Submitted by Ricardo Cedraz Duque Moliterno (ricardo.moliterno@uefs.br) on 2018-09-17T22:06:41Z No. of bitstreams: 1 Disserta??o de Mestrado - Washington.pdf: 1852656 bytes, checksum: 454b1394baec4b0dbe61bc22fcf2e3ee (MD5) / Made available in DSpace on 2018-09-17T22:06:41Z (GMT). No. of bitstreams: 1 Disserta??o de Mestrado - Washington.pdf: 1852656 bytes, checksum: 454b1394baec4b0dbe61bc22fcf2e3ee (MD5) Previous issue date: 2018-02-06 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The fungi Lasiodiplodia theobromae and Neofusicoccum parvum are pathogens responsible for reducing mango yield due to dieback, which both cause in the culture. Inheritance studies and variability in accessions for fungal diseases in mango, in general, are quite limited, despite their importance for the development of cultivars. The goals of this study were1) to evaluate the inheritance of resistance to dieback disease caused by L. theobromae and N. parvumin three crossings, and 2) evaluate 75mango accessions for resistance to these fungi to support mango breeding programs. Young branches containing young leaves were selected, and suspension of 103 conidia/mL of L. theobromae or N. parvum was sprayed, separately. After inoculation, leaves were protected in a plastic wet chamber for 48 hours. The infection was estimated by taking the branches without symptoms/total number of inoculated branches. Plants without disease symptoms in more than 90% of the brancheswere considered as resistant. Monogenic resistance, conferred by recessive alleles, was observed for L. theobromae for the three crossings evaluated, despite the 13:3 segregation detected for 'Keitt' x 'Tommy Atkins'. For N. parvum, recessive resistance with 3:1 monogenic segregation was observed in the crossing 'Keitt' x 'Tommy Atkins', and 9:7 epistatic segregation was observed in the crossing ?Haden? x ?Tommy Atkins? and ?Espada x ?Tommy Atkins?. ?Apple DCG?, ?Ataulfo?, ?Da porta?, ?Dashehari?, ?Espada?, ?Haden?, ?Haden 2H?, ?Heidi?, ?Irwin?, ?Keitt?, ?Kent?, ?N?ldica?, ?Recife? and ?Smith? were resistant to both pathogens, being promising for breeding programs. 'Rosa' are resistant to L. theobromae, and susceptible to N. parvum. 'Tommy Atkins', the most grown cultivar in the S?o Francisco valley, was susceptible to both pathogens.The mono or digenic inheritance observed in the present study indicates the possibility of selection in the F2 generation and identification of plants with double resistance to L. theobromae and N. parvum in mango trees. / Os fungos Lasiodiplodia theobromae e Neofusicoccum parvum s?o pat?genos que podem limitar a produ??o da mangueira devido a doen?a morte-descendente, que ambos ocasionam na cultura. Estudos de heran?a da resist?ncia e da variabilidade em acessos para doen?as f?ngicasem mangueira, em geral, s?o bastante limitados, apesar da sua import?ncia para o desenvolvimento de cultivares. Este estudo teve como objetivos 1) avaliar a heran?a da resist?ncia ? doen?a morte-descendente causadas por L. theobromae e N. parvumem tr?s cruzamentos, e 2) avaliar 75acessos de mangueira para resist?ncia aos dois fungos, para apoiar programas de melhoramento da cultura. Foram selecionados ramos jovens contendo folhas jovens para aplica??o, com aux?lio de pulverizador manual, de suspens?o de con?dios de 103 con?dios/mL de L. theobromae ou N. parvum. Ap?s a inocula??o, as folhas foram protegidas em c?mara ?mida de pl?stico durante 48 horas. A infec??o foi estimada tomando-se ramos sem sintomas / n?mero total de ramos inoculados. As plantas sem sintomas de doen?a em mais de 90% dos ramos, em diferentes per?odos de inocula??o, foram consideradas resistentes.Foi observada resist?ncia monog?nica conferida por alelos recessivos para L. theobromae, para os tr?s cruzamentos avaliados, apesar da segrega??o 13:3 para ?Keitt? x ?Tommy Atkins?. Para N. parvum foi observada resist?ncia recessiva com segrega??o monog?nica 3:1 no cruzamento ?Keitt? x ?Tommy Atkins? e segrega??o epist?tica 9:7 nos cruzamentos ?Haden? x ?Tommy Atkins? e ?Espada x ?Tommy Atkins?. Os acessos ?Apple DCG?, ?Ataulfo?, ?Da porta?, ?Dashehari?, ?Espada?, ?Haden?, ?Haden 2H?, ?Heidi?, ?Irwin?, ?Keitt?, ?Kent?, ?N?ldica?, ?Recife? e ?Smith?apresentaram resist?ncia aos dois pat?genos, sendo bastante promissoras para programas de melhoramento. ?Rosa? apresentou resist?ncia a L. theobromae, sendo considerada suscet?vel ao N. parvum. ?Tommy Atkins?, a mais cultivada no vale do S?o Francisco, foi suscept?vel a ambos os pat?genos. A heran?a mono ou dig?nica observada no presente estudo indica a possibilidade de sele??o na gera??o imediata aos cruzamentos, bem como a possibilidade da identifica??o de plantas com resist?ncia dupla a L. theobromae e N. parvum em mangueira.

Contre la pourriture grise et les maladies du bois (MdBs), qui sont des maladies cryptogamiques majeures de la vigne, la lutte biologique a un potentiel de développement considérable dans le contexte actuel de réduction des intrants chimiques en viticulture.L’objectif de cette thèse est de sélectionner et d'étudier des souches bactériennes antagonistes de Botrytis cinerea (Pourriture grise) et de deux champignons pathogènes clefs liés aux MdBs: Phaeomoniella chlamydospora et Neofusicoccum parvum. Les expériences de screening principales sont réalisées in vivo et in planta sur 46 souches bactériennes isolées dans le vignoble bordelais. Le niveau de protection par les souches antagonistes dépend significativement de la souche bactérienne, de l’espèce de champignon pathogène ciblée, du tissu ou organe végétal hôte, mais aussi pour N. parvum, du mode d’application de la souche bactérienne et, pour B. cinerea, du génotype lié aux transposons : transposa ou vacuma.Une réduction significative de 40 à 64% de la taille des nécroses dues à P. chlamydospora et/ou N. parvum est induite par trois souches bactériennes Pantoea agglomerans (S1), Paenibacillus sp. (S19) et Bacillus pumilus (S32) sur des boutures de vigne non greffées. Ces souches ont fait l'objet d'investigations approfondies pour déterminer leurs principaux modes d’action : Antibiose, production de composés volatils qui ont été identifiés et/ou induction de différents gènes de défense de la vigne.Concernant B. cinerea, les souches Enterobacter cowanii (S22), Enterobacter sp. (S23), Bacillus ginsengihumi (S38) et Bacillus sp. (S43, S46) présentent un pouvoir antagoniste important par production de composés volatils et diffusibles anti-Botrytis, ainsi que par compétition pour les nutriments par E. cowanii (S22). / Biological control of gray mold and grapevine trunk diseases (GTDs), which are major fungal diseases of grapevine, has a considerable potential development in the current context of reduction of chemical input in viticulture.The aim of this study was to select and study bacterial strains for antagonism against Botrytis cinerea, the causal agent of gray mold, and two key pathogens involved in GTDs: Phaeomoniella chlamydospora and Neofusicoccum parvum. The main screening experiments for antagonistic activity of 46 bacterial strains, isolated from Bordeaux vineyards, have been carried out under different in vivo and in planta conditions. The efficacy of protection by the antagonistic strains significantly depended on the bacterial strain, the targeted pathogen species, the host plant tissue or organ and, for N. parvum, also on the application mode of the bacterial strain and, for B. cinerea, on the transposon genotype: transposa or vacuma.A significant reduction in length of necrosis due to P. chlamydospora and/or N. parvum, ranging between 40 and 64% in non-grafted vine cuttings, resulted from three bacterial strains: Pantoea agglomerans (S1), Paenibacillus sp. (S19) and Bacillus pumilus (S32). These strains were thoroughly further investigated to determine their major modes of action by i) Antibiosis ii) production of antifungal volatile organic compounds, which have been identified, and/or iii) induction of different grapevine defense genes. Concerning B. cinerea, Enterobacter cowanii (S22), Enterobacter sp. (S23) Bacillus ginsengihumi (S38), Bacillus sp. (S43, S46) were of prime importance in the biocontrol by producing anti-Botrytis volatile and diffusible compounds or by competing for nutrients (case of E. cowanii S22).

La réduction des effets secondaires des produits phytosanitaires sur l’environnement est un enjeux majeur du Grenelle de l’environnement. L’utilisation prolongée du cuivre comme fongicide a entrainé son accumulation dans les sols viticoles. Le cuivre étant phytotoxique,son accumulation dans la plante pourrait modifier la morphologie, la biochimie ou la physiologie des organes végétaux et perturber les interactions vigne-bio-agresseur. Nousavons donc évalué les effets de la contamination cuprique des sols viticoles sur la sensibilitéde la vigne à un cortège de bio-agresseurs par une approche plus intégrée de la culture et deson patho-système.Pour cela, nous avons mesuré les teneurs en cuivre total et disponible des parcelles viticoles.Nous avons exposé au cuivre trois cépages puis nous avons mesuré la distribution du cuivre et ses effets dans la plante. Enfin nous avons évalué l’impact induit pour trois bio-agresseurs de la vigne. Nos résultats montrent que la teneur en cuivre disponible du sol dépend surtout de la teneur total en cuivre (qui résulte de l’historique parcellaire) mais aussi des teneurs en carbone total et en particules fines du sol. L’absorption du cuivre et son transfert vers les parties aériennes varient selon les cépages. Lorsque la contamination cuprique augmente, les interactions vigne-bio-agresseurs sont modifiées : si les effets semblent négatifs pourScaphoideus titanus et Plasmopara viticola, la contamination cuprique semble favorable au développement de Neofusicoccum Parvum. Ces résultats soulignent la nécessité d’avoir une vision la plus intégrée possible du système, et d’intensifier les interactions de recherches entre l’agronomie et l’écologie évolutive. / Following the ‘Grenelle of the environment’ conference, one of the current critical issues in agriculture is reducing the side effects of pesticides on the environment. Prolonged use of copper as a fungicide has led to copper accumulation in vineyard soils. Copper can induce aphytotoxicity. Its accumulation in plants may modify the morphology, biochemistry orphysiology of plant organs and may disturb the plant-pest interactions. The effects of coppercontamination of vineyard soils on the vine sensitivity to the pests received until now a verylimited attention. The present work investigates such effects.To carry out this work, we measured the levels of total and available copper in vineyard soils.We contaminated three grapevine cultivars with copper and measured the copper distributionand its effects on plant. Then, we evaluated the impact induced on three pests of the vine. Ourresults show that the available copper content in the soil is related to the total copper content(which results from the history of the plot), but also to total carbon and fine soil particlescontents. The copper absorption and the copper transferred to the aerial parts of the vine varyaccording to the cultivar. When copper contamination increases, the vine-pests interactionsare modified: a negative effects appears for Scaphoideus titanus and Plasmopara viticola,while important copper contaminations appears to promote the development ofNeofusicoccum parvum.These results underscore the need for a more integrated system approach, and to intensifyinteractions between agronomy and evolutionary ecology research.

Elevados valores de incidência da podridão apical da goiaba vêm sendo observados em levantamentos recentes no Estado de São Paulo. Entretanto, pouco se conhece sobre a etiologia e influência das variáveis ambientais no monociclo dessa doença. O objetivo desse trabalho foi identificar as espécies de Botryosphaeriaceae que causam a podridão apical e estudar as condições ambientais favoráveis, in vivo e in vitro, para o desenvolvimento do monociclo desses patógenos. Para a identificação das espécies, 56 isolados monospóricos foram obtidos das principais regiões produtoras de goiabas do Estado de São Paulo. Com auxílio da caracterização morfológica e de análises filogenéticas realizadas com dados de sequências de DNA da região ITS e β-tubulina foram identificadas as espécies Fusicoccum aesculi, Neofusicoccum parvum e Neofusicoccum ribis. O crescimento micelial dessas espécies foi avaliado sob as temperaturas de 10, 15, 20, 25, 30, 35 e 40 ºC. A germinação de conídios foi avaliada sob temperaturas de 10, 15, 20, 25, 30, 35 e 40 ºC com períodos de molhamentos de 4, 6, 12, 24 e 48 horas. Nos experimentos in vivo goiabas cv. Kumagai foram inoculadas com suspensões de conídios das três espécies identificadas com auxílio de ferimentos artificiais. Esses frutos foram mantidos sob as temperaturas de 15, 20, 25, 30 e 35 ºC com períodos de molhamento após a inoculação de 6, 12, 24 e 48 horas. Avaliou-se o período de incubação, o diâmetro da lesão ao sétimo dia após a inoculação e taxa de crescimento da lesão. A temperatura ótima para o crescimento micelial de N. ribis foi de 28,2 ºC e não houve diferença entre as temperaturas ótimas de N. parvum e F. aesculi, que foram de aproximadamente 31 ºC. A temperatura ótima para germinação dos conídios foi de 30 ºC para as três espécies. Houve incremento na germinação com o aumento do período de molhamento e na faixa de temperatura ótima encontram-se valores superiores a 70% de germinação a partir do molhamento de 6 horas. A espécie F. aesculi alcança elevadores valores de germinação a 40 ºC em períodos de molhamento superiores a 12 horas. A temperatura ótima para o crescimento da lesão foi de aproximadamente 30 ºC para as três espécies e o período molhamento de 48 horas proporcionou um aumento na severidade da doença quando comparado com o molhamento de 6 horas. Os menores períodos de incubação foram obtidos na temperatura de 30 ºC e com períodos de molhamento de 48 horas após a inoculação. Não houve diferença entre os valores das taxas de crescimento da lesão das três espécies e média das taxas das três espécies foi de 0,4. A doença se desenvolveu melhor em condições de temperaturas elevadas e períodos prolongados de umidade. Esse é o primeiro relato de Neofusicoccum parvum e Neofusicoccum ribis associados à podridão apical no Brasil. / Increased incidence of stylar-end rot in guava fruits have been observed in the São Paulo State - Brazil. However, little is known about its etiology and the influence of environment variables on the monocycle of this disease. This study aimed to identify Botryosphaeriaceae species that cause stylar-end rot in guava and to analyze the favorable environment conditions, in vitro and in vivo, for the monocycle development of this pathogen. We used 56 monosporic isolates from diseased fruit from guava producing regions from the São Paulo State - Brazil. The morphological and phylogenetic analyses performed with sequence data of the ITS region and β- tubulin allowed to identify the species Fusicoccum aesculi, Neofusicoccum parvum and Neofusicoccum ribis, as causal agents of the disease. The mycelial growth was evaluated at 10, 15, 20, 25, 30, 35 and 40 °C. Conidial germination was evaluated at 10, 15, 20, 25, 30, 35 and 40 °C with wetness periods of 4, 6, 12, 24 and 48 hours. In the in vivo experiments, guavas cv. Kumagai were inoculated with conidial suspensions of the three species identified by artificial wounds. The fruits were kept at 15, 20, 25, 30 and 35 °C with wetness periods of 6, 12, 24 and 48 hours after inoculation. We evaluated the incubation period, the lesion diameter on the seventh day after inoculation and the lesion growth rate. The optimum temperature for mycelial growth of N. ribis was 28.2 °C and there was no difference between the optimal temperatures (approximately 31 ºC) for N. parvum and F. aesculi. The optimal temperature for conidial germination was 30 ºC for the three species. There was an increase in germination with the increase of the wetness period. Within the optimal temperature range, values were higher 70% of germination after a wetness period of 6 hours. F. aesculi reached high germination values at 40 °C in wetness periods exceeding 12 hours. The optimal temperature for lesion growth rate was approximately 30 °C for the three pathogens and the wetness period of 48 hours caused higher disease severity as compared with the wetness period of 6 hours. The shortest incubation periods were obtained at 30 ºC with wetness periods of 48 hours. There was no difference between lesion growth rate for the three species and the average rate for three pathogens was 0.4/day. The best conditions for the disease development are high temperatures and prolonged periods of moisture. This is the first report of Neofusicoccum parvum and Neofusicoccum ribis associated with stylarend rot of guava fruits in Brazil.

Elevados valores de incidência da podridão apical da goiaba vêm sendo observados em levantamentos recentes no Estado de São Paulo. Entretanto, pouco se conhece sobre a etiologia e influência das variáveis ambientais no monociclo dessa doença. O objetivo desse trabalho foi identificar as espécies de Botryosphaeriaceae que causam a podridão apical e estudar as condições ambientais favoráveis, in vivo e in vitro, para o desenvolvimento do monociclo desses patógenos. Para a identificação das espécies, 56 isolados monospóricos foram obtidos das principais regiões produtoras de goiabas do Estado de São Paulo. Com auxílio da caracterização morfológica e de análises filogenéticas realizadas com dados de sequências de DNA da região ITS e β-tubulina foram identificadas as espécies Fusicoccum aesculi, Neofusicoccum parvum e Neofusicoccum ribis. O crescimento micelial dessas espécies foi avaliado sob as temperaturas de 10, 15, 20, 25, 30, 35 e 40 ºC. A germinação de conídios foi avaliada sob temperaturas de 10, 15, 20, 25, 30, 35 e 40 ºC com períodos de molhamentos de 4, 6, 12, 24 e 48 horas. Nos experimentos in vivo goiabas cv. Kumagai foram inoculadas com suspensões de conídios das três espécies identificadas com auxílio de ferimentos artificiais. Esses frutos foram mantidos sob as temperaturas de 15, 20, 25, 30 e 35 ºC com períodos de molhamento após a inoculação de 6, 12, 24 e 48 horas. Avaliou-se o período de incubação, o diâmetro da lesão ao sétimo dia após a inoculação e taxa de crescimento da lesão. A temperatura ótima para o crescimento micelial de N. ribis foi de 28,2 ºC e não houve diferença entre as temperaturas ótimas de N. parvum e F. aesculi, que foram de aproximadamente 31 ºC. A temperatura ótima para germinação dos conídios foi de 30 ºC para as três espécies. Houve incremento na germinação com o aumento do período de molhamento e na faixa de temperatura ótima encontram-se valores superiores a 70% de germinação a partir do molhamento de 6 horas. A espécie F. aesculi alcança elevadores valores de germinação a 40 ºC em períodos de molhamento superiores a 12 horas. A temperatura ótima para o crescimento da lesão foi de aproximadamente 30 ºC para as três espécies e o período molhamento de 48 horas proporcionou um aumento na severidade da doença quando comparado com o molhamento de 6 horas. Os menores períodos de incubação foram obtidos na temperatura de 30 ºC e com períodos de molhamento de 48 horas após a inoculação. Não houve diferença entre os valores das taxas de crescimento da lesão das três espécies e média das taxas das três espécies foi de 0,4. A doença se desenvolveu melhor em condições de temperaturas elevadas e períodos prolongados de umidade. Esse é o primeiro relato de Neofusicoccum parvum e Neofusicoccum ribis associados à podridão apical no Brasil. / Increased incidence of stylar-end rot in guava fruits have been observed in the São Paulo State - Brazil. However, little is known about its etiology and the influence of environment variables on the monocycle of this disease. This study aimed to identify Botryosphaeriaceae species that cause stylar-end rot in guava and to analyze the favorable environment conditions, in vitro and in vivo, for the monocycle development of this pathogen. We used 56 monosporic isolates from diseased fruit from guava producing regions from the São Paulo State - Brazil. The morphological and phylogenetic analyses performed with sequence data of the ITS region and β- tubulin allowed to identify the species Fusicoccum aesculi, Neofusicoccum parvum and Neofusicoccum ribis, as causal agents of the disease. The mycelial growth was evaluated at 10, 15, 20, 25, 30, 35 and 40 °C. Conidial germination was evaluated at 10, 15, 20, 25, 30, 35 and 40 °C with wetness periods of 4, 6, 12, 24 and 48 hours. In the in vivo experiments, guavas cv. Kumagai were inoculated with conidial suspensions of the three species identified by artificial wounds. The fruits were kept at 15, 20, 25, 30 and 35 °C with wetness periods of 6, 12, 24 and 48 hours after inoculation. We evaluated the incubation period, the lesion diameter on the seventh day after inoculation and the lesion growth rate. The optimum temperature for mycelial growth of N. ribis was 28.2 °C and there was no difference between the optimal temperatures (approximately 31 ºC) for N. parvum and F. aesculi. The optimal temperature for conidial germination was 30 ºC for the three species. There was an increase in germination with the increase of the wetness period. Within the optimal temperature range, values were higher 70% of germination after a wetness period of 6 hours. F. aesculi reached high germination values at 40 °C in wetness periods exceeding 12 hours. The optimal temperature for lesion growth rate was approximately 30 °C for the three pathogens and the wetness period of 48 hours caused higher disease severity as compared with the wetness period of 6 hours. The shortest incubation periods were obtained at 30 ºC with wetness periods of 48 hours. There was no difference between lesion growth rate for the three species and the average rate for three pathogens was 0.4/day. The best conditions for the disease development are high temperatures and prolonged periods of moisture. This is the first report of Neofusicoccum parvum and Neofusicoccum ribis associated with stylarend rot of guava fruits in Brazil.