Orientador: Luiz Antonio Graciolli / Banca: Ana Maria Rodrigues Cassiolato / Banca: Eustáquio Souza Dias / Resumo: O presente trabalho teve por objetivo evidenciar os principais componentes químicos que influenciam o cultivo do cogumelo Pleurotus florida, bem como avaliar a composição química dos corpos de frutificação cultivados em diferentes resíduos agrícolas. Para tanto, foram utilizados seis resíduos agrícolas: palha de arroz (PA), palha de feijão (PF), palha de trigo (PT), palha de sorgo (PS), folha de bananeira (FB) e sabugo de milho (SM). Estes resíduos, coletados na Fazenda de Ensino, Pesquisa e Extensão da UNESP, Campus de Ilha Solteira, foram analisados quanto aos teores de macro (N, P, K, Ca, Mg) e micronutrientes (Fe, Mn e Zn), lignina, celulose, hemicelulose, cinzas e relação C/N. Foram avaliados: o tempo necessário para a colonização do substrato (corrida micelial), o início da formação de primórdios, o tempo total de cultivo, o número de cogumelos, a produção e a eficiência biológica utilizando a fórmula: EB = (peso fresco dos cogumelos/peso seco do substrato inicial) x 100. Além disso, os cogumelos foram avaliados quanto aos teores de macro (N, P, K, Ca e Mg) e micronutrientes (Fe, Mn e Zn) e o teor de proteínas. O substrato PF apresentou resultados semelhantes para produção (189,8 g/kg-1), EB (89,2%) e número de cogumelos (12) à PA, substrato utilizado tradicionalmente no cultivo de Pleurotus em escala comercial. Não foi possível atribuir apenas a um fator químico as altas produções e EB observadas em PA e PF e muito menos para a baixa produção em PS (77,8 g/kg-1) e SM (53,2 g/kg-1). No geral, substratos com conteúdo de N ao redor de 1,0%, relação C/N em torno de 45%, baixo teor lignina, alto conteúdo de cinzas e maiores teores de P, K e Ca foram os melhores para o cultivo de P. florida. Entre os macronutrientes analisados, P. florida apresentou maiores teores de K, seguido por P. O Ca e o Mg estiveram presentes em pequenas quantidades nos corpos... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This study aimed to show the main chemical components that influence the cultivation of the mushroom Pleurotus florida, and evaluate the chemical composition of fruiting bodies grown on different agricultural residues. For that, six agricultural residues were used: rice straw (RS), bean straw (BS), wheat straw (WS), sorghum straw (SS), banana leaf (BL) and cob of maize (CM). These wastes were collected at Teaching, Research and Extention Farm of UNESP, Campus of Ilha Solteira, analyzed for the levels of macro (N, P, K, Ca, and Mg) and micronutrients (Fe, Mn and Zn), lignin, cellulose, hemicellulose, ash and the C/N ratio. Was evaluated: spawn run time, earliness, the total time of cultivation and mushroom number, production and biological efficiency using the formula: EB = (fresh weight of mushrooms/dry weight of initial substrate) x 100. Moreover, the mushrooms were evaluated for levels of macro (N, P, K, Ca and Mg) and micronutrients (Fe, Mn and Zn) and protein content. The substrate BS showed similar results for yield (189,8 g/kg-1), EB (89,2%) and number of mushrooms (12) as RS substrate traditionally used for the cultivation of Pleurotus in commercial scale. Unable to allocate only the chemical factors and the high yields and EB observed in BS and RS, much less for the yield low in SS (77,8 g/kg-1) and CM (53,2 g/kg-1). In general, substrates with N content of around 1.0%, C/N ratio around 45%, low lignin content, high ash content, increased by higher levels of P, K, Ca were the best for the cultivation of P. florida. Among the macronutrients analyzed, P. florida with higher contents of K, followed by P. The Ca and Mg were present in small amounts in fruiting bodies. Among the micronutrients, Zn was present in high amount, followed by Fe and Mn. P. florida showed high ability to accumulate Zn. The mushrooms obtained in this work could not be considered a source of minerals... (Complete abstract click electronic access below) / Mestre

In der vorliegenden Arbeit wurde eine Beeinflussung der Aktivität der humanen neutrophilen Elastase (EC 3.4.21.37) durch wässrige und Dichlormethan-Extrakte von 15 Basidiomyceten festgestellt. Durch aktivitätsgeleitete Fraktionierung (mehrfache SC, GC-MS) der Dichlormethan-Extrakte von Heterobasidion annosum (Fr.) Bref. und Lactarius deterrimus Grög. wurden Fraktionen freier langkettiger Fettsäuren als ein wirksames Prinzip der Elastase-Hemmung und auch der Kollagenase-Hemmung (Clostridium histolyticum Kollagenase, EC 3.4.24.3) isoliert und identifiziert. Das Screening von 17 freien langkettigen Fettsäuren zeigte, dass einfach ungesättigte Fettsäuren eine stärkere Hemmung der Elastase-Aktivität bewirkten als ihre gesättigten bzw. mehrfach ungesättigten Homologa: Ölsäure (C18:1 cis-9): IC50 5µM; Stearin-(C18:0), Linolsäure (C18:2 cis-9,12): IC50 10µM; alpha- (C18:3 cis-9,12,15), gamma-Linolensäure (C18:3 cis-6,9,12): IC50 15µM. Inhibitorisch am stärksten wirksam war Erucasäur! e (C22:1 cis-13): IC50 450nM. Für Kollagenase wurde hingegen gezeigt, dass die gesättigten Fettsäuren eine erheblich stärkere Hemmaktivität als ihre ungesättigten Homologa aufwiesen. Aktivste Verbindungen waren Palmitin- (C16:0), Heptadecan- (C17:0), Stearin- und Nonadecansäure (C19:0) mit IC50-Werten von 20-45µM. Die Untersuchung von 9 ausgewählten Fettsäuren bezüglich der Hemmung der Aktivität der MMP-9 (EC 3.4.24.35) zeigte als aktivste Verbindungen Palmitolein- (16:1 cis-9), alpha- und gamma-Linolensäure. Die wirksamen Konzentrationen (250µM) lagen jedoch sehr hoch. Zytotoxizitätsuntersuchungen (ECV-304) der Extrakte von H. annosum und L. deterrimus sowie der freien Fettsäuren schlossen sich ebenso wie Untersuchungen zur Proteaseaktivität der Zelllinien ECV-304, MCF-7 und MDA-MB 231 an. Die Proteaseaktivität der Zellen nahm in der Reihenfolge ECV-304 < MCF-7 < MDA-MB 231 zu. Die einzig untersuchte Fettsäure gamma-Linolensäure zeigte keine reproduzierbare Beeinflussung d! er Proteaseaktivität. / In the present paper it was established that the activity of humane neutrophil elastase (EC 3.4.21.37) is affected by aqueous and dichloromethane extracts of 15 basidiomycetes. Bioassay-guided fractionation (repeated CC, GC-MS) of dichloromethane extracts of Heterobasidion annosum (Fr.) Bref. and Lactarius deterrimus Grög. led to isolation and identification of fractions of free fatty acids as one active principle of elastase inhibition as well as collagenase inhibition (Clostridium histolyticum collagenase, EC 3.4.24.3). By testing 17 free fatty acids for elastase inhibition it was shown that the inhibition rate of unsaturated acids was much higher than the rate of the saturated ones: oleic acid (C18:1 cis-9): IC50 5µM; stearic acid (C18:0), linoleic acid (C18:2 cis-9,12): IC50 10µM; linolenic acid (C18:3 cis-9,12,15), gamma-linolenic acid (C18:3 cis-6,9,12): IC50 15µM. The highly active erucic acid with an IC50 value of 450nM is remarkable. As a result for collagenase we can assume that the saturated fatty acids were more potent than the unsaturated ones. Palmitic acid (C16:0), heptadecanoic acid (C17:0), stearic acid, and nonadecanoic acid (C19:0) were the most potent fatty acids with IC50 values of 20-45µM. 9 selected fatty acids were investigated for their ability to inhibit the activity of MMP-9 (EC 3.4.24.35). Palmitoleic acid (16:1 cis-9), linolenic acid, and gamma-linolenic acid were the most potent fatty acids but their inhibiting concentrations were very high (250µM). Investigation of cytotoxicity of the extracts of H. annosum, L. deterrimus, and free fatty acids as well as investigation of protease activity of ECV-304, MCF-7, and MDA-MB 231 cells followed. Protease activity of cells increased in the following manner: ECV-304 < MCF-7 < MDA-MB 231. The only investigated fatty acid gamma-linolenic acid did not influence protease activity reproducibly.

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Resumo: O mal-do-fio (ou queima-do-fio) é uma doença causada pelo fungo Basidiomiceto Ceratobasidium sp., que afeta diversas plantas frutíferas nativas ou cultivadas. A doença ocorre com mais freqüência em zonas de alta precipitação e temperaturas elevadas, típicas de regiões de florestas tropicais como a Amazônia e a Mata Atlântica. Em São Paulo, recentemente detectou-se a ocorrência do mal-do-fio, em caquizeiro na região de Mogi das Cruzes. Essa doença pode- se tornar importante com a expansão do cultivo de fruteiras no Estado. A maioria das pesquisas sobre o patossistema focalizou a epidemiologia e o controle do fungo. Entretanto a etiologia do patógeno ainda não está totalmente definida, especialmente para populações do fungo infectando caqui e chá no Estado de São Paulo. Há também pouca informação sobre a divergência genética entre populações do patógeno de hospedeiros distintos como caqui e chá. O primeiro objetivo deste trabalho foi determinar o posicionamento filogenético global de populações de Ceratobasidium sp. do caqui e do chá, em relação a espécies de Ceratobasidium sp. descritas no mundo. Foram analisadas seqüências de DNA da região ITS-5.8S do rDNA, inferindo-se a história dos alelos ou haplótipos deste lócus, por meio de métodos filogenéticos, cladísticos e coalescentes. Observou-se que uso de C. noxium é apropriado para denominar espécies associadas ao mal-do- fio em caqui e chá, apesar de C. noxium do caqui e do chá constituírem populações filogeneticamente independentes, as quais denominamos de Grupo Diospyrus e Grupo Camellia. Este estudo trouxe uma contribuição importante para a compreensão das relações filogenéticas e biologia de populações de C. noxium em caqui e chá. Uma vez esclarecidas as questões filogenéticas, o segundo objetivo deste trabalho foi testar a patogenicidade cruzada de... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The white-thread blight is a disease caused by the Basidiomycete fungus Ceratobasidium sp. that affects several native or cropped tree fruits. This disease frequently occurs in zones of high precipitation and high temperatures typical of the tropical forest regions such as the Amazon and the Atlantic Forest. In São Paulo, the occurrence of the white-thread blight was detected only recently on kaki orchards closer to Mogi das Cruzes. That disease can become important with the expansion of the fruit trees cropping areas in the State. Most of the researches about the pathosystem has focused on the epidemiology and control of fungus. However the etiology of the pathogen is not totally defined yet, especially for the fungus populations infecting kaki and tea in São Paulo State. There is also little information available about the biological and genetic divergence between pathogen populations from distinct hosts, such as kaki and tea. The first objective of this research was to determine the global phylogenetic placement of populations of Ceratobasidium from kaki and tea, considering the species of Ceratobasidium described throughout the world. Sequences of the ITS-5.8S region of the rDNA were analyzed, inferring the alleles or haplotypes history for this locus, by phylogenetics, cladistisc and coalescent methods. We observed that the use of C. noxium is appropriate to denominate the fungus species associate with the white-thread blight on kaki and tea, despite the fact that C. noxium from kaki and tea constitutes phylogenetically independent populations, which we denominate Diospyrus and Camellia groups. This study brought an important contribution for the understanding of the phylogenetics and population biology of C. noxium infecting kaki and tea. Once the phylogenetics subjects have been cleared, the second objective of this work was to test the cross-pathogenicity... (Complete abstract, access electronic address below) / Orientador: Paulo Cezar Ceresini / Coorientador: Alcebíades Ribeiro Campos / Banca: Adriana Zanin Kronka / Banca: Cézar Junior Bueno / Mestre

One of the visual modification of wood is the formation of dark zone lines (ZLs) via interaction of fungi. The result is called spalted wood, which has hitherto been produced mainly in small batches. The main goal of the present study is to further develop techniques for rapid formation of ZLs in hardwoods. Various white rot and brown rot fungi were tested to this purpose. Initially, interactions of 148 combinations of 17 basidiomycetes in malt extract agar were evaluated and their antagonistic interactions were characterised in order to identify fungal pairs capable of rapidly forming high-quality ZLs. Six types of interactions were observed, among others; antibiosis and inhibition in contact, which differ in terms of variables including mycelial overgrowth and zone line formation. Furthermore, 23 pairs of ZL forming fungi on malt extract agar were identified. Then the interactions of five selected pairs of fungi grown on the hardwood species Acer pseudoplatanus L., Betula pendula Roth. and Populus nigra L. were examined to assess their utility for controlled mycological wood modification, also in terms of a possible substrate dependency of their interactions. The results indicate that Lentinus tigrinus fungus is one of the best and quickest producer of ZLs in mycological wood modification.

Of concern in this research were the ecological parameters associated with the establishment of a model invasive plant species, Imperata cylindrica, across a scale of ecological organization. Specifically, the study addressed the species’ ability to: differentially respond to abiotic and biotic constraints during seedling establishment, exhibit a novel underground competitive interference mechanism, and alter the decomposition dynamics in newly invaded ecosystems. Finally, the last portion of the research was centered around creating a predictive habitat model that will provide information on the most important variables responsible for creating habitat for this species. The population level seedling study indicated that soil characteristics and light availability play a significant role in seedling establishment. There were large trends in biomass allocation attributable to soil type with seedlings performing best in high nutrient soils representative of the Mississippi Alluvial Valley physiographic region. I. cylindrica seedlings also showed a positive response to increased seedling density during the initial stages of seedling establishment. The community level research examining a hypothesized novel interference mechanism deployed by I. cylindrica showed a significant and robust pattern of I. cylindrica damaging its own belowground tissue more often than that of its surrounding neighbors. Therefore, it is highly unlikely that I. cylindrica gains a competitive advantage by exposing the native plant assemblage to pathogen invasion (via ruptured tissue) as the plant would expose itself to these pathogens (to which it is evolutionarily naive) at much higher volumes. The ecosystem level examination of this globally important invasive species indicated that I. cylindrica invasion into native systems will significantly accelerate ambient rates of decomposition. Furthermore, fungal community composition in invaded areas was drastically altered as well as bacterial community functional activity in relation to several key enzymes responsible for the decomposition of plant tissue which were produced more abundantly in invaded areas.The landscape-scale analyses and modeling work validated decades of anecdotal evidence and indicated that anthropogenic disturbance factors associated with road maintenance and construction (soil disturbance and vegetation removal) are the principal factors responsible for creating habitat suitable for invasion by this species.

This thesis is focused on environmental preferences of wood-decaying fungi and their relationships with forest structure and development. Relationships of fungi to properties of wood and forest stands were studied on the basis of field observations in Central-European mountain spruce forests. Plot-based approach was used to reveal a general pattern in the diversity of fungi within a single forest stand and between different stands. The analysis of stand structure provided a background for plot-based approach. Substrate-based approach was used to study single species preferences and their communities. In addition, the influence of wood properties (including fungi and their rots) on the regeneration of spruce on logs was studied.