A doença da bananeira \'mal-do-Panamá\', causada pelo fungo Fusarium oxysporum f. sp. cubense (Foc) é uma das doenças mais destrutivas da bananeira e é considerada uma das seis doenças economicamente mais importante da história da humanidade. Algumas cultivares resistentes, como a \'BRS Platina\', foram lançadas pela Embrapa, porém para a sustentabilidade da resistência é necessário entender os mecanismos moleculares envolvidos na resposta de resistência e defesa. O objetivo deste estudo foi caracterizar o processo de infecção pelo Foc raça 1 em três cultivares contrastantes para a resistência e analisar o padrão transcricional no início da interação. A análise histopatológica indicou que o Foc raça 1 penetra pela raízes laterais e principal, colonizando os espaços inter e intracelular do córtex nas três cultivares. Foram visualizadas, hifas \'globosas\' na cultivar suscetível \'Maçã\' com a formação de estruturas de resistência, como clamidósporos. Na cultivar resistente \'BRS Platina\', foi observado por microscopia óptica no período inicial da interação (24 horas após inoculação) a indução de respostas de defesa da planta, como formação de zona de cicatrização, e aos 15 dias após inoculação, formação de tilose, presença de cristais de oxalato de cálcio e deposição de calose. Foi utilizada a tecnologia Illumina para sequenciamento massal de RNA e abordagens de bioinformática para identificar genes diferencialmente expressos (DE) relacionados com a resposta de defesa de bananeira em interações compatíveis e incompatíveis. O sequenciamento paired-end gerou um total de 113.632.486 fragmentos (reads) com alta qualidade. Do total de reads alinhados no genoma referência (\'DH-Pahang\'), 55.555.480 alinharam-se com genes conhecidos e anotados no genoma referência, sendo utilizados para a análise DE inoculado x não inoculado, permitindo detectar 2.307 genes para as três cultivares. Os genes anotados de cada cultivar foram comparados, sendo identificados quatro genes comuns para as três cultivares, dez compartilhados entre \'Maçã\' e \'Prata-anã\', 21 compartilhados entre \'BRS Platina\' e \'Maçã\', 114 compartilhados entre \'BRS Platina\' e \'Prata-anã\', além de 75 serem exclusivos de \'Maçã\', 599 de \'BRS Platina\' e 1484 de \'Prata-anã\'. O mecanismo de resistência/defesa ao Foc em \'BRS Platina\', ocorre em nível de percepção precoce na presença do patógeno desencadeando resposta de defesa inexistente em \'Maçã\', e com cinética distinta da cultivar com resposta intermediária (\'Prata-anã\'). Dessa forma, os resultados permitiram propor um modelo da resposta de defesa/resistência ao Foc raça 1 em bananeira, baseando-se no nível de indução de genes que codificam para proteínas de reconhecimento do patógeno (receptor like kinase), fatores de transcrição (WRKY e MYB); reforço e síntese de parede celular, degradação da parede celular do fungo (quitinase e glucanases), heat shocks, enzimas antioxidantes e na resposta visualizada pela histologia na cultivar \'BRS Platina\'. Sendo assim, este trabalho fornece novas perspectivas para estudos de análise funcional, identificação e anotação de novos genes relacionados a resposta de defesa e resistência ao Foc raça 1. / The banana Panama disease, caused by fungus Fusarium oxysporum f. sp. cubense (Foc), is one of the most destructive disease of the industry, and it is considered one of the six most economically important of all times. A few cultivars, such as \'BRS Platina\', were released, but it is still necessary to understand molecular mechanisms involved in defense response and resistance. The objective of this study was to characterize the infection process by Foc in three banana cultivars contrasting for resistance to Foc and to analyze the transcriptional profile at the beginning of interaction. In this way, Foc race 1 penetrated the main and lateral roots, colonizing inter- and intracellular spaces of the root cortex in the three cultivars. Hyphae were globose in the susceptible cultivar \'Maçã\' with the formation of resilience structure, such as chlamydospores. In the resistant cultivar \'BRS Platina\', during the initial period of interaction (24 hours after inoculation), induced of plant defense responses, such as a healing zone, tylosis formation, presence of calcium oxalate and callose deposition. The Illumina technology were applied to sequence RNA, followed by bioinformatic tools to identify genes differentially expressed (DE) related to resistance and defense response in the compatible and incompatible interactions. Pair-end sequencing generated a total of 113,632,486 reads with high quality. From the total of aligned reads to the banana reference genome (\'DH-Pahang\'), 55,555,480 aligned with gene models annotated in the reference genome. The aligned contigs were analysed for DE, comparing inoculated x non-inoculated, enabling the detection of 2307 genes for the three cultivars. Each annotated gene from each cultivar was compared: four common genes to the three cultiars; 10 genes were shared between \'Maçã\' and \'Prata-anã\'; 21 shared between \'BRS Platina\' and \'Maçã\'; 114 shared between \'BRS Platina\' and \'Prata-anã\', plus 75 exclusive to \'Maçã\'; 599 exclusive to \'BRS Platina\' and 1,484 to \'Prata-anã\'. The mechanism of resistance/defense in \'BRS Platina\', level of perception occurs early in the presence of the pathogen defense response triggering nonexistent in \'Maçã\' and with kinetics distinct cultivar with intermediate response (\'Prata-anã\'). Thus, the results have provided a model of defense response/resistance to Foc race 1 in banana, based on the level of gene induction that encode recognition proteins (Receptor-like Kinase, RLK), transcription factors (WRKY and MYB), cell wall synthesis and reinforcement, degradation of fungal cell wall (chitinases and glucanases), heat shocks , proteins;anto-oxidative enzymes and visualized by histologcal in response cultivar \'BRS Platina\'. The present work offer new perspectives to functional analyses, identification and annotation of new genes related to resistance and defense response to Foc race 1.

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.

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

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.

A doença da bananeira \'mal-do-Panamá\', causada pelo fungo Fusarium oxysporum f. sp. cubense (Foc) é uma das doenças mais destrutivas da bananeira e é considerada uma das seis doenças economicamente mais importante da história da humanidade. Algumas cultivares resistentes, como a \'BRS Platina\', foram lançadas pela Embrapa, porém para a sustentabilidade da resistência é necessário entender os mecanismos moleculares envolvidos na resposta de resistência e defesa. O objetivo deste estudo foi caracterizar o processo de infecção pelo Foc raça 1 em três cultivares contrastantes para a resistência e analisar o padrão transcricional no início da interação. A análise histopatológica indicou que o Foc raça 1 penetra pela raízes laterais e principal, colonizando os espaços inter e intracelular do córtex nas três cultivares. Foram visualizadas, hifas \'globosas\' na cultivar suscetível \'Maçã\' com a formação de estruturas de resistência, como clamidósporos. Na cultivar resistente \'BRS Platina\', foi observado por microscopia óptica no período inicial da interação (24 horas após inoculação) a indução de respostas de defesa da planta, como formação de zona de cicatrização, e aos 15 dias após inoculação, formação de tilose, presença de cristais de oxalato de cálcio e deposição de calose. Foi utilizada a tecnologia Illumina para sequenciamento massal de RNA e abordagens de bioinformática para identificar genes diferencialmente expressos (DE) relacionados com a resposta de defesa de bananeira em interações compatíveis e incompatíveis. O sequenciamento paired-end gerou um total de 113.632.486 fragmentos (reads) com alta qualidade. Do total de reads alinhados no genoma referência (\'DH-Pahang\'), 55.555.480 alinharam-se com genes conhecidos e anotados no genoma referência, sendo utilizados para a análise DE inoculado x não inoculado, permitindo detectar 2.307 genes para as três cultivares. Os genes anotados de cada cultivar foram comparados, sendo identificados quatro genes comuns para as três cultivares, dez compartilhados entre \'Maçã\' e \'Prata-anã\', 21 compartilhados entre \'BRS Platina\' e \'Maçã\', 114 compartilhados entre \'BRS Platina\' e \'Prata-anã\', além de 75 serem exclusivos de \'Maçã\', 599 de \'BRS Platina\' e 1484 de \'Prata-anã\'. O mecanismo de resistência/defesa ao Foc em \'BRS Platina\', ocorre em nível de percepção precoce na presença do patógeno desencadeando resposta de defesa inexistente em \'Maçã\', e com cinética distinta da cultivar com resposta intermediária (\'Prata-anã\'). Dessa forma, os resultados permitiram propor um modelo da resposta de defesa/resistência ao Foc raça 1 em bananeira, baseando-se no nível de indução de genes que codificam para proteínas de reconhecimento do patógeno (receptor like kinase), fatores de transcrição (WRKY e MYB); reforço e síntese de parede celular, degradação da parede celular do fungo (quitinase e glucanases), heat shocks, enzimas antioxidantes e na resposta visualizada pela histologia na cultivar \'BRS Platina\'. Sendo assim, este trabalho fornece novas perspectivas para estudos de análise funcional, identificação e anotação de novos genes relacionados a resposta de defesa e resistência ao Foc raça 1. / The banana Panama disease, caused by fungus Fusarium oxysporum f. sp. cubense (Foc), is one of the most destructive disease of the industry, and it is considered one of the six most economically important of all times. A few cultivars, such as \'BRS Platina\', were released, but it is still necessary to understand molecular mechanisms involved in defense response and resistance. The objective of this study was to characterize the infection process by Foc in three banana cultivars contrasting for resistance to Foc and to analyze the transcriptional profile at the beginning of interaction. In this way, Foc race 1 penetrated the main and lateral roots, colonizing inter- and intracellular spaces of the root cortex in the three cultivars. Hyphae were globose in the susceptible cultivar \'Maçã\' with the formation of resilience structure, such as chlamydospores. In the resistant cultivar \'BRS Platina\', during the initial period of interaction (24 hours after inoculation), induced of plant defense responses, such as a healing zone, tylosis formation, presence of calcium oxalate and callose deposition. The Illumina technology were applied to sequence RNA, followed by bioinformatic tools to identify genes differentially expressed (DE) related to resistance and defense response in the compatible and incompatible interactions. Pair-end sequencing generated a total of 113,632,486 reads with high quality. From the total of aligned reads to the banana reference genome (\'DH-Pahang\'), 55,555,480 aligned with gene models annotated in the reference genome. The aligned contigs were analysed for DE, comparing inoculated x non-inoculated, enabling the detection of 2307 genes for the three cultivars. Each annotated gene from each cultivar was compared: four common genes to the three cultiars; 10 genes were shared between \'Maçã\' and \'Prata-anã\'; 21 shared between \'BRS Platina\' and \'Maçã\'; 114 shared between \'BRS Platina\' and \'Prata-anã\', plus 75 exclusive to \'Maçã\'; 599 exclusive to \'BRS Platina\' and 1,484 to \'Prata-anã\'. The mechanism of resistance/defense in \'BRS Platina\', level of perception occurs early in the presence of the pathogen defense response triggering nonexistent in \'Maçã\' and with kinetics distinct cultivar with intermediate response (\'Prata-anã\'). Thus, the results have provided a model of defense response/resistance to Foc race 1 in banana, based on the level of gene induction that encode recognition proteins (Receptor-like Kinase, RLK), transcription factors (WRKY and MYB), cell wall synthesis and reinforcement, degradation of fungal cell wall (chitinases and glucanases), heat shocks , proteins;anto-oxidative enzymes and visualized by histologcal in response cultivar \'BRS Platina\'. The present work offer new perspectives to functional analyses, identification and annotation of new genes related to resistance and defense response to Foc race 1.

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.

La fusariose des épis est l’une des principales maladies des céréales, majoritairement causée par le champignon pathogène et toxinogène, Fusarium graminearum (Fg). Lors son développement in planta, le champignon produit des mycotoxines dommageables pour la santé humaine et animale, dont le déoxynivalénol (DON). De nombreux loci à effet quantitatif sur la résistance à Fg ont été identifiés chez le blé tendre. Certains d’entre eux ont été corrélés à la capacité à détoxifier le DON, en particulier par glucosylation sous l’action d’UDP-glucosyltransférases (UGT). Une UGT d’orge impliquée dans la conjugaison du DON a été identifiée en système hétérologue. Brachypodium distachyon (Bd) a récemment émergé comme modèle d’étude pour les céréales. Ce travail à l’aide d’approches transcriptomique et métabolomique a mis en évidence que lors de l’interaction avec Fg, Bd met en place des réponses macroscopiques, moléculaires et métaboliques similaires à celles connues chez le blé et l’orge. La recherche d’UGTs candidates capables de conjuguer le DON en DON-3-O-glucoside (D3G) chez Bd a permis l’identification d’un candidat. L’analyse fonctionnelle du gène correspondant a été conduite par des approches de mutagenèse et de surexpression. Ceci a montré une sensibilité accrue des lignées mutantes à la toxine et à l’agent pathogène. A l’inverse les lignées surexpresseurs ont montré une tolérance et résistance quantitative à la toxine et l’agent pathogène. Ces résultats ont été corrélés par la détection in planta de DON et D3G, dans des proportions variables selon les lignées. Ces résultats démontrent le rôle majeur que joue la glucosylation du DON dans l’établissement de la résistance observée chez Bd en réponse à Fg. / Fusarium head blight is a major cereal disease, mostly caused by the pathogenic and toxin-producing fungus, Fusarium graminearum (Fg). During its development in planta, the fungus produces mycotoxins harmful to human and animal health, including deoxynivalenol (DON). Many quantitative trait loci exhibiting an effect on resistance to Fg have been identified in wheat. Some of them were correlated with the ability to detoxify DON, particularly by glucosylation by UDP-glycosyltransferases (UGT). A barley UGT involved in the conjugation of DON was identified in a heterologous system. Brachypodium distachyon (Bd) has recently emerged as a model species for cereals. Using transcriptomic and metabolomic approaches, we show that when interacting with Fg, Bd implements macroscopic, molecular and metabolic responses similar to those known in wheat and barley. The search for UGT candidates able to conjugate DON into DON-3-O-glucoside (D3G) in Bd resulted in the identification of the Bradi5g03300 gene. Functional analyses of this gene showed increased sensitivity of the mutant lines to the toxin and to the pathogen. Conversely the overexpressor lines showed a tolerance to the toxin and quantitative resistance to Fg. These results were correlated with the detection of differential amounts of DON and D3G in the different lines. These results demonstrate the important role of DON glucosylation in the resistance establishment of Bd observed in response to Fg.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

La fusariose de l’épi est une maladie fongique des cultures céréalières au Québec. Les objectifs de ce projet étaient de vérifier l’influence de quatre facteurs (météorologie, cultivar, date de semis, fongicide) sur les rendements et la qualité du blé et d’évaluer les performances de neuf modèles à prédire l’incidence de cette maladie. Pendant deux ans, à quatre sites expérimentaux au Québec, des essais de blé de printemps et d’automne ont été implantés pour amasser un jeu de données météorologiques, phénologiques et épidémiologiques. L’application d’un fongicide a réduit la teneur en désoxynivalénol des grains dans neuf essais sur douze et a augmenté les rendements dans sept essais. De plus, les modèles prévisionnels américains et argentins ont eu de meilleures performances que les modèles canadiens et italiens quand leur seuil de décision était ajusté et que le développement du blé était suivi au champ. / Fusarium head blight (FHB) is a fungal disease of cereal crops in Quebec. The objectives of this project were to check the influence of four factors (weather, cultivar, seeding date, fungicide application) on wheat yield and quality, and to evaluate the performance of nine forecasting models of FHB. During two years, at four experimental sites in Quebec, trials of spring and winter wheat have been implemented to establish a set of meteorological, phenological and epidemiological data. A fungicide application reduced deoxynivalenol content in grains in nine out of twelve trials and increased yields in seven trials. In addition, the American and Argentinean forecasting models performed better than the Canadian and Italian models when their decision threshold was adjusted and the development of wheat was observed in the field.