Rozprawy doktorskie na temat „BLACK FUNGUS”

The black truffle (Tuber melanosporum Vitt.) industry in Australia is relatively new and has enormous potential but some truffières (truffle farms) fail to meet anticipated harvest projections. Inappropriate soil conditions and climate, and the management of such factors are suggested as the primary reasons for inadequate yield. In addition, requirements for ascocarp initiation and development and the role of the host plant in such processes are unknown. This study examines interactions between European hazel (Corylus avellana L.) and the ectomycorrhizal (ECM) black truffle symbiont in a commercial truffière (Hazel Hill) in south-western Australia. Specific studies were initiated to examine the interactions of host physiology, mycorrhizal infection and the interaction with abiotic factors. The study related specific physiological processes of the host plant to the known life cycle of the black truffle to determine the role of the host plant in ascocarp production, if any. The work also examined the effect of silvicultural treatments intended to increase truffle production. A review of existing literature was undertaken to determine the key soil and climatic factors required for successful truffle production. Climatic conditions appeared more important than soil chemistry and structure in Western Australia, with significant seasonal variation in air and soil temperatures required plus irrigation to supplement summer rainfall. This information was used to define areas with potential for truffle production in the south-west of Western Australia: the cooler, high rainfall regions (>1000 mm annual rainfall) where there is sufficient seasonal variation in soil temperature and availability of adequate quantities of quality water for irrigation. Subsurface soil acidification and salinity, as well as groundwater salinity, are constraining factors. Lime amendment is necessary to create sufficiently high pH and CaCO3 levels required by the truffle fungus. A field trial was established to monitor the seasonal C dynamics of European hazel in the context of the life cycle of the black truffle. Maximum translocation of sucrose in the phloem sap coincided with the period of anticipated rapid growth of the truffle ascocarp implicating the use of current photosynthate in C nutrition of the ascocarp. Sampling of non-structural carbohydrates (NC) of above and belowground plant material indicated maximum storage of C in the host coincides with maturation of the ascocarp. These observations provide evidence of a synchronous growth habit of the plant host and the ascocarp. The C allocation patterns of European hazel in response to liming a loamy soil, taken from near the Hazel Hill truffière, and inoculation with ECM fungi (T. melanosporum, Hebeloma sp. and Scleroderma sp.) were examined in a glasshouse pot trial. Liming increased biomass allocation to the shoot and induced deficiencies of phosphorus and manganese. Colonisation by ECM fungi significantly increased net photosynthesis, indicating the sink strength of these fungi, but there was no relationship between the level of mycorrhizal infection and fine root NC. The maximum rate (40 g lime kg-1 soil) reduced infection by Hebeloma and Scleroderma and had no impact on T. melanosporum. Further, infection rates of T. melanosporum did not increase in response to lime suggesting lime is not necessary for ECM development in this soil type. Fertiliser is widely used in commercial truffières in Australia but the consequences for truffle production are unknown. In a field trial, the growth and physiological response of European hazel to forms of phosphorus (34 and 68 kg ha-1 apatite-P and 68 kg ha-1 triple super phosphate -P) and nitrogen (50 kg ha-1 of NO3- and NH4-N) were examined as well as the mycorrhizal response to fertiliser. Apatite-P increased phloem sap sucrose concentrations which was attributed to increased root biomass and associated sink capacity. Fertiliser application did not change fine root NC concentrations suggesting no increase in allocation of C to ECM structures. The highest rate of apatite-P decreased mycorrhizal infection rates of T. melanosporum and, most likely, was the result of increased infection rates of Hebeloma. In contrast to the literature relating to indigenous Australian ECM fungi, the highest rate of soluble-P did not decrease ECM infection rates in T. melanosporum. Nitrogen treatments increased foliar N content and improved gas exchange efficiency of plants, and had no adverse impact on the level of ECM infection. Fertilisation with N significantly increased soil respiration rates suggesting N limits mineralisation at this site. Some truffières manage the canopies of the host tree to ensure maximum exposure of the soil surface in order to increase soil temperatures. As there are no published data on the effect of pruning on black truffle production, a field trial was established to document the impact of canopy pruning on host physiology and soil temperature. The removal of 65% of canopy leaf area reduced phloem sap sucrose concentrations, soil respiration rate and the soluble: insoluble NC ratio of fine roots in the short term (1-3 weeks). There was no compensatory response of leaf gas exchange parameters as a result of pruning. Generally, there was no long term impact on plant physiological parameters as a result of pruning. Long term effects on soil temperature were observed as a result of pruning. Mean annual temperature and amplitude increased significantly beneath pruned trees and spring, summer and autumn soil temperatures increased as did diurnal variation as a result of pruning. Pruning did not increase winter soil temperatures and therefore would probably not impact on ascocarp maturation during this period. This research has provided insight into the C physiology of hazel associated with the black truffle and the consequences for truffle production. The results provide anecdotal evidence of direct C transfer between the host and the developing truffle, contrary to the existing paradigm that the ascocarp is saprotrophic for the majority of its growth and development. There is a need to validate this finding as there are consequences for management of commercial truffières. Liming of loam duplex soils can reduce the abundance of the most common competitor ECM fungi and should be encouraged in commercial truffières. Applying phosphorus and nitrogen to commercial truffières will improve growth rates of planted trees without adversely impacting on ECM infection by black truffle fungi, although the impact on truffle production remains unknown. It is anticipated truffle production will improve in the longer term as a result of pruning and prudent canopy management. Management options should include tree removal to reduce planting density and increase soil exposure in truffières. There is a need for longer term trials to be established to determine the C nutrition of the truffle ascocarp and to clearly define the key stages of the black truffle life cycle in Western Australia.

The black truffle (Tuber melanosporum Vitt.) industry in Australia is relatively new and has enormous potential but some truffieres (truffle farms) fail to meet anticipated harvest projections. Inappropriate soil conditions and climate, and the management of such factors are suggested as the primary reasons for inadequate yield. In addition, requirements for ascocarp initiation and development and the role of the host plant in such processes are unknown. This study examines interactions between European hazel (Corylus avellana L.) and the ectomycorrhizal (ECM) black truffle symbiont in a commercial truffiere (Hazel Hill) in south-western Australia. Specific studies were initiated to examine the interactions of host physiology, mycorrhizal infection and the interaction with abiotic factors. The study related specific physiological processes of the host plant to the known life cycle of the black truffle to determine the role of the host plant in ascocarp production, if any. The work also examined the effect of silvicultural treatments intended to increase truffle production. A review of existing literature was undertaken to determine the key soil and climatic factors required for successful truffle production. Climatic conditions appeared more important than soil chemistry and structure in Western Australia, with significant seasonal variation in air and soil temperatures required plus irrigation to supplement summer rainfall. This information was used to define areas with potential for truffle production in the south-west of Western Australia: the cooler, high rainfall regions (>1000 mm annual rainfall) where there is sufficient seasonal variation in soil temperature and availability of adequate quantities of quality water for irrigation. Subsurface soil acidification and salinity, as well as groundwater salinity, are constraining factors. Lime amendment is necessary to create sufficiently high pH and CaCO3 levels required by the truffle fungus. A field trial was established to monitor the seasonal C dynamics of European hazel in the context of the life cycle of the black truffle. Maximum translocation of sucrose in the phloem sap coincided with the period of anticipated rapid growth of the truffle ascocarp implicating the use of current photosynthate in C nutrition of the ascocarp. Sampling of non-structural carbohydrates (NC) of above and belowground plant material indicated maximum storage of C in the host coincides with maturation of the ascocarp. These observations provide evidence of a synchronous growth habit of the plant host and the ascocarp. The C allocation patterns of European hazel in response to liming a loamy soil, taken from near the Hazel Hill truffiere, and inoculation with ECM fungi (T. melanosporum, Hebeloma sp. and Scleroderma sp.) were examined in a glasshouse pot trial. Liming increased biomass allocation to the shoot and induced deficiencies of phosphorus and manganese. Colonisation by ECM fungi significantly increased net photosynthesis, indicating the sink strength of these fungi, but there was no relationship between the level of mycorrhizal infection and fine root NC. The maximum rate (40 g lime kg-1 soil) reduced infection by Hebeloma and Scleroderma and had no impact on T. melanosporum. Further, infection rates of T. melanosporum did not increase in response to lime suggesting lime is not necessary for ECM development in this soil type. Fertiliser is widely used in commercial truffieres in Australia but the consequences for truffle production are unknown. In a field trial, the growth and physiological response of European hazel to forms of phosphorus (34 and 68 kg ha-1 apatite-P and 68 kg ha-1 triple super phosphate -P) and nitrogen (50 kg ha-1 of NO3- and NH4-N) were examined as well as the mycorrhizal response to fertiliser. Apatite-P increased phloem sap sucrose concentrations which was attributed to increased root biomass and associated sink capacity. Fertiliser application did not change fine root NC concentrations suggesting no increase in allocation of C to ECM structures. The highest rate of apatite-P decreased mycorrhizal infection rates of T. melanosporum and, most likely, was the result of increased infection rates of Hebeloma. In contrast to the literature relating to indigenous Australian ECM fungi, the highest rate of soluble-P did not decrease ECM infection rates in T. melanosporum. Nitrogen treatments increased foliar N content and improved gas exchange efficiency of plants, and had no adverse impact on the level of ECM infection. Fertilisation with N significantly increased soil respiration rates suggesting N limits mineralisation at this site. Some truffieres manage the canopies of the host tree to ensure maximum exposure of the soil surface in order to increase soil temperatures. As there are no published data on the effect of pruning on black truffle production, a field trial was established to document the impact of canopy pruning on host physiology and soil temperature. The removal of 65% of canopy leaf area reduced phloem sap sucrose concentrations, soil respiration rate and the soluble: insoluble NC ratio of fine roots in the short term (1-3 weeks). There was no compensatory response of leaf gas exchange parameters as a result of pruning. Generally, there was no long term impact on plant physiological parameters as a result of pruning. Long term effects on soil temperature were observed as a result of pruning. Mean annual temperature and amplitude increased significantly beneath pruned trees and spring, summer and autumn soil temperatures increased as did diurnal variation as a result of pruning. Pruning did not increase winter soil temperatures and therefore would probably not impact on ascocarp maturation during this period. This research has provided insight into the C physiology of hazel associated with the black truffle and the consequences for truffle production. The results provide anecdotal evidence of direct C transfer between the host and the developing truffle, contrary to the existing paradigm that the ascocarp is saprotrophic for the majority of its growth and development. There is a need to validate this finding as there are consequences for management of commercial truffieres. Liming of loam duplex soils can reduce the abundance of the most common competitor ECM fungi and should be encouraged in commercial truffieres. Applying phosphorus and nitrogen to commercial truffieres will improve growth rates of planted trees without adversely impacting on ECM infection by black truffle fungi, although the impact on truffle production remains unknown. It is anticipated truffle production will improve in the longer term as a result of pruning and prudent canopy management. Management options should include tree removal to reduce planting density and increase soil exposure in truffieres. There is a need for longer term trials to be established to determine the C nutrition of the truffle ascocarp and to clearly define the key stages of the black truffle life cycle in Western Australia.

The black truffle (Tuber melanosporum Vitt.) industry in Australia is relatively new and has enormous potential but some truffieres (truffle farms) fail to meet anticipated harvest projections. Inappropriate soil conditions and climate, and the management of such factors are suggested as the primary reasons for inadequate yield. In addition, requirements for ascocarp initiation and development and the role of the host plant in such processes are unknown. This study examines interactions between European hazel (Corylus avellana L.) and the ectomycorrhizal (ECM) black truffle symbiont in a commercial truffiere (Hazel Hill) in south-western Australia. Specific studies were initiated to examine the interactions of host physiology, mycorrhizal infection and the interaction with abiotic factors. The study related specific physiological processes of the host plant to the known life cycle of the black truffle to determine the role of the host plant in ascocarp production, if any. The work also examined the effect of silvicultural treatments intended to increase truffle production. A review of existing literature was undertaken to determine the key soil and climatic factors required for successful truffle production. Climatic conditions appeared more important than soil chemistry and structure in Western Australia, with significant seasonal variation in air and soil temperatures required plus irrigation to supplement summer rainfall. This information was used to define areas with potential for truffle production in the south-west of Western Australia: the cooler, high rainfall regions (>1000 mm annual rainfall) where there is sufficient seasonal variation in soil temperature and availability of adequate quantities of quality water for irrigation. Subsurface soil acidification and salinity, as well as groundwater salinity, are constraining factors. Lime amendment is necessary to create sufficiently high pH and CaCO3 levels required by the truffle fungus. A field trial was established to monitor the seasonal C dynamics of European hazel in the context of the life cycle of the black truffle. Maximum translocation of sucrose in the phloem sap coincided with the period of anticipated rapid growth of the truffle ascocarp implicating the use of current photosynthate in C nutrition of the ascocarp. Sampling of non-structural carbohydrates (NC) of above and belowground plant material indicated maximum storage of C in the host coincides with maturation of the ascocarp. These observations provide evidence of a synchronous growth habit of the plant host and the ascocarp. The C allocation patterns of European hazel in response to liming a loamy soil, taken from near the Hazel Hill truffiere, and inoculation with ECM fungi (T. melanosporum, Hebeloma sp. and Scleroderma sp.) were examined in a glasshouse pot trial. Liming increased biomass allocation to the shoot and induced deficiencies of phosphorus and manganese. Colonisation by ECM fungi significantly increased net photosynthesis, indicating the sink strength of these fungi, but there was no relationship between the level of mycorrhizal infection and fine root NC. The maximum rate (40 g lime kg-1 soil) reduced infection by Hebeloma and Scleroderma and had no impact on T. melanosporum. Further, infection rates of T. melanosporum did not increase in response to lime suggesting lime is not necessary for ECM development in this soil type. Fertiliser is widely used in commercial truffieres in Australia but the consequences for truffle production are unknown. In a field trial, the growth and physiological response of European hazel to forms of phosphorus (34 and 68 kg ha-1 apatite-P and 68 kg ha-1 triple super phosphate -P) and nitrogen (50 kg ha-1 of NO3- and NH4-N) were examined as well as the mycorrhizal response to fertiliser. Apatite-P increased phloem sap sucrose concentrations which was attributed to increased root biomass and associated sink capacity. Fertiliser application did not change fine root NC concentrations suggesting no increase in allocation of C to ECM structures. The highest rate of apatite-P decreased mycorrhizal infection rates of T. melanosporum and, most likely, was the result of increased infection rates of Hebeloma. In contrast to the literature relating to indigenous Australian ECM fungi, the highest rate of soluble-P did not decrease ECM infection rates in T. melanosporum. Nitrogen treatments increased foliar N content and improved gas exchange efficiency of plants, and had no adverse impact on the level of ECM infection. Fertilisation with N significantly increased soil respiration rates suggesting N limits mineralisation at this site. Some truffieres manage the canopies of the host tree to ensure maximum exposure of the soil surface in order to increase soil temperatures. As there are no published data on the effect of pruning on black truffle production, a field trial was established to document the impact of canopy pruning on host physiology and soil temperature. The removal of 65% of canopy leaf area reduced phloem sap sucrose concentrations, soil respiration rate and the soluble: insoluble NC ratio of fine roots in the short term (1-3 weeks). There was no compensatory response of leaf gas exchange parameters as a result of pruning. Generally, there was no long term impact on plant physiological parameters as a result of pruning. Long term effects on soil temperature were observed as a result of pruning. Mean annual temperature and amplitude increased significantly beneath pruned trees and spring, summer and autumn soil temperatures increased as did diurnal variation as a result of pruning. Pruning did not increase winter soil temperatures and therefore would probably not impact on ascocarp maturation during this period. This research has provided insight into the C physiology of hazel associated with the black truffle and the consequences for truffle production. The results provide anecdotal evidence of direct C transfer between the host and the developing truffle, contrary to the existing paradigm that the ascocarp is saprotrophic for the majority of its growth and development. There is a need to validate this finding as there are consequences for management of commercial truffieres. Liming of loam duplex soils can reduce the abundance of the most common competitor ECM fungi and should be encouraged in commercial truffieres. Applying phosphorus and nitrogen to commercial truffieres will improve growth rates of planted trees without adversely impacting on ECM infection by black truffle fungi, although the impact on truffle production remains unknown. It is anticipated truffle production will improve in the longer term as a result of pruning and prudent canopy management. Management options should include tree removal to reduce planting density and increase soil exposure in truffieres. There is a need for longer term trials to be established to determine the C nutrition of the truffle ascocarp and to clearly define the key stages of the black truffle life cycle in Western Australia.

In recent years two new diseases, brown spot and black pit, have been observed on potatoes in South Africa. Brown spot symptoms appear on the foliage as small brown lesions, whereas black pit symptoms appear on the tubers as small dark sunken lesions. In this study the causal organism of brown spot and black pit of potatoes in South Africa was determined. During initial isolation, one fungus and two bacteria were isolated, which were included in the trial. Only the treatments where the fungal isolate was used in inoculation resulted in the development of brown spot lesions. The causal organism of brown spot and black pit were identified as Alternaria alternata which is consistent with other research. To better understand the spread of Alternaria alternata between the plants and/or tubers a pot trial was conducted. It was observed that when planting an inoculated seed tuber brown spot may develop on foliage. But the daughter tubers harvested from plants infected with A. alternata will not necessarily develop black pit. Daughter tubers are most likely infected by A. alternata during harvesting and black pit lesions develop in high humidity in storage. Cultivar resistance is one of the most important measures in controlling plant diseases. Cultivar susceptibility of thirteen South African potato cultivars (Avalanche, Buffelspoort, BP1, Fabula, Fianna, Frodo, Hertha, Labadia, Lanorma, Mondial, Pentland Dell, Up-To-Date and Van Der Plank) was evaluated. Pot trials showed that all the evaluated cultivars are susceptible to infection by Alternaria alternata. Various crops (tomatoes, cabbage, mustard, wheat, oats, tobacco and maize) were assessed to determine the host range of Alternaria alternata (potato pathotype) in rotation crops in South Africa potato growing regions. Of the crops evaluated, the pathogen was able to infect only tomato crops. Only wheat, maize and oats can safely be used in the crop rotation in South Africa, as various potato pathogens attack cabbage, mustard and tobacco plants. This study will lead to a better understanding of brown spot and black pit diseases of potatoes in South Africa and globally. The study emphasise the need for further research which will help to reduce brown spot and black pit diseases of potato.
Dissertation (MSc Agric)--University of Pretoria, 2013.
gm2014
Microbiology and Plant Pathology
unrestricted

A mancha preta dos citros (MPC) é uma doença que limita a exportação de laranja brasileira para o Japão e países da Europa. Exceto para Citrus aurantium e seus híbridos, todas as outras variedades são susceptíveis ao patógeno. O fungo Guignardia citricarpa, descoberto por Kiely em 1948 em New South Wales na Austrália, é o estádio sexual do agente causal da MPC e a sua fase imperfeita é Phyllosticta citricarpa. Uma importante característica da MPC é seu longo período de latência. A infecção pode ser efetuada por ascósporos e picnidiósporos. A lesão nos frutos fica restrita ao flavedo (epicarpo), sendo que o fungo não infecta o albedo (mesocarpo). O albedo é rico em celulose, carboidratos solúveis, pectinas, compostos fenólicos (flavonóides), aminoácidos e vitaminas. Os compostos fenólicos presentes nas plantas são produtos do metabolismo secundário, e podem ser resultantes da interação planta-ambiente e podem sintetizados como resposta ao ataque de fitopatógenos. Os fenólicos presentes nos citros incluem flavonóides, antocianinas, coumarinas e psorolenos entre outros. Estes compostos podem exibir ação antimicrobiana e antiviral e podem contribuir controle da MPC. A aplicação de fungicidas é o método de controle da MPC. Uma outra possibilidade de controle seria a ativação de fatores de resistência através do uso de indutores bióticos (Saccharomyces cerevisiae) e abióticos (Bion e ácido salicílico). Deste modo, este trabalho teve como objetivos estudar os efeitos do uso de extratos aquosos, metanólicos e etanólicos de albedo de laranja na germinação formação de apressório e crescimento micelial de P. citricarpa in vitro, bem como avaliar o uso da indução de resistência para o controle da MPC através dos indutores S. cerevisiae, Bion e ácido salicílico em pós e pré-colheita em frutos de laranja ‘Pêra-Rio’ e folhas de limão ‘Siciliano’. Os resultados experimentais mostraram que os extratos de albedo, nas concentrações de 10 e 100 mg / mL de água, foram capazes de inibir em 100% a germinação, formação de apressório e o crescimento micelial de P. citricarpa. Foi observado que os extratos, dependendo da concentração, podem ter ação fungicida ou fungistática. No tocante a utilização de S. cerevisiae, Bion e ácido salicílico, aplicados em pós-colheita, foi observado que estes agentes não foram capazes de impedir o desenvolvimento de novas lesões em frutos de laranja ‘Pêra-Rio’. Também foi constatado que S. cerevisiae e Bion, aplicados em pré-colheita, não foram capazes de induzir resistência contra P. citricarpa em folhas de limão ‘Siciliano’ inoculadas previamente a campo com G. citricarpa. Nesse sentido, sugere-se que outros estudos sejam conduzidos, principalmente no que se refere ao uso potencial de extratos do albedo de laranja como medida alternativa de controle da MPC.
Black spot of citrus (CBS) has been a limiting factor in the export of brazilian oranges to Japan and European countries and Japan. Except for Citrus aurantium and its hybrids, all commercially growing Citrus spp. are susceptible to the pathogen. The fungus Guignardia citricarpa, discovered by Kiely in 1948 in New South Wales, is the sexual stage of the causal agent of CBS and Phyllosticta citricarpa is the imperfect stage. An important characteristic of CBS is the long latent period after infection. The infection is carried out by ascospores and pycnidiospores. The fungicidal application is the most important method of control of CBS. The CBS lesion in citrus fruits is limited to the flavedo, since P. citricarpa does not infect the albedo. The albedo is rich in cellulose, soluble carbohydrates, pectin, phenolic compounds, amino acids and vitamins. The phenolics present in the plants are secondary metabolic products and are believed to be produced as a result of the plant interaction with the enviromment and synthesized as a response to attempted phytopathogen attacks. The phenolics that occur in Citrus include flavonoids, anthocyanins, coumarins and psorolens. These compounds may exhibit antiviral and antimicrobial activities, and may contribute to the control of CBS disease. An another possibility to the CBS control is the activation of factors resistance by the use of abiotics (Bion and salicylic acid) and biotics (Saccharomyces cerevisiae) “plant defence activator” (inducers). Therefore, the objectives of this paper were to study the in vitro effects of aqueous, ethanolic and methanolic albedo orange extracts on the germination, appressorium formation and mycelial growth of P. citricarpa as well as to evaluate the use of the S. cerevisiae, Bion and salicylic acid as “plant defence activator” at post and preharvest conditions in fruit of “Pêra-Rio” and leaves of ’Siciliano’ lemon. The results showed that the use of albedo extracts, 10 and 100 mg per mL of water, inhibited 100 % the germination, appressorium formation and mycelial growth of P. citricarpa. It was also observed that the extracts of albedo, depending upon the concentration, exhibited fungicidal or fungistatic activity. The use of S. cerevisiae, Bion and salicylic acid at postharvest conditions did not affect the development of new lesions of CBS in ‘Pêra-Rio’ orange fruit. It was also observed that the use of S. cerevisiae and Bion at preharvest conditions, did not induce resistance against P. citricarpa in leaves of ‘Siciliano’ lemon naturally infected with G. citricarpa under field conditions. Thus, it is suggested that other studies be carried out, mainly regarding the potential of orange albedo’s extracts as a alternative method for CBS control.

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The Black-Litterman model calculates the expected market returns as a combination of a set of investor views and a neutral reference point. The model uses Bayesian approach to blend both sources of information. The results from the Black-Litterman model, in contrast to the traditional approach, are quite intuitive, stable and consistent with the investors views. The purpose of this thesis is to provide a detailed analysis of each component of the Black-Litterman model and verify if the use of the Black-Litterman model, introducing the views of the market based on the Central Bank report, FOCUS, outperforms brasilians Hegde Funds.
O modelo Black-Litterman calcula os retornos esperados de mercado como uma combinação de um conjunto de expectativas específicas de cada investidor e um ponto de referência neutro. A combinação dessas duas fontes de informações são feitas pelo modelo utilizando a abordagem bayesiana. Os resultados obtidos a partir do modelo Black-Litterman, ao contrário da abordagem tradicional, são bastante intuitivos, estáveis e consistentes em relação as expectativas dos investidores. O objetivo dessa dissertação é fazer uma análise detalhada de cada um dos componentes do modelo Black-Litterman e verificar se a utilização o modelo de Black-Litterman, introduzindo as opiniões de mercado com base no relatório FOCUS do Banco Central, supera o retorno dos fundos multimercados brasileiros.

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CAPEs
Piedra preta é uma micose superficial assintomática, caracterizada pela formação de nódulos rígidos e enegrecidos localizados ao longo do fio capilar. É uma micose considerada rara, inócua, podendo acometer tanto o gênero feminino quanto o masculino em qualquer faixa etária. Piedraia hortae, agente etiológico desta micose, é um fungo filamentoso, demáceo que produz o pigmento melanina, caracterizando uma coloração enegrecida na parede celular e nas estruturas fúngicas. Este pigmento, por sua vez, é caracterizado como um potente fator de virulência, favorecendo, desse modo, seu potencial em causar parasitismo no cabelo. Na literatura, não há referências citando que outros fungos demáceos, que formam ascostroma, ascos e ascosporos, semelhantes ao P. hortae, possam ser agente etiológico dessa feohifomicose. Assim, o objetivo desse trabalho foi avaliar a capacidade de fungos demáceos de formarem nódulos na porção extrafolicular de fios de cabelo humano, semelhantes aos da piedra preta. Foram obtidos onze isolados de fungos demáceos, sendo dez da Coleção de Cultura Micoteca URM, UFPE, os quais estavam preservados sob óleo mineral e posteriormente semeados em meio Àgar Batata Dextrose para estimulação do crescimento; e um isolado proveniente do Laboratório de Micologia Médica. Foram preparadas suspensões dos isolados dos fungos demáceos em 2,0 ml de água destilada esterilizada, ajustada para concentração final de 106 células/mL. Em seguida, vertidos separadamente 0,5 mL de cada suspensão sobre os fios de cabelo contidos nas placas de Petri previamente esterilizada e mantidas a temperatura de 28°C e 37°C. O experimento foi conduzido através de observações macroscópicas e microscópicas dos cabelos durante 40 dias e acompanhado em intervalo de 5 dias. Dois isolados (970 e 3334) mostraram um elevado potencial capaz de causar infecção no fio capilar. Entretanto, oito isolados apresentaram apenas uma colonização. Exophiala dermatitidis e Cladosporium tenuissimo foram capazes de formar nódulos semelhantes ao da piedra preta, porém estruturas de reprodução como ascostroma, ascos e ascosporos, não foram vizualizadas.Todavia, podemos inferir que esses fungos apresentam um potencial capaz de parasitar os fios de cabelo, degradando e destruindo a queratina e os componentes cutiulares, sendo o potente agente de tricomicose.
Black piedra is a superficial mycosis asymptomatic, characterized by the formation of hard lumps and blackened located along the capillary yarn. It is a mycosis considered rare, innocuous and can affect both the female and the male at any age. Piedraia hortae, the etiologic agent of this mycosis, is a filamentous fungus, dematiaceous that making the pigment melanin, featuring a blackish color in the cell wall and the fungal structures. This pigment, in turn, is characterized as a potent virulence factor, facilitating there by the potential to cause parasitic hair. In the literature, there are no references citing that other demáceos fungi that form ascostroma, asci and ascospores, similar to P. hortae, may be etiologic agent of this phaeohyphomycosis. The objective of this paper is to demáceos fungal ability to form nodules in extrafollicular portion of human hair, similar to those of black piedra. Eleven isolates of fungi demáceos, obtained of the Culture Collection URM, UFPE, which were preserved under mineral oil and then plated on potato dextrose agar medium for growth promotion; and one isolated from the Medical Mycology Laboratory. Fungal isolates demáceos suspensions were prepared in 2.0 ml of sterile distilled water, adjusted to a final concentration of 106cells / ml. Then separately poured into 0.5 mL of each suspension over the strands of hair contained in the previously sterile Petri dishes and kept at 28 °C and 37 °C. The experiment was conducted by macroscopic and microscopic observations of the hair for 40 days and monitoring in the range of 5 days. Two isolates (970 and 3334) showed a high potential capable of causing infection in the capillary. However, eight isolates showed only colonization. Exophiala dermatitidis and Cladosporium tenuissimo was capable of forming nodules were similar to those of black piedra; however ascostroma structures such as playback, asci and ascosporos were not displayed. However, we can infer that these fungi have the potential able to parasitize the hairs, degrading and destroying the keratin and cuticulares components, the powerful Trichomycosis agent.

O licor negro, cuja composição majoritária é de lignina, é o principal subproduto gerado após a obtenção de polpa celulósica. Este polímero natural é caracterizado pela elevada massa molecular (MM) e composição aromática. Processos físico-químicos são empregados na fragmentação da lignina com o objetivo de obter compostos aromáticos, cuja fonte não renovável é oriunda de derivados do petróleo. A biodegradação da lignina, causada pela ação de enzimas ligninolíticas fúngicas pode prover compostos aromáticos de baixo peso molecular, como fenóis e antioxidantes, utilizados em diversos segmentos industriais. O objetivo deste trabalho foi estudar a modificação estrutural da lignina causada por fungos basidiomicetos coletados de mata nativa e campos de reflorestamento de eucalipto. Os ensaios de biodegradação foram inicialmente conduzidos utilizando-se 10 espécies de fungos, cultivados em duas concentrações de licor negro (10% e 15%, v/v). No ensaio com licor menos concentrado, utilizaram-se 10 espécies, e no ensaio com licor 15% utilizou-se somente a espécie Pycnoporus sanguineus. As análises realizadas para o estudo da modificação estrutural da lignina foram: espectroscopia na região do infravermelho (FTIR/ATR), cromatografia líquida de gel filtração e espalhamento de raios-X a baixo ângulo (SAXS). P. sanguineus e espécies de fungos do gênero Trametes possuem elevada capacidade de consumir a lignina presente no licor 10%, após 14 dias. A atividade de enzimas ligninolíticas de P. sanguineus foi investigada nas duas concentrações de licor. No licor 10%, observou-se elevada atividade das enzimas ligninolíticas manganês peroxidase (MnP), lacase e peroxidases; somente a MnP foi detectada com maior atividade no licor 15%. Análises de Demanda Bioquímica de Oxigênio (DBO) indicaram que o licor negro 10% biodegradado por P. sanguineus possui elevada carga de DBO, sendo tóxico a Daphia magna e Hydra attenuatta. Análises de metabólitos produzidos pelos fungos no licor negro 10% por cromatografia gasosa acoplada à espectrometria de massas (GC-MS) indicaram, principalmente, a produção de açúcares e álcoois em concentrações não muito diferentes dos seus respectivos controles. Embora os fungos sejam capazes de reduzir a massa molecular da lignina presente no licor negro, compostos derivados da sua fragmentação podem sofrer reações de condensação ou repolimerização, contribuindo, eventualmente, para elevar sua massa molecular
Black liquor which the bulk content is lignin, is the main product generated after obtaining cellulose pulp. This natural polymer is characterized by high molecular weight (MW) and aromatic compounds. Physical-chemical processes can be employed to fragment lignin and to produce aromatic compounds to replace those from non-renewable sources such as petroleum. Biodegradation of lignin can provide low molecular weight compounds, such as phenols and antioxidants, which may be employed in various industrial processes. The aim of this work was to investigate the structural modification of lignin caused by basidiomycetes fungi, obtained from native forest and Eucalyptus fields. The biodegradation assay was done with10 fungi species cultivated with two concentrations of black licor (10% and 15%, v/v). The assay with 10% liquor, 10 species of basidiomycete fungi were used, and in the assay with 15% liquor, it was used only Pycnoporus sanguineus. The analyses carried out to determine the structural modification of lignin were: infrared spectroscopy (FTIR/ATR), liquid chromatography gel filtration and small angle X-ray scattering (SAXS). P. sanguineus and fungi of Trametes genera had high capacity to consume the lignin content in the 10% liquor, after 14 days of cultivation. After that, P. sanguineus was selected for assays of ligninolytic enzymes activities in the two liquor concentrations. High activities of manganese peroxidase (MnP), laccase and peroxidase were detected in the 10% liquor, whereas in the concentration of 15% only high MnP activity was detected. Biochemical Oxygen Demand, analysis (BOD) indicated that the biodegraded liquor had high BOD load, and it was toxic to Daphia magna and Hydra attenuata. Metabolites produced by the fungi in the liquor 10% were analysed by gas chromatography-mass spectrometry (GC-MS), which indicated mainly the production of sugars and alcohols in concentrations not far from their respective controls. Although the fungus is able to decrease molecular weight of lignin, its fragments derivatives can undergo condensation or repolymerization reactions, contributing, eventually, to increase its molecular weight

Neste trabalho testou-se o potencial antagonista de 16 fungos endofíticos isolados de videiras (Vitis vinifera L.), de castas representativas do Alentejo produzidas em modo de proteção integrada e em modo biológico, contra Guignardia bidwellii. Os isolados identificados após ITS-PCR e sequenciação pertencem aos géneros Epicoccum, Alternaria, Botrytis, Athelia, Phoma e Gibberella. Os isolados testados mostraram atividade antagonista contra G. bidwellii quer por inibição direta, quer através da produção de compostos voláteis, à exceção dos dois isolados de B. cinerea. No entanto, todos os isolados produziram alguns compostos voláteis com reconhecida atividade antimicrobiana, tais como benzaldeído, 3-metil-1-butanol e derivados de ácido propanoico. Foi ainda observado que seis dos isolados produziram também metabolitos não voláteis com capacidade de inibir o crescimento de G. bidwellii. Os resultados obtidos vêm mostrar o potencial dos fungos endofíticos como agentes de luta biológica no controlo de G. bidwellii, podendo constituir novas alternativas no âmbito de Proteção de Plantas; ABSTRACT: Endophytic fungi present in grapevines (Vitis vinifera L.) with the ability to inhibit the growth of the causal agent of black rot (Guignardia bidwellii) In this work the antagonistic potential of 16 endophytic grapevine fungi isolates (Vitis vinifera L.), from representative cultivars of the Alentejo region produced either under integrated pest management or organic mode, was tested against Guignardia bidwellii. Isolates were identified through ITS-PCR and sequencing, as belonging to the genera Epicoccum, Alternaria, Botrytis, Athelia, Phoma and Gibberella. Isolates showed antagonist activity against G. bidwellii either by direct inhibition or through the production of volatile compounds, with the exception of two isolates of B. cinerea. Nevertheless, all isolates produced volatile compounds with known antimicrobial activity such as benzaldehyde, 3-methyl-1-butanol and propionic acid derivatives. Additionally, six isolates produced non-volatile metabolites with the ability to inhibit G. bidwellii growth. These results show the potential that endophytic fungi have as agents for biological control of G. bidwellii, opening new options in the field of Plant Protection.

Mestrado em Engenharia Agronómica - Instituto Superior de Agronomia
Black foot of grapevine is an important disease caused primarily by fungi of the genus Ilyonectria. These pathogens affect grapevine nurseries and young vineyards, causing the decline and death of plants. In the nursery, the infections are mainly attributed to infected propagation material and soilborne inoculum, thus hampering the selection of healthy plant material. The aim of this study was to detect/identify possible sources of inoculum for Ilyonectria spp. and "Cylindrocarpon" pauciseptatum, also associated with the disease, throughout the different nursery stages. Samples of plant material (rootstock and scion mother plants, grafted-cuttings and rooted graftlings), soil (from mother-fields, nurseries and plots in rotation), callusing-medium (before and after utilization), air chambers and warehouses, water from hydration tanks and tap water, cutting tools and paraffin, were analysed by classical and molecular techniques (nested-PCR and multiplex nested-PCR). Multiplex nested-PCR revealed to be more sensitive and fast than classical technique, allowing the simultaneous detection of I. liriodendri and/or I. macrodidyma and/or "C." pauciseptatum and the identification of unknown inoculum sources for these fungi (eg. callusing-medium and water samples). These studies should be pursued in order to confirm the viability of the inoculum detected.

A citricultura brasileira concentra-se principalmente no Estado de São Paulo que contribui com 80,4 % da produção nacional, sendo o Brasil um dos maiores produtores mundiais de citros. Um dos problemas enfrentados pela citricultura é a sua vulnerabilidade a pragas e doenças, devido principalmente a baixa diversidade genética nas variedades comerciais utilizadas, associada ao sistema de plantio em áreas extensas. Uma das doenças que vem causando crescentes prejuízos para a citricultura brasileira é a pinta preta ou mancha preta dos citros causada pelo fungo Guignardia citricarpa Kiely. O uso de conhecimentos de biologia molecular e métodos biotecnológicos devem ser considerados como importante alternativa para a produção de plantas geneticamente modificadas expressando genes de resistência. Para se obter plantas de citros resistentes a doenças, se faz necessário identificar genes que estejam relacionados com os mecanismos de defesa da planta. Na tentativa de identificar estes genes, o objetivo geral deste trabalho foi a identificação de genes in silico no banco de dados do Projeto Millenium CitEST e a análise de expressão diferencial de genes envolvidos na defesa. Mais de 7600 sequencias foram identificadas nas buscas no CitEST com similaridade aos genes R e genes envolvidos na HR e defesa, MAPKs e SNF1. Destes, foram selecionados 273 sequencias para experimentos de macroarranjo para análise da interação Citrus sinensis-Guignardia citricarpa. A análise estatística revelou que 171 genes (62,63%) apresentaram expressão diferencial significativa ao nível de 5% de probabilidade. Destes, 80 apresentaram expressão diferencial significativa maior do que duas vezes, dos quais 38 genes foram induzidos e 42 foram reprimidos no tecido infectado. Entre os genes induzidos estão MAPKs, genes de resistência (R), genes envolvidos na resposta de hipersensibilidade (HR) e na defesa da planta. Entre os transcritos reprimidos, há quatro similares a peroxidases e cinco similares a catalases, o que era esperado já que catalases e algumas peroxidases são capazes de remover H2O2, e assim a planta produz espécies reativa de oxigênio capaz de desencadear a ativação de genes de defesa. Os dados do macroarranjo foram validados via transcrição reversa seguida de PCR quantitativo em tempo real (RT-PCRq) de 9 genes. As análises confirmaram a expressão diferencial de 8 deles sendo que somente um apresentou resultado contrastante ao macroarranjo, o que demonstra a eficiência da metodologia de macroarranjos para estudo de muitos genes simultaneamente. Os genes diferencialmente expressos identificados na interação C. sinensis-G. citricarpa são de grande importância, pois são fortes candidatos para serem utilizados na transformação genética de plantas com o objetivo de obter novas variedades de plantas com resistência a patógenos.
The Brazilian citrus industry is concentrated mainly in the State of Sao Paulo which contributes with 80.4% of national production, with Brazil being a leading world producer of citrus. One of the problems facing the citrus industry is its vulnerability to pests and diseases, mainly due to low genetic diversity of the commercial varieties used, linked to the system of planting in extensive areas. A disease that is causing increasing damage to the brazilian citrus industry is the black spot of citrus caused by the fungus Guignardia citricarpa Kiely. The use of knowledge of molecular biology and biotechnological methods should be considered as an important alternative for the production of genetically modified plants expressing genes for resistance. In order to obtain citrus plants resistant to diseases it is necessary to identify genes that are related to the defense mechanisms of the plant. In an attempt to identify these genes, the general aim of this study was to identify genes in silico in the database of the Millennium CitEST Project and to perform differential expression analysis of genes involved in the defense mechanisms. More than 7600 reads were identified in the CitEST search with similarity to R genes, genes involved in HR and defense, MAPKs and SNF1. It was selected 273 reads for macroarray experiments to analysis of Citrus sinensis-Guignardia citricarpa interaction. Statistical analysis revealed that 171 genes (62.63%) showed significant differential expression at the level of 5% probability. From these, 80 showed significant differential expression higher than two fold, in which 38 genes were induced and 42 were repressed in infected tissue. Among the induced genes are MAPKs, resistance (R) genes, genes involved in hypersensitivity response (HR) and plant defense. Among the suppressed transcripts, there are four similar to peroxidases and five similar to catalases, which is expected because catalases and some peroxidases are able to remove H2O2, and so the plant produces reactive oxygen species capable of triggering the activation of defense genes. The macroarray data were validated by reverse transcription followed by quantitative real-time PCR (RT-PCRq) of 9 genes. The analysis confirmed the differential expression of 8 of them, and only one presented different result of macroarray which demonstrate the efficiency of the macroarray methodology to analyze several genes simultaneously. The genes differentially expressed in the interaction of C. sinensis x Guignardia citricarpa identified are of great importance because they are strong candidates for use in genetic transformation of plants with the objective of obtaining new varieties of plants resistant to pathogens.

This work sets out to answer what constitutes successful black political activity in the context of European Structural Funds. It proposes that much analysis of black political activity in Britain focuses on matters of 'race' and racism. This work argues that black political activity is broader than this and that by analysing black activists in an environment where matters of 'race' and racism are not pre-eminent, the range and diversity of black political activity would become apparent. The work uses a framework derived from political opportunities theorising and policy network literature to examine the proposition. The work explores different aspects of black political and community activity over time arguing there has a existed a diversity of black actor and organisation operating in difficult circumstances and that these actors have been instrumental in changing the way they have been treated in racialised societies. The work then goes on to look at the policy environment in which Structural Funds operate and the role of black organisations and individuals in this process. It found that Structural Funds was a very restrictive process that was not amenable to black engagement on the basis of 'race'. The work then considered the experiences of unsuccessfull non-participants and successful black organisations involved in the process. Successful organisations could put their achievements down to a range of factors including: dynamic leadership, flexible use of racialised identities, partnership working and organisational capability. Unsuccessful and non-participants lacked some of these traits or were constrained by the process. It is argued that the processes and networks surrounding Structural Funds were important in determining success, with Objective 1 having a slightly more open structure than the Objective 2 programme. The work shows that black organisational and political activity is varied, that the institutional arrangements, presence of elite alliances, and how 'race' issues are perceived in the policy environment combine with the activities of activists to constitute successful and unsuccessful activity. It shows that although racialisation is important in determining the nature of black political activity, even where 'race' is not pre-eminent it is not all consuming. There is scope to admire and look more closely at the diversity and range of black activists operating in and outside policy making networks.

Dissertation (PhD (Agric))--University of Stellenbosch, 2005.
ENGLISH ABSTRACT: During the past few years a drastic reduction has been noted in the survival rate of grafted grapevines in nurseries, as well as in young vineyards in the Western Cape Province of South Africa. Circumstantial evidence suggested that Cylindrocarpon spp., which cause black foot disease of grapevine, were associated with this decline. Black foot disease of grapevine is a relatively new, and as yet poorly known disease affecting vines in various countries where grapevines are cultivated. Primary aims of this research have been (1) to conduct nursery surveys in order to determine which fungi are involved in the decline phenomenon, with special reference to the involvement of Cylindrocarpon spp., (2) to identify and characterise the organisms believed to be the causal organisms of black foot disease, and (3) the development of control and/or management strategies to prevent or eradicate Cylindrocarpon infections. Nursery grapevines were sampled at different stages from three commercial nurseries in the Wellington area of the Western Cape Province and were investigated during the 19992000 season by means of destructive sampling. The first samples were taken in September from callused cuttings prior to planting in nurseries. After planting, asymptomatic rooted cuttings were selected from nurseries after 3, 6 and 9 months. Isolation studies clearly demonstrated that different “Cylindrocarpon spp.” infected cuttings from nursery soils. These species rarely occurred in rootstock propagation material prior to planting. At the time of planting, the susceptible basal ends (especially the pith area) of most of the nursery cuttings are partly or even fully exposed. Callus roots also break during the planting process, resulting in small wounds susceptible to infection by soilborne pathogens. The isolation studies revealed that the first infections occurred in the roots, followed by infections of the rootstocks. These infections increased progressively during the course of the growing season. Substantial variation in cultural and morphological characters was observed among the Cylindrocarpon isolates obtained from the nursery survey, as well as from isolations that were made from diseased grapevines. Morphological and phylogenetic studies were conducted to identify these “Cylindrocarpon spp.” and to establish their association with black foot disease. Sequences of the partial nuclear large subunit ribosomal DNA (LSU rDNA), internal transcribed spacers 1 and 2 of the rDNA including the 5.8S rDNA gene (ITS), and partial β-tubulin gene introns and exons were used for phylogenetic inference. Phylogenetic analyses confirmed the diversity observed among the isolates and four Cylindrocarpon-like species were identified. One of these species was initially identified as Cylindrocarpon destructans. However, further research revealed C. destructans to represent a species complex. Grapevine isolates of “C. destructans” proved to be identical to the ex-type strain of Cylindrocarpon liriodendri, which also produced a teleomorph, Neonectria liriodendri in culture. A second species was newly described in this study as Cylindrocarpon macrodidymum (Neonectria macrodidyma). The two remaining Cylindrocarpon-like species were placed in a new genus, Campylocarpon. The two species were named Campylocarpon fasciculare and Campylocarpon pseudofasciculare. Pathogenicity studies confirmed that all four species were able to reduce root and shoot mass significantly. Knowledge obtained pertaining to the disease cycle of black foot disease suggest that suitable management strategies should focus on prevention of primary infection in nurseries. However, at present, no fungicides are registered for control of this disease in South African vineyards or nurseries. Thirteen fungicides were screened in vitro for mycelial inhibition of these pathogens. Prochloraz manganese chloride, benomyl, flusilazole and imazalil were the most effective fungicides tested, and were subsequently included in semi-commercial field trials. Basal ends of grafted cuttings were dipped (1 min) in various chemical and biological treatments prior to planting in open-rooted nurseries. Black foot pathogens were not isolated from grafted cuttings prior to planting in nurseries. Additional treatments involved soil amendments with Trichoderma formulations and hot water treatment (50°C for 30 min) of dormant nursery grapevines. Field trials were evaluated after a growing season of eight months. The incidence of black foot pathogens was not significantly and/or consistently reduced by the majority of chemical or biological treatments. However, these pathogens were not isolated from uprooted plants that were subjected to hot water treatment. It is therefore recommended that hot water treatment of dormant nursery plants be included in an integrated strategy for the proactive management of black foot disease in grapevine nurseries.
AFRIKAANSE OPSOMMING: Gedurende die afgelope paar jaar is ‘n drastiese afname waargeneem in die sukses van geënte wingerdplante in kwekerye, sowel as jong wingerde van die Wes-Kaap. Omstandigheidsgetuienis dui daarop dat Cylindrocarpon spp., wat die wingerdsiekte swartvoet veroorsaak, geassosieer word met hierdie agteruitgang. Swartvoet is ‘n relatiewe nuwe siekte waarvan daar baie min inligting bekend is, alhoewel dit voorkom in verskeie lande waar wingerd verbou word. Die primêre doel van navorsing was (1) om opnames in wingerdkwekerye uit voer om te bepaal watter swamme betrokke is by die verskynsel van agteruitgang, met spesiale verwysing na die betrokkenheid van Cylindrocarpon spp., (2) om die organismes te identifiseer en te karakteriseer wat daarvan verdink word dat hulle die siekte swartvoet veroorsaak, en (3) om beheer en/of bestuurspraktyke te ontwikkel om Cylindrocarpon infeksies te voorkom of uit te wis. Kwekeryplantjies in drie kommersiële kwekerye in die Wellington omgewing van die Wes-Kaap is gedurende verskillende tye gedurende die groeiseisoen gemonitor. Die opnames het plaasgevind gedurende die 19992000 seisoen deur middel van destruktiewe monsterneming. Die eerste monsters is geneem in September nadat die stokkies geënt en gekallus is en voordat dit in die kwekery geplant is. Na plant is asimptomatiese, gewortelde plante vanuit die kwekerye na 3, 6 en 9 maande uitgehaal. Isolasiestudies dui duidelik daarop dat verskillende “Cylindrocarpon spp.” plante vanuit die kwekerygrond geïnfekteer het. Hierdie spesies het selde voorgekom in onderstok-voortplantingsmateriaal voor plant. Tydens plant is die vatbare basale gedeelte, veral die pit, van die meeste geënte stokkies gedeeltelik of selfs volledig blootgestel. Kalluswortels breek ook tydens plant wat wonde laat vir infeksie deur grondgedraagde siektes. Die isolasiestudies dui ook daarop dat die eerste infeksies in die wortels plaasgevind het, gevolg deur infeksies van die onderstokke. Hierdie infeksies het toenemend voorgekom gedurende die verloop van die groeiseisoen. Substansiële variasie in kultuur- en morfologiese eienskappe is waargeneem in die Cylindrocarpon isolate wat tydens die kwekeryopnames versamel is, sowel as van isolasies wat gemaak is uit siek plante. Morfologiese en filogenetiese studies is uitgevoer om hierdie “Cylindrocarpon spp.” te identifiseer en hul betrokkenheid by die siekte swartvoet uit te klaar. Gedeeltelike DNS volgordes van die groot ribosomale subeenheid (“LSU rDNA”), interne getranskribeerde spasiëerderarea (“ITS1, “ITS2”), insluitend die 5.8S rRNS geen, en gedeeltelike β-tubilien geen introns and eksons is gebruik vir filogenetiese analise. Filogenetiese analises het die diversiteit wat waargeneem is tussen die verskillende isolate bevestig deurdat vier Cylindrocarpon-agtige spesies geïdentifiseer is. Een van hierdie spesies is aanvanklik geïdentifiseer as Cylindrocarpon destructans. Verdere navorsing het egter daarop gedui dat C. destructans ‘n spesie-kompleks verteenwoordig. “C. destructans” afkomstig van wingerd blyk identies te wees aan die ex-tipe isolaat van Cylindrocarpon liriodendri, wat ook ’n teleomorf, Neonectria liriodendri in kultuur vorm. ’n Tweede spesie is nuut beskryf in hierdie studie as Cylindrocarpon macrodidymum (Neonectria macrodidyma). Die twee oorblywende Cylindrocarpon-agtige spesies is geplaas in ‘n nuwe genus, Campylocarpon. Die twee spesies staan bekend as Campylocarpon fasciculare en Campylocarpon pseudofasciculare. Patogenisiteitstudies het bevestig dat al vier spesies die vermoë het om wortel- en lootmassa van wingerdplant drasties te verlaag. Kennis wat opgedoen is rakende die lewensiklus van swartvoet dui daarop dat bestuurspraktyke daarop moet fokus om primêre infeksies in wingerdkwekerye te voorkom. Op die oomblik is daar egter geen fungisiedes geregistreer vir die beheer van die siekte in Suid- Afrikaanse wingerde of kwekerye nie. Dertien fungisiedes is in vitro geëvalueer om te bepaal of dit miseliumgroei van hierdie swamme kan inhibeer. Prochloraz mangaan chloried, benomyl, flusilasool en imazalil was die effektiefste fungisiedes wat ondersoek is, en is gevolglik ingesluit in semi-kommersiële veldproewe. Die basale gedeelte van geënte stokkies is gedoop (1 min) in verskeie chemies en biologiese behandelings voordat dit geplant is in die kwekerye. Patogene wat geassosieer word met swartvoet is nie vanuit geënte stokkies geïsoleer voordat dit in die kwekerye geplant is nie. Addisionele behandelings het bestaan uit grondtoevoegings met Trichoderma formulasies, sowel as warmwaterbehandeling (50°C vir 30 min) van dormante kwekeryplante. Die veldproewe is geëvalueer na ‘n groeiseisoen van 8 maande. Die voorkoms van swartvoet patogene is nie betekenisvol/konstant verlaag deur die meeste chemies en biologiese behandelings nie. Hierdie patogene is egter nie vanuit plante geïsoleer wat na uithaal aan warmwaterbehandeling blootgestel is nie. Dit word dus aanbeveel dat warmwaterbehandeling van dormante kwekeryplante deel word van ‘n geïntegreerde strategie vir die pro-aktiewe beheer van swartvoet in wingerdkwekerye.

Fungi with dark-coloured mycelia - also known as black fungi - form a ubiquitous fraction of microbial communities on rock surfaces all over the world. These organisms show a high capacity to adapt to a wide range of ecological conditions, including those peculiar of extreme environments. Their high tolerance to multiple abiotic stresses, such as solar radiation and osmotic stress is assured by the production of melanin pigments and osmoprotective compounds. The cosmopolitan genera Lichenothelia and Saxomyces were taken as iconic representatives of polyextremotolerant rock-inhabiting black fungi. I investigated their phylogenetic relationships with an extended taxon sampling within the class Dothideomycetes, the most diverse and life-style rich fungal class in Ascomycota. The three loci phylogenetic inference I set up, considering both environmental samples and culture isolates, highlighted the paraphyly of the two genera. The integrative taxonomy approach based on morphological as well as phylogenetic evidences allowed the taxonomical revision of the genera and the description of three new taxa. Rock inhabiting fungi often share substrata with green algae and cyanobacteria and some of them are associated with lichen thalli. Lichenothelia is of a particular interest because it includes lichen parasites and species which are loosely associated with algae or which grow independently on rock. Given their life style plasticity, we chose it for an in vitro culture experiment, studying the development of three Lichenothelia species when co-cultured with two different subaerial algae isolated from lichens (i.e., lichen photobionts). The results showed that the presence of algae neither influence the growth rate of fungi nor the formation of any lichen-like structure. However, this standardized approach proved suitable for future investigations on fungal-algal interactions in other systems. Previous multi-locus phylogenies of Dothideomycetes have investigated evolutionary relationships at order and family level within the class but they often failed to resolve the early diverging nodes, which were generating inconsistent placements of some clades. Here, I applied a phylogenomic approach to resolve relationships in Dothideomycetes, adding the newly sequenced Lichenothelia and Saxomyces genomes, to a wide dataset comprised of 238 individuals. I explored the influence of tree inference methods, supermatrix vs. coalescent-based species tree, and the impact of varying amounts of genomic data. The phylogenomic reconstructions, based on up to three thousand genes, provide well-supported topologies for Dothideomycetes, recovering Lichenothelia and Saxomyces among the earliest diverging lineages in the class together with other rock inhabiting fungi and lichens and thus, suggesting the rock-inhabiting life style as ancestral in the class. Further studies will be necessary to shed light on the molecular bases of stress tolerance and latent capacity of establishing symbiosis of these fungi.

Background: Since 1998 Sweden has individual accounts as a part of both public and occupational schemes (Sundén 2006). Yearly, 2,5% of the pensionable income is set aside to the premium pension (The Swedish Pension Agency 2021) Individuals are able to choose how the premiums should be paid in the system and in what way the money should be invested, either by choosing from the fund market or by refraining from making an active choice and instead let the Swedish pension agency management their money in the passive alternative called AP7 Såfa. The passive alternative AP7 Såfa is a portfolio which adapts to the age of the investor and is built to fit a long-term pension investment.  Purpose: This study will focus on evaluating if the passive alternative AP7 Såfa has an excess risk-adjusted return compared to given portfolios in the premium pension selection system, or if the investor would benefit from managing the portfolio more actively. The study will also search for benefits using the Treynor-Black model to check the optimal allocation between this actively managed portfolio versus the passive alternative AP7 Såfa.  Conclusion: This thesis has shown that there might be superior strategies rather than the index fund to find risk-adjusted excess return in the premium pension system. However, those strategies require professional analysts in order to forecast securities. For households choosing between active management themselves or the passive index fund AP7 Såfa, the most beneficial investment is to be passive and to not actively manage the portfolio. The optimum strategy is found to be the Treynor-Black model with a combined portfolio of the index fund and the active portfolio.

O Brasil é líder absoluto no comércio internacional de suco de laranja concentrado congelado participando com 82% do volume comercializado no mundo. Guignardia citricarpa é um fungo Ascomiceto agente causal da Mancha Preta dos Citros (MPC), uma doença importante no contexto da Citricultura, causando lesões negras em frutos tornando-os impróprios para a exportação, já que não são aceitos na União Européia pois o patógeno é classificado como quarentenário. O presente trabalho teve como objetivo principal estabeler a metodologia de transformação genética de G. citricarpa para futuramente auxiliar no entendimento dos mecanismos de patogenicidade desta espécie, visando diminuir perdas na citricultura brasileira devido à MPC. Além disso, foi realizada uma análise do gene da enzima endopoligalacturonase, a qual está associada à capacidade de patógenos em colonizar plantas. Para elucidar esse fenômeno, foi realizada a busca de genes relacionados à patogenicidade em outros fungos fitopatogênicos previamente descritos e confirmados participarem no processo de doenças em diversas plantas. Considerando que esses genes possuem uma região conservada para esse fim, após o alinhamento dessas sequências foram construídos primers e o gene de endopoligalacturonase foi identificado no gênero Guignardia sp. Outra espécie pertencente a esse gênero é G. mangiferae, conhecida como endófito de citros, ou seja, coloniza os tecidos internos da planta hospedeira sem causar dano. Em análises enzimáticas foi observado que a quantidade de endopoligalacturonase produzida pela espécie patogênica é superior à da espécie endofítica, mostrando que essa enzima pode participar do processo de patogenicidade de G. citricarpa. Estudos para comprovar a participação de genes nos mecanismos de patogenicidade em diversas espécies utilizando reconhecimento de genes e genômica funcional, expressão e knockout de genes estão sendo realizados, permitindo uma visão geral da organização genômica do sistema patogênico. Pensando nisso, esse trabalho descreve pela primeira vez a metodologia de transformação genética de G. citricarpa via micélio e a obtenção de transformantes expressando a proteína verde fluorescente (GFP). Micélios do fungo foram transformados pelo sistema via Agrobacterium tumefaciens com o plasmídeo pFAT-gfp, contendo os genes de resistência à higromicina B (hgr) e da GFP. A otimização do protocolo de agrotransformação foi realizada a partir do teste de diferentes condições como: tipo de membrana, concentração de agente indutor e tempo de cocultivo. A melhor condição incluiu a utilização de membrana de éster de celulose; 200 PM de AS e 96 horas de co-cultivo. Os transformantes apresentaram alta estabilidade mitótica (82%) e tiveram a inserção do gene hgr confirmada por PCR e do gfp observada em microscopia óptica de epifluorescência. Além disso, foi acompanhado o desenvolvimento do fungo inoculado em frutos, mostrando a interação planta-patógeno. O estabelecimento do sistema de transformação por Agrobacterium para G. citricarpa possibilita o uso dessa ferramenta para estudos de mutagênese insercional e interrupção gênica visando a identificação de genes importantes, como os envolvidos com os mecanismos de patogenicidade utilizados por esse fungo.
Brazil is the world leader in the international trade of frozen orange juice concentrate, taking part with around 82% of the traded volume. Guignardia citricarpa (anamorph Phyllosticta citricarpa) is a fungal pathogen of citrus plants, being described as the causal agent of citrus black spot (CBS), one of the most important fungal diseases of citrus worldwide. Its symptoms are black lesions on fruit, making them unsuitable for the international fresh market, since they are included in the quarantine list of the European Plant Protection Organization (EPPO). Moreover, when the disease is severe it may cause extensive premature fruit drop that reduces yields of fruit for processing. Taking this into consideration, the current work aimed to improve the understanding on the pathogenic mechanisms of this fungus. Firstly, in an attempt to elucidate this phenomenon, it was performed a search for pathogenicity genes previously reported to some pathogenic fungi and confirmed to participate in the process of infection in many plants , especially endopolygalacturonase. Primers were designed using the conserved regions of the genes and allowed the identification of the Guignardia spp. endopolygalacturonase gene for the first time. This enzyme has been described as playing important role in the process of fungal diseases in plants. In the present work, enzymatic analysis showed that the pathogen G. citricarpa produced significantly greater amounts of endopolygalacturonase when compared to G. mangiferae, a closely related fungus described as a citrus endophyte. This result suggests that this enzyme may participate in the process of pathogenicity, characteristic of the pathogenic species. Genetic transformation methods have been used to prove the involvement of genes in pathogenic mechanisms, however, a suitable methodology for G. citricarpa has not been described yet. In this way, this study describes for the first time a methodology for genetic transformation of G. citricarpa via mycelia and the successful generation of transformants expressing the green fluorescent protein (GFP) and resistant to the hygromycin B antibiotic. Mycelia of the fungus were genetically transformed via Agrobacterium tumefaciens hosting the plasmid pFAT-gfp, which carries the genes for resistance to hygromycin B (hph) and for GFP (gfp). The protocol was optimized through different test conditions (type of membrane, concentration of the inducing agent acetosyringone and duration of the co-cultivation period). The higher transformation efficiencies were observed using cellulose ester membrane, 200 PM of acetosyringone and 96 hours of cocultivation. The transformants showed high mitotic stability (82%) and the insertion of the T-DNA was confirmed by PCR and GFP expression through epifluorescence microscopy observation. Moreover, it was observed the development of the fungus in inoculated oranges, showing the plant-pathogen interaction observed by epifluorescense microscopy. The establishment of the Agrobacterium-mediated transformation system for G. citricarpa represents an important step on the search for unveiling important genes of this fungus, such as those involved in the pathogenic mechanisms.

Neem cake is a product of the cold pressing from the neem kernels to obtain neem oil. Bio-active substances from neem cake extracted solutions were evaluated for their potential to control the root knot nematodes and other pests of plants. In this study different concentrations of the solution extracted from neem cake was tested against the second stage juveniles of the plant parasitic nematode Meloidogyne spp. and four phytopathogenic fungi: Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytopthora capsici. Toxicity of neem cake extractions is represented by the EC50 value for the second-stage juvenile (J2) of Meloidogyne spp. and the four phytopathogenic fungi via Probit analysis. A 5% dilution of the solvent extracting from neem cake already caused 100% larval mortality after 24 hours exposure. Undiluted neem cake extraction effectively inhibited the growth of the four phytopathogenic fungi. The EC50 value of neem cake on J2-larvae of Meloidogyne nematode and on the fungi Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytophthora capsici was 0.51, 0.74, 0.30, 0.51 and 4.33%, respectively
Bánh dầu neem là sản phẩm của quá trình ép nhân hạt neem để lấy dầu. Các hoạt chất sinh học từ dịch chiết bánh dầu neem đã được đánh giá có tiềm năng lớn trong phòng trừ tuyến trùng nốt sưng và các loài dịch hại khác của nhiều loại cây trồng. Trong nghiên cứu này các nồng độ dịch chiết khác nhau của bánh dầu neem đã được thử nghiệm khả năng diệt tuyến trùng (ấu trùng tuổi 2 thuộc giống Meloidogyne spp.) và ức chế 4 loài nấm bệnh như: Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. và Phytopthora capsici. Độc tính của dịch chiết bánh dầu neem được biểu diễn bởi giá trị EC50 đối với ấu trùng tuổi 2 của tuyến trùng Meloidogyne spp. và các loài nấm bệnh thông qua phân tích Probit. Dịch chiết bánh dầu neem ở nồng độ 5% đã làm chết 100% cá thể IJ2 của Meloidogyne spp sau 24 giờ phơi nhiễm. Dịch nguyên chất bánh dầu neem ức chế cả 4 loài nấm bệnh. Giá trị EC50 của bánh dầu neem lên ấu trùng tuổi 2 của Meloidogyne spp và các loài nấm bệnh Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytophthora capsici tương ứng là 0.51, 0.74, 0.30, 0.51 và 4.33%

The purpose of this ethnography was to examine the elements of my personal goals and aspirations of being an elementary school teacher with the real experience of teaching students within my classroom. Through the lens of an ethnography, and grounded in the research components of culturally inclusive education and, this thesis provides a critical and needed pedagogical approach to how teachers can make a difference in the lives of their students, and in the process learn that they are really the ones being taught.

Neem cake is a product of the cold pressing from the neem kernels to obtain neem oil. Bio-active substances from neem cake extracted solutions were evaluated for their potential to control the root knot nematodes and other pests of plants. In this study different concentrations of the solution extracted from neem cake was tested against the second stage juveniles of the plant parasitic nematode Meloidogyne spp. and four phytopathogenic fungi: Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytopthora capsici. Toxicity of neem cake extractions is represented by the EC50 value for the second-stage juvenile (J2) of Meloidogyne spp. and the four phytopathogenic fungi via Probit analysis. A 5% dilution of the solvent extracting from neem cake already caused 100% larval mortality after 24 hours exposure. Undiluted neem cake extraction effectively inhibited the growth of the four phytopathogenic fungi. The EC50 value of neem cake on J2-larvae of Meloidogyne nematode and on the fungi Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytophthora capsici was 0.51, 0.74, 0.30, 0.51 and 4.33%, respectively.
Bánh dầu neem là sản phẩm của quá trình ép nhân hạt neem để lấy dầu. Các hoạt chất sinh học từ dịch chiết bánh dầu neem đã được đánh giá có tiềm năng lớn trong phòng trừ tuyến trùng nốt sưng và các loài dịch hại khác của nhiều loại cây trồng. Trong nghiên cứu này các nồng độ dịch chiết khác nhau của bánh dầu neem đã được thử nghiệm khả năng diệt tuyến trùng (ấu trùng tuổi 2 thuộc giống Meloidogyne spp.) và ức chế 4 loài nấm bệnh như: Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. và Phytopthora capsici. Độc tính của dịch chiết bánh dầu neem được biểu diễn bởi giá trị EC50 đối với ấu trùng tuổi 2 của tuyến trùng Meloidogyne spp. và các loài nấm bệnh thông qua phân tích Probit. Dịch chiết bánh dầu neem ở nồng độ 5% đã làm chết 100% cá thể IJ2 của Meloidogyne spp sau 24 giờ phơi nhiễm. Dịch nguyên chất bánh dầu neem ức chế cả 4 loài nấm bệnh. Giá trị EC50 của bánh dầu neem lên ấu trùng tuổi 2 của Meloidogyne spp và các loài nấm bệnh Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytophthora capsici tương ứng là 0.51, 0.74, 0.30, 0.51 và 4.33%.

This research examines whether Corporate Social Responsibility (CSR) initiatives impact fund financial performance of 46 Nordic mutual funds during the past five years (2016-2020). Previous studies have focused on which of the two alternatives generates a rewarding financial result. To complement previous research, this study focuses on the development of CSR - whether the gap formed between CSR initiatives and fund financial performance is moving closer towards each other based on the increased focus of ESG factors during the recent years. Two categories, strong and weak ESG performance, are juxtapositioned to find out at what stage the development is. Furthermore, analysis on which of the three categories: environmental, social or governance individually, is the driving factor of the development of CSR initiatives. Based on the matched pair methodology and hypothesis testing, funds with CSR initiatives have reached the point, in which they perform equally well to their peers without such motives. The underlying driver behind the development are examined.

A citricultura é uma atividade rural extremamente importante no contexto sócio-econômico brasileiro. A laranja representa 49% de toda a produção brasileira de frutas. Entretanto, anualmente milhares de toneladas de frutos são perdidas devido à ação de fitopatógenos. A Mancha Preta dos Citros (MPC) tem sido responsável por grandes prejuízos em várias regiões produtoras de citros no mundo, sendo designada como barreira fitossanitária, principalmente no mercado Europeu. Para o consumo in natura, a aparência dos frutos é fator essencial, onde a MPC compromete a comercialização de frutas frescas. O controle químico de patógenos de plantas é geralmente o mais utilizado para minimizar os danos causados pela MPC, apesar do uso de tais produtos implicar em altos custos, tanto para os agricultores com para o meio ambiente, causando contaminação do solo e da água e aumentando a pressão de seleção sobre a população do patógeno. Dessa forma, o controle biológico torna-se uma alternativa atrativa, como uma estratégia que permite um menor impacto ambiental aliado à proteção da planta contra o patógeno. No entanto, para sua aplicação, são necessários estudos relacionados à utilização de técnicas de biocontrole, como por exemplo, a utilização de microrganismos produtores de enzimas hidrolíticas. Tais enzimas, como as quitinases, endoglicanases e β- glicosidases têm a capacidade de digerir a parede celular de fungos e bactérias. Neste trabalho, 24 isolados de G. citricarpa foram avaliados quanto à sensibilidade aos fungicidas usados em campo para controle da doença: piraclostrobina e carbendazim, nas dosagens de 0,5; 1 e 2 mg i.a./mL, a fim de verificar o efeito da pressão de seleção causada pelo uso contínuo destes compostos. Dois dos isolados apresentaram resistência ao carbendazim em todas as dosagens testadas, indicando que o uso desse agroquímico pode selecionar mutantes resistentes, resultando na não eficiência deste produto no controle da MPC. Uma alternativa para minimizar este tipo de efeito seria a utilização deste composto em combinação com outros princípios ativos. As atividades celulolíticas e quitinolíticas de 96 isolados de fungos filogeneticamente distintos foram avaliadas para a seleção de potenciais agentes de controle biológico. Os quatro isolados que apresentaram maior atividade de cada uma destas enzimas, além de duas linhagens de Trichoderma, foram testados como potencial de biocontrole de G. citricarpa em um experimento com folhas laranja ‘Valência’, comparando a ação destes com a de fungicidas comerciais. Apesar de o melhor controle do patógeno ter sido observado em folhas tratadas com piraclostrobina, dois isolados fúngicos mostraram eficiência similar ao fungicida, inibindo o desenvolvimento de G. citricarpa, o que sugere a possibilidade de futura utilização de métodos de controle biológico para a Mancha Preta dos Citros.
Citriculture is an extreme important rural activity in social and economical national context in Brazil. Orange is 49% of total brazilian fruit production. However, thousands of tons are lost due to the action of phytopathogens annually. The Black Spot of Citrus (BSC) is responsible for great lost in various citrus producers regions all around the world, being already designed as a phytosanitary barrier, mainly in European market. For in natura fruit consuming, the fruit esthetic is a limiting factor, where BSC compromises the market of fresh affected fruits. Chemical control of plant pathogens is the most commonly way used to minimize damages in citriculture by BSC, although the application of such products implies in high costs, not only for farmers but also for environment, causing soil and water contamination and increasing the selection pressure on pathogen population. On this way, the biocontrol became an attractive way, as a strategy that permits a minor environmental impact besides the plant protection against phytopathogens. For this application, are necessary researches based on utilization of biocontrol techniques, as for example, the using of microorganism producers of hydrolytic enzymes. Such enzymes, like quitinases, endoglicanases and β-glicosydases are able to digest the fungal and bacterial cell wall. In this work, 24 strains of G. citricarpa were evaluated about the sensibility to fungicides used in field for BSC control: piraclostrobin and carbendazin, in dosages of 0,5, 1,0 and 2,0 mg a.i./mL, aiming to verify the effect of selection pressure caused by continuous use of this compounds. Two of these strains presented resistance to carbendazim in all evaluated dosages, showing that the use of this agrochemical may select resistant individuals, resulting a non-efficiency of this compound for BSC control. An alternative to minimize this kind of effect must be the application of this compound in combination to others active principles. The celullolitic and chitinollitic activities of 96 fungi strains widely spread phylogenetically were evaluated for selection of potential biological control agents. Four strains that presented major activity of each enzyme, besides two Trichoderma lineages, were tested as potential biological control of G. citricarpa in an experiment with ‘Valência’ orange leaves, comparing the action of these biocontrolers with commercial fungicides. Although a better pathogen control was achieved in leaves treated with piraclostrobin, two fungi strains revealed to have the similar efficiency to fungicide, inhibiting the development of G. citricarpa, suggesting the possible future utilization of biocontrol methods to Black Spot of Citrus.

Devido à percepção dos consumidores sobre o impacto da utilização de pesticidas sobre o ambiente e saúde humana, além da aquisição de resistência por parte dos fitopatógenos, a sociedade tem exercido pressões que levaram ao estabelecimento de políticas governamentais que restringem a utilização de fungicidas levando agricultores e pesquisadores a considerar a aplicação de técnicas de controle biológico de fitopatógenos fúngicos. Guignardia citricarpa é o agente causal da pinta preta dos citros, que é de grande importância econômica, pois interfere na produção e causa depreciação estética dos frutos, acarretando em prejuízos principalmente na comercialização de frutos in natura para o mercado externo. Neste contexto, o objetivo deste trabalho foi a avaliação in vitro do potencial de linhagens de Saccharomyces cerevisiae, utilizadas em processos fermentativos, como agentes de biocontrole contra G. citricarpa. Através de ensaio em placa, foi evidenciado que entre as linhagens de S. cerevisiae testadas (BG-1, CR-1, CAT-1, KD-1, K-1 e PE-2), a CR-1 demonstrou a maior atividade antagônica contra o fitopatógeno, causando 73% de inibição do crescimento micelial. Também foi demonstrado que as linhagens são capazes de produzir voláteis de ação fungistática inibindo em até 83% o desenvolvimento do patógeno. O filtrado de cultura autoclavado e não autoclavado, bem como as células inativadas termicamente, obtidas a partir do crescimento da linhagem CR-1 por 24 h em meio YEPD, não causaram redução do crescimento vegetativo do fungo. A produção de enzimas hidrolíticas extracelulares (quitinases, β-1,3-glucanases e proteases) pela levedura não foi detectada em meio YEPD contendo glicose ou preparação de parede celular de G. citricarpa, nos tempos de cultivo avaliados. Baseado nas informações obtidas, foi possível constatar que as linhagens de S. cerevisiae, em especial a linhagem CR-1, são potenciais antagonistas para o controle de G. citricarpa. O possível mecanismo utilizado para inibição pela levedura é a produção de voláteis, no entanto, outros mecanismos não podem ser descartados. Desta forma, o presente trabalho mostra o potencial de S. cerevisiae no controle de G. citricarpa em frutos de laranja na pós-colheita.
Due to the consumers perception about the impact caused by pesticides utilization over the environment and human health, besides the acquisition of resistance for part of the phytopathogens, the society has exercised pressures that had led to the establishment of governmental politics that restrict the use of fungicides leading agriculturists and researchers to consider the application of techniques of biological control of plant pathogenic fungi. Guignardia citricarpa is the causal agent of citrus black spot that has a great economic importance, therefore interfering in production and causing aesthetic depreciation of the fruits that can interfere with commercialization of fresh-fruit in the external market. In this context, the aim of this work was to evaluate in vitro the potential of Saccharomyces cerevisiae strains, used in fermentative process, as biocontrol agents against G. citricarpa. Through plate assay it was evidenced that among the tested strains of S. cerevisiae (BG-1, CR-1, CAT-1, KD-1, K-1 and PE-2), the strain CR-1 was the one that demonstrated the greatest antagonic activity against the phytopathogen, causing 73% of micelial growth inhibition. It was also demonstrated that the strains were able to produce volatile compounds with fungistatic action inhibiting up to 83% the development of the pathogen. The autoclaved and not autoclaved culture filtrate, as well as the termical inactivated cell obtained from the growth of strain CR-1 in YEPD medium for 24 h, did not cause reduction in the fungal vegetative growth. The production of extracellular hydrolytic enzymes (chitinases, β-1,3-glucanases and proteases) by the yeast was not detected in YEPD medium with glucose or cell wall preparation of G. citricarpa at the evaluated times. Based upon the obtained information it was possible to evidence that the strains of S. cerevisiae, specially the strain CR-1, are potentials antagonists for the control of G. citricarpa. The possible mechanism used for inhibition by yeast is the volatile production, however other mechanisms cannot be discarded. Thus, the present work shows the potential of S. cerevisiae to control G. citricarpa in orange fruits in postharvest.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
A efetiva produção de etanol lignocelulósico depende de leveduras capazes de fermentar a fração hemicelulósica da biomassa lignocelulósica que é composta em sua maioria por xilose. Há uma grande dificuldade em encontrar esses microrganismos já que a capacidade de utilizar xilose como fonte de carbono não é apresentada pela maioria das leveduras. Neste trabalho leveduras isoladas de diferentes materiais da zona rural,foram testadas quanto à assimilação de xilose e glicose, e fermentação de xilose a 30°C, onde observou-se diferentes comportamentos entre diferentes linhagens da mesma espécie. As linhagens de leveduras isoladas foram identificadas pela técnica molecular PCR-RFLP empregando-se as regiôes ITS 5,8 S e D1/D2 do DNA ribossômico. Três leveduras identificadas como espécies de Cândida tropicalis foram avaliadas quanto a capacidade de produção de xilitol e etanol a partir de meio basal com xilose como única fonte de carbono e do licor negro da extração da celulose hidrolisado por enzimas do fungo Thermoascus aurantiacus. A produção de xilitol foi semelhante nos dois substratos, entretanto a produção de etanol foi maior quando o licor negro foi empregado, esses resultados mostraram que a quantidade e a natureza dos açúcares presentes nos meios foram um interferente maior no metabolismo da levedura que os compostos tóxicos presentes no licor negro
The effective lignocellulosic ethanol production depends on yeasts that are able to ferment the hemicellulosic biomass fraction, which is mostly composed by xylose. There is a big difficulty in finding such microorganisms because most yeasts do not use xylose as a carbon source. In this work, yeasts isolated from different materials were tested for xylose and glucose assimilation and xylose fermentation at 30°C ,where it was observed different behaviors between strains of the same species. The strains isolated were identified by PCR-RFLP molecular technique employing the ITS 5,8 and D1/D2 regions of ribosomal DNA. Three yeasts were identified as Cândida tropicalis species and their ability to produce xylitol and ethanol from xylose and the black liquor from the cellulose extraction hydrolyzed by enzymes of the fungus Thermoascus aurantiacus was evaluated. The xylitol production was similar in both media, but the ethanol production was higher when the black liquor was used, these results shows that the quantity and nature of the sugars present in the media have a bigger interference in the metabolism of yeast to toxic compounds present in black liquor

Thesis (MScAgric)--University of Stellenbosch, 2002.
ENGLISH ABSTRACT: Historically root diseases have been a production-limiting problem for the strawberry industry worldwide. In the Western Cape Province of South Africa the most serious root disease is black root rot, which causes losses of up to 30%, annually. The aims of this study were to investigate aspects of the etiology and epidemiology of this disease in the Western Cape, and to provide information that can be incorporated in an integrated disease management strategy. In Chapter I a summary of published information on this disease is presented. Disease symptoms include severe stunting of plants, which have black, rotted, reduced root systems. Even though this disease is of great economic importance, the etiology remains unresolved. However, soilborne fungal root pathogens, particularly Pythium and Rhizoctonia spp. have been implicated as major role players. Control of this disease, as well as the other root diseases affecting strawberries, has relied on soil fumigation with broad spectrum chemical fumigants, in particular methyl bromide. However, due to the ozone depleting action of methyl bromide it was decided at the signing of the Montreal Protocol to remove this chemical from the market. This action has caused great demand for alternative measures to control root diseases on many crops including strawberries. Development of integrated disease management strategies is dependent upon a more complete understanding of the etiology, biology and ecology of the disease complex. In Chapter 2 fungal pathogens associated with diseased plants were isolated and Koch's postulates were carried out. The most frequently isolated fungal pathogens were Pythium irregulare, Rhizoctonia spp. and Cylindrocarpon destructans. Two morphotypes of Rhizoctonia were isolated viz. a brown and a white type. Pythium irregulare was isolated more frequently in July than in September, and was not isolated at all in November. Rhizoctonia spp. were present at all sampling dates but were more frequently isolated in November than at the other times. All the fungi that were tested were pathogenic and caused root lesions. Cylindrocarpon destructans and Coniella fragariae did not have a stunting effect on the plants. These results confirm a major role for Pythium and Rhizoctonia in the black root rot complex and suggest that there is a complimentary seasonal variation in occurrence between these two pathogens. The Rhizoctonia species and anastomosis groups of isolates obtained from diseased strawberries in the Western Cape Province were determined, and their pathogenicity and relative virulence assessed. Both binucleate and multinucleate types were recovered from diseased roots and identified as R. fragariae and R. so/ani, respectively. All isolates of R. solani were members of anastomosis group (AG) 6, whereas three AG types were identified among isolates of R. fragariae, viz. AG-A, AG-G and AG-I at a relative occurrence of 69%, 25%, 6% respectively. All Rhizoctonia isolates were pathogenic to strawberry, but R. solani (AG 6) was the most virulent causing severe stunting of plants. This is the first species confirmation and AG type identification of Rhizoctonia taxa causing root rot of strawberries in South Africa. An assessment of the presence and quantity of black root rot pathogens associated with soils prior to fumigation and post fumigation with methyl bromide, was made in Chapter 4. Isolations were also made from nursery plants to determine whether any black root rot pathogens were in the plants before transplanting. Results demonstrated that after fumigation the soil was free of all pathogenic fungi associated with the disease. However, the main pathogens involved in black root rot, viz. Rhizoctonia fragariae, R. solani and Pythium spp. were isolated from nursery plants. The fact that the plants are already infected with these pathogens renders the prospects for control of this disease difficult. Further studies are urgently required to develop production practices that can be included in disease management programmes. In vitro studies were carried out to determine the ECso values of different fungicides for isolates of Pythium irregulare, Rhizoctonia fragariae AG-A, AG-G and AG-I and R. solani AG 6. Benomyl, fludioxonil and tolc1ofos-methyl were used in these tests. Field trials were also conducted using these fungicides. In Chapter 5 it is shown that in general application of fungicides improved the yield and did not affect the survival rate of strawberry plants. Fludioxonil showed potential for short-term use. Applications of fungicides that targeted specific fungal genera were not sufficient to control the disease. Seasonal fluctuation of Pythium and Rhizoctonia spp. became apparent with the occurrence of Pythium being relatively high early in the season but low late in the season. Conversely, the occurrence of Rhizoctonia was low at the beginning of the season but high late in the season. In the short-term there is potential for fungicide applications as part of an integrated disease management strategy, but the economic feasibility of this practice needs to be assessed. In this study the major pathogens causing black root rot were identified in the Western Cape Province of South Africa, and important information regarding the epidemiology of the disease was reported. These results can be incorporated in an integrated management strategy to reduce losses of strawberry production, attJibutable to black root rot.
AFRIKAANSE OPSOMMING: Wortelsiektes is wêreldwyd 'n produksie-beperkende probleem vir die aarbeibedryf. . Swartwortelvrot, wat jaarliks verliese van tot 30% veroorsaak, is die belangrikste wortelsiekte in die Wes-Kaap Provinsie van Suid-Afrika. Die doelwitte van hierdie studie was om aspekte van die etiologie en epidemiologie van die siekte in die Wes- Kaap te ondersoek en inligting wat in geïntegreerde siektebestuurstrategië ingesluit kan word, te voorsien. In Hoofstuk 1 word 'n opsomming van gepubliseerde inligting aangaande die siekte uiteengesit. Siektesimptome sluit ernstige verdwerging van plante met swart verotte en verkleinde wortelstelsels in. Alhoewel die siekte van groot ekonomiese belang is, is die etiologie grootliks onbekend. Grondgedraagde wortelpatogene swamme, spesifiek Pythium en Rhizoctonia spp., is egter as belangrike rolspelers geïdentifiseer. Tot dusver het die beheer van hierdie siekte sowel as ander wortelsiektes van aarbeie berus op grondberoking met breë spektrum chemiese berokingsmiddels, spesifiek metielbromied. As gevolg van die osoonafbrekende aksie van metielbromied is daar egter tydens die ondertekening van die Montreal Protocol besluit om dié middel van die mark te verwyder. Hierdie besluit het 'n groot aanvraag na alternatiewe beheermaatreëls vir wortelsiektes van verskeie gewasse, insluitende aarbeie, veroorsaak. Die ontwikkeling van geïntegreerd siektebestuurstrategieë is egter afhanklik van 'n meer volledige begrip van die etiologie, biologie en ekologie van die siektekompleks. In Hoofstuk 2 is die patogene swamme wat met die siekte geassosieer word, geïsoleer, en is Koch se postulate uitgevoer. Die mees algemeen geïsoleerde patogene swamme was Pythium irregulare, Rhizoctonia spp. en Cylindrocarpon destructans. Twee morfotipes van Rhizoctonia is geïsoleer, nl. 'n bruin tipe en 'n wit tipe. Pythium irregulare is meer dikwels in Julie as in September geïsoleer, maar glad nie in November nie. Rhizoetonia het tydens alle monstertye voorgekom, maar is meer dikwels in November geïsoleer. Al die swamme wat getoets is, was patogenies en het letsels op die wortels veroorsaak. Cylindroearpon des true tans en Coniella fragariae het nie'n verdwergingseffek op plante gehad nie. Hierdie resultate bevestig die dominante rol van Pythium en Rhizoctonia in die swartwortelvrot kompleks en dui op 'n komplementêre seisoenale variasie in die voorkoms van hierdie twee patogene. Die Rhizoctonia spesies en anastomose groepe (AG) van die isolate geisoleer vanaf siek aarbeiplante in die Wes-Kaap Provinsie is bepaal, en die patogenisiteit en relatiewe virulensie is beraam. Sowel tweekernige as multikernige tipes is vanaf siek wortels geïsoleer en respektiewelik as R. fragariae en R. so/ani geïdentifiseer. Alle isolate van R. so/ani was lede van anastomose groep 6, terwyl drie AG tipes, nl. AGA, AG-G en AG-I onder die R. fragariae isolate geïdentifiseer is met relatiewe voorkomste van 69%, 25%, 6% respektiewelik. Alle Rhizoctonia isolate was patogenies op aarbeie, maar R. so/ani (AG 6) was die mees virulente en het ernstige verdwerging van plante veroorsaak. Hierdie is die eerste bevestiging van spesies en identifisering van AG tipes van Rhizoctonia taksa wat wortelvrot van aarbeie in Suid Afrika veroorsaak. In Hoofstuk 4 is 'n beraming van die voorkoms en hoeveelheid swartwortelvrot patogene geassosieer met grond voor, en na beroking met metielbromied, gemaak. Isolasies is ook vanaf kwekeryplante gemaak om te bepaal of enige swartwortelvrot patogene voor oorplanting in die plante teenwoordig was. Die resultate het getoon dat grond na beroking vry was van alle patogeniese swamme geassosieër met die siekte. Die hoof patogene betrokke in die swartwortelvrot kompleks, nl. Rhizoctonia fragariae, R. so/ani en Pythium spp. was egter in die kwekery plante teenwoordig. Die feit dat plante reeds met hierdie patogene geïnfekteer is, maak die vooruitsigte vir die beheer van hierdie siekte moeilik. Verdere studies word dringend benodig vir die ontwikkeling van produksiepraktyke wat by siektebestuursprogramme ingesluit kan word. In vitro studies om die ECso waardes van die isolate van Pythium irregulare, Rhizoctonia fragariae AG-A, AG-G en AG-I en R. so/ani AG 6 vir die fungisiedes benomyl, fludioxonil en tolclofos-metiel te bepaal, is uitgevoer. Hierdie fungisiedes is ook in veldproewe getoets. In Hoofstuk 5 is getoon dat aanwending van fungisiedes die opbrengs verbeter het en nie die oorlewing van aarbeiplante beïnvloed het nie. Fludioxonil het potensiaal vir korttermyn gebruik getoon. Die aanwending van fungisiedes wat spesifieke swamgenera teiken, was nie voldoende om die siekte te beheer nie. Seisoenale fluktuasies van Pythium en Rhizoctonia spp. het duidelik geword met die relatief hoë voorkoms van Pythium vroeg in die seisoen, maar lae voorkoms laat in die seisoen, terwyl die voorkoms van Rhizoctonia laag was aan die begin van die seisoen, maar hoog later in die seisoen. In die korttermyn is daar potensiaal vir fungisiedtoedienings as deel van 'n geïntegreerde siektebestuurstrategie, maar die ekonomiese haalbaarheid van hierdie praktyk moet bepaal word. In hierdie studie is die hoof patogene wat swartwortelvrot van aarbeie in die Wes-Kaap Provinsie van Suid-Afrika veroorsaak geïdentifiseer, en belangrike inligting rakende die epidemiologie van die siekte is aangeteken. Hierdie resultate kan in 'n geïntegreerde bestuurstrategie geïnkorporeer word om verliese van aarbeiproduksie, toeskryfbaar aan swartwortelvrot te, verminder.

Kyoto University (京都大学)
0048
新制・課程博士
博士(農学)
甲第22477号
農博第2381号
新制||農||1074(附属図書館)
学位論文||R2||N5257(農学部図書室)
京都大学大学院農学研究科森林科学専攻
(主査)准教授 舘野 隆之輔, 教授 北島 薫, 教授 德地 直子
学位規則第4条第1項該当

A pinta preta dos citros tem como agente causal o fungo G. citricarpa. A doença causa depreciação estética dos frutos e praticamente inviabiliza a exportação de laranja in natura principalmente para a União Européia, por ser considerada uma doença quarentenária. Frutos colhidos contendo infecções quiescentes, embora sem sintomas, podem desenvolvê-los durante o processo de exportação. A utilização de fungicidas é o principal método de controle utilizado em pré e pós-colheita. No entanto, o custo do controle químico é significativamente alto e já foi relatado o surgimento de isolados resistentes. A levedura S. cerevisiae é capaz de inibir o desenvolvimento in vitro de G. citricarpa e o antagonismo se deve a produção de compostos orgânicos voláteis de ação fungistática. Em virtude da procura por novos métodos de controle menos danosos à saúde humana e meio ambiente, dos prejuízos causados pela doença e do potencial dos compostos voláteis no controle da doença na pós-colheita, o trabalho visou identificar os compostos voláteis produzidos pela levedura e elucidar os mecanismos envolvidos na inibição do fitopatógeno. Esse conhecimento é fundamental para o desenvolvimento de técnicas inovativas de controle. A produção de compostos voláteis antimicrobianos por S. cerevisiae foi dependente do substrato utilizado para o seu cultivo, sendo a glicose, sacarose e maltose fontes de carbono favoráveis. Foi verificado através da análise em SPME-GC-MS que a levedura produziu em meio BDA, principalmente compostos pertencentes ao grupo dos álcoois, constituindo 95% da composição total, além de ésteres em menor proporção. A mistura artificial de voláteis mimetizou os efeitos dos voláteis produzidos pela levedura, sendo os valores de MIC50 e MIC100 de 0,48 µL mL-1 e 2,84 µL mL-1, respectivamente. A exposição do fitopatógeno aos compostos voláteis reduziu a síntese de proteínas e inibiu a atividade de enzimas associadas à morfogênese (quitinase, b-1,3-glucanase, lacase e tirosinase). Os compostos voláteis desencadearam o processo de estresse oxidativo no fungo, fato observado através da elevação da atividade de enzimas (superóxido dismutase e catalase) associadas a mecanismos de detoxificação de espécies ativas de oxigênio. A análise das proteínas diferencialmente expressas de G. citricarpa, através de eletroforese bidimensional, indicou que os compostos voláteis alteraram a expressão de 40 proteínas, sendo 29 inibidas e 11 superexpressas. A mistura artificial de voláteis também foi ativa contra fungos de diversos grupos taxonômicos, no entanto, não apresentou atividade sobre as bactérias testadas. Os compostos 3-metil-1-butanol e 2- metil-1-butanol foram os mais ativos quando utilizados isoladamente contra Colletotrichum gloesporioides, Penicillium digitatum, Sclerotinia sclerotiorum e Fusarium oxysporum. Sementes de feijão fumigadas com a mistura artificial de voláteis exibiram redução na incidência de S. sclerotiorum. A mistura também exibiu efeito nematicida sobre Meloidogyne javanica. Portanto, os resultados demonstram que os voláteis identificados em S. cerevisiae afetam a síntese de proteínas e a atividade de enzimas associadas ao crescimento vegetativo de G. citricarpa. Finalmente, fumigação utilizando compostos voláteis apresenta potencial no controle de G. citricarpa em pós-colheita, bem como de outros organismos de importância agronômica. O trabalho também contribuiu com novos elementos sobre o papel dos voláteis na interação entre microrganismos na natureza.
The citrus black spot has as causal agent the fungus G. citricarpa. The disease causes aesthetic depreciation of fruits and practically makes them unviable for exportation mainly to European Union where the disease is considered a quarantine pest. Harvested fruits can have quiescent infections although without symptoms and can develop them during the exportation process. The use of fungicides is the main chemical control method used in pre and post-harvest, however the cost is significantly high and it was already described the development of fungicide resistant strains. The yeast S. cerevisiae is able to inhibit the in vitro G. citricarpa growth, and the antagonism is due to production of volatile organic compounds (VOCs) of fungistatic effect. Because of the search for new control methods less harmful for human health and environment, the losses caused by the disease and the potential of the VOCs in the control of the pathogen in the post-harvest, the aim of this work was to identify the VOCs produced by the yeast and to elucidate the mechanisms involved in the inhibition of the fungus. This knowledge is essential for the development of innovative control techniques. The results showed that the production of antimicrobial VOCs by S. cerevisiae was dependent on the substrate used, being glucose, sucrose and maltose favorable carbon sources. It was verified through analysis in SPME-GC-MS that the yeast, grown on PDA medium, produced mainly compounds belonging to the group of alcohols (95% of the total composition), besides esters in smaller proportion. An artificial volatile mixture reproduced the effects of the VOCs produced by the yeast, being the values of MIC50 and MIC100 of 0.48 µL mL-1 and 2.84 µL mL-1, respectively. The phytopathogen exposition to the VOCs reduced protein synthesis and inhibited the activity of morphogenesis associated enzymes like chitinase, b-1,3-glucanase, laccase and tyrosinase. The VOCs initiated the oxidative stress process, which was observed through the increase in the activity of superoxide dismutase and catalase, enzymes associated to the detoxification of active oxygen species. The analysis of the differentially expressed proteins of G. citricarpa, through two-dimensional gel electrophoresis, indicated that the VOCs modified the expression of 40 proteins, being 29 down regulated and 11 up regulated. The artificial mixture of VOCs was also active against fungi of several taxonomic groups, however, there were no activity against bacteria. The compounds 3-methyl-1-butanol and 2-methyl-1-butanol were the most active when tested alone against Colletotrichum gloesporioides, Penicillium digitatum, Sclerotinia sclerotiorum and Fusarium oxysporum. Bean seeds fumigated with the artificial mixture of VOCs showed reduction in S. sclerotiorum incidence. In addition, the VOCs exhibited nematicidal effect against Meloidogyne javanica. The results demonstrate that the VOCs produced by S. cerevisiae affect protein synthesis and the activity of enzymes associated to the vegetative growth in G. citricarpa. Thus, the fumigation using VOCs presents potential in the control of citrus black spot disease in post-harvest as well as of other organisms of agronomic importance. The present workalso contributed with new information regarding the role of the VOCs in the interactions among microorganisms in nature.

Since the beginning of the 1990s, there has been a sharp rise in anti-tobacco activism, adverse public opinion, litigation, and new legislation to counter the tobacco industry and reduce use. Despite this sharp rise in activism, the role of African Americans in this advocacy process has mostly escaped the analysis of the political science research community. This includes 1999 when payments to the states began from the historically significant Master Settlement Agreement (MSA), which was signed in 1998 between 46 Attorneys General and the tobacco industry. This research project analyzed the dynamics in the state tobacco coalitions in Arkansas and Georgia. It delved into the roles African Americans played in an effort to leverage resources for the black community. These funds represented needed resources for building capacity and infrastructure. The research used both primary and secondary data. The primary data were gathered by semi-structured interviews with state health officials, coalition members, and policy-makers all intimately involved in the allocation process. Secondary data were gathered from journals, newspaper articles, by-laws and program reports. Information was also gathered from publications and websites of reputable organizations working in tobacco prevention. These included the Campaign for Tobacco-free Kids and Americans for Non Smokers’ Rights.Through the lens of Interest Group Theory research analyzed the role African Americans played in the initial allocation of Tobacco Master Settlement Agreement funds in Arkansas and Georgia. It was found that African Americans in leadership roles are important to the initial allocation process. Despite Arkansas’ success in securing 15% of State Tobacco Prevention funds allocated through an Historically Black College or University for minority communities, blacks in leadership positions were no guarantee that resources would be allocated to black communities.

Resumo: A efetiva produção de etanol lignocelulósico depende de leveduras capazes de fermentar a fração hemicelulósica da biomassa lignocelulósica que é composta em sua maioria por xilose. Há uma grande dificuldade em encontrar esses microrganismos já que a capacidade de utilizar xilose como fonte de carbono não é apresentada pela maioria das leveduras. Neste trabalho leveduras isoladas de diferentes materiais da zona rural,foram testadas quanto à assimilação de xilose e glicose, e fermentação de xilose a 30°C, onde observou-se diferentes comportamentos entre diferentes linhagens da mesma espécie. As linhagens de leveduras isoladas foram identificadas pela técnica molecular PCR-RFLP empregando-se as regiôes ITS 5,8 S e D1/D2 do DNA ribossômico. Três leveduras identificadas como espécies de Cândida tropicalis foram avaliadas quanto a capacidade de produção de xilitol e etanol a partir de meio basal com xilose como única fonte de carbono e do licor negro da extração da celulose hidrolisado por enzimas do fungo Thermoascus aurantiacus. A produção de xilitol foi semelhante nos dois substratos, entretanto a produção de etanol foi maior quando o licor negro foi empregado, esses resultados mostraram que a quantidade e a natureza dos açúcares presentes nos meios foram um interferente maior no metabolismo da levedura que os compostos tóxicos presentes no licor negro
Abstract: The effective lignocellulosic ethanol production depends on yeasts that are able to ferment the hemicellulosic biomass fraction, which is mostly composed by xylose. There is a big difficulty in finding such microorganisms because most yeasts do not use xylose as a carbon source. In this work, yeasts isolated from different materials were tested for xylose and glucose assimilation and xylose fermentation at 30°C ,where it was observed different behaviors between strains of the same species. The strains isolated were identified by PCR-RFLP molecular technique employing the ITS 5,8 and D1/D2 regions of ribosomal DNA. Three yeasts were identified as Cândida tropicalis species and their ability to produce xylitol and ethanol from xylose and the black liquor from the cellulose extraction hydrolyzed by enzymes of the fungus Thermoascus aurantiacus was evaluated. The xylitol production was similar in both media, but the ethanol production was higher when the black liquor was used, these results shows that the quantity and nature of the sugars present in the media have a bigger interference in the metabolism of yeast to toxic compounds present in black liquor
Orientador: Roberto da Silva
Coorientador: Daniela Alonso Bocchini Martins
Banca: Eleni Gomes
Banca: Sandra Regina Ceccato Antonini
Mestre

Muscodor albus Worapong, Strobel & Hess, isolate CZ-620 (MA) is an endophytic fungus that produces volatile organic compounds (VOCs) and non-volatile antimicrobial compounds. The use of these VOCs to inhibit or kill a wide range of microorganisms is termed mycofumigation. This dissertation focuses on parameters of MA mycofumigation including: production and bioactivity of previously un-described water-soluble antimicrobial compounds produced by MA; distribution of antimicrobial compounds from a MA point source in three soil types as measured by effects on Verticillium dahliae and Colletotrichum coccodes; control of V. dahliae and C. coccodes on potato; the ability of MA to colonize soil; and the effects of mycofumigation on ectomycorrhizal fungi (EMF) in vitro and on the colonization of onion roots by the arbuscular mycorrhizal (AM) fungus Glomus intraradices. The bioactivity of water-soluble compounds produced in potato dextrose broth was significantly increased as measured in growth reduction of C. coccodes, V. dahliae, and Rhizoctonia solani. No reduction was observed for Aphanomyces cochlioides and Pythium ultimum. Antimicrobial compounds from a MA colonized barley point source reduced V. dahliae and C. coccodes populations in soils by 60-100% at distances up to 9 cm from the inoculation source depending on soil type. Mortality rate ranging from 70-100% was observed within a 3 cm radius from the inoculation source. In both field and greenhouse trials, MA colonized barley formulation reduced Verticillium wilt and black dot root rot severity and reduced populations of both pathogens in potato tissue as measured by real-time quantitative PCR and serial dilution. Planting directly into mycofumigated soil previously infested with V. dahliae or C. coccodes resulted in equal control of the pathogens when compared to a one-week mycofumigation period prior to planting. After six weeks of incubation MA did not colonize sterile soil further than 0.5 cm away from a MA inoculation point. In vitro experiments showed that most of the tested EMF were inhibited in the presence of MA VOCs, but were able to resume growth when removed from VOCs. Incorporating MA into soil had no negative but supportive effect on onion root colonization by the AM fungus G. intraradices.

Die Elektronisierung der Finanzmärkte ist in den letzten Jahren weit vorangeschritten. Praktisch jede Börse verfügt über ein elektronisches Handelssystem. In diesem Kontext beschreibt der Begriff Algorithmic Trading ein Phänomen, bei dem Computerprogramme den Menschen im Wertpapierhandel ersetzen. Sie helfen dabei Investmententscheidungen zu treffen oder Transaktionen durchzuführen. Algorithmic Trading selbst ist dabei nur eine unter vielen Innovationen, welche die Entwicklung des Börsenhandels geprägt haben. Hier sind z.B. die Erfindung der Telegraphie, des Telefons, des FAX oder der elektronische Wertpapierabwicklung zu nennen. Die Frage ist heute nicht mehr, ob Computerprogramme im Börsenhandel eingesetzt werden. Sondern die Frage ist, wo die Grenze zwischen vollautomatischem Börsenhandel (durch Computer) und manuellem Börsenhandel (von Menschen) verläuft. Bei der Erforschung von Algorithmic Trading wird die Wissenschaft mit dem Problem konfrontiert, dass keinerlei Informationen über diese Computerprogramme zugänglich sind. Die Idee dieser Dissertation bestand darin, dieses Problem zu umgehen und Informationen über Algorithmic Trading indirekt aus der Analyse von (Fonds-)Renditen zu extrahieren. Johannes Gomolka untersucht daher die Forschungsfrage, ob sich Aussagen über computergesteuerten Wertpapierhandel (kurz: Algorithmic Trading) aus der Analyse von (Fonds-)Renditen ziehen lassen. Zur Beantwortung dieser Forschungsfrage formuliert der Autor eine neue Definition von Algorithmic Trading und unterscheidet mit Buy-Side und Sell-Side Algorithmic Trading zwei grundlegende Funktionen der Computerprogramme (die Entscheidungs- und die Transaktionsunterstützung). Für seine empirische Untersuchung greift Gomolka auf das Multifaktorenmodell zur Style-Analyse von Fung und Hsieh (1997) zurück. Mit Hilfe dieses Modells ist es möglich, die Zeitreihen von Fondsrenditen in interpretierbare Grundbestandteile zu zerlegen und den einzelnen Regressionsfaktoren eine inhaltliche Bedeutung zuzuordnen. Die Ergebnisse dieser Dissertation zeigen, dass man mit Hilfe der Style-Analyse Aussagen über Algorithmic Trading aus der Analyse von (Fonds-)Renditen machen kann. Die Aussagen sind jedoch keiner technischen Natur, sondern auf die Analyse von Handelsstrategien (Investment-Styles) begrenzt.
During the last decade the electronic trading on the stock exchanges advanced rapidly. Today almost every exchange is running an electronic trading system. In this context the term algorithmic trading describes a phenomenon, where computer programs are replacing the human trader, when making investment decisions or facilitating transactions. Algorithmic trading itself stands in a row of many other innovations that helped to develop the financial markets technologically (see for example telegraphy, the telephone, FAX or electronic settlement). Today the question is not, whether computer programs are used or not. The question arising is rather, where the border between automatic, computer driven and human trading can be drawn. Conducting research on algorithmic trading confronts scientists always with the problem of limited availability of information. The idea of this dissertation is to circumnavigate this problem and to extract information indirectly from an analysis of a time series of (fund)-returns data. The research question here is: Is it possible to draw conclusions about algorithmic trading from an analysis of (funds-)return data? To answer this question, the author develops a complete definition of algorithmic trading. He differentiates between Buy-Side and Sell-Side algorithmic trading, depending on the functions of the computer programs (supporting investment-decisions or transaction management). Further, the author applies the multifactor model of the style analysis, formely introduced by Fung and Hsieh (1997). The multifactor model allows to separate fund returns into regression factors that can be attributed to different reasons. The results of this dissertation do show that it is possible to draw conclusions about algorithmic trading out of the analysis of funds returns. Yet these conclusions cannot be of technical nature. They rather have to be attributed to investment strategies (investment styles).

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O Brasil é o maior produtor de laranja e o maior exportador mundial de suco concentrado. A pinta preta dos citros, causada por Guignardia citricarpa, é uma doença de grande importância econômica, principalmente para o Estado de São Paulo. O interesse no controle biológico de fitopatógenos, como alternativa de controle e como forma de reduzir os problemas ocasionados pelo uso intensivo de fungicidas, tem levado ao desenvolvimento de técnicas alternativas para uma agricultura mais sustentável. O trabalho teve por objetivo avaliar o efeito de agentes de controle biológico (Bacillus subtilis e Trichoderma harzianum) e produtos alternativos (biofertilizantes e leite) no controle da pinta preta dos frutos cítricos em cultivos convencional e orgânico. Além disso, foi avaliado o efeito de Bacillus subtilis, Paenibacillus lentimorbus e Streptomyces sp. no controle de G. citricarpa e Penicillium digitatum em pós-colheita. Os experimentos em campo foram realizados em pomar de laranja`Valência´ e `Pêra´, localizadas nos municípios de Conchal e Santa Eudóxia, SP, respectivamente. Em cultivo convencional de `Valência´ foram avaliados dois biofertilizantes (Microgeo® e Bio2), nas safras 2003/2004 e 2004/2005, com 15 repetições por tratamento, sendo uma planta por repetição. As árvores foram pulverizadas em intervalos de 28 dias, sendo o início em 08/12/03, para a safra 2003/2004 e 08/11/2004, para a safra 2004/2005. Na safra 2003/2004 as concentrações utilizadas do biofertilizante Microgeo® foram 0; 10; 20; 30 e 40%. Na safra 2004/2005 foram repetidos os mesmos tratamentos dasafra anterior e incluído tratamento com o biofertilizante Bio2 nas concentrações de 0; 2,5; 5,0; 7,5 e 10%. Para as avaliações foram utilizadas uma escala de notas de 1=0,5% a 6=49% da área do fruto atacada. Os biofertilizantes apresentaram comportamento...
Brazil is the worldwide biggest producer and exporter of orange and orange juice. The citrus black spot (CBS), caused by Guignardia citricarpa, is a disease of great economic importance, mainly for the São Paulo State. The interest in biological control of plant pathogens, as mitigation of the problems caused by intensive use of fungicides, has led to development of alternative techniques for a more sustainable agriculture. The objective of this work was to evaluate the effects of biocontrol agents (Bacillus subtilis and Trichoderma sp.) and other biocompatible products (cow milk and biofertilizers) for managing CBS in organic and conventional systems. Besides, the effect of B. subtilis, Paenibacillus lentimorbus and Streptomyces sp. for control of G. citricarpa and Penicillium digitatum, in post-harvest was also evaluated. The filed experiments were carried through in trees of `Valencia' and `Pera´, located in Conchal and Santa Eudóxia, SP, respectively. In `Valencia´ conventional system were evaluated two biofertilizers (Microgeo® and Bio2). The experiments were conducted at harvests 2003/2004 and 2004/2005. The trees were sprayed in 28 days intervals, and the first was in 08 December 2003 in 2003/2004 harvest, and 08 November 2004 in 2004/2005 harvest.

O Brasil é considerado o maior produtor de citros e o maior exportador de suco de laranja. Doenças de pós-colheita representam uma grande perda na citricultura, sendo que para muitos frutos a serem exportados, existe uma exigência para que os mesmos estejam isentos de resíduos químicos. Em relação a alguns patógenos de importância em pós-colheita podemos destacar, Guignardia citricarpa (Mancha-pretados- citros), Penicillium digitatum (Bolor-verde) e Colletotrichum gloeosporioides (Antracnose). Dada a importância econômica que representa esse complexo de doenças dos frutos cítricos, tanto em termos de comprometimento da qualidade dos frutos, limitações às exportações e dificuldade de controle, a busca de alternativas adicionais que possam viabilizar a capacidade produtiva dos produtores e garantir a obtenção de frutos com excelentes padrões de qualidade torna-se imprescindível. Nesse contexto, pode-se inserir o emprego de medidas de controle alternativas que não englobam o controle químico clássico. Sob esse ponto de vista inclui-se o controle através do uso de agentes bióticos e abióticos e a indução de resistência em plantas. Portanto neste trabalho, foi estudada a viabilidade do controle de doenças pós-colheita em citros, envolvendo a ação direta sobre os patógenos através do uso dos extratos etanólicos de albedo (mesocarpo) de laranja doce (Citrus sinensis var. Valência) e flavedo (exocarpo ou epicarpo) de limão-Tahiti (Citrus aurantifolia Swing var. Tahiti). O extrato do albedo apresentou efeito antifúngico sobre G. citricarpa e o flavedo do limão “Tahiti” sobre C. gloeosporioides, além de se demonstrar a existência de compostos voláteis com efeito tóxico. A segunda parte envolveu o controle e a indução de resistência em frutos, através do uso dos agentes bióticos Lentinula edodes e Agaricus blazei e do agente abiótico ácido jasmônico. Foi possível observar que o extrato aquoso do albedo (C. sinensis), flavedo (C. aurantifolia), L. edodes e A. blazei diminuíram o aparecimento de novas lesões causadas por G. citricarpa, porém não apresentaram efeitos sobre P. digitatum e C. gloeosporioides em frutos de C. sinensis var. Valência quando tratados em pós-colheita. Dessa maneira, no presente trabalho demonstrou-se a viabilidade de um possível controle alternativo de doenças pós-colheita em citros, buscando-se novos agentes que atuem como indutores de resistência ou de controle direto sobre os fitopatógenos.
Brazil is considered the biggest citrus producer and the biggest orange juice exporter. Post-harvest diseases represent a great loss in the citriculture, and for many fruits to be exported they should be free of chemical residues. In relation to some pathogens of importance in post-harvest it can be mentioned Guignardia citricarpa (black-spot-ofcitrus), Penicillium digitatum (green-mold) and Colletotrichum gloeosporioides (anthracnose). Because of the economical importance that represents this disease complex in citric fruits, in terms of compromising fruit quality, limitations to the exports and control difficulties, the search for alternative control measures that can make possible improve the producing capacity of the producers and the obtaining of fruits with excellent quality are indispensable. Thus, in this context it can be included measures of alternative control that do not include the chemical control. Under this point of view, control include the use of biotic and abiotic agents and the resistance induction in plants. Therefore, it was studied the viability of the control of post-harvest diseases in citrus, involving the direct action on the patogens by using ethanolic extracts of albedo (mesocarp) of sweet orange (Citrus sinensis var. Valência) and flavedo (exocarp or epicarp) of lemon-Tahiti (Citrus aurantifolia Swing var. Tahiti). The results showed that the extract of the albedo exhibited antifungal activity on G. citricarpa and the flavedo of the "Tahiti" lemon on C. gloeosporioides, and it was also demonstrated the existence of volatile compounds toxic to the fungus. The second part involved the control and resistance induction in the fruits, by using the biotic agents Lentinula edodes and Agaricus blazei and the abiotic agent jasmonic acid. It was possible to observe that the aqueous extracts from the albedo (C. sinensis), flavedo (C. aurantifolia), L. edodes and A. blazei reduced the formation of new lesions caused by G. citricarpa, however they did not exhibit effects on P. digitatum and C. gloeosporioides in fruits of C. sinensis var. Valência when treated in post-harvest. Thus, in the present work it was demonstrated the viability of possible alternative control measures of diseases in post-harvest of citrus, indicating the need of looking for new agents to act as resistance inducers or agents to directly control on the phytopathogens.

A mancha preta dos citros causada pelo fungo Guignardia citricarpa Kiely [Phyllosticta citricarpa (McAlp.) Van der Aa], possui duas formas de infecção, conídios e ascósporos. Informações a respeito da importância dos conídios na epidemiologia da doença são escassas ou controversas. Visando uma maior compreensão sobre o patossistema citros-G. citricarpa (P. citricarpa), os objetivos desta dissertação foram: elaborar e validar uma escala diagramática para avaliação da severidade da mancha preta em folhas de citros; verificar o efeito da temperatura e da duração do período de molhamento na formação de apressórios formados a partir de conídios; observar por meio de microscopia eletrônica de varredura a germinação de conídios e a formação de apressórios sobre folhas destacadas de limão 'Siciliano' submetidas a diferentes temperaturas e períodos de molhamento. A escala diagramática com níveis de severidade de 1; 3; 6; 12; e 24% de área foliar lesionada foi validada por dois grupos de avaliadores, com e sem experiência na quantificação de doenças. Comparada com a avaliação sem escala, o uso da escala proporcionou melhor precisão e acurácia tanto para avaliadores experientes como inexperientes, quando considerada a estimativa média dos mesmos. Na maioria dos casos, os desvios entre estimativas e severidade atual da doença foram mais evidentes para os níveis de severidade entre 5 e 15%. A reprodutibilidade das avaliações resultou em valores de R 2 mais uniformes para a maioria dos avaliadores experientes. Diferenças consideráveis de precisão foram observadas entre avaliadores inexperientes. O efeito da temperatura (10 o C – 40 o C) e da duração do molhamento (4 – 48 h) na formação de apressórios formados a partir de conídios de G. citricarpa (P. citricarpa) foi avaliado sob condições "in vitro" e sobre a superfície de folhas de limão 'Siciliano'. A formação de apressórios ocorreu em todas as temperaturas a partir de 12 horas de molhamento, sendo os extremos de temperatura (10 o C e 40 o C) menos favoráveis à formação de apressórios. A temperatura mínima para formação de apressórios, estimada pela função beta generalizada foi de 3 o C e a máxima de 48,4 o C, ambas para 48 horas de molhamento. A formação de apressórios foi consideravelmente favorecida pela duração do período de molhamento, com o máximo de apressórios formados a 24 horas de molhamento, para a maioria das temperaturas. O período de molhamento constituído de 48 horas foi essencial para que os esporos submetidos a temperaturas de 10 o C e 40 o C, formassem apressórios. A superfície de resposta obtida pela multiplicação das funções beta generalizada e monomolecular apresentou um ajuste satisfatório para os dados observados na estimativa da porcentagem relativa de apressórios formados (R 2 =0,75). As amostras observadas em microscopia eletrônica de varredura possibilitaram a aquisição de imagens de conídios e apressórios sobre a superfície de folhas de limão 'Siciliano' em todas as combinações de temperatura e molhamento avaliadas.
Citrus black spot caused by Guignardia citricarpa Kiely [Phyllosticta citricarpa (McAlp.) van der Aa] presents two infection forms, conidia and ascospores. Information regarding the importance of the conidia in the epidemiology of the disease is scarce and controversial. Seeking a better understanding on the pathosystem citrus-G. citricarpa (P. citricarpa), the objectives of this dissertation were: elaborate and validate a diagrammatic scale for assessments of the citrus black spot; verify the effect of the temperature and of the wetness duration in the appressorium formation; observe through scanning electron microscopy the germination and formation of appressorium on outstanding lemon 'Siciliano' leaves submitted to different temperatures and wetness duration. The diagrammatic scale with severity levels of 1; 3; 6; 12; and 24% of diseased leaf area was validated by two groups of raters, with experience and without experience in the quantification of diseases. The scale provided better precision and accuracy for both experienced and inexperienced raters, considering the estimates average of them. In the majority of cases, the bias between estimated and actual disease severity were more evident for disease severity levels between 5 and 15%. The reproducibility of assessments resulted in R 2 with more uniforms values for the majority of the experienced raters, considerable differences of precision were observed among inexperienced raters. The effect of the temperature (10 o C - 40 o C) and of the wetness duration (4 – 48 h) in the germination of conidia and appressoria formation of G. citricarpa (P. citricarpa), was assessed "in vitro" and on the surface of lemon 'Siciliano' leaves. The appressoria formation occurred in all the temperatures starting from 12 hours of wetness. The extreme temperatures (10 o C and 40 o C) were less favorable to the apressorium formation. The minimum temperature for appressorium formation, estimated by generalized beta function was of 3 o C and the maximum of 48,4 o C, both for 48 hours of wetness. The appressorium formation was favored considerably by the wetness duration period, with the maximum of apressoria formed at 24 hours of wetness, for majority of the temperatures. The wetness duration period constituted of 48 hours was essential so that the spores submitted to temperatures of 10 o C and 40 o C, formed appressorium. The response surface obtained by the multiplication of the generalized beta and monomolecular functions provided a close fit to observed data in the estimate of the relative percentage of formed appressorium (R 2 =0,75). The samples observed in scanning electron microscopy made possible the acquisition of images of conidia and appressoria on the surface of lemon 'Siciliano' leaves in all the temperature combinations and wetness evaluated.

Els fongs micorízics formen associacions simbiòtiques amb les plantes i juguen un paper important en els processos de l'ecosistema. Els fongs micorízics són rellevants per la seva funció d'obtenció de compostos de carboni al mateix temps que aporten nutrició mineral a la seva planta hoste simbiòtica, però també els fongs micorízics tenen un interès econòmic, al proporcionar esporocarps comestibles. Entre ells, Tuber melanosporum és un dels fongs més apreciats en el mercat i el seu cultiu i producció silvestre és important per a l'economia i el paisatge en les zones rurals del sud d'Europa. L'objectiu principal d'aquesta tesi va ser analitzar: i) la situació actual de el sector de la tòfona a l'àrea mediterrània; ii) la dependència de la productivitat de T. melanosporum de la variabilitat de les precipitacions; iii) les interaccions de T. melanosporum amb altres fongs del sòl en plantacions; i iv) la diversitat de tipus de compatibilitat sexual o "mating types" a les plantacions de T. melanosporum. Amb l'objectiu de dilucidar la situació actual del sector de la tòfona a l'àrea mediterrània, un total de 17 panelistes experts van participar en una enquesta Delphi on van respondre un qüestionari en dues rondes. Es van analitzar tres registres continus de 49 anys de la producció anual de T. melanosporum de les principals regions productores de tòfona del sud d'Europa al nord-est d'Espanya, el sud de França i el nord i centre d'Itàlia per comprendre la dependència de la producció de T. melanosporum de les condicions climàtiques. Finalment, es van dur a terme dos experiments en una plantació experimental d'1 ha consistent en 249 plantes de Quercus ilex de cinc anys inoculats amb T. melanosporum plantats en una pastura recentment abandonada envoltada de bosc situada al Prepirineu oriental; i en 29 plantacions de T. melanosporum de Terol (Espanya) de 3, 5, 7, 10, 14 i 20 anys. Utilitzant diverses tècniques moleculars com la PCR en temps real (qPCR), la seqüenciació massiva d'ADN (PacBio RS II) i un assaig de qPCR patentat (WO2012 / 032.098) per quantificar els dos tipus de compatibilitat sexual, descrivim els canvis de la composició de la comunitat fúngica i la distribució del tipus de compatibilitat sexual. Els resultats obtinguts a través de les dues rondes Delphi ens van permetre tenir una descripció completa de la cadena de valor actual de la tòfona i confirmem una evolució del sector a causa de l'èxit del cultiu de la tòfona. A causa de l'augment de la producció de T. melanosporum en els últims anys, s'ha observat una tendència a la baixa dels preus de la tòfona que han alertat el sector. Per reforçar el sector de la tòfona, es van identificar accions consensuades per desenvolupar en el futur. Pel que fa a la part ecològica d'aquesta tesi, revela com la producció de T. melanosporum, depèn significativament dels totals de precipitació anteriors de juny a agost, mentre que massa precipitació a la tardor afecta negativament la collita d'hivern. Una de les principals preocupacions d'aquesta tesi ha estat si altres fongs ectomicorízics podrien desplaçar T. melanosporum a les plantacions i afectar la producció de tòfones. Trobem una major abundància relativa de fongs ECM diferents de T. melanosporum associats amb arbres més propers a bosc, però els arbres amb major diàmetre del coll de l'arrel van tenir major biomassa de miceli de T. melanorporum i van ser aquells on la comunitat fúngica es va veure menys afectada per la distància a bosc. No trobem associació entre el miceli de T. melanosporum a terra i la distància a bosc o l'abundància de fongs ECM diferents de T. melanosporum. En l'estudi de cronoseqüència, trobem que T. melanosporum es va desenvolupar de manera constant al llarg dels anys, ja que el miceli extraradical de T. melanosporum no es va correlacionar amb canvis en altres fongs ECM. La freqüència del tipus de compatibilitat sexual no va canviar al llarg dels anys. En resum i amb base a aquests resultats, es pot concloure que (i) hi ha la necessitat de promoure el consum de tòfones, donat l'augment de la producció a causa del cultiu de tòfones; (ii) la producció de T. melanosporum depèn significativament de les pluges de l'estiu anterior, mentre que massa precipitació de tardor té efectes negatius; (iii) encara que el veïnatge del bosc influirà en la comunitat de fongs del sòl, això pot tenir un efecte limitat sobre el creixement del miceli de T. melanosporum; (iv) T. melanosporum pot mantenir la colonització dels arbres a llarg termini; (v) l'inici de la reproducció sexual no està limitat per una presència desproporcionada de qualsevol dels tipus de compatibilitat sexual, ni pel desplaçament per fongs ECM diferents de T. melanosporum.
Los hongos micorrícicos forman asociaciones simbióticas con las plantas y juegan un papel importante en los procesos del ecosistema. Los hongos micorrícicos son relevantes por su función de obtención de compuestos de carbono a la vez que aportan nutrición mineral a su planta huésped simbiótica, pero también los hongos micorrícicos tienen un interés económico, al proporcionarnos esporocarpos comestibles. Entre ellos, Tuber melanosporum es uno de los hongos más apreciados en el mercado y su cultivo y producción silvestre es importante para la economía y el paisaje en las zonas rurales del sur de Europa. El objetivo principal de esta tesis fue analizar: i) la situación actual del sector de la trufa en el área mediterránea; ii) la dependencia de la productividad de T. melanosporum de la variabilidad de las precipitaciones; iii) las interacciones de T. melanosporum con otros hongos del suelo en plantaciones; y iv) la diversidad de tipos de compatibilidad sexual o “mating types” en las plantaciones de T. melanosporum. Con el objetivo de dilucidar la situación actual del sector de la trufa en el área mediterránea, un total de 17 panelistas expertos participaron en una encuesta Delphi donde respondieron un cuestionario en dos rondas. Se analizaron tres registros continuos de 49 años de la producción anual de T. melanosporum de las principales regiones productoras de trufa del sur de Europa en el noreste de España, el sur de Francia y el norte y centro de Italia para comprender la dependencia de la producción de T. melanosporum de las condiciones climáticas. Finalmente, se llevaron a cabo dos experimentos en una plantación experimental de 1 ha consistente en 249 plantas de Quercus ilex de cinco años inoculados con T. melanosporum plantados en un pastizal recientemente abandonado rodeado de bosque ubicado en el Prepirineo oriental; y en 29 plantaciones de T. melanosporum de Teruel (España) de 3, 5, 7, 10, 14 y 20 años. Utilizando varias técnicas moleculares como la PCR en tiempo real (qPCR), la secuenciación masiva de ADN (PacBio RS II) y un ensayo de qPCR patentado (WO2012 / 032098) para cuantificar los dos tipos de compatibilidad sexual, describimos los cambios de la composición de la comunidad fúngica y la distribución del tipo de compatibilidad sexual. Los resultados obtenidos a través de las dos rondas Delphi nos permitieron tener una descripción completa de la cadena de valor actual de la trufa y confirmamos una evolución del sector debido al éxito del cultivo de la trufa. Debido al aumento de la producción de T. melanosporum en los últimos años, se ha observado una tendencia a la baja de los precios de la trufa que han alertado al sector. Para reforzar el sector de la trufa, se identificaron acciones consensuadas para desarrollar en el futuro. En cuanto a la parte ecológica de esta tesis, revela cómo la producción de T. melanosporum, depende significativamente de los totales de precipitación anteriores de junio a agosto, mientras que demasiada precipitación otoñal afecta negativamente la cosecha de invierno. Una de las principales preocupaciones de esta tesis ha sido si otros hongos ectomicorrícicos podrían desplazar a T. melanosporum en las plantaciones y afectar la producción de trufas. Encontramos una mayor abundancia relativa de hongos ECM distintos de T. melanosporum asociados con árboles más cercanos al bosque, pero los árboles con mayor diámetro de cuello de la raíz tuvieron mayor biomasa de micelio de T. melanorporum y fueron aquellos cuya comunidad fúngica se vio menos afectada por la distancia al bosque. No encontramos asociación entre el micelio de T. melanosporum en el suelo y la distancia al bosque o la abundancia de hongos ECM distintos de T. melanosporum. En el estudio de cronosecuencia, encontramos que T. melanosporum se desarrolló de manera constante a lo largo de los años, ya que el micelio extraradical de T. melanosporum no se correlacionó con cambios en otros hongos ECM. La frecuencia del tipo de compatibilidad sexual no cambió a lo largo de los años. En resumen y con base en estos resultados, se puede concluir que (i) existe la necesidad de promover el consumo de trufas, dado el aumento de la producción debido al cultivo de trufas; (ii) la producción de T. melanosporum depende significativamente de las lluvias del verano anterior, mientras que demasiada precipitación otoñal tiene efectos negativos; (iii) aunque la vecindad del bosque influirá en la comunidad de hongos del suelo, esto puede tener un efecto limitado sobre el crecimiento del micelio de T. melanosporum; (iv) T. melanosporum puede mantener la colonización de los árboles a largo plazo; (v) el inicio de la reproducción sexual no está limitado por una presencia desproporcionada de cualquiera los tipos de compatibilidad sexual, ni por el desplazamiento por hongos ECM distintos de T. melanosporum.
The mycorrhizal fungi form symbiotic associations with plants and play an important role in ecosystem processes. Mychorrizal fungi are relevant for their function for obtaining carbon compounds while providing mineral nutrition to their symbiotic host plant, but also, mycorrhizal fungi have an economic interest, by providing us with edible sporocarps. Among them, T. melanosporum is one of the most appreciated fungi in the market and its wild production and cultivation is important for rural livelihoods and landscapes in southern Europe. The main objective of this thesis was to analyse: i) the current situation of the truffle sector in the Mediterranean area; ii) the dependency of T. melanosporum productivity on rainfall variability; iii) the interactions of T. melanosporum with other soil fungi in plantations; and iv) the mating types diversity in T. melanosporum plantations. Aiming to elucidate the current situation of the truffle sector in the Mediterranean area, a total of 17 expert panellist participated in a Delphi survey where they answered a questionnaire in two of rounds. Three continuous, 49 year-long records of the annual T. melanosporum production from southern Europe’s main truffle producing regions in northeastern Spain, southern France and northcentral Italy was analysed to understand the dependency of T. melanosporum production on climate conditions. Finally, two experiments were carried out in a 1-ha experimental plantation consisting of 249 five-year-old Q. ilex inoculated with T. melanosporum planted in a recently abandoned pasture surrounded by forest located in the eastern Pre-Pyrenees; and in 29 T. melanosporum plantations from Teruel (Spain) with 3, 5, 7, 10, 14 and 20 years-old. Using several molecular techniques such as real-time PCR (qPCR), high-throughput DNA sequencing (PacBio RS II), and a patented qPCR essay (WO2012/032098) to quantify the two mating types, we describe the compositional changes of the fungal community and mating type distribution. The results obtained through the two Delphi rounds allowed us to have a complete description of the current truffle value chain and we confirmed an evolution of the sector due to the cultivation success of truffles. Due to the increase in T. melanosporum production in recent years, a trend of truffle prices decrease has been observed in the last years that have alerted the sector. To reinforce the truffle sector, highly agreed actions to develop in the future were identified. Regarding the ecological part of this thesis, it reveals how T. melanosporum production, significantly relies on previous June-August precipitation totals, while too much autumnal precipitation negatively affects winter harvest. One main concern of this thesis is if other ectomycorrhizal fungi could displace T. melanosporum in plantations and impair truffle production. We found a higher relative abundance of non-T. melanosporum ECM fungi associated with trees closer to the forest, but trees with larger root collar diameter had greater biomass of T. melanorporum mycelium and were those whose fungal community was less affected by the distance to the forest. We did not find association between T. melanosporum mycelium in the soil and distance to the forest or the abundance of non-T. melanosporum ECM fungi was observed. In the chronosequence study, we found that T. melanosporum developed steadily over the years as extraradical mycelium and T. melanosporum was not correlated with changes in other ECM fungi. Mating type frequency did not change across the years. In summary and based on these results, it can be concluded that (i) there is a need to promote the consumption of truffles, given the increase in production due to the cultivation of truffles; (ii) the T. melanosporum production significantly depends on previous summer rainfall, whereas too much autumnal precipitation has negative effects; (iii) even though the forest vicinity will influence the soil fungal community, this may have a limited effect on the growth of truffle mycelium; (iv) T. melanosporum is able to maintain colonization of the trees over the long term; (v) the initiation of the sexual reproduction is not limited by a disproportionate presence of either mating type, nor by displacement by non-T. melanosporum ECM fungi.