Foram realizados experimentos em laboratório, casa-de-vegetação e campo visando os seguintes objetivos: (i) avaliar a eficiência de diferentes métodos de detecção do fungo Fusarium graminearum em sementes de trigo (Triticum aestivum L.); (ii) estudar o efeito do patógeno na germinação das sementes, bem como na emergência de plântulas e verificar a sua transmissão da semente para a plântula; (iii) avaliar o efeito de diferentes doses de nitrogênio (0, 30, 60, 90, 120 e 180 kg.ha-1), cultivares (IAC – 24, IAC – 350 e IAC – 370), aplicação de mistura de fungicidas (triciclazole + tebuconazol: três aplicações de 0,75 L.ha-1) na parte aérea das plantas na incidência de F. graminearum; (iv) avaliar o efeito do tratamento de sementes de trigo na incidência do patógeno empregando, individualmente, os seguintes fungicidas com as respectivas dosagens (g i.a.100 kg–1 de sementes): captana (150,0), tiofanato metílico (75,0), triflumizole (45,0), triticonazole (45,0), triadimenol (13,5), tolyfluanida (75,0), tebuconazole (5,0), fludioxonil (5,0), difeconazole (30,0) e thiabendazole (30,0). Na detecção de F. graminearum em sementes de trigo da cultivar BR 23 foram utilizados os seguintes métodos: meio semi-seletivo (MSS); MSS + 2,4- D (0,02%); MSS + KCl (-0,8MPa); papel de filtro (PF) com e sem o congelamento das sementes; PF + 2,4 – D (0,02%); restritores hídricos (KCl, NaCl, manitol e sacarose) nos potenciais osmóticos de –0,4, -0,6, -0,8 e –1,0 MPa. A maior incidência do patógeno foi observada com a utilização de MSS + 2,4 – D e a menor percentagem de sementes germinadas foi observada com o uso de PF com congelamento. A germinação de 30 diferentes genótipos de trigo foi significativamente influenciada pela quantidade de sementes mortas com a presença de F. graminearum, contudo não foi encontrada correlação entre a incidência do patógeno e a germinação das sementes. Também foi constatado que o fungo não afeta a emergência das plântulas, contudo verificou-se que a sua taxa de transmissão para diferentes partes da plântula (raiz, colo e haste) é variável de acordo com a com o genótipo infectado. O inóculo da semente pode contribuir para o desenvolvimento de epidemias no campo. Verificou-se que maiores doses de N promovem o aumento da incidência do patógeno nas sementes, enquanto que a aplicação da mistura de fungicidas promove a redução. Não houve diferenças significativas entre as cultivares IAC – 250, IAC – 350 e IAC – 370 na incidência do patógeno. No tratamento químico de sementes de trigo da cultivar BR 18 Terena, as menores incidências do fungo foram observadas empregando-se os fungicidas tiofanato metílico e tebuconazole. Não houve influência dos fungicidas tanto na germinação quanto na emergência e velocidade de emergência de plântulas. Por outro lado, fungicidas como o captana, triticonazole, tebuconazole e triadimenol afetaram negativamente a altura de plântulas, interferindo no desempenho das mesmas. / Experiments were carried out in laboratory, green house and field aiming at the following objectives: (i) to evaluate the efficiency of different methods of detection of Fusarium graminearum; (ii) to study the effect of the pathogen in the germination of wheat seeds, as well as in the emergence of seedlings and to verify the transmission of the seed to the seedling; (iii) to evaluate the effect of different dosages of nitrogen (0, 30, 60, 90, 120 and 180 kg.ha-1), in the incidence of cultivars (IAC - 24, IAC - 350 and IAC - 370) and the application of a mixture of fungicides (triciclazole + tebuconazol: three applications of 0,75 L.ha-1) in the aerial part of the plants in the incidence of F. graminearum; (iv) to evaluate the effect of the treatment of wheat seeds in the incidence of the pathogen employed individually, the following fungicides, with the respective dosages (g i.a.100 kg-1 of seeds): captana (150,0), methylic tiophanate (75,0), triflumizole (45,0), triticonazole (45,0), triadimenol (13,5), tolyfluanida (75,0), tebuconazole (5,0), fludioxonil (5,0), difeconazole (30,0) and thiabendazole (30,0). In the detection of F. graminearum in wheat seeds of the cultivar BR 23 the following methods were used: semiselective media (MSS); MSS + 2,4-D (0,02%); MSS + KCl (-0,8MPa); blotter (PF) with and without the freezing of the seeds; 2,4 PF + - D (0,02%); hidric restrictors (KCl, NaCl, manitol and sacarose) in the osmotic potentials of -0,4, -0,6, -0,8 and -1,0 MPa. The highest incidence of the pathogen was observed with the use of MSS + 2,4 - D and the lowest percentage of germinated seeds was observed with the use deep freezing blotter method of PF with freezing. The germination of 30 different wheat genotypes was significantly influenced by the quantity of dead seeds with the presence of F. graminearum. However, correlation between the incidence of the pathogen and the germination of the seeds was not found. It was found that the fungus does not affect the emergence of seedlings, however it was verified that its rate of transmission for different parts of seedling is variable in accordance with the infected genotype. It was also verified that higher doses of N promote the increase of the incidence of the pathogen in the seeds, and that the application of the mixture of fungicides promotes the reduction. There was no effect of cultivars IAC – 250, IAC – 350 e IAC – 370 on the incidence of pathogen. Regarding the chemical treatment of wheat seeds of cultivar BR 18 Terena, the lowest incidences of the fungus were observed using the fungicides methylic tiophanate and tebuconazole. There was no effect of fungicides both on the germination and on the seedlings emergence and emergence velocity. On the other hand, fungicides such as the captan, triticonazole, tebuconazole and triadimenol affected the height of plântulas negatively, interfering with their performance.

Submitted by Gabriela Lopes (gmachadolopesufpel@gmail.com) on 2016-10-03T17:35:05Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Taiane_Viana.pdf: 904492 bytes, checksum: 30037558592bcb3fdf60ab01d643c574 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2016-10-06T20:46:17Z (GMT) No. of bitstreams: 2 Dissertacao_Taiane_Viana.pdf: 904492 bytes, checksum: 30037558592bcb3fdf60ab01d643c574 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-10-06T20:46:17Z (GMT). No. of bitstreams: 2 Dissertacao_Taiane_Viana.pdf: 904492 bytes, checksum: 30037558592bcb3fdf60ab01d643c574 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2013-12-04 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / O cultivo do trigo no Brasil é marcado pelo elevado número de doenças capazes de causar danos à produção. Entre as principais doenças que afetam o trigo está a giberela ou fusariose da espiga, causada pelo agente etiológico Gibberella zeae (Schw.) Petch, (anamorfo Fusarium graminearum Schwabe) que causa impacto negativos não somente à produtividade mas também a qualidade dos grãos produzidos. As condições ambientais para que a infecção ocorra carac terizam-se com temperaturas de 20-25°C e duração do molhamento foliar de, no mínimo, 48 horas consecutivas. O controle químico é realizado via aplicação de fungicidas a partir do inicio da floração estendendo-se até o final do florescimento, sendo que este tipo de controle caracteriza-se pela sua menor eficiência quando comparado com o controle de doenças foliares. Caso as condições climáticas impeçam a realização das aplicações de fungicidas no período indicado, o controle fica muito dificultado, por outro lado, se não ocorrer clima favorável à infecção, não se justifica o tratamento. A ineficiência dos tratos culturais no controle de giberela, associado à natureza quantitativa da característica de resistência, e o fato de sofrer alta influência do ambiente resultam na necessidade de identificar fontes efetivas de resistência que possam ser utilizadas em programas de melhoramento de trigo. De fato, existem culturas onde o controle das doenças mais importantes ocorre, quase que exclusivamente, por meio da resistência sendo que a identificação de genotipos que contenham resistência é um dos primeiros passos para selicioná-los. Desta forma, é fundamental o conhecimento de cultivares que apresentem níveis de resistência a giberela e métodos que possam identificar estas cultivares. Frente ao exposto o objetivo deste trabalho foi: i) separar genótipos suscetíveis e que contenham diferentes níveis de resistência utilizando avaliações morfológicas e moleculares; ii) verificar a resistência à giberela nas cultivares Frontana e Pampeano. Para isso foram utilizadas avaliações de características morfológicas citadas na literatura e associadas a resistência à giberela, assim como, o meprego de primers descritos na literatura significativamente associados à QTL de resistência à giberela. Como resultado do estudo espera-se auxiliar no desenvolvimento de ferramentas que possam ser utilizadas em programas de melhoramento de trigo visando facilitar a identificação de fontes de resistência. / The cultivation of wheat in Brazil is marked by the large number of diseases that can damage the production. Among the major diseases affecting wheat is wheat scab or Fusarium head blight (FHB), caused by the agent Gibberella zeae (Schw.) Petch (anamorph Fusarium graminearum Schwabe ) that causes negative impact not only on productivity but also the quality of the grain produced. The environmental conditions for infection are temperatures between 20-25°C and 48 consecutive hours of leaf wetness. Chemical control is performed via application of fungicides from the start extending until the end of flowering, and this type of control is characterized by its low efficiency when compared to the disease control. If the weather conditions prevent the aplication of fungicide applications within this period, there is no possibility control, on the other hand, if it does not favorable environment for infec tion treatment is not justified. The inefficiency of cultural control of FHB, associated with the quantitative trait (QTL- Quantitative traits loci)of resistance, and the fact of suffering high influence of the environment results in the need to identify effective sources of resistance that can be used in wheat breeding programs. In fact, there are cultures where control of major diseases occurs almost exclusively through resistance where the identification of genotypes containing resist ance is one of the first steps in their selection. Thus it is crucial to understand cultivar present levels of resistance to FHB and methods that can identify these cultivars.The objective of this study was: i) separate susceptible and containing different levels of resistance using molecular and morphological evaluations; ii) determine FHB resistance using cultivars Frontana and Pampeano as a source of resistance. Using morphological assessments cited in the literature and associated with resistance to FHB, as, primers as described in the literature as significantly associated with QTL for resistance to FHB. As a result of the study one expects to develop tools that can be used to facilitate the identification of sources of resistance.

La fusariose de l’épi est une maladie fongique qui touche toutes les cultures de céréales à paille à travers le monde entrainant des baisses de rendements et de la qualité des grains. La fusariose pose également un problème pour la sécurité alimentaire lié à la contamination des grains infectés par des mycotoxines. Le développement de variétés résistantes est considéré comme la méthode la plus efficace et la plus durable pour réduire les dommages causés par la maladie et pour limiter la contamination par les mycotoxines. L’amélioration de la résistance à la fusariose chez le blé dur (Triticum durum Desf.) demeure un défi du fait de son extrême sensibilité à la maladie et de la faible variabilité génétique disponible pour ce caractère. L’objectif principal de cette thèse a été d’évaluer l’effet de Fhb1, le QTL majeur de résistance à la fusariose chez le blé tendre (Triticum aestivumL.), au sein de fonds génétiques de blé dur élite. Pour cela, trois populations de cartographie, comprenant chacune environ 100 F7-RIL (lignées pures recombinantes ou « recombinant inbred lines »), ont été développées à partir de croisements entre la lignée expérimentale de blé dur DBC-480, portant une introgression de Fhb1, et les cultivars de blé dur Karur, Durobonus et SZD1029K. Les lignées ont été évaluées au champ, sur trois saisons, pour leur résistance globale à la fusariose après inoculation en spray de Fusarium culmorum. Des notations morphologiques (date de floraison, hauteur des plantes) ont également été réalisées afin d'évaluer leur influence sur l'infestation. Les lignées ont été génotypées à l’aide de marqueurs SSR et de marqueurs GBS (génotypage par séquençage ou « genotyping-by-sequencing ») développés par DArTseq. L’analyse de liaison a permis d’identifier des QTL de résistance sur les bras des chromosomes 2BL, 3BS, 4AL, 4BS, 5AL et 6AS. DBC-480 contribuait à l’allèle de résistance à tous ces loci. Le QTL sur 3BS a été détecté au sein des trois populations centré sur l’intervalle de Fhb1, confirmant, pour la première fois, son introgression dans le blé dur. L’évaluation de la résistance à la propagation après inoculation ponctuelle, réalisé au sein d’une des trois populations, a également permis de valider l’effet de Fhb1 sur la résistance de type 2 chez le blé dur. La hauteur des plantes influe fortement sur la résistance globale à la fusariose et, en particulier, l’allèle de nanisme Rht-B1b est associé à une plus grande sensibilité à la maladie dans les trois populations. Cependant, l’effet négatif de Rht-B1b sur la résistance est largement compensé dans les lignées possédant Fhb1. Des lignées semi-naines avec un meilleur niveau de résistance ont été sélectionnées et favoriseront le développement de cultivars de blé dur résistants à la fusariose. / Fusarium head blight (FHB) is a devastating disease affecting small-grain cereals worldwide causing yield and quality losses. FHB affects food safety due to the contamination of infected grains by mycotoxins. Host plant resistance is considered the most efficient and sustainable approach to contain FHB and mycotoxin contaminations. In durum wheat (Triticum durum Desf.) breeding for FHB resistance remains a challenge due to its extreme susceptibility and to lack of genetic variation available in the primary durum wheat gene pool. The primary goal of this thesis was to evaluate the effect of Fhb1, the major common wheat (Triticum aestivum L.) FHB resistance QTL, in elite durum wheat background. Three F7-RIL (recombinant inbred lines) mapping populations of about 100 lines were developed from crosses between the durum wheat experimental line DBC-480, harboring Fhb1, and the durum wheat cultivars Karur, Durobonus and SZD1029K. The RILs were tested under field conditions by artificial spray inoculation with Fusarium culmorum in three seasons. Morphological traits (flowering date, height) were also recorded to assess their influence on FHB infestation. Genotyping of the lines was performed with SSR and genotyping-by-sequencing (GBS) DArTseq markers. QTL analysis identified genomic regions associated with FHB resistance on chromosome arms 2BL, 3BS, 4AL, 4BS, 5AL and 6AS. DBC-480 contributed the resistant allele at all loci. Fhb1 was detected in all three populations, demonstrating for the first time its successful deployment in durum wheat. The effect of Fhb1 on FHB resistance in durum wheat was further verified by evaluating type 2 resistance in one of the three populations. Plant height had a strong influence in modulating FHB severity. Although the semi-dwarf allele Rht-B1b was associated with increased FHB susceptibility, its negative effect was efficiently counterbalanced in lines carrying Fhb1. Semi-dwarf lines with enhanced levels of resistance were selected and will assist the development of FHB resistant cultivars.

No description available.

The fungal plant pathogen Fusarium graminearum Schwabe (teleomorph Gibberella zeae¬) produces a dangerous trichothecene mycotoxin called deoxynivalenol (DON) and causes a devastating disease of barley (Hordeum vulgare L.) called Fusarium head blight (FHB). Food and feed products derived from barley, such as dried distillers grains with solubles (DDGS), may be contaminated with DON and pose a threat to the health of humans and domestic animals. New methods to mitigate the threat of DON in barley need to be developed and implemented. TRI101 and TRI201 are trichothecene 3-O-acetyltransferases that modify DON and reduce its toxicity. The first objective of this research was to isolate unique TRI101 and TRI201 enzymes that modify DON efficiently. We hypothesized that TRI101/TRI201 enzymes from different species of Fusarium would have varying rates and abilities to modify DON. Using degenerate primers, an internal portion of TRI101 or TRI201 was identified in 54 strains of Fusarium. Full-length sequences of seven TRI101 or TRI201 genes were cloned and expressed in yeast. All seven genes acetylated DON, but at different rates. The second objective of this research was to utilize transformed yeast expressing TRI101 or TRI201 to reduce DON levels in barley mashes and ultimately in DDGS. We hypothesized that DON levels would be reduced in DDGS derived from mashes prepared with transformed yeast. Five different barley genotypes were used to prepare the fermentation mashes and DON levels were reduced in all DDGS samples derived from mashes prepared with transformed yeast. The third objective of this study was to characterize barley genotypes developed at Virginia Tech for resistance to FHB and DON. We hypothesized that significant differences in resistance would be observed among barley genotypes and FHB resistance would be associated with reduced DON accumulation. From 2006 to 2010, FHB resistance was assessed in hulled (22 to 37) and hulless (13 to 32) barley genotypes by measuring incidence and index, and DON resistance was determined by quantifying DON levels in ground grain using gas chromatography-mass spectrometry. Our study showed that FHB and DON resistance is significantly determined by genotype. The final objective of this study was to develop a robust tissue culture system necessary for future development of transformed barley plants with FHB resistance gene(s). We hypothesized that callus production would vary among barley genotypes. In our analysis of 47 Virginia barley genotypes, 76% (36/47) of the genotypes produced callus tissue and there were significant differences in callus size. Our work sets the stage for identifying and characterizing DON detoxification genes in the future. The development of commercial barley lines that do not accumulate DON and that are resistant to FHB will directly impact growers and producers of small grains in the eastern U.S. / Ph. D.

La fusariose des épis des céréales est due à un champignon pathogène (Fusarium spp.) qui entraine non seulement une perte directe de rendement en interrompant le remplissage des grains, mais également une perte indirecte par la production de mycotoxines, responsables de perturbations de procédés industriels et d'intoxications alimentaires. Parmi les différents moyens de lutte, la création de variétés résistantes semble la stratégie la plus efficace et la plus durable. Plusieurs variétés exotiques ont ainsi été identifiées comme résistantes mais présentent la particularité d'être difficilement cultivables sous nos latitudes. L'obtention de variétés résistantes adaptées à nos climats passe donc par une meilleure connaissance des mécanismes de résistance disponibles. La détection de QTL (Quantitative Trait Loci) chez le blé tendre a déjà permis de localiser certains des facteurs génétiques impliqués dans des mécanismes de résistance. Cependant l'utilisation de ces résultats en amélioration des plantes se heurte à deux contraintes majeures : la position des QTL reste le plus souvent imprécise et ces QTL, dérivant de populations biparentales, ne représentent qu'une fraction de la diversité génétique potentiellement intéressante pour ce caractère. L'objectif de la thèse était d'étudier la résistance du blé à la fusariose en réalisant une approche de génétique d'association. L'utilisation de génotypes faiblement apparentés et d'une densité élevée de marqueurs a permis de détecter des associations entre le polymorphisme génétique et des variations phénotypiques. Après avoir réalisé une synthèse sur le pathogène et sur l'approche par génétique d'association, le premier travail a été de réaliser une carte génétique fiable afin de pouvoir localiser les associations mises en évidence. Un panel de 195 variétés de blé tendre élites a été évalué pour leur résistance globale à la fusariose, pour leur résistance à la progression des symptômes et pour leur résistance à l'accumulation des mycotoxines. Des notations morphologiques (précocité, taille, aristation, nombre d'épillets par épi et extrusion des anthères) ont également été réalisées afin d'évaluer leur influence dans l'infestation. Ces variétés ont été génotypées avec 3016 marqueurs SNP, 200 marqueurs SSR et 1400 marqueurs DArT. Préalablement à la réalisation des tests d'association, l'étude de la structure a mis en évidence trois origines géographiques (française, anglaise et allemande) des variétés du panel. De même, l'étude du déséquilibre de liaison a montré que celui-ci était conservé sur une distance comprise entre 2 et 6 cM. L'étude d'association réalisée sur les notations d'infestation a permis d'identifier plusieurs zones associées, dont plusieurs sont colocalisées avec des Meta-QTL et/ou avec des zones identifiées avec les notations morphologiques. La densification en marqueurs de 6 zones associées avec les différentes notations d'infestation, localisées sur les chromosomes 1A, 1D, 2A, 2B, 5A et 7A, ont permis de confirmer ces régions, de restreindre fortement l'intervalle d'intérêt et de mettre en évidence de potentiels gènes candidats. Cette analyse a ainsi permis de mieux comprendre les mécanismes génétiques et morphologiques impliqués dans la résistance à la fusariose des épis. La décomposition en trois partie de cette résistance (résistance globale, résistance à la progression des symptômes, et résistance à l'accumulation des mycotoxines) montre l'existence de mécanismes génétiques indépendants et donc complémentaire à prendre en compte dans une stratégie de création de variétés résistantes à la fusariose des épis. / Fusarium head blight (FHB), caused by the fungal pathogen Fusarium spp., is a major cereals disease that not only cause direct yield losses, by interrupting grain filling, but also indirect quality losses by the production of mycotoxins, responsible of industrial processes disturbance and food poisoning. Most wheat breeding programs in FHB-affected areas of the world have been screening germplasm to identify sources of improved tolerance. Unfortunately, these sources of genetic resistance are often of exotic origin and not adapted to West European growing conditions. The selection of adapted varieties with improved tolerance therefore needs a better characterization of resistance mechanisms. Several QTL for FHB resistance in wheat have been identified in European germplasm, but the use of these information in marker-assisted selection is constrained by the precision of the QTL and the low diversity tested by using biparental population.The aim of the present study is to use association mapping, also called linkage disequilibrium mapping, to identify loci involved in the resistance to FHB. This method refers to the analysis of statistical associations between genotypes determined in a collection of individuals, and the phenotypes of the same individuals. A dense genetic map was compiled to localize precisely the association results. Resistance to F. graminearum was studied in a panel of 195 elite wheat varieties by the evaluation of three components: resistance to global infection, resistance to symptom progression and resistance to accumulation of mycotoxins. Morphological factors (plant height, heading date, awnedness, spikelets per ears, anther extrusion), known to influence resistance to FHB, were also recorded. All the varieties have been genotyped with around 3300 SNP markers, 200 SSR markers and 1400 DArT markers. We first investigated the structure of the panel, which could generate bias in the estimate of allele effects, if not included explicitly in the association models. We showed that the structure is based on geographical origin (French, German and UK). Study of the linkage disequilibrium (LD) showed an extent of LD between 2 and 6 cM. Results of association studies permitted to identify several loci for each of the evaluated components of resistance. Some of these loci colocalized with the results of the MetaQTL analysis and/or with loci associated with morphological traits. We selected more specifically 6 loci, located on chromosome 1A, 1D, 2A, 2B, 5A and 7A. Marker saturation of the regions, allowed to confirm the genome wide association results and to increase the accuracy of the loci of interest. This analysis allowed to better understand the many factors that influenced FHB resistance, whether genetic or morphological. Results show that the genetic mechanisms are independent between the three components and therefore, information obtained for each component are to be used complementary to create varieties with increased resistance to FHB.

La fusariose des épis est l’une de maladies les plus dommageables des céréales tempérées et est principalement causée par le champignon toxinogène Fusarium graminearum (Fg). Ces dix dernières années, de nombreuses études ont rapporté l’induction transcriptionnelle de gènes de la plante codant pour des cytochromes P450 (P450) en réponse l’infection par Fg. Les P450s constituent une famille enzymatique impliquée dans de nombreuses voies métaboliques, certaines avec des intérêts potentiels dans la résistance face aux maladies. Nous avons utilisé la petite graminée modèle Brachypodium distachyon (Bd) pour caractériser fonctionnellement le premier gène codant pour un P450 induit chez la plante au cours de la fusariose des épis par l’utilisation de lignées altérées dans la séquence ou l’expression du gène Bradi1g75310 codant le P450 BdCYP711A29. Nous avons montré qu’en plus d’être un facteur de sensibilité à la maladie, le gène Bradi1g75310 est impliqué dans une voie de biosynthèse hormonale chez Bd, celle des strigolactones (SLs). En effet, en plus de complémenter génétiquement les phénotypes aériens de la lignée mutante max1-1 d’Arabidopsis thaliana altérée dans le gène homologue MAX1 (AtCYP711A1), une lignée de Bd surexprimant Bradi1g75310 (lignée OE) exsude davantage d’orobanchol, une SL spécifique, que la lignée sauvage ou mutante. Une analyse préliminaire de l’impact direct de l’orobanchol sur la croissance de Fg semble indiquer une activation des étapes précoces du développement du champignon (germination) qui pourrait être à l’origine de l’induction plus rapide de gènes de défenses observée chez une lignée OE de Bradi1g75310. Nous avons également montré que les 4 paralogues de Bradi1g75310 chez Bd, qui codent également pour des CYP711A, sont tous capable de complémenter la lignée max1-1 et avons généré du matériel végétal fondamental pour la poursuite de l’étude de la diversification des SLs chez la plante monocotylédone modèle Bd. Au global, ce projet constitue une première étape dans la caractérisation de l’implication des P450 dans la réponse de la plante face à l’infection par Fg en plus de donner de nouveaux indices concernant le rôle des SLs dans les interactions plante-pathogène. Les résultats obtenus au cours de ce travail de thèse pourront permettre l’amélioration de caractères tant développementaux que de résistance à la fusariose chez les céréales cultivées. / Fusarium Head Blight (FHB) is one of the most important diseases of temperate cereals and is mostly caused by the toxin producing-fungus Fusarium graminearum (Fg). This last decade, several studies reported the transcriptional activation of cereal cytochrome P450-encoding genes (P450s) in response to Fg infection. P450s constitute an enzymatic family participating in very diverse metabolic pathways with potential interest for disease resistance. We used the model temperate cereal Brachypodium distachyon (Bd) to functionally characterize the first FHB-induced P450- encoding gene using Bd lines altered in the locus or gene expression of the Bradi1g75310 gene encoding the BdCYP711A29 P450. We showed that in addition to be a plant susceptibility factor towards the disease, the Bradi1g75310 gene is involved in the hormonal biosynthetic pathway of strigolactones (SLs) in Bd. Indeed, in addition to genetically complement the shoot phenotypes of the Arabidopsis thaliana mutant line for the homologous gene MAX1 (AtCYP711A1, max1-1 line), a Bd linewhich overexpresses the Bradi1g75310 gene (OE) exudes more orobanchol, a specific SL, compared to wild-type or mutant lines. Preliminary analysis of the direct impact of orobanchol on Fg growth suggests an activation of early fungal development (germination) likely to induce faster induction of defense-related genes during FHB, observed in Bradi1g75310 OE line. We showed that the four paralogs of Bradi1g75310 encoding BdCYP711A P450s are all able to genetically complement max1-1 line and provide important plant material for studying SLs diversification in the model monocot B. distachyon. Overall, this project constitutes a first step in the characterization of P450s involvement in plant response towards Fg infection in addition to give new evidences about the role of SLs in plant-pathogen interactions. Results obtained during this Ph.D. project will allow the improvement of both developmental and FHB-related traits in cereal crops.

La fusariose est une des maladies les plus importantes altérant le blé en Bourgogne. L’espèce fongique Fusarium graminearum est l’un des agents principaux de la maladie. L’interaction hôte-pathogène peut entrainer la production de mycotoxines toxiques pour l’homme et l’animal.La seule alternative pour prévenir le développement de la maladie est de contrôler l’inoculum primaire dans son habitat naturel : les adventices, le sol et les résidus de culture. En raison de la tendance à la réduction du travail du sol, une attention particulière doit être portée au rôle des résidus de culture dans la survie et le développement de F. graminearum. Dans ce travail de thèse, deux approches ont été choisies pour mieux comprendre le développement saprotrophe du champignon et ses conséquences. La première, à travers un essai en champ dans le contexte pédoclimatique Bourguignon, avait pour but de définir la part relative des différentes sources d’inoculum dans le développement de la fusariose et l’accumulation des mycotoxines dans les grains. Cet essai devait en outre permettre de déterminer si des indicateurs précoces du développement de la maladie sur épi et de l’accumulation de mycotoxines pouvaient être identifiés. La seconde, à travers un essai en microcosmes, avait pour but de suivre le développement de F. graminearum dans le sol et les résidus.Ce travail a permis de souligner l’importance de la gestion des résidus de culture dans le développement de la fusariose du blé. Favoriser une décomposition biologique rapide des résidus de culture et/ou introduire des cultures intermédiaires assainissantes constituent des perspectives de recherche sur lesquelles doivent porter nos efforts / Fusarium Head Blight (FHB), mainly caused by the fungal species Fusarium graminearum, is one of the most important disease altering wheat crops in Burgundy. Moreover the plant-pathogen interaction leads to the production of mycotoxins potentially toxic for humans and animals.The only alternative to date to prevent the development of the disease is to control the saprotrophic development of F. graminearum in its natural habitat, i.e. weeds, soil and crops residues. Due to the trend of reduced tillage, special attention should be paid to the role of crop residues in the survival and development of F. graminearum.Two approaches were chosen to better understand the saprotrophic development of F. graminearum and its consequences towards FHB. i) The first through a field experiment in the Burgundian pedoclimatic context aimed at defining the relative importance of the different sources of inoculum in the development of FHB and the accumulation of mycotoxins in grains. The field experiment was also to determine whether early indicators of disease development on ears and accumulation of mycotoxins could be identified. ii) The second, through test microcosms, was to follow the development of F. graminearum in the soil and crop residues.This work highlighted the importance of crop residues management in the development of FHB and gave new understanding about the survival of the fungus on these residues. Improve the biological decomposition of crop residues at the soil surface or/and using suppressive intermediate crops could be the next prospective to investigate to limit the soil inoculum potential of saprotrophic F. graminearum

No description available.

Triticum aestivum (bread wheat), one of the most globally important cereal crops, is vulnerable to fusarium head blight (FHB). The disease is mainly associated with the pathogen Fusarium graminearum and generates yield losses and mycotoxin contaminated grains with low quality. One possible solution to overcome this problem is the production of FHB resistant wheat varieties by crossing with strongly resistant germplasm from either wheat or closely related species. Thinopyrum elongatum is a wild grass that carries genetic resistance to FHB on the long arm of its chromosome 7E (7EL). In the first part of this research project, five Th. elongatum accessions were characterized for their response to F. graminearum infection. In the second part, BC1F4 progeny derived from the cross CS-ph1b × CS-7E(7D) were characterized to better define the 7E fragments introgressed into the 7D chromosome. Progeny were screened with a series of known 7E-specific genetic markers and for their FHB resistance. Among the 43 wheat plants tested, twelve FHB resistant progeny were shown to carry a similar, smaller 7EL introgressed fragment based on genetic marker screening. To characterize further the introgressed 7EL fragments, additional 7EL-specific markers as well as 7DL-specific markers for homoeologous wheat sequences were designed. As neither wheat nor Th. elongatum genomes were fully sequenced at the time, this made the designing procedure challenging; a cross-walking strategy between wheat and Th. elongatum draft genomic sequences was used. Twelve pairs of markers for homoeologous sequence regions of 7EL and 7DL chromosomes plus six individual 7EL- and four 7DL-specific markers were successfully designed. Nine novel 7EL-specific markers were associated with the smallest 7EL fragment carrying FHB resistance. That smallest introgressed 7EL fragment replaced approximately half of the 7DL chromosome, based on the absence of 7DL markers in some progeny. The novel 7EL- and 7DL-specific markers as well as the proposed genetic order for novel and previously designed markers contributed greatly to the characterization of the introgressed 7EL fragments in the 7DL chromosome. Further analysis of progeny from the next generations of these plants and from other families will be required to confirm the results and possibly obtain much smaller 7EL fragments.