La brûlure foliaire causée par Alternaria dauci est la maladie foliaire la plus dommageable pour les cultures de carottes, entravant la récolte mécanique. Seuls des cultivars partiellement résistants sont connus et commercialisés, mais leurs niveaux de résistance sont insuffisants. Les mécanismes de la résistance quantitative des plantes aux agents pathogènes sont mal caractérisés. Nous avons choisi d'étudier ces mécanismes dans l'interaction A. dauci-carotte. Auparavant, plusieurs résultats expérimentaux convergents ont montré que la résistance aux toxines fongiques entre en jeu dans cette interaction. Les tests de toxicité effectués avec des suspensions cellulaires de carotte ont révélé une corrélation entre la résistance des carottes à A.dauci et la résistance des cellules de carotte aux exsudats du champignon. Ces résultats nous ont incités à étudier les toxines impliquées dans le pouvoir pathogène d'A. dauci et afin de pouvoir étudier la réponse de la plante à celles –ci. En utilisant les profils HPLC de la phase organique d'exsudats de différentes souches de champignons, nous avons découvert une corrélation entre la production de toxines et l’agressivité de ces souches suggérant que la production de toxines joue un rôle majeur dans l’interaction A. dauci-carotte. Nous avons effectué l'extraction, la purification et la caractérisation de l'une des molécules candidates que nous avons nommé aldaulactone. Nous avons démontré sa toxicité grâce à un nouveau protocole de quantification de cellules mortes et vivantes. Un transcriptome d’A. dauci et une étude de l’expression des gènes en fonction de la production d’aldaulactone ont été utilisées pour étudier sa voie de biosynthèse. / Alternaria leaf blight, caused by the necrotrophic fungus Alternaria dauci, is the most damaging foliar disease of carrots, especially because it hampers leaf-pull harvesting. Only partially – and insufficiently – resistant cultivars exist. In general, partial resistance mechanisms are poorly understood, so we chose to study them in this interaction. Previous results obtained in the lab highlighted a correlation between plant resistance to the fungus and plant cell resistance toward fungal toxins. It was also shown using carrot cell suspensions that fungal exudates’ toxicity was only present in the organic phase. These results led us to better characterize the toxins produced by A. dauci, in order to get a deeper understanding of carrot cell resistance mechanisms toward those toxins. HPLC analysis of the exudates from different fungal strain uncovered a correlation between toxin production and the aggressiveness of the fungal strains, suggesting that toxin production is an important component of said aggressiveness. We extracted, purified and characterize one of these candidates, and named it aldaulactone. Using a new image analysis protocol, we demonstrated the toxicity of Aldaulactone on carrot cell suspensions. Transcriptomic data from Alternaria dauci were used to explore the biosynthesis pathway of Aldaulactone. Candidate Genes were selected and their level of expression compared with aldaulactone production in various A. dauci cultures.

Submitted by Socorro Pontes (socorrop@ufersa.edu.br) on 2017-07-04T13:48:44Z No. of bitstreams: 1 JoseMC_TESE.pdf: 3634440 bytes, checksum: 6ac32b0fb28add3a21ce28608edd3cdf (MD5) / Approved for entry into archive by Vanessa Christiane (referencia@ufersa.edu.br) on 2017-07-04T16:47:51Z (GMT) No. of bitstreams: 1 JoseMC_TESE.pdf: 3634440 bytes, checksum: 6ac32b0fb28add3a21ce28608edd3cdf (MD5) / Made available in DSpace on 2017-07-04T16:57:57Z (GMT). No. of bitstreams: 1 JoseMC_TESE.pdf: 3634440 bytes, checksum: 6ac32b0fb28add3a21ce28608edd3cdf (MD5) Previous issue date: 2017-02-24 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The objective of the present work was to identify sources of resistance in melon accessions to Liriomyza sativae; Alternaria spp., and Acidovorax avenae. Two experiments were performed in a greenhouse and in the Laboratory of Applied Entomology (UFERSA) in a completely randomized design, with ten replicates, each plant being a plot. In the first experiment 38 accessions and two hybridswere evaluated. The infestation was held in cages coated with antiophidic screen. The second experiment was performed with the accessions that revealed to be promising and some were susceptible in the first trial. In the evaluations, the number of mines, larvae, pulps and adult flies were determined. In some materials, leaf colorimetry, number, size of stomata and trichomes were analyzed. The accessions AM 55 and Nantais Oblong as well as the Ruthenium hybrid are resistant to Liriomyza sativa. The number of mines, larvae and adult flies correlated negatively with the number of trichomes and positively with the size of the stomata. Regarding the Alternaria leaf blight, fifty-eight accessions were initially evaluated in a randomized block design (DBC) with three replications under field conditions. In a second experiment, twelve accessions were also evaluated under field conditions, in both tests the evaluations were accomplished 50 days after the transplanting, as far as the severity and incidence of plants with Alternaria. It was concluded that there is variability in the germplasm of melon for reaction to the Alternaria leaf blight, being the accessions C-AC-04, C-AC-08, C-AC-26, C-AC-31, I 162, I 173 e MR-1 with lower severity averages and incidence of diseased plants, being evaluated as promising for use in breeding programs of melon for resistance to the pathogen. For the reaction to A. avenae, fifty accessions were evaluated in greenhouse and in field conditions. There has been accomplished inoculations with bacterial suspension in the concentration os 1,0 x 108 UFC ml-1. There were evaluated leaves (in a controlled environment), leaves and fruits (in field) with bacteriose symptoms. The C-AC-12 accession has intermediate resistance Acidovorax avenae / Os objetivos do presente trabalho foram: identificar fontes de resistência em acessos de meloeiro a Liriomyza sativae, Alternaria spp. e Acidovorax avenae. Dois experimentos foram realizados em casa de vegetação e no Laboratório de Entomologia Aplicada da (UFERSA) em delineamento inteiramente casualizado, com dez repetições, sendo cada planta uma parcela. No primeiro experimento foram avaliados 38 acessos e dois híbridos de meloeiro. A infestação foi realizada em gaiolas revestidas com tela antiafídica. O segundo experimento foi realizado com os acessos que se mostraram promissores e alguns suscetíveis no primeiro ensaio. Nas avaliações, determinou-se o número de minas, larvas, pupários e moscas adultas, em alguns materiais fez-se analise de colorimetria das folhas, número, tamanho de estômatos e tricomas. Os acessos AM 55 e Nantais Oblong bem como o híbrido Rutênio são resistentes a Liriomyza sativae. O número de minas, larvas e moscas adultas correlacionaram negativamente com o número de tricomas e positivamente com o tamanho dos estômatos. Com relação a mancha de alternaria, inicialmente foram avaliados 58 acesso sem delineamento em blocos casualizados (DBC) com três repetições em condições de campo. Em um segundo experimento, foram avaliados 12 acessos também em condições de campo, em ambos os ensaios as avaliações foram realizadas aos 50 dias pós o transplantio, quanto a severidade e incidência de plantas com alternariose.Concluiu-se que, existe variabilidade no germoplasma de meloeiro para reação a mancha de alternaria, sendo os acessos C-AC-04, C-AC-08, C-AC-26, C-AC-31, I 162, I 173 e MR-1 com menores médias de severidade e incidência de plantas doentes, sendo avaliados como promissores para uso em programas de melhoramento genético do meloeiro visando resistência ao patógeno. Para a reação a A. avenae, foram avaliados 50 acessos de meloeiro em estufa e em condições de campo. Realizou-se inoculações com suspensão bacteriana na concentração de1, 0 x 108 UFC ml-1.Avaliou-se folhas (em ambiente controlado), folhas e frutos (em campo) com os sintomas da bacteriose. O acesso C-AC-12 possui resistência intermediaria a Acidovorax avenae / 2017-07-04