Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2017-04-12T17:59:45Z No. of bitstreams: 1 texto completo.pdf: 2243894 bytes, checksum: ab4e44704d5ce4a8b8eed861c68ee7fc (MD5) / Made available in DSpace on 2017-04-12T17:59:45Z (GMT). No. of bitstreams: 1 texto completo.pdf: 2243894 bytes, checksum: ab4e44704d5ce4a8b8eed861c68ee7fc (MD5) Previous issue date: 2015-02-23 / Fundação de Amparo à Pesquisa do Estado de Minas Gerais / A goiabeira (Psidium guajava) é uma cultura de grande importância, destacando-se como uma atividade econômica e social de grande expressão. Nativa da América Tropical, a goiabeira, é amplamente cultivada em todas as regiões do Brasil. A produção da goiabeira tem sido limitada por vários fatores, dentre eles, destacam-se as doenças, com especial ênfase às que ocorrem em pós-colheita. A pinta-preta da goiaba é a doença pós-colheita de maior incidência. As goiabas acometidas pela doença apresentam sintomas que constituem-se, inicialmente, de pontos deprimidos, os quais evoluem rapidamente na superfície dos frutos, de forma concêntrica e tornam-se lesões de coloração escura com diversos pontos negros correspondendo aos sinais do patógeno. Vários trabalhos associam a pinta-preta da goiaba ao fungo Phyllosticta psidiicola entretanto, ainda não existem trabalhos mais abrangentes com estudos taxonômicos mais apurados que investiguem qual o real agente etiológico dessa doença. Além da pinta-preta da goiaba, existem outras doenças de pós-colheita muito comuns causadas por fungos da ordem Botryosphaeriales como a podridão parda associada ao fungo Dothiorella dominicana e a podridão de Lasiodiplodia associada à Lasiodiplodia theobromae. Entretanto, também existe uma carência de estudos mais abrangentes associados a essas doenças. Nos últimos anos novas espécies têm sido propostas a partir de estudos moleculares, evidenciando a existência de um complexo de espécies. Assim, os objetivos deste trabalho foram estudar a etiologia da pinta-preta da goiaba baseada na combinação de características morfológicas e moleculares, verificar o relacionamento das espécies causadoras dessa doença com as espécies endofíticas, estabelecer o posicionamento filogenético das espécies encontradas, bem como comprovar a patogenicidade das espécies associadas. Foram realizadas coletas de goiabas com sintomas de podridões pós- colheita, frutos secos e mumificados nos mercados do município de Viçosa – Minas Gerais e em uma área de cultivo no município de Piraúba – Minas Gerais. Folhas, ramos e frutos sadios foram coletados nesta mesma área, a fim de obter os isolados endofíticos correspondentes a área com elevada ocorrência da doença na pós- colheita. Isolados monospóricos foram obtidos e armazenados. Estes tiveram o DNA extraído e a região TEF1-α amplificada e sequenciada. A partir dos resultados das análises filogenéticas, um isolado representativo de cada espécie foi selecionado para a caracterização morfológica e testes de patogenicidade. Seis espécies de Botryosphaeriales foram identificadas, entre elas, três pertencentes ao gênero Neofusicoccum, uma pertencente ao gênero Phyllosticta e duas espécies de Lasiodiplodia. Entre estas espécies, duas serão propostas como novas (Neofusicoccum sp. e Lasiodiplodia sp.). Todas as espécies foram comparadas morfológica e filogeneticamente, com exceção de Lasiodiplodia sp. que não esporulou em meio de cultura. Todas as espécies tiveram patogenicidade comprovada. Os resultados deste trabalho serão fundamentais para futuros estudos envolvendo medidas de manejo da doença, programas de quarentena e especialmente, para o desenvolvimento de variedades de goiabas resistentes à pinta- preta e outras podridões de frutos. / The guava (Psidium guajava) is an fruit crop of great importance, standing out as an economic and social activity of great expression. Native to tropical America, the guava, is widely cultivated in all regions of Brazil. The production of guava has been limited by several factors, specially the diseases, with special emphasis to those that occur in post-harvest. The guava black spot is a post-harvest higher incidence disease. Diseased guavas presents symptoms that are initially depressed points, which are changing rapidly on the surface of the fruit, concentrically and become dark-colored with many black spots corresponding to the signs of the pathogens in the lesions. Several studies associate the black guava spot with the fungus Phyllosticta psidiicola however, there are no more comprehensive work with more accurate taxonomic studies investigating the real causative agent of this disease. Besides the black guava spot, there are other very common post-harvest fungal diseases caused by the order Botryosphaeriales as the brown fruit rot disease associated with Dothiorella dominicana and the Lasiodiplodia rot associated with Lasiodiplodia theobromae. However there is also a lack of more comprehensive studies associated with these diseases. In recent years new species have been proposed based specially on molecular studies, showing the existence of a complex of species. The objectives of this study were to study the etiology of black spot of guava based on a combination of morphological and molecular characteristics, verify the relationship of the species that cause this disease with the endophytic species, establish the phylogenetic position of the species found, as well as prove the pathogenicity of the associated species. Guavas samples were collected with symptoms of post-harvest decay as also dried and mummified fruit disease in Viçosa local markets - Minas Gerais and in a farming area with a high occurrence of the disease after harvesting in the city of Piraúba - Minas Gerais. Healthy leaves, branches and fruits were collected in the same area, in order to get the endophytic isolates from this high incidence area. Single spore isolates were obtained and stored. These has the DNA extracted and TEF1-α region amplified and sequenced. Based on the results of the phylogenetic analysis one isolate of each species was selected for morphological characterization and pathogenicity test. Six species belonging to the Botryosphaeriales were identified, in which three belongs to Neofusicoccum genera, one belongs to the genera Phyllosticta and two species of Lasiodiplodia. Among these species, two will be proposed as new (Neofusicoccum sp. and Lasiodiplodia sp.). All species were compared morphologically and phylogenetically except Lasiodiplodia sp. since it was not sporulating in culture. All species proved to be pathogenic. Despite its importance, the etiology of black spot and other post-harvest fruit rot diseases has been neglected and remained obscure for a long time. The results of this work will be critical for future studies involving disease management, quarantine programs and especially for the development of guava varieties resistant to the black spot and others post-harvest fruit rot diseases.

Macrophoma rot is a general term for fruit rots of Vitis spp. caused by the fungus Neofusicoccum ribis (syn. Botryosphaeria ribis) or closely related or renamed taxa, including Botryosphaeria dothidea. While mainly observed as a fruit pathogen of muscadine grape, the disease has recently been observed on bunch grapes in Virginia. Isolates (N = 835) were collected from Petit Manseng fruit clusters from seven Virginia vineyards in 2018 and 2019. A subset of these isolates was sequenced using three primer sets (ITS, RPB2, and EF). The preliminary result showed a single taxonomic strain of N. ribis. A controlled inoculation study of Petit Manseng clusters verified that infection could occur anytime between bloom and 2 weeks post-veraison; however, both the mean cluster incidence and the severity of Macrophoma rot did not differ from each other at any growth stage during the season. A season-long cluster exposure experiment showed that any amount of sun exposure significantly increased Macrophoma rot severity compared to shaded clusters, and that full sun exposure was associated with greatest rot severity. This finding contravenes current management recommendations for Macrophoma rot, and it raises yet unanswered questions as to why exposed clusters are more susceptible to Macrophoma rot than are shaded clusters. An in vitro fungicide assay study using nine fungicides identified captan, thiophanate-methyl, and tetraconazole as potential candidates for management of Macrophoma rot which need to be investigated further. / Master of Science in Life Sciences / Macrophoma rot is a general term for fruit rots of grapes caused by the pathogenic fungi in the family Botryosphaeriaceae. The rot is mainly observed on Muscadine grapes, but recently more cases were found on a wine grape cultivar Petit Manseng in Virginia. Macrophoma rot symptoms begin as dark brown, circular lesions on the surface of the berry and look similar to sunburn and other fruit rots. As the disease progresses, the lesion envelopes the entire berry and black fruiting bodies develop. Severe cases may lead to crop loss. The same group of pathogens is also associated with rots on other crops including apple, pear, olive, and kiwis. Very little is known about the disease cycle and the control of Macrophoma rot, therefore, an investigation into this fungal pathogen was needed. Multiple studies with the wine grape variety Petit Manseng were conducted during the 2018-2019 growing seasons, including a survey, leaf removal trial, and an inoculation study. Results showed that a species called Neofusicoccum ribis was found in vineyards across northern and central Virginia based on the genetic identification of fungal isolates collected at seven vineyards in those areas. Macrophoma symptoms were observed to be more prevalent and severe in more exposed clusters based on a leaf removal experiment. An artificial inoculation experiment revealed that grape clusters are susceptible to Neofusicoccum ribis at any time during the season. Based on the screening of nine fungicides, three chemicals (captan, thiophanate-methyl, and tetraconazole) showed promising results as possible management tools for Macrophoma rot. The knowledge collected will lead to an increase in understanding of this fungal pathogen and to further studies to manage Macrophoma rot.

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.

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.

Memoria para optar al título profesional de Ingeniero Agrónomo / Las enfermedades de la madera de la vid son cada vez más recurrentes en los viñedos y parrales de Chile y del mundo, infectando y mermando de manera considerable la producción. Se sabe que son enfermedades muy agresivas y que pueden provocar la muerte de la planta. Las investigaciones realizadas han demostrado la efectividad de agentes biológicos para prevenir y controlar plagas y enfermedades. En este estudio se determinó la eficacia de diferentes biocontroladores de distinto origen frente a Neofusicoccum australe, hongo asociado a la enfermedad del brazo muerto descrita como “Black dead arm”. Los ensayos se realizaron bajo condiciones de laboratorio y se usaron sarmientos provenientes de plantas sanas de uva de mesa cv. Thompson Seedless. Los sarmientos fueron inoculados con micelio del patógeno sobre una herida realizada en el centro de la estaca.

[EN] Almond is one of the main crops of Majorca Island. Since 2008, symptoms of severe decline of almond trees have been observed in several orchards from different areas of the Island. Disease symptoms are similar to those described by different authors in other parts of the world caused by fungal trunk pathogens. In order to study the etiology of this problem, surveys were conducted on almond orchards distributed throughout the main growing regions in Majorca for six consecutive years (2009-2014). Based on morphological and molecular identification, 14 fungal species were recovered from almond wood samples: Collophora hispanica, Diplodia olivarum, D. seriata, Eutypa lata, E. leptoplaca, Fomitiporia mediterranea, Neofusicoccum luteum, N. mediterraneum, N. parvum, Omphalotus olearius, Phaeoacremonium amygdalinum, Pm. iranianum, Phellinus pomaceus and Pleurostomophora richardsiae and two species were recovered from one apricots orchard near almonds orchards: : Pm. minimum and Pm. venezuelense. Based on the DNA sequence analyses and morphological features, C. hispanica and Pm. amygdalinum proved distinct from all known species, and have been described. The most common species recovered from almond samples were P. richardsiae and D. seriata, followed by other species belonging to the family Botryosphaeriaceae and C. hispanica. The most frequently species isolated were also widely distributed and present in more regions. Subsequently, two pathogenicity tests were carried on almond trees by using representative isolates of some of the most frequent species. The first one was held for two consecutive years (2013 and 2014) with five species of Botryosphaeriaceae (D. olivarum, D. seriata, N. luteum, N. mediterraneum and N. parvum) and two species of Diatrypaceae (E. lata and E. leptoplaca). Fungi were inoculated on 1-2 years old almond trees of four different cultivars ('Jordi', 'Ferragnes', 'Pons' and 'Vivot') under field conditions. Nine months after inoculation, the total length of internal necrosis was evaluated. All species were pathogenic on almond. Neofusicoccum luteum caused the longest average lesion during the first year, and N. mediterraneum and N. parvum caused the longest lesion during the second year. Eutypa leptoplaca caused the shortest lesion length in both years of study. In addition, fungal lesion length varied depending on the variety of almond evaluated. In the first year of study, the more tolerant variety was 'Jordi', while in the second year, 'Ferragnes' and 'Vivot' varieties showed the highest degree of tolerance to fungal infection. In the second trial, almond seedlings variety 'Ferragnes' were inoculated with C. hispanica, Pm. amygdalinum, Pm. iranianum and P. richardsiae. Six months after inoculation the lesion length was evaluated. All species inoculated were pathogenic on almond, being P. richardsiae the most virulent species. Finally, the ability of some commercial fungicides to protect pruning wounds from infection by four species of Botryosphaeriaceae (D. seriata, N. luteum, N. mediterraneum and N. parvum) was evaluated. This study was conducted in two phases, an initial in vitro evaluation (mycelial growth assay) with 10 fungicides, followed by an evaluation of five fungicides, which proved to be effective in the in vitro trial, applied on pruning wounds at 1 and 7 days after inoculation. Internal lesion length and the percentage of re-isolation of the pathogen were calculated. tebuconazole and pyraclostrobin were the most effective fungicides in the in vitro evaluation, followed by cyproconazole and thiophanate-methyl. Thiophanate-methyl was the most effective fungicide to protect pruning wounds from infections caused by species of Botryosphaeriaceae. / [ES] Desde el año 2008, en parcelas de diferentes zonas de la isla se ha constatado la presencia síntomas de decaimiento de ramas y muerte de almendros, que recuerdan a los descritos por diferentes autores en otras zonas del mundo causados por hongos patógenos de la madera en diversos cultivos. Para estudiar su etiología se realizaron prospecciones en parcelas de almendros de la isla durante seis años (2009-2014). Se caracterizaron los síntomas y se tomaron muestras que se analizaron en laboratorio. En los análisis se obtuvieron 14 especies fúngicas de muestras de almendro: Collophora hispanica, Diplodia olivarum, D. seriata, Eutypa lata, E. leptoplaca, Fomitiporia mediterranea, Neofusicoccum luteum, N. mediterraneum, N. parvum, Omphalotus olearius, Phaeoacremonium amygdalinum, Pm. iranianum, Phellinus pomaceus, Pleurostomophora richardsiae y dos especies encontradas sólo en muestras de una parcela de albaricoqueros junto a parcelas de almendros: Pm. minimum y Pm. venezuelense. Collophora hispanica y Pm. amygdalinum son dos nuevas especies fúngicas. Las especies más frecuentes en las parcelas de almendro estudiadas fueron P. richardsiae y D. seriata, seguidas por las otras especies pertenecientes a la familia Botryosphaeriaceae y por C. hispanica. Las especies que se aislaron con mayor frecuencia, fueron a su vez las que tuvieron una distribución más amplia; presentes en más comarcas. Posteriormente, se estudió la patogenicidad a almendro de algunas de las especies detectadas. Se realizaron dos ensayos de patogenicidad, el primero se realizó dos años (2013 y 2014) con las cinco especies de Botryosphaeriaceae (D. olivarum, D. seriata, N. luteum, N. mediterraneum y N. parvum) y las dos de Diatrypaceae (E. lata y E. leptoplaca) aisladas, que se inocularon en árboles de 1 a 2 años de cuatro variedades de almendro ('Jordi', 'Ferragnes', 'Pons' y 'Vivot') en una parcela experimental. El ensayo se evaluó a los nueve meses de la inoculación, midiendo la longitud de las necrosis internas producidas. En este ensayo se demostró la patogenicidad a almendro de las siete especies que se ensayaron. Las especies que causaron las lesiones de mayor longitud fueron N. luteum el primer año de ensayo, y N. parvum y N. mediterraneum el segundo año. Ambos años, la especie que causó las lesiones de menor longitud fue E. leptoplaca. La dimensión de la lesión producida por el hongo inoculado dependía de la variedad de almendro evaluada. La variedad 'Jordi' fue la menos sensible el primer año de estudio, y 'Vivot' y 'Ferragnes' lo fueron el segundo año. En el segundo ensayo se estudió la patogenicidad de las especies C. hispanica, Pm. amygdalinum, Pm. iranianum y P. richardsiae en plantones de almendro de la variedad 'Ferragnes' en invernadero. Nuevamente, la evaluación se realizó a los seis meses de la inoculación, midiendo la longitud de las lesiones internas. Todas las especies inoculadas resultaron patógenas a almendro, siendo P. richardsiae la especie que causó la mayor longitud de lesión. Finalmente, se evaluaron fungicidas para la protección de heridas de poda frente a la infección por cuatro especies de Botryosphaeriaceae (D. seriata, N. luteum, N. mediterraneum y N. parvum). Este estudio se llevó a cabo en dos fases; en primer lugar, una evaluación in vitro (reducción del crecimiento miceliar) con diez fungicidas y, posteriormente, una evaluación de cinco de estos fungicidas, elegidos entre los más efectivos in vitro, aplicándolos en heridas de poda uno o siete días tras el corte y la inoculación. Como en los casos anteriores, para evaluar este estudio se midió la longitud de la lesión, pero además también se calculó el porcentaje de reaislamiento del patógeno inoculado en cada caso. Los fungicidas tebuconazol y piraclostrobin, seguidos de ciproconazol y metil tiofanato, se mostraron como los más efectivos en la evaluación in vitro, mientras que el fungicida más efectivo para la protección de / [CA] Des de l'any 2008, en parcel¿les de diferents zones de Mallorca s'han observat símptomes de decaïment de branques i mort d'ametllers, que recorden als que diferents autors han descrit en altres zones del món causats per fongs de fusta en diversos cultius. Per estudiar la seua etiologia es van realitzar prospeccions en parcel¿les d'ametllers de l'illa durant sis anys consecutius (2009-2014). En aquestes prospeccions es van caracteritzar símptomes i es van prendre mostres que es van analitzar al laboratori. En les anàlisis de laboratori es van obtenir 14 espècies fúngiques de mostres d'ametller: Collophora hispanica, Diplodia olivarum, D. seriata, Eutypa lata, E. leptoplaca, Fomitiporia mediterranea, Neofusicoccum luteum, N. mediterraneum, N. parvum, Omphalotus olearius, Phaeoacremonium amygdalinum, Pm. iranianum, Phellinus pomaceus i Pleurostomophora richardsiae i dues espècies trobades només a una parcel¿la d'albercoquers situada a prop de parcel¿les d'ametllers: Pm. minimum i Pm. venezuelense. Collophora hispanica i Pm. amygdalinum són dues espècies noves. Les espècies més freqüents en les parcel¿les d'ametller estudiades van ser P. richardsiae i D. seriata, seguides per les altres espècies pertanyents a la família Botryosphaeriaceae i per C. hispanica. Les espècies que es van aïllar amb més freqüència, van ser també les que van tenir una distribució més àmplia; presents en més comarques. Posteriorment, es va estudiar la patogenicitat d'algunes de les espècies detectades. Concretament, es van realitzar dos assajos de patogenicitat. El primer es va dur a terme dos anys (2013 i 2014), amb les cinc espècies de Botryosphaeriaceae (D. olivarum, D. seriata, N. luteum, N. mediterraneum i N. parvum) i les dues de Diatrypaceae (E. lata i E. leptoplaca) aïllades, que es van inocular en arbres d'1 a 2 anys de quatre varietats d'ametller ('Jordi', 'Ferragnes', 'Pons' i 'Vivot') en una parcel¿la experimental. L'assaig es va avaluar als nou mesos des de la inoculació, mesurant la longitud de les necrosis internes produïdes. En aquest assaig es va demostrar la patogenicitat respecte l'ametller de les set espècies que es van assajar. Les espècies que van causar les lesions de major longitud van ser N. luteum el primer any d'assaig, i N. parvum i N. mediterraneum el segon any. En els dos anys dels assajos l'espècie que va causar lesions de menor longitud va ser E. leptoplaca. Es va observar que la dimensió de la lesió causada pel fong inoculat depenia de la varietat d'ametller avaluada. La varietat 'Jordi' va ser la menys sensible el primer any d'estudi, i 'Vivot' i 'Ferragnes' ho van ser el segon any. En el segon assaig es va estudiar la patogenicitat de les espècies C. hispanica, Pm. amygdalinum, Pm. iranianum i P. richardsiae en plançons d'ametller de la varietat 'Ferragnes' en hivernacle. L'avaluació es va realitzar als 6 mesos de la inoculació, mesurant la longitud de les lesions necròtiques internes. Totes les espècies van resultar patògenes d'ametller, sent P. richardsiae l'espècie que va causar les lesions més extenses. Finalment, es va realitzar un estudi d'avaluació de fungicides per a la protecció de ferides de poda enfront de la infecció per quatre espècies de Botryosphaeriaceae (D. seriata, N. luteum, N. mediterraneum i N. parvum). Aquest estudi es va dur a terme en dues fases, una avaluació in vitro (reducció del creixement micelià) amb 10 fungicides i, posteriorment, una avaluació de cinc d'aquests fungicides, elegits entre els més efectius in vitro, aplicant-los en ferides de poda a 1 o 7 dies després del tall i la inoculació. Novament per avaluar aquest estudi es va mesurar la longitud de la lesió, però a més també es va calcular el percentatge de reaïllament del patogen inoculat en cada cas. Els fungicides tebuconazol i piraclostrobin, seguits de ciproconazol i metil tiofanat, es van mostrar com els més efectius in vitro, mentre que el fungicida més efect / Olmo García, D. (2016). Etiología y control de las enfermedades fúngicas de la madera del almendro en la isla de Mallorca [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/60158

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.