lines, with a reduced abundance of OTUs from the Ruminococcacea UCG-014 and Faecalibacterium genera, and an increased abundance of OTUs from the Eisenbergiella genus. The main association between the microbiota and the immune phenotypes implied TCRϒδ expression on TCRϒδ+ T cells, which especially shared negative associations with OTUs from the Escherichia-Shigella genus. Finally, opposing the HR and LR groups, we identified 3 OTUs more abundant in the HR group, from the Tyzzeralla family, the Angelakisella genus, and to a lower extent the Flavonifractor family. Conversely, an OTU from the CAG-352 genus was more abundant in the LR group. ConclusionsWe assessed that the caecal microbiota composition is associated with the IBV vaccine response in laying chickens, and that TCRϒδ expression on TCRϒδ+ T cells appears as one of the main components potentially involved in this interaction.

Background Salmonella Enteritidis (SE) is one of the major causes of human foodborne intoxication through the consumption of contaminated poultry products. Genetic selection of animals more resistant to Salmonella carriage and the modulation of gut microbiota are two promising ways of decreasing individual Salmonella carriage. This study aims to identify the main genetic and microbial factors controlling the individual levels of Salmonella carriage in chickens (Gallus gallus) in controlled experimental conditions. Two-hundred and forty animals from the White Leghorn inbred lines, N and 61, were infected by SE at 7 days of age. After infection, animals were kept in isolators to reduce the recontamination of birds by Salmonella. Caecal contents were sampled at 12 days post-infection and used for DNA extraction. Microbiota DNA was used to measure individual counts of SE by digital PCR and to determine the bacterial taxonomic composition through a 16S rRNA gene high-throughput sequencing approach. Results Results confirmed that the N line is more resistant to Salmonella carriage than the 61 line, and that intra-line variability is higher for the 61 line. Furthermore, the 16S analysis showed strong significant differences in microbiota taxonomic composition between the two lines. Out of 617 Operational Taxonomic Units (OTUs), over 390 were differentially abundant between the two lines. Furthermore, within the 61 line, we found a difference in the microbiota taxonomic composition between high and low Salmonella carriers, with 39 differentially abundant OTUs. Finally, via metagenome functional prediction based on 16S data, we identified several metabolic pathways potentially associated to microbiota taxonomic differences (e.g. butyrate metabolism) between high and low carriers. Conclusions Overall, this study demonstrates that the caecal microbiota composition of the N and 61 lines is influenced by the host genetics, which could be one of the reasons why these lines differ for their Salmonella carriage in experimental infection conditions.

Improving the digestive efficiency of broiler chickens (Gallus gallus) could reduce organic waste, increase the use of alternative feed not used for human consumption and reduce the impact of feed in production costs. By selecting chicken lines divergently for their digestive efficiency, we showed previously that digestive efficiency is under genetic control and that the two resulting divergent lines, D+ (high digestive efficiency or "digestibility +") and D- (low digestive efficiency or "digestibility -"), also differ for the abundance of specific bacteria in their caeca. Here we perform a more extensive census of the bacteria present in the digestive microbiota of 60 chickens selected for their low apparent metabolizable energy corrected for nitrogen balance (AMEn-) or high (AMEn+) digestive efficiency in a [D+ x D-] F8 progeny of 200 individuals. We sequenced the 16S rRNA genes of the ileal, jejunal and caecal microbiotas, and compared the compositions and predicted functions of microbiotas from the different intestinal segments for 20 AMEn+ and 19 AMEn- birds. The intestinal segment of origin was the main factor structuring the samples. The caecal microbiota was the most impacted by the differences in digestive efficiency, with 41 bacterial species with abundances differing between highly and poorly efficient birds. Furthermore, we predicted that the caecal microbiota of efficient birds might be enriched in genes contributing to the degradation of short chain fatty acids (SCFA) from non-starch polysaccharides. These results confirm the impact of the genetic selection led on digestibility on the caecal microbiota taxonomic composition. They open the way toward the identification of specific, causal genes of the host controlling variations in the abundances of bacterial taxons.

International audience

National audience; Le microbiote intestinal des animaux d’élevage, dont le porc et le poulet, forme une symbiose avec son hôte. Il remplit de multiples fonctions essentielles à sa santé : notamment digestion et production de métabolites essentiels, effet barrière contre les pathogènes intestinaux, développement du tractus digestif et du système immunitaire et régulation de l’immunité tout au long de la vie de l’animal. Par ailleurs des variations de composition du microbiote intestinal sont associées aux variations de nombreux caractères d’intérêt des animaux d’élevage, en particulier la santé. Moduler la composition du microbiote intestinal pour améliorer la santé des animaux par différents leviers, notamment l’alimentation, les conditions d’élevage et la génétique est une voie de recherche prometteuse. Cette présentation expliquera ce qu’apportent les méthodes de séquençage à haut débit à la connaissance du microbiote intestinal du porc et du poulet et présentera les perspectives appliquées qui sont envisageables à l’heure actuelle pour améliorer leur santé par la modulation du microbiote intestinal.

The increasing cost of conventional feedstuffs has bolstered interest in genetic selection for digestive efficiency (DE), a component of feed efficiency, assessed by apparent metabolisable energy corrected to zero nitrogen retention (AMEn). However, its measurement is time-consuming and constraining, and its relationship with metabolic efficiency poorly understood. To simplify selection for this trait, we searched for indirect metabolic biomarkers through an analysis of the serum metabolome using nuclear magnetic resonance (1H NMR). A partial least squares (PLS) model including six amino acids and two derivatives from butyrate predicted 59% of AMEn variability. Moreover, to increase our knowledge of the molecular mechanisms controlling DE, we investigated 1H NMR metabolomes of ileal, caecal, and serum contents by fitting canonical sparse PLS. This analysis revealed strong associations between metabolites and DE. Models based on the ileal, caecal, and serum metabolome respectively explained 77%, 78%, and 74% of the variability of AMEn and its constitutive components (utilisation of starch, lipids, and nitrogen). In our conditions, the metabolites presenting the strongest associations with AMEn were proline in the serum, fumarate in the ileum and glucose in caeca. This study shows that serum metabolomics offers new opportunities to predict chicken DE.

International audience

La sélection des pondeuses commerciales repose sur un ensemble de critères où les paramètres de qualité de l’œuf prennent une importance croissante. Les caractères tels que le poids de l’œuf, la solidité de coquille (mesurée directement ou indirectement) la proportion de jaune et la qualité du blanc (unités Haugh) présentent une assez forte variabilité génétique (héritabilité de 0,30 à 0,60). Malgré une corrélation négative entre le poids d’œuf et le nombre d’œufs, la combinaison de la sélection en lignée pure et du croisement a permis d’améliorer le nombre d’œufs tout en stabilisant le poids d’œuf à un niveau élevé (62 g en moyenne). La variabilité génétique de la couleur de coquille, de la proportion de jaune et de la composition en lipides du jaune a été explorée, chez la poule comme chez la caille, par la sélection expérimentale et l’étude de races locales. Cependant, les gènes contrôlant les caractères de ponte et de la qualité technologique de l’œuf sont encore mal connus. Quelques protocoles de détection de QTL ont permis d’identifier des régions chromosomiques contrôlant une part significative de la variabilité de ces caractères, chez la poule et la caille. Récemment des gènes majeurs ont été identifiés comme l’enzyme FMO3 responsable de l’odeur de poisson d’œufs de poule nourries au tourteau de soja. De nombreuses études portent sur la la qualité bactériologique des oeufs, avec une priorité donnée au portage de salmonelles, pour laquelle plusieurs gènes ont été identifiés (SLC11A1, TLR4). Ces perspectives sont encourageantes pour répondre aux demandes croissantes du consommateur en matière de qualité des produits.

Fire blight, caused by the bacterial pathogen Erwinia amylovora, is one of the most destructive diseases of apple (Malus x domestica). In order to analyse the genetic determinism of resistance to fire blight in apple, a quantitative trait analysis (QTL) approach was used. A F1 progeny of 164 individuals derived from a cross between the apple cultivars `Prima¿ and `Fiesta¿ was inoculated in greenhouse conditions. Seven copies per genotype were used. The length of the necrosis observed on shoots was scored 7 and 14 days after inoculation. The MapQTL software was used for QTL analyses, using two previously built maps of the parents, and the symptoms scored on shoots. Digenic interactions between all pairwise combinations of genetic markers were tested using a two-way ANOVA model with the SAS software. QTL were detected at the same locations both 7 and 14 days after inoculation. Two weak effect QTL deriving from `Prima¿ were detected on linkage groups (LG) 3 and LG16. One strong effect QTL deriving from `Fiesta¿ was detected on LG7 that explained 46.6% of the phenotypic variation observed in the progeny. Two additional significant (P

Le développement de systèmes d’élevage avicoles durables passe par la recherche d’animaux plus résilients et capables d’assimiler des aliments de moindre qualité ou en tout cas de qualité plus variable. Alors que l’efficacité métabolique des animaux a déjà été largement améliorée, leur efficacité digestive a été moins explorée et exploitée. Si les travaux de génétique ont montré les possibilités de sélection pour l’efficacité digestive, les gènes et les mutations à l’origine de la variabilité de ce caractère complexe restent méconnus. Leur connaissance permettrait d’améliorer l’efficacité de la sélection génétique, en intégrant dans le choix des animaux une information moléculaire. Par ailleurs, disposer de tels biomarqueurs permettrait d’étudier plus finement l’influence des facteurs d’élevage et de développer des stratégies ciblées sur l’amélioration de l’efficacité digestive. Les travaux de génomique positionnelle ont d’ores et déjà permis d’identifier plusieurs régions du génome (QTL) contrôlant l’efficacité digestive et les caractères associés. Ils vont s’enrichir d’approches de génomique fonctionnelle pour préciser les gènes et les voies métaboliques impliqués et de métagénomique pour caractériser les éventuelles relations entre variation d’efficacité digestive et de composition du microbiote.

absent

Background: Improving digestive efficiency is a major goal in poultry production, to reduce production costs, make possible the use of alternative feedstuffs and decrease the volume of manure produced. Since measuring digestive efficiency is difficult, identifying molecular markers associated with genes controlling this trait would be a valuable tool for selection. Detection of QTL (quantitative trait loci) was undertaken on 820 meat-type chickens in a F2 cross between D- and D+ lines divergently selected on low or high AMEn (apparent metabolizable energy value of diet corrected to 0 nitrogen balance) measured at three weeks in animals fed a low-quality diet. Birds were measured for 13 traits characterizing digestive efficiency (AMEn, coefficients of digestive utilization of starch, lipids, proteins and dry matter (CDUS, CDUL, CDUP, CDUDM)), anatomy of the digestive tract (relative weights of the proventriculus, gizzard and intestine and proventriculus plus gizzard (RPW, RGW, RIW, RPGW), relative length and density of the intestine (RIL, ID), ratio of proventriculus and gizzard to intestine weight (PG/I); and body weight at 23 days of age. Animals were genotyped for 6000 SNPs (single nucleotide polymorphisms) distributed on 28 autosomes, the Z chromosome and one unassigned linkage group. Results: Nine QTL for digestive efficiency traits, 11 QTL for anatomy-related traits and two QTL for body weight at 23 days of age were detected. On chromosome 20, two significant QTL at the genome level co-localized for CDUS and CDUDM, i.e. two traits that are highly correlated genetically. Moreover, on chromosome 16, chromosome-wide QTL for AMEn, CDUS, CDUDM and CDUP, on chromosomes 23 and 26, chromosome-wide QTL for CDUS, on chromosomes 16 and 26, co-localized QTL for digestive efficiency and the ratio of intestine length to body weight and on chromosome 27 a chromosome-wide QTL for CDUDM were identified. Conclusions: This study identified several regions of the chicken genome involved in the control of digestive efficiency. Further studies are necessary to identify the underlying genes and to validate these in commercial populations and breeding environments.

Le microbiote digestif est un symbionte dont on a démontré l’implication essentielle dans la régulation de la physiologie de l’hôte qui l’héberge pour la digestion, le métabolisme et l’immunité. Ce microbiote a en particulier un rôle majeur, et jusqu’ici sous-estimé, sur la santé de son hôte et participe à son adaptabilité à des environnements changeants. Les technologies de séquençage à haut débit devraient faciliter l’inclusion de cet écosystème complexe dans le phénotypage des animaux d’élevage. Il faut pour ce faire prendre en compte sa composition, sa diversité, sa stabilité au cours du temps et ses variations selon différents facteurs comme le génotype de l’hôte, l’alimentation et les conditions d’élevage. Ces connaissances devraient permettre de définir les propriétés d’un microbiote digestif « normal » associé à une bonne santé, d’en préciser les dysfonctionnements et de comprendre leurs répercussions sur l’état de santé de l’animal. Cela conduira à identifier des marqueurs de diagnostic et de pronostic de ces dysfonctionnements, fournissant des moyens de contrôle nouveaux de la santé en élevage, appréhendée en terme d’optimisation des rendements de production, de résistance aux maladies et de maintien du bien-être. A terme, le contrôle du microbiote digestif par l’alimentation, la génétique ou les pratiques d’élevage permettra de favoriser l’adaptation des animaux à leur environnement d’élevage en améliorant leur robustesse, de limiter l’utilisation d’antibiotiques et de préserver la sécurité sanitaire des aliments en évitant le développement de bactéries pathogènes pour l’Homme. L’ampleur des données à générer et à analyser pour les différentes espèces d’intérêt reste le défi à relever., The gastrointestinal microbiota is a symbiont deeply involved in the physiology of its host through the regulation of digestion, metabolism and immunity and consequently in the host adaptability to changing environments. In particular, the role of the gastrointestinal microbiota in maintaining host health is important and has been under-estimated until now. It is an inherent component of the animal that should be described when phenotyping animals. The rapid development of high throughput sequencing technologies facilitates the description of this complex ecosystem in livestock species. For each animal species, its description should include the measure of its composition, diversity and stability over time and the measure of its variability according to different factors such as genotype, nutrition and breeding conditions. This knowledge should allow defining the properties of a ‘normal’ gastrointestinal microbiota associated with health, identifying the microbiota dysfunctions and understanding their consequences on host physiology. It will lead to the identification of diagnostic and prognostic markers for gastrointestinal dysbiosis thus furnishing new tools for promoting health. Finally, the manipulation of the gastrointestinal microbiota will constitute a way to better adapt animals to their environment by improving their robustness, limit the use of antibiotics and improve food safety by preventing the development of bacteria threatening human health. Due to the high quantity of data to be acquired and analyzed, achieving these objectives for each species of interest remains a challenge.

International audience; Increasing feed efficiency of broiler chickens by selective breeding could lead to decreased feed cost and reduced environmental impact of poultry production. At INRA, two broiler chicken lines (D+/D-) were divergently selected for their digestive efficiency. Strong differences were shown between both lines for the anatomy and histology of the digestive tract, and for the intestinal microbiota composition. In the present study, we investigated whether this selection also had an effect on susceptibility to colibacillosis, which is one of the main causes of economic losses in poultry production. The broiler lines D+/D- were challenged with an avian pathogenic Escherichia coli strain. A first experiment was conducted to assess the 50% lethal dose by subcutaneous infection of hatchlings, whereas a second experiment reproduced colibacillosis by infecting air sacs of 23-day-old chicks. The 50% lethal dose was very low for both lines. However, the line with the higher digestive efficiency (D+) was the less susceptible to colibacillosis. This result is interesting for selection purposes and opens the way to integrative genetic studies of the interactions between digestion efficiency and resistance to colibacillosis.

absent

absent

absent

SummaryIn higher plants, soluble sugars are mainly present as sucrose, glucose, and fructose [1]. Sugar allocation is based on both source-to-sink transport and intracellular transport between the different organelles [2, 3] and depends on actual plant requirements [4]. Under abiotic stress conditions, such as nitrogen limitation, carbohydrates accumulate in plant cells [5]. Despite an increasing number of genetic studies [6, 7], the genetic architecture determining carbohydrate composition is poorly known. Using a quantitative genetics approach, we determined that the carrier protein SWEET17 is a major factor controlling fructose content in Arabidopsis leaves. We observed that when SWEET17 expression is reduced, either by induced or natural variation, fructose accumulates in leaves, suggesting an enhanced storage capacity. Subcellular localization of SWEET17-GFP to the tonoplast and functional expression in Xenopus oocytes showed that SWEET17 is the first vacuolar fructose transporter to be characterized in plants. Physiological studies in planta provide evidence that SWEET17 acts to export fructose out of the vacuole. Overall, our results suggest that natural variation in leaf fructose levels is controlled by the vacuolar fructose transporter SWEET17. SWEET17 is highly conserved across the plant kingdom; thus, these findings offer future possibilities to modify carbohydrate partitioning in crops.

Session : Génétique, sélection et reproduction; Améliorer l'efficacité digestive des poulets est un enjeu majeur qui permettrait non seulement de limiter les coûts de production mais aussi de valoriser efficacement des matières premières alternatives tout en réduisant le volume des rejets. La mesure de l'efficacité digestive étant relativement lourde, disposer de marqueurs génétiques impliqués dans la variabilité de ce caractère serait un outil précieux pour la sélection. Une première recherche de QTL est présentée, impliquant 820 poulets de chair issus de 6 familles dans un croisement de type F2 entre les lignées D- et D+, sélectionnées de façon divergente sur l'énergie métabolisable apparente corrigée pour un bilan azoté nul (EMAn), mesurée à 3 semaines d'âge avec un régime contenant un blé de mauvaise qualité (variété Rialto). Cinq critères de digestibilité ont été mesurés : l'EMAn, des coefficients de digestibilité de la matière sèche (CUDMS), de l'amidon (CUDAmidon), des protéines (CUDProtéines) et des lipides (CUDLipides). Les animaux ont tous été génotypés avec 6000 marqueurs SNP, choisis parmi 57636 marqueurs comme les plus informatifs pour notre dispositif. Au total, les marqueurs sont répartis sur 28 autosomes, 1 groupe de liaison et le chromosome Z. En utilisant le logiciel QTLMap (développé à l'INRA) pour des analyses de liaison en cartographie d'intervalle, 9 QTL (locus de caractère quantitatif) ont été détectés. Sur le chromosome 20, deux QTL significatifs au niveau du génome ont été observés à la même position pour CUDAmidon et CUDMS, deux caractères fortement corrélés. De même, des QTL significatifs au niveau du chromosome pour EMAn, CUDAmidon, CUDMS, CUDProtéines sont localisés dans la même région du chromosome 16. Sur les chromosomes 23 et 26, des QTL significatifs au niveau du chromosome ont été identifiés pour CUDAmidon, celui porté par le chromosome 26 étant situé à 1 cM d'un QTL contrôlant le rapport de la longueur de l'intestin au poids vif détecté sur ce même dispositif. Sur le chromosome 27, un QTL significatif au niveau du chromosome est détecté pour CUDMS.

Chantier qualité GA; Salmonella enterica serotypes Enteritidis and Typhimurium and Campylobacter jejuni are responsible for most cases of food poisoning in Europe. These bacteria do not cause severe disease symptoms in chicken, but they are easily propagated by symptomless chicken carriers which cannot be easily isolated. This animal tolerance is detrimental to food safety. In this particular case, increasing animal's resistance is not sufficient, since some animals considered as resistant are able to carry bacteria during several weeks without displaying disease symptoms. We review studies aimed at evaluating the resistance of chicken to Salmonella and Campylobacter intestinal colonization, either a few days or several weeks after infection. While studies of the genetic control of Campylobacter colonization are only beginning, mostly due to technical difficulties in infection protocols, genetic studies of Salmonella colonization have been conducted for now more than 20 years. They have initially reported an estimation of the genetic parameters associated with resistance to Salmonella colonization and are now aimed at identifying the genomic regions controlling variation of this trait in experimental lines and commercial populations. With the advent of high-throughput genomics, we are closer than ever to identify the true genes controlling resistance to Enterobacteria colonization in chicken. The comparison of genes involved in early resistance to intestinal colonization with genes controlling resistance to bacteria persistence several weeks after infection (i.e., carrier-state) should soon highlight the differences between the molecular mechanisms underlying those two distinct phenotypes. It will also be highly interesting to compare the genes or genomic regions controlling Campylobacter and Salmonella, in order to evaluate the feasibility of a selection conducted on both bacteria simultaneously.

Background Salmonella propagation by apparently healthy chicken and subsequent food security concerns could be decreased by the selection and use of chicken lines more resistant to carrier-state. In the present study we applied the first steps of the genomic selection methodology to assess the interest of including genetic markers for the genetic evaluation of hen lines infected with Salmonella Enteritidis. Methods We studied commercial laying hen lines divergently selected for resistance to Salmonella carrier-state at two different ages. A total of 600 animals were typed with 1536 SNP markers and artificially infected with S. Enteritidis. Phenotypes were collected four weeks (young animals) or five weeks (adults) later. Two types of variance component analyses, including or not including SNP data, were performed and compared. All variance components were estimated by Bayesian methods and Gibbs sampling. Results The comparison of both genetic analyses shows that SNP are efficient in capturing genetic variation, although none of them captures a large affect on the traits studied. Average accuracies do not change between analyses, showing that using SNP data does not really increase information. Conclusions These preliminary results show that genomic selection for Salmonella carrier-state resistance in laying hens is promising, although a denser SNP coverage of the genome on a higher number of animals is needed to assess its feasibility and efficiency compared to classical pedigree evaluation.

Synthèse; absent

Chantier qualité GA; International audience; Salmonella propagation by apparently healthy chickens could be decreased by the selection and use of chicken lines that are more resistant to carrier state. Using a reduced set of markers, this study investigates, for the first time to the authors’ knowledge, the feasibility of a genomic selection approach for resistance to carrier state in hen lines. In this study, commercial laying hen lines were divergently selected for resistance to Salmonella carrier state at 2 different ages: young chicks and adults at the peak of lay. A total of 600 birds were typed with 831 informative SNP markers and artificially infected with Salmonella Enteritidis. Phenotypes were collected 28 d (389 young animals) or 38 d (208 adults) after infection. Two types of variance component analyses, including SNP data or not, were performed and compared. The set of SNP used was efficient in capturing a large part of the genetic variation. Average accuracies from mixed model equations did not change between analyses, showing that using SNP data does not increase information in this data set. These results confirm that genomic selection for Salmonella carrier state resistance in laying hens is promising. Nevertheless, a denser SNP coverage of the genome on a greater number of animals is still needed to assess its feasibility and efficiency.

Chantier qualité GA; International audience; Chicken's ability to carry Salmonella without displaying disease symptoms leads to an invisible propagation of Salmonella in poultry stocks. Using chicken lines more resistant to carrier state could improve both animal health and food safety. Previous studies identified several QTL for resistance to carrier state. To improve genome coverage and QTL detection power we produced a new set of 480 informative SNP markers and genotyped a larger number of animals. Ten additional microchromosomes could be covered when compared with previous studies. These new data led to the identification of 18 QTL significant at the chromosome-wide level. The only QTL significant at the genome-wide level were identified on microchromosomes 14 and 22 and have never been identified previously. Using a higher number of animals improved the power and the precision of QTL detection. Some of the QTL newly identified are located close to candidate genes or microsatellite markers previously identified for their involvement in the genetic control of resistance to Salmonella, which confirms their interest for selection purposes.

Chantier qualité GA; International audience; Improving the fowl's natural ability to clear Salmonella from their body is important in reducing disease prevalence in poultry flocks, as recommended by a recent regulation of the European Commission. It may be efficient, as expected from estimation of heritability coefficients : 0.16 in chicks and 0.18 for global contamination of hens. The animal's age has to be considered since the genetic correlation between resistances at the two ages is negative. Selecting two series of divergent lines for increased or decreased resistance, after inoculation at one week of age (chick resistance) or at the peak of lay (adult resistance) confirmed the efficiency at least of selection for the adult resistance. In parallel, genes controlling variations to Salmonella resistance were researched and several QTLs identified in crosses between experimental lines and, for some of them, confirmed in commercial lines. Thanks to the derivation of a model of Salmonella propagation within a flock, it has been shown that a combination of vaccination and genetic selection can result in very low percentage of contamination.

National audience

Chantier qualité GA; The ability of chickens to carry Salmonella without displaying disease symptoms is responsible for Salmonella propagation in poultry stocks and for subsequent human contamination through the consumption of contaminated eggs or meat. The selection of animals more resistant to carrier state might be a way to decrease the propagation of Salmonella in poultry stocks and its transmission to humans. Five QTL controlling variation for resistance to carrier state in a chicken F(2) progeny derived from the White Leghorn inbred lines N and 6(1) had been previously identified using a selective genotyping approach. Here, a second analysis on the whole progeny was performed, which led to the confirmation of two QTL on chromosomes 2 and 16. To assess the utility of these genomic regions for selection in commercial lines, we tested them together with other QTL identified in an [Nx6(1)] x N backcross progeny and with the candidate genes SLC11A1 and TLR4. We used a commercial line divergently selected for either low or high carrier-state resistance both in young chicks and in adult hens. In divergent chick lines, one QTL on chromosome 1 and one in the SLC11A1 region were significantly associated with carrier-state resistance variations; in divergent adult lines, one QTL located in the major histocompatibility complex on chromosome 16 and one in the SLC11A1 region were involved in these variations. Genetic studies conducted on experimental lines can therefore be of potential interest for marker-assisted selection in commercial lines.

Salmonellosis is a frequent disease in poultry stocks, caused by several serotypes of the bacterial species Salmonella enterica and sometimes transmitted to humans through the consumption of contaminated meat or eggs. Symptom-free carriers of the bacteria contribute greatly to the propagation of the disease in poultry stocks. So far, several candidate genes and quantitative trait loci (QTL) for resistance to carrier state or to acute disease have been identified using artificial infection of S. enterica serovar Enteritidis or S. enterica serovar Typhimurium strains in diverse genetic backgrounds, with several different infection procedures and phenotypic assessment protocols. This diversity in experimental conditions has led to a complex sum of results, but allows a more complete description of the disease. Comparisons among studies show that genes controlling resistance to Salmonella differ according to the chicken line studied, the trait assessed and the chicken's age. The loci identified are located on 25 of the 38 chicken autosomal chromosomes. Some of these loci are clustered in several genomic regions, indicating the possibility of a common genetic control for different models. In particular, the genomic regions carrying the candidate genes TLR4 and SLC11A1, the Major Histocompatibility Complex (MHC) and the QTL SAL1 are interesting for more in-depth studies. This article reviews the main Salmonella infection models and chicken lines studied under a historical perspective and then the candidate genes and QTL identified so far.

Posttranscriptional regulation of a variety of mRNAs by small 21- to 24-nucleotide RNAs, notably the microRNAs (miRNAs), is emerging as a novel developmental mechanism. In legumes like the model Medicago truncatula, roots are able to develop a de novo meristem through the symbiotic interaction with nitrogen-fixing rhizobia. We used deep sequencing of small RNAs from root apexes and nodules of M. truncatula to identify 100 novel candidate miRNAs encoded by 265 hairpin precursors. New atypical precursor classes producing only specific 21- and 24-nucleotide small RNAs were found. Statistical analysis on sequencing reads abundance revealed specific miRNA isoforms in a same family showing contrasting expression patterns between nodules and root apexes. The differentially expressed conserved and nonconserved miRNAs may target a large variety of mRNAs. In root nodules, which show diverse cell types ranging from a persistent meristem to a fully differentiated central region, we discovered miRNAs spatially enriched in nodule meristematic tissues, vascular bundles, and bacterial infection zones using in situ hybridization. Spatial regulation of miRNAs may determine specialization of regulatory RNA networks in plant differentiation processes, such as root nodule formation.

Le but du programme est de combler les déficits en marqueurs observés pour trois espèces aviaires : la caille, le canard et la poule. La stratégie choisie est l'obtention, à partir de plusieurs individus de lignées d'intérêt, de SNP (Single Nucleotide Polymorphism, polymorphisme d'un nucléotide) par une nouvelle technologie de séquençage à haut débit (séquenceur 454 GS-FLX, Roche). Nous séquençons des représentations réduites du génome, en sélectionnant d'une part des fragments de restriction d'ADN génomique - les mêmes chez tous les individus - et d'autre part les transcrits qui représentent globalement la partie du génome correspondant aux gènes exprimés. Ces expérimentations sont réalisées à partir d'échantillons d'ADN ou d'ARN issus d'individus de lignées à l'origine de croisements existants, pour chacune des trois espèces. Les données générées par plusieurs "runs" de séquence seront traitées in silico : contigage à haut débit, recherche de SNP, comparaison avec les banques de séquences connues...En plus de l'intérêt que représente la production d'un très grand nombre de SNP nouveaux, cette technologie devrait permettre de mieux séquencer les régions riches en (G+C) correspondant aux plus petits des microchromosomes pour lesquels il n'y a pas de séquence chez la poule. La comparaison des séquences des transcrits obtenues chez la caille et le canard avec la séquence du génome de la poule permettra d'établir une "cartographie virtuelle" des SNP obtenus, grâce à la grande conservation de synténie existant entre ces trois espèces.

Des QTL aux gènes de résistance au portage de salmonelles chez le poulet

RÉSUMÉ Le but du programme est de combler les déficits en marqueurs observés pour trois espèces aviaires : la caille, le canard et la poule. La stratégie choisie est l'obtention, à partir de plusieurs individus de lignées d'intérêt, de SNP (Single Nucleotide Polymorphism, polymorphisme d'un nucléotide) par une nouvelle technologie de séquençage à haut débit (séquenceur 454 GS-FLX, Roche). Nous séquençons des représentations réduites du génome, en sélectionnant d'une part des fragments de restriction d'ADN génomique - les mêmes chez tous les individus - et d'autre part les transcrits qui représentent globalement la partie du génome correspondant aux gènes exprimés. Ces expérimentations sont réalisées à partir d'échantillons d'ADN ou d'ARN issus d'individus de lignées à l'origine de croisements existants, pour chacune des trois espèces. Les données générées par plusieurs "runs" de séquence seront traitées in silico : contigage à haut débit, recherche de SNP, comparaison avec les banques de séquences connues... En plus de l'intérêt que représente la production d'un très grand nombre de SNP nouveaux, cette technologie devrait permettre de mieux séquencer les régions riches en (G+C) correspondant aux plus petits des microchromosomes pour lesquels il n'y a pas de séquence chez la poule. La comparaison des séquences des transcrits obtenues chez la caille et le canard avec la séquence du génome de la poule permettra d'établir une "cartographie virtuelle" des SNP obtenus, grâce à la grande conservation de synténie existant entre ces trois espèces. ABSTRACT The aim of the project is to fill the lack in markers observed for three avian species (quail, duck and poultry). The chosen strategy is to obtain, by using individuals from several lines of interest, SNP (Single Nucleotide Polymorphism) by a high-throughput sequencing technology (sequencer 454 GS-FLX, Roche). Two reduced representations of the genome are sequenced: size-selected digested genomic DNA and transcriptome, which globally represents the part of the genome corresponding to the expressed genes. These experiments are realized from samples of DNA or RNA from individuals of lines from existing crosses, for each species. The data generated by several sequencing runs will be in silico analyzed. Besides the interest of the production of a very large number of new SNP, this technology should allow to sequence GC rich regions corresponding to the smallest microchromosomes for which there is no sequence in chicken. The comparison of the transcriptome sequences in quail and duck with the chicken genome assembly will allow to establish a "virtual cartography" of the obtained SNP, thanks to the synteny conservation existing between these three avian species.

Increasing resistance to acute Salmonellosis (that is, contamination level shortly after infection) is not sufficient to reduce the risk for consumers to be contaminated by Salmonella. Indeed, animals may remain contaminated at a low level for weeks or months. Increased resistance to the Salmonella carrier state, i.e., animals' ability to clear bacteria, is needed; it involves measuring bacterial contamination several weeks after inoculation with a low dose. To study such resistance traits, three convergent approaches were used. A quantitative trait loci (QTL) study was performed, taking advantage of inbred lines differing in resistance. Several QTLs controlling resistance at a younger age were identified and are currently being confirmed in a new cross before finer mapping, using advanced intercross lines. These inbred lines are also presently being compared using functional genomics. In parallel, a selection experiment for increased or decreased resistance at a younger and a later age was undertaken. Besides providing genetic models differing in their levels of resistance, it underlined the importance of the choice of selection criterion, whether marker assisted or not. Indeed, genes controlling resistance are strongly dependant on age; selecting for resistance at a younger age might result in increased susceptibility at an older age. Finally, the results of this experiment were used in a model of the intra-flock propagation of Salmonella. It showed that introducing a proportion of resistant animals within a flock of susceptible hens could dramatically change the evolution of contamination. Moreover, it demonstrated the magnitude of synergy between selection and vaccination, which should enhance the interest of increased resistance. The results show that selection for increased resistance to the Salmonella carrier state may be efficient, providing that the appropriate criteria of selection are used.

International audience; Woolly apple aphid (WAA; Eriosoma lanigerum Hausm.) can be a major economic problem to apple growers in most parts of the world, and resistance breeding provides a sustainable means to control this pest. We report molecular markers for three genes conferring WAA resistance and placing them on two linkage groups (LG) on the genetic map of apple. The Er1 and Er2 genes derived from 'Northern Spy' and 'Robusta 5',respectively, are the two major genes that breeders have used to date to improve the resistance of apple rootstocks to this pest. The gene Er3, from 'Aotea 1' (an accession classified as Malus sieboldii), is a new major gene for WAA resistance. Genetic markers linked to the Er1 and Er3 genes were identified by screening random amplification of polymorphic deoxyribonucleic acid (DNA; RAPD) markers across DNA bulks from resistant and susceptible plants from populations segregating for these genes. The closest RAPD markers were converted into sequence-characterized amplified region markers and the genome location of these two genes was assigned to LG 08 by aligning the maps around the genes with a reference map of 'Discovery' using microsatellite markers. The Er2 gene was located on LG 17 of 'Robusta 5' using a genetic map developed in a M.9 x 'Robusta 5' progeny. Markers for each of the genes were validated for their usefulness for marker-assisted selection in separate populations. The potential use of the genetic markers for these genes in the breeding of apple cultivars with durable resistance to WAA is discussed

International audience; Most agronomic traits of importance, whether physiological ( such as nutrient use efficiency) or developmental ( such as flowering time), are controlled simultaneously by multiple genes and their interactions with the environment. Here, we show that variation in sulfate content between wild Arabidopsis thaliana accessions Bay- 0 and Shahdara is controlled by a major quantitative trait locus that results in a strong interaction with nitrogen availability in the soil. Combining genetic and biochemical results and using a candidate gene approach, we have cloned the underlying gene, showing how a single - amino acid substitution in a key enzyme of the assimilatory sulfate reduction pathway, adenosine 5'- phosphosulfate reductase, is responsible for a decrease in enzyme activity, leading to sulfate accumulation in the plant. This work illustrates the potential of natural variation as a source of new alleles of known genes, which can aid in the study of gene function and metabolic pathway regulation. Our new insights on sulfate assimilation may have an impact on sulfur fertilizer use and stress defense improvement.

Besides being a metabolic fuel, carbohydrates play important roles in plant growth and development, in stress responses, and as signal molecules. We exploited natural variation in Arabidopsis (Arabidopsis thaliana) to decipher the genetic architecture determining carbohydrate content. A quantitative trait locus (QTL) approach in the Bay-0 × Shahdara progeny grown in two contrasting nitrogen environments led to the identification of 39 QTLs for starch, glucose, fructose, and sucrose contents representing at least 14 distinct polymorphic loci. A major QTL for fructose content (FR3.4) and a QTL for starch content (ST3.4) were confirmed in heterogeneous inbred families. Several genes associated with carbon (C) metabolism colocalize with the identified QTL. QTLs for senescence-related traits, and for flowering time, water status, and nitrogen-related traits, previously detected with the same genetic material, colocalize with C-related QTLs. These colocalizations reflect the complex interactions of C metabolism with other physiological processes. QTL fine-mapping and cloning could thus lead soon to the identification of genes potentially involved in the control of different connected physiological processes.

Publication Inra prise en compte dans l'analyse bibliométrique des publications scientifiques mondiales sur les Fruits, les Légumes et la Pomme de terre. Période 2000-2012. http://prodinra.inra.fr/record/256699; International audience; Powdery mildew, caused by the ascomycete fungus Podosphaera leucotricha, is one of the most damaging diseases of apple worldwide. Polygenically determined resistance might contribute to a significant increase of resistance to this disease in new cultivars. A quantitative trait locus (QTL) analysis was performed in an F, progeny derived from a cross between the apple cultivar Discovery and the apple hybrid TN10-8. Powdery mildew incidence was assessed during four years (five seasons) in spring and/or autumn in a French local orchard. Seven additive and/or dominant QTLs were detected over the five seasons, with effects (R-2) ranging from 7.5% to 27.4% of the progeny phenotypic variation. Two QTLs, on linkage groups (LGs) 2 and 13, were consistently identified and accounted together from 29% to 37% of the phenotypic variation according to the year of assessment. The other QTLs were identified during one (LGs 1, 14), two (LG10), or three (LGs 8, 17) seasons. Their instability indicated a changing genetic determinism according to the year of assessment, for which several hypotheses may be put forward. The QTLs on LGs 2 and 8 mapped close to clusters of resistance gene analogs (RGAs) and major genes for resistance to mildew or apple scab previously identified. The stable QTLs identified on LGs 2 and 13, together with the strong effect QTL located on LG 8, are of special interest for breeding purposes, especially if combined with other major resistance genes