Submitted by (lucia.rodrigues@ufrpe.br) on 2017-03-21T13:49:42Z No. of bitstreams: 1 Luiz Gustavo de Lima Melo.pdf: 741943 bytes, checksum: 78412f9372d9818a63e65dee4cf2dfdb (MD5) / Made available in DSpace on 2017-03-21T13:49:42Z (GMT). No. of bitstreams: 1 Luiz Gustavo de Lima Melo.pdf: 741943 bytes, checksum: 78412f9372d9818a63e65dee4cf2dfdb (MD5) Previous issue date: 2012-03-02 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Lasiodiplodia Theobromae is responsible for numerous diseases in many economically important crops that may be infected in both pre and postharvest, may cause different symptoms. In grapevine, the pathogen infects the berries, causing Lasiodiplodia rot, causing injuries that impair or prevent the marketing of the fruit. Given the importance of this pathogen and the high levels of infection in the Lower Basin producing area of Submédio do Vale São Francisco, a region important in the production of fine table grapes, this study aimed to evaluate the epidemiological parameters of Lasiodiplodia rot in berries of grape cvs. Italy Muscat and Thompson Seedless. We conducted pathogenicity and aggressiveness of 10 isolates of L. theobromae (L1, L2, L3, L4, L5, L7 L6, L8, L9 and L10), inoculated on wounds produced by a punch with eight needles 2 mm deep on cv. Italy Muscat at a concentration of 106 spores/mL. All isolates tested were pathogenic to grape. In the test of aggression L1 isolate was more aggressive, with higher average compared to other injuries, which is used in subsequent tests. We studied the influence of region inoculation inoculum (equatorial and stalk), methods of inoculation (spray and drop) in addition to epidemiological parameters: concentration of inoculum (103, 104, 105, 106 and 107 spores/ml), a period of wetting (0, 12, 24, 36, 48h) and temperature (2, 5, 10, 15, 20, 25 ° C). Regarding to the inoculum concentration, for both cvs. Italy Muscat and Thompson Seedless, 107 conidia/ml was the best, using 10 μl applied on the equatorial region by droplet inoculation method promoting the highest development of lesions on grape berries. Concerning to the epidemiological parameters, temperature and wetness, there has been a further development of lesions in about 25 ° C under a 48 h wetting period. / Lasiodiplodia theobromae é responsável por inúmeras doenças em diversas culturas de importância econômica que podem vir a ser infectadas tanto em pré como em pós-colheita, podendo causar diferentes sintomas. Na videira, o patógeno infecta as bagas, causando a podridão por lasiodiplodia, provocando lesões que prejudicam ou inviabilizam a comercialização do fruto. Diante da relevância deste fitopatógeno e dos altos níveis de infecção na região produtora do Submédio São Francisco, região importante na produção de uvas finas de mesa, este trabalho buscou avaliar os parâmetros epidemiológicos da podridão por lasiodiplodia em bagas de uvas das cultivares Itália Muscat e Thompson Seedless. Realizou-se teste de patogenicidade e agressividade com 10 isolados de L. theobromae (L1, L2, L3, L4, L5, L6 L7, L8, L9 e L10), inoculados sobre ferimentos produzidos por um furador com oito agulhas de 2 mm de comprimento em uvas da cv. Itália Muscat na concentração de 106 conídios/mL. Todos os isolados testados foram patogênicos a uva. O isolado L1 foi o mais agressivo, apresentando maiores médias de lesões em comparação aos demais, sendo este utilizado nos testes posteriores. Estudou-se a influência da região de deposição do inoculo (equatorial e peduncular), métodos de inoculação (gota e pulverização) além dos parâmetros epidemiológicos: concentração de inóculo (103, 104, 105, 106 e 107 conídios/mL), período de molhamento (0, 12, 24, 36, 48 h), e temperatura (2, 5, 10, 15, 20, 25ºC). Em relação à concentração de inóculo foi verificado que tanto para a cv. Itália Muscat quanto na cv Thompson Seedless a melhor concentração de 107 conídios/mL, aplicado 10 μL na região equatorial pelo método de inoculação de gota foi o que proporcionou maior desenvolvimento de lesão sobre as bagas de uva. Em relação aos aspectos epidemiológicos, temperatura e período de molhamento, verificou-se um maior desenvolvimento de lesão em torno de 25°C sob um período de 48 horas de molhamento.

MELO, J. G. M. Diversidade genética e patogenicidade de Lasiodiplodia theobromae associado ao cajueiro (Anacardium occidentale). 2010. 60 f. Dissertação (Mestrado em Agronomia/Fitotecnia) - Centro de Ciências Agrárias, Universidade Federal do Ceará, Fortaleza, 2010. / Submitted by Francisco Lacerda (lacerda@ufc.br) on 2014-07-18T19:23:53Z No. of bitstreams: 1 2010_dis_jgmmelo.pdf: 3168052 bytes, checksum: 25bb3844a3f6ad3a76bec38956b5b761 (MD5) / Approved for entry into archive by José Jairo Viana de Sousa(jairo@ufc.br) on 2014-07-21T18:39:42Z (GMT) No. of bitstreams: 1 2010_dis_jgmmelo.pdf: 3168052 bytes, checksum: 25bb3844a3f6ad3a76bec38956b5b761 (MD5) / Made available in DSpace on 2014-07-21T18:39:42Z (GMT). No. of bitstreams: 1 2010_dis_jgmmelo.pdf: 3168052 bytes, checksum: 25bb3844a3f6ad3a76bec38956b5b761 (MD5) Previous issue date: 2010 / The cashew has a major socio-economic and cultural development for the Northeast region, by generating employment for farmers and is frequently their only source of income. Thus, research on this fruit has increased greatly in recent years, aimed primarily to reduce damage and increase plant productivity. Diseases, as in other crops, cause significant damage. Among cashew diseases, gummosis and black stem rot, both caused by the fungus Lasiodiplodia theobromae, stand out among the major diseases in some regions of the semi-arid. However, there are few studies on the biology, morphology, genetic characterization of this pathogen, a fact that burden their management. This study aimed to characterize the genetic and pathogenic diversity of a population of 22 isolates of L. theobromae associated with cashew, using microsatellite markers and inoculations in cashew plants. For molecular characterization, DNA was extracted from mycelium of each isolated of the fungus grown in liquid medium. Each DNA sample was subjected to a polymerase chain reaction (PCR) with 15 pairs of primers designed to flank specific regions containing simple repeated sequences (microsatellites) in this species. The amplified products were observed in gel electrophoresis stained with ethidium bromide, and the data recorded on a binary and analyzed by grouping unbalanced based on arithmetic mean (UPGMA) using the program MVSP. Genetic similarities were estimated by the coefficient of Nei and Li The results indicated a large genetic variability of the population, represented by different groups of similarity ranging from 22.2% to 100%. Three of these isolates showed 100% genetic similarity with the markers tested. However, they were morphologically differences and one of them was originated from different location than the others. The data showed that the genetic similarity alone should not be applied as the only factor of diversity among isolates. To test the pathogenicity, each isolate was inoculated on cashew seedlings. Lesion development size and recovery of the fungus were recorded. The results showed no statistical differences between the isolates, which demonstrates that, regardless of place of origin, all isolates were pathogenic, and its growth rate decreases, i. e, the plant must produce some form of defense against pathogen, which hinders their infection. / O cajueiro tem uma grande importância sócio-econômica e cultural para a região Nordeste, por gerar emprego para os agricultores, sendo por muitas vezes a sua única fonte de renda. Assim, as pesquisas com esta fruteira têm aumentado bastante nos últimos anos, visando primariamente, reduzir os danos fitossanitários e elevar a produtividade. As doenças, como nas demais espécies cultivadas, causam significativos danos. Dentre as doenças do cajueiro, a resinose e a podridão preta das hastes, ambas causadas pelo fungo Lasiodiplodia theobromae, se destacando entre as principais doenças em algumas regiões do semi-árido nordestino. Porém, ainda há poucos estudos sobre a biologia, morfologia, caracterização genética desse patógeno, fato que dificulta o seu manejo. Este estudo objetivou caracterizar a diversidade genética e patogênica de uma população com 22 isolados de L. theobromae associada ao cajueiro, utilizando marcadores microssatélites e inoculações em plantas de cajueiro. Para a caracterização molecular, extraiu-se o DNA de cada isolado a partir do micélio do fungo crescido em meio líquido. Cada amostra foi submetida à reação de polimerase em cadeia (PCR) com 15 pares de iniciadores desenvolvidos para flanquear regiões especificas contendo sequências simples repetidas (microssatélites) dessa espécie. Os produtos amplificados foram observados em gel de eletroforese corados com brometo de etídio, e os dados tabulados em planilha binária e analisados pelo método de agrupamento não balanceado baseado na média aritmética (UPGMA), utilizando o programa MVSP. As similaridades genéticas foram estimadas pelo coeficiente de Nei e Li. Os resultados indicaram uma grande variabilidade genética da população avaliada, representada por diversos grupos de similaridade variando de 22,2% a 100%. Três destes isolados apresentaram entre si, 100% de similaridade genética com os marcadores testados. No entanto, eram morfologicamente diferentes e um deles era procedente de localidade diferente que os demais. Os dados mostraram que a similaridade genética por si só não deve ser aplicado como único fator de diversidade entre isolados. Para se testar a patogenicidade, os isolados foram inoculados em mudas de cajueiro, analisando-se o tamanho da lesão e a taxa de crescimento do fungo. Os resultados mostraram não haver diferenças estatísticas entre os isolados, fato que demonstra que, independente do local de origem, todos os isolados mostraram-se patogênicos, e a sua taxa de crescimento decresce, isto é, a planta deve produzir algum tipo de defesa contra o patógeno, o que dificulta a sua infecção.

LIMA, J. S. Diversidade cultural, morfológica e patogênica de isolados de Lasiodiplodia theobromae associados a frutíferas tropicais. 2011. 57 f. Dissertação (Mestrado em Agronomia/Fitotecnia) - Centro de Ciências Agrárias, Universidade Federal do Ceará, Fortaleza, 2011. / Submitted by Francisco Lacerda (lacerda@ufc.br) on 2014-07-17T20:42:49Z No. of bitstreams: 1 2011_dis_jslima.pdf: 799039 bytes, checksum: f02184b16bd28dc37fcf64844fac7c7b (MD5) / Approved for entry into archive by José Jairo Viana de Sousa(jairo@ufc.br) on 2014-07-21T18:41:57Z (GMT) No. of bitstreams: 1 2011_dis_jslima.pdf: 799039 bytes, checksum: f02184b16bd28dc37fcf64844fac7c7b (MD5) / Made available in DSpace on 2014-07-21T18:41:57Z (GMT). No. of bitstreams: 1 2011_dis_jslima.pdf: 799039 bytes, checksum: f02184b16bd28dc37fcf64844fac7c7b (MD5) Previous issue date: 2011 / Lasiodiplodia theobromae is an ubiquitous, polyphagous and opportunistic fungus with a reduced pathogenic ability. Nevertheless, it may infect several plant species over tropical and temperate regions, causing many different kinds of symptoms. The increasingly expansion of diseases caused by L. theobromae in tropical fruit plants has been imposing severe losses both at orchard level and in post-harvest at market, threaten the fruit crop industry in Northeast Region of Brazil. Therefore there is an urgent need for research pursuing basic knowledge on population biology of the fungus and host-pathogen interactions. This study aimed to characterize a L. theobromae population which has been associated to tropical plant species growing under different ecosystems in northeastern Brazil. Colony growth in culture, color and size of conidia and ability to cause disease upon inoculation on cashew nut (Anacardium occidentale), soursop (Annona muricata), yellow mombin (Spondias mombin) and Brazil plum (Spondias tuberosa) were evaluated. He study was carried out at Plant Pathology Lab and screenhouse of Embrapa Agroindustria Tropical in Fortaleza, Ceará State. Results showed a high diversity of morphology and hyphal growth among fungus isolates. Also, a very high variability on disease expression upon inoculation into four plant species was observed. However, it was found a lack of specificity of isolates as to infect cashew plants, since all isolates were able to infect cashew with similar high aggressiveness, which demonstrated a high degree of susceptibility of cashew clone used (CCP 76). Similar results were found also for soursop plants as host. Brazil plum showed a very high resistance to all isolates. The data points out for the existence of morphological and pathogenic interactions within L. theobromae population studied. According with the results, altitude and region of isolate origin has no effect on the studied features. / Lasiodiplodia theobromae é um fungo cosmopolita, polífago e oportunista, com reduzida especialização patogênica, que infecta espécies de plantas em regiões tropicais e temperadas, causando os mais variados sintomas. A crescente expansão das doenças causadas por L. theobromae em frutíferas tropicais vem causando inestimáveis perdas, tanto no sistema produtivo como em pós-colheita, representando uma ameaça à fruticultura no Nordeste. Daí a necessidade de conhecimentos básicos sobre a biologia populacional e a interação do patógeno com as plantas hospedeiras. Este estudo teve como objetivo caracterizar isolados de L. theobromae associados a frutíferas tropicais de diferentes regiões, avaliando o aspecto cultural, morfológico e patogênico. Foram avaliados o crescimento micelial, coloração da colônia, dimensões dos conídios e patogenicidade dos isolados em mudas de cajazeira, cajueiro, gravioleira e umbuzeiro. O trabalho foi realizado na Casa de Vegetação e no Laboratório de Fitopatologia da Embrapa Agroindústria Tropical. Os dados de caracterização morfocultural mostraram haver alta diversidade na população do patógeno. As inoculações realizadas nas quatro diferentes espécies hospedeiras apontaram alta variabilidade patogênica entre os isolados do fungo. Em mudas de cajueiro CCP 76 não foi possível observar especificidade patogênica, pois todos os isolados apresentaram similar nível de agressividade, demonstrando a suscetibilidade deste clone ao patógeno, suscetibilidade essa, também observada em gravioleira. O umbuzeiro foi a espécie que apresentou maior resistência ao fungo. Os dados mostraram que existe interação entre as características morfoculturais e a agressividade dos isolados de L. theobromae. De acordo com os resultados, a altitude dos locais de origem dos isolados não influencia em suas características morfoculturais e patogênicas.

MOREIRA, R. C. Reação de clones de cajueiro comum à resinose. 2011. 50 f. Dissertação (Mestrado em Agronomia/Fitotecnia) - Centro de Ciências Agrárias, Universidade Federal do Ceará, Fortaleza, 2011. / Submitted by Francisco Lacerda (lacerda@ufc.br) on 2014-07-29T20:47:43Z No. of bitstreams: 1 2011_dis_rcmoreira.pdf: 1025018 bytes, checksum: e1b7f6949d2d3421c19394408a8c619f (MD5) / Approved for entry into archive by José Jairo Viana de Sousa(jairo@ufc.br) on 2014-08-06T20:28:35Z (GMT) No. of bitstreams: 1 2011_dis_rcmoreira.pdf: 1025018 bytes, checksum: e1b7f6949d2d3421c19394408a8c619f (MD5) / Made available in DSpace on 2014-08-06T20:28:35Z (GMT). No. of bitstreams: 1 2011_dis_rcmoreira.pdf: 1025018 bytes, checksum: e1b7f6949d2d3421c19394408a8c619f (MD5) Previous issue date: 2011 / The cashew nut (Anacardium occidentale) represents a social and economic tradition in Brazilian northeastern as one of the most important source of income and labor for its people. The releasing of early-dwarf clones contributed to increase yield and nut and apple quality, allowing gain in profit by small growers, in spite of increase genetic vulnerability of orchards. Cashew gummosis is presently the most important disease of this species in semi-arid regions of northeast. This study aimed to evaluate reactions of common selected clones of cashew to gummosis under high disease pressure of semi-arid region. Twenty clones selected by the cashew breeding program of Embrapa Agroindústria Tropical were evaluated for three consecutive years. Clones named CNPAT 06, CNPAT 08, CNPAT 11, CNPAT 12 and CNPAT 13 attained the highest levels of resistance, while CNPAT 07, CNPAT 09, CNPAT 14, COMUM 05 and COMUM 31 otherwise were the most susceptible ones. It was also found that clones CNPAT 08, CNPAT 11 and CNPAT 15 are able to heal gummosis cankers, which points to a new kind of resistance to be considered. Finally, it was shown that monitoring gummosis incidence it is possible to estimate disease severity. / O cajueiro (Anacardium occidentale) no nordeste brasileiro caracterizando-se como uma das mais importantes fontes de emprego e renda das populações. O lançamento de clones de cajueiro tipo anão-precoce aumentou a produtividade e qualidade das castanhas e pendúculos, permitindo uma exploração comercial mais vantajosa aos produtores, porem a uniformidade genética deixou os pomares mais vulneráveis. A resinose hoje é considerada a principal doença do cajueiro no semiárido nordestino. Esse estudo teve por objetivo avaliar a reação de genótipos de cajueiro do tipo comum à resinose sob condições de elevada incidência no semiárido. Vinte clones de cajueiro-comum previamente selecionados pelo programa de melhoramento genético da Embrapa Agroindústria Tropical foram usados no estudo e dessa forma foi possível concluir que há fontes de resistência para uso em programas de melhoramento. Os clones CNPAT 06, CNPAT 08, CNPAT 11, CNPAT 12 e CNPAT 13 se apresentara como os mais resistentes a resinose, enquanto que os clones CNPAT 07, CNPAT 09, CNPAT 14, COMUM 05 e COMUM 31 forma os que obtiveram maiores notas de severidade de resinose. Outro importante resultado obtido foi que os clones CNPAT 08, CNPAT 11 e CNPAT 15 apresentaram capacidade de recuperação da resinose. Por fim, sugere-se que com a avaliação da incidência é possível estimar a severidade de resinose em pomares de cajueiro.

Although the molecular mechanisms underlying the onset and progression of cancer has been unraveled to a great extend, cancer continues to remain a leading cause of death around the world. Clinical efficacy of the existing anticancer drugs are largely compromised by the inherent and acquired resistance of cancer cell types and the severe side effects evoked by chemotherapeutic agents. Hence, the search for novel anticancer drugs with minimum side effects remains an active area of cancer research. Although molecular targeted drugs are preferred over the conventional cytotoxic chemotherapy, the screening of natural compounds with cytotoxic potentialities continues as they can serve as lead structures for the development of tumor selective anticancer drugs. Plants and microorganisms have been the prominent sources of therapeutic agents. Microorganisms being readily renewable, inexhaustible sources of diverse bioactive secondary metabolites are preferred over plants as sources of bioactive compounds. Endophytes are microorganisms that reside within the living tissue of host plant and they enhance the survival value of the host plant by mediating various stress tolerance mechanisms. Endophytic fungi have gained attention as potential sources of bioactive secondary metabolites following the discovery of a taxol producing endophytic fungus Taxomyces adrenae, from Taxus brevifolia. Moreover, endophytes occupy a unique biological niche in which they maintain a balanced interaction with the host organism and other co-inhabiting microorganisms. All these factors contribute to the chemical diversity of the metabolites they produce. Plants restricted to extreme or unique habitats or those with ethnobotanical value are likely to lodge endophytes that possess a unique hoard of secondarymetabolites. Saraca asoca is a traditionalmedicinal plant with its occurrence restricted to countries such as India, Sri Lanka, Burma and Malaysia. The purpose of the present study is to explore the endophytic fungal population associated with S. asoca in search of novel anticancer lead structures. S. asoca was found to house a diverse endophytic fungal population belonging to 37 different species. Identification of the fungal isolates was based on ITS (internal transcribed spacer region) sequence analysis as well as colony and spore characteristics. The organic extracts of all fungal species were assessed for their in vitro cytotoxicities in three human cancer cell lines, HeLa, HepG2 and PC3 byMTT assay. 18 species exhibited remarkable cytotoxic activities, among which Pestalotiopsis sp. 6 exhibited themost significant cytotoxicity. The strain with second highest activity was Lasiodiplodia theobromae. In order to identify the active principle present in the organic extracts of Pestalotiopsis sp. 6 and L. theobromae, the organic extracts were chromatographed on TLC plates and individual compounds were recovered by scraping off from the TLC plates and extracting with methanol. The cytotoxicity assay of the TLC purified compounds suggested the cytotoxic activity of Pestalotiopsis sp.6 to be a synergetic effect of two or more compounds whereas the cytotoxicity of L. theobromae organic extract was largely due to a single compound. Hence the active principle present in L. theobromae organic extract was purified by bioassay - guided column chromatography. Repeated chromatography of the crude extract using three silica gel columns resulted in the isolation of anticancer compound. Based on the analysis of ESI-MS, IR, NMR and UV spectral data, the isolated compound was identified as a novel steroidal saponin, cholestan-3-O-¯-Dglucopyranoside (cholestanol glucoside - CG). The in vitro cytotoxic effects of CG towards seven human cancer cell lines, HeLa, HepG2, PC3, U251,MCF 7, OVCAR3 and A549 were examined. Among the cell lines screened, HeLa cells weremost vulnerable to CG treatment, with an IC50 value of 3.2 ¹M. Hence themode of cell death induction in HeLa cells by CG was further investigated. Analysis of cell cycle progression by propidium iodide (PI) staining revealed that CG arrests the cells in S phase of cell cycle prior to the induction of cell death. The morphological and biochemical features of apoptosis were investigated by nuclear staining, DNA fragmentation assay and Annexin V-FITC/ PI dual staining. All these results suggested that CG effectively induced apoptosis in HeLa cells in a concentration dependent manner. It was also found that CG treatment induced remarkable ROS generation and mitochondrial membrane potential loss. The pretreatment of cells with an antioxidant, N-acetyl cysteine (NAC), blocked CG induced ROS generation, mitochondrialmembrane depolarization and apoptotic cell death. Hence it could be concluded that CG kills the cancer cells by augmenting their basal oxidative stress and hence is less likely to be toxic to normal cells. Moreover, a high Bax to Bcl-2 ratio, high levels of Apaf-1 and p53, activation of procaspase-3 and procaspase-9 and cleavage of PARP were observed in CG treated HeLa cells. Taken together, our results suggested that CG induced apoptosis in HeLa cells via ROS mediated mitochondria dependent pathway. Biosynthesis of secondarymetabolites by filamentous fungi is influenced by the availability of nutrient factors. Therefore, it is essential to optimize the culturemedium components to ensure a maximum and consistent yield of desired metabolite by the fungal isolate. We designed a chemically defined production medium for CG production by L. theobromae. Carbon source, nitrogen source and microelements in the production medium were further optimized in stationary flask cultures to improve the mycelial growth and yield of CG by L. theobromae. The conventional one-factor at a time (OFAT)method was employed for the optimization of carbon and nitrogen sourceswhose contribution effects towards the final yield are large. Response surface methodology (RSM) was employed for the optimization of microelements. Optimization of culturemedium enhanced the yield of CG from 10mg L¡1 to 50mg L¡1. Various secondarymetabolites are produced by organisms in response to different stress conditions. This knowledge has been exploited in plant cell culture systems to increase the yield of particular secondary metabolites by artificial implementation of stress conditions. We investigated the effect of oxidative, osmotic and heat shock stresses on the production of CG by L. theobromae. Heat shock and osmotic stresses in liquid cultures were found to enhance the yield of CG by 1.2-fold, relative to the controls. Oxidative stress by both menadione and H2O2 enhanced the yield by 1.8-fold compared to the controls. Thus oxidative stress proved to be an efficient enhancer of CG production by L. theobromae. These findings ensure a large scale, cost-effective production of CG.

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-03-27T15:47:09Z No. of bitstreams: 1 Valeria Sandra de Oliveira Costa.pdf: 1029280 bytes, checksum: 7057060a8446db15d9f71e5c654fa2be (MD5) / Made available in DSpace on 2017-03-27T15:47:09Z (GMT). No. of bitstreams: 1 Valeria Sandra de Oliveira Costa.pdf: 1029280 bytes, checksum: 7057060a8446db15d9f71e5c654fa2be (MD5) Previous issue date: 2009-02-02 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Dieback and stem-end rot in trees and fruits of mango have constituted a serious problem for the agricultural regions of Brazil, and its etiology and control is still a challenge. In the first study, the diversity of isolates of Botryosphaeriaceae from plants and fruits of mango was evaluated in semi-arid region of Northeast Brazil. Morphological characteristics and DNA sequence (5.8 S rDNA, ITS-1 e ITS-2) were used to identify a species already known and to report the presence of two new species of Botryosphaeriaceae in this region. The three species of Botryosphaeriaceae were found always in their anamorphic stage: Botryosphaeria dothidea, Neofusicoccum parvum and Lasiodiplodia theobromae. This is the first occurrence of B. dothidea and N. parvum associated with dieback and stem-end rot on mango in Brazil.Botryosphaeria dothidea and N. parvum species were prevalent in the Vale São Francisco, while L. theobromae prevailed in the Vale Assú. When inoculated in mango fruits ‘Tommy Atkins’, L. theobromae and N. parvum showed to be moreaggressive if compared to B. dothidea. In the second study, it was evaluated the influence of humidity (0 and 72 h in moist chamber) and temperature (25, 30 and 35º C) in the severity of stem-end rot in fruits of mango and the susceptibility of fruits of species different (mango, papaya, avocado and banana) to the species of Botryosphaeriaceae associated with this disease. Mango fruits were inoculated with 15 isolates, five isolates of each of the three species of Botryosphaeriaceae. The humidity and temperature significantly influenced the severity of stem-end rot in fruits of mango. The fruits of mango inoculated with L. theobromae developed symptoms of stem-end rot regardless of the presence of a moist chamber and fruits of mango inoculated with B. dothidea and N. parvum showed symptoms only when they were exposed to a period of 72 h in a moist chamber. The greatest lesions caused by both L. theobromae and by N. parvum wereobserved in fruits kept at temperatures of 25 and 30 ºC. It was not observed thedevelopment of symptoms in the fruits inoculated with B. dothidea when they were maintained at the three temperatures evaluated during the evaluation period. The isolates of L. theobromae and N. parvum were pathogenic when inoculated in healthy fruit of mango, papaya, avocado and banana. The isolates of B. dothidea were not pathogenic on fruits of banana. / A morte descendente e a podridão peduncular em árvores e frutos de mangueira vêm se constituindo em um sério problema para as regiões agrícolas do Brasil, sendo o seu controle e etiologia ainda um desafio. No primeiro estudo, a diversidade de isolados de Botryosphaeriaceae de plantas e frutos de mangueira foi avaliada na região Semi-Árida do Nordeste do Brasil. Características morfológicas e seqüência de DNA (5.8 S rDNA, ITS-1 e ITS-2) foram usados para identificar uma espécie já conhecida e relatar a presença de duas novas espécies de Botryosphaeriaceae nesta região. As três espécies de Botryosphaeriaceae foram encontradas sempre em seu estádio anamórfico: Botryosphaeria dothidea, Neofusicoccum parvum e Lasiodiplodia theobromae. Essa é a primeira ocorrência de B. dothidea e N. parvum associados à morte descendente e podridão peduncular em mangueira no Brasil. Botryosphaeria dothidea e N. parvum foram asespécies prevalentes no Vale do São Francisco, enquanto L. theobromae prevaleceu no Vale do Assú. Quando inoculadas em frutos de mangueira ‘Tommy Atkins’, L. theobromae e N. parvum mostraram maior agressividade quando comparadas com B. dothidea. No segundo estudo, foi avaliada a influência da umidade (0 e 72 h de câmara úmida) e temperatura (25, 30 e 35ºC) na severidade da podridão peduncular em frutos de mangueira e a suscetibilidade de frutos de diferentes (manga, mamão, abacate e banana) às espécies de Botryosphaeriaceae associadas a esta doença. Os frutos de mangueira foram inoculados com 15 isolados, sendo cinco isolados de cada espécie de Botryosphaeriaceae. A umidade e a temperatura influenciaram significativamente a severidade da podridão peduncular em frutos de mangueira. Os frutos de mangueira inoculados com L. theobromae desenvolveram sintomas de podridão independente da presença de câmara úmida e os frutos de mangueira inoculados com B. dothidea e N. parvum apenas quando osmesmos foram expostos a um período de 72 h de câmara úmida. As maiores lesões causadas tanto por L. theobromae como por N. parvum foram observadas nos frutos mantidos nas temperaturas de 25 e 30 ºC. Não foi constatado o desenvolvimento de sintomas nos frutos inoculados com B. dothidea quando eles foram mantidos nas três temperaturas avaliadas durante o período de avaliação. Os isolados de L. theobromae e N. parvum foram patogênicos quando inoculados em frutos sadios de mangueira, mamoeiro, abacateiro e bananeira. Os isolados de B. dothidea só não foram patogênicos em frutos de bananeira.

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / Lasiodiplodia theobromae à um fungo cosmopolita, polÃfago e oportunista, com reduzida especializaÃÃo patogÃnica, que infecta espÃcies de plantas em regiÃes tropicais e temperadas, causando os mais variados sintomas. A crescente expansÃo das doenÃas causadas por L. theobromae em frutÃferas tropicais vem causando inestimÃveis perdas, tanto no sistema produtivo como em pÃs-colheita, representando uma ameaÃa à fruticultura no Nordeste. Daà a necessidade de conhecimentos bÃsicos sobre a biologia populacional e a interaÃÃo do patÃgeno com as plantas hospedeiras. Este estudo teve como objetivo caracterizar isolados de L. theobromae associados a frutÃferas tropicais de diferentes regiÃes, avaliando o aspecto cultural, morfolÃgico e patogÃnico. Foram avaliados o crescimento micelial, coloraÃÃo da colÃnia, dimensÃes dos conÃdios e patogenicidade dos isolados em mudas de cajazeira, cajueiro, gravioleira e umbuzeiro. O trabalho foi realizado na Casa de VegetaÃÃo e no LaboratÃrio de Fitopatologia da Embrapa AgroindÃstria Tropical. Os dados de caracterizaÃÃo morfocultural mostraram haver alta diversidade na populaÃÃo do patÃgeno. As inoculaÃÃes realizadas nas quatro diferentes espÃcies hospedeiras apontaram alta variabilidade patogÃnica entre os isolados do fungo. Em mudas de cajueiro CCP 76 nÃo foi possÃvel observar especificidade patogÃnica, pois todos os isolados apresentaram similar nÃvel de agressividade, demonstrando a suscetibilidade deste clone ao patÃgeno, suscetibilidade essa, tambÃm observada em gravioleira. O umbuzeiro foi a espÃcie que apresentou maior resistÃncia ao fungo. Os dados mostraram que existe interaÃÃo entre as caracterÃsticas morfoculturais e a agressividade dos isolados de L. theobromae. De acordo com os resultados, a altitude dos locais de origem dos isolados nÃo influencia em suas caracterÃsticas morfoculturais e patogÃnicas. / Lasiodiplodia theobromae is an ubiquitous, polyphagous and opportunistic fungus with a reduced pathogenic ability. Nevertheless, it may infect several plant species over tropical and temperate regions, causing many different kinds of symptoms. The increasingly expansion of diseases caused by L. theobromae in tropical fruit plants has been imposing severe losses both at orchard level and in post-harvest at market, threaten the fruit crop industry in Northeast Region of Brazil. Therefore there is an urgent need for research pursuing basic knowledge on population biology of the fungus and host-pathogen interactions. This study aimed to characterize a L. theobromae population which has been associated to tropical plant species growing under different ecosystems in northeastern Brazil. Colony growth in culture, color and size of conidia and ability to cause disease upon inoculation on cashew nut (Anacardium occidentale), soursop (Annona muricata), yellow mombin (Spondias mombin) and Brazil plum (Spondias tuberosa) were evaluated. He study was carried out at Plant Pathology Lab and screenhouse of Embrapa Agroindustria Tropical in Fortaleza, Cearà State. Results showed a high diversity of morphology and hyphal growth among fungus isolates. Also, a very high variability on disease expression upon inoculation into four plant species was observed. However, it was found a lack of specificity of isolates as to infect cashew plants, since all isolates were able to infect cashew with similar high aggressiveness, which demonstrated a high degree of susceptibility of cashew clone used (CCP 76). Similar results were found also for soursop plants as host. Brazil plum showed a very high resistance to all isolates. The data points out for the existence of morphological and pathogenic interactions within L. theobromae population studied. According with the results, altitude and region of isolate origin has no effect on the studied features.

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-02-16T14:45:19Z No. of bitstreams: 1 Alice Maria Goncalves Santos.pdf: 828943 bytes, checksum: e2e058b935319e6ca5c5d01db52c323c (MD5) / Made available in DSpace on 2017-02-16T14:45:19Z (GMT). No. of bitstreams: 1 Alice Maria Goncalves Santos.pdf: 828943 bytes, checksum: e2e058b935319e6ca5c5d01db52c323c (MD5) Previous issue date: 2009-02-25 / In postharvest, the mango is predisposed to several diseases, including the complex known as stem-end rot fungus that has Lasiodiplodia theobromae as a major player in addition to rot by fusicocco (Fusicoccum parvum) and anthracnose caused by Colletotrichum gloeosporioides. The mangicultura is of great economic importance for Brazil, but has been losing ground with the losses that occur in post-harvest disease incidence mainly during transportation and storage, there is need to develop technologies that demonstrate effectiveness in delaying the senescence of fruits improving the marketing capability. Among the technologies we highlight the physical methods such as the use of gamma irradiation, cold storage and use of modified atmosphere. This study aimed to evaluate the three methods of physical control over the development of post-harvest mango and on the physico-chemical properties held for the treatments. For this, the fruit were inoculated with different firopatógenos and after 24 hours applied to gamma radiation doses of 0, 0.25, 0.35, 0.45 kGy, and as the Cobalt 60 source. The fruits were packed with PVC film with 380 mm x 10 microns in trays and kept under refrigeration at 13 º C for 15 days and after that period kept at room temperature, 25 ° C without the plastic film. We observed a reduction of disease development for the three fungi studied, reaching the full control of the disease when evaluating the development of anthracnose in mangoes that were subjected to higher dose of 0.45 kGy. Mangoes irradiated showed no significant differences for some parameters (ascorbic acid, total acidity (TTA), pH, total soluble solids (TSS), ratio (TSS / TTA), firmness and flesh color). Inoculated mangoes like C. gloeosporioides showed no significant changes to the physical and chemical properties, except for spots on the peel. The mangoes inoculated with L. theobromae and F. parvum showed no significant differences between the treatments that compromise the marketing thereof, thus demonstrating the efficiency of these technologies in disease control and not causing changes in properties of the sleeves / Na pós-colheita, a manga está predisposta a diversas doenças, entre elas o complexo conhecido como podridão peduncular que tem o fungo Lasiodiplodia theobromae como um dos principais agentes, além da podridão por fusicoccum (Fusicoccum parvum) e a antracnose causada por Colletotrichum gloeosporioides. A mangicultura é de grande importância econômica para o Brasil, mas vem perdendo espaço com as perdas que acontecem na pós-colheita, principalmente pela incidência de doenças durante o transporte e armazenamento, havendo necessidade de desenvolver tecnologias que demonstrem eficiência em retardar a senescência das frutas, melhorando a capacidade de comercialização. Entre as tecnologias destacam-se os métodos físicos como a utilização da radiação gama, o armazenamento refrigerado e o uso da atmosfera modificada. O presente trabalho objetivou avaliar os três métodos de controle físico sobre o desenvolvimento pós-colheita de manga e sobre as características físico-químicas conservadas durante os tratamentos. Para isso, as frutas foram inoculadas com os diferentes firopatógenos, e após 24 horas aplicada a radiação gama nas doses de 0; 0,25; 0,35; 0,45 kGy, tendo como fonte o Cobalto60. As frutas foram embaladas com filme plástico de PVC com 380 mm x 10 microns em bandejas de isopor e mantidas sob refrigeração a 13ºC por 15 dias e após esse período conservadas sob temperatura ambiente, de 25ºC sem o filme plástico. Foi observada uma redução do desenvolvimento da doença para os três fungos avaliados, chegando ao controle total da doença quando avaliado o desenvolvimento da antracnose nas mangas que foram submetidas à dose mais alta de 0,45 kGy. As mangas irradiadas não demonstraram diferenças significativas para alguns dos parâmetros avaliados (ácido ascórbico, acidez titulável total (ATT), pH, sólidos solúveis totais (SST), razão (STT/ATT), firmeza e cor da polpa). As mangas inoculadas como o C. gloeosporioides não apresentaram alterações significativas para as propriedades físico-químicas, exceto manchas nas cascas. As mangas inoculadas com L. theobromae e F. parvum não apresentaram diferenças significativas entre ao tratamentos aplicados que comprometesse a comercialização das mesmas, demonstrando assim a eficiência dessas tecnologias no controle das doenças e não causando alterações das propriedades das mangas.

The dioxygenase-cytochrome P450 fusion proteins (DOX-CYP) comprise a heme-containing enzyme family that shares structural and catalytic properties with mammalian prostaglandin H (PGH) synthases. 7,8-Linoleate diol synthase (7,8-LDS) of Gaeumannomyces graminis was first characterized, and DOX-CYP enzymes are of mechanistic and biological interest. The growing number of fungal genome sequences has revealed DOX-CYP homologues in medically and economically important species. The aim of this thesis was to identify novel members of the DOX-CYP fusion protein family. The devastating rice pathogen Magnaporthe oryzae contains two DOX-CYP genes. The fungus synthesizes 7S,8S-dihydroxyoctadecadienoic acid (7,8-DiHODE) by dioxygenation of linoleic acid to 8R-hydroperoxyoctadecadienoic acid (8R-HPODE), and subsequent isomerisation to the diol. 7,8-LDS of M. oryzae was identified by gene deletion, but the infection and reproduction processes of the Δ7,8-LDS strain were not altered. A mutant with constitutive protein kinase A activity profoundly changed the oxygenation profile, possibly due to post-translational modification. The human pathogens Aspergillus fumigatus and A. clavatus contain three DOX-CYP, designated psi producing oxygenase A (ppoA), ppoB, and ppoC, and form three oxylipins: 5S,8R-DiHODE, 8R,11S-DiHODE, and 10R-hydroxyoctadecadienoic acid.  PpoA was identified as 5,8-LDS, and ppoC as 10R-DOX. The 8,11-linoleate hydroperoxide isomerase activity was reduced by two imidazole-containing P450 inhibitors, miconazole and 1-benzylimidazole. PpoB could not be linked to the biosynthesis of 8,11-DiHODE for the following reasons: First, the 8,11-hydroperoxide isomerase activity was retained in A. fumigatus ΔppoB strains. Second, the P450 domain of the deduced ppoB of A. clavatus lacks a heme-thiolate cysteine ligand, presumably essential for hydroperoxide isomerase activity. Linoleate 9R-DOX activities of Aspergillus terreus and Lasiodiplodia theobromae were discovered. 9R-HPODE was further converted into unstable allene oxides, as judged by the accumulation of their hydrolysis products, α- and γ-ketols. These allene oxide synthase activities were specific for 9R-hydroperoxides. The 9R-DOX and AOS were found to have unique characteristics. In conclusion, novel DOX-CYP enzymes were identified in human and plant pathogenic fungi. These enzymes might be involved in biological processes, and show interesting catalytic similarities to human PGH synthase and thromboxane synthase (CYP5A).