Agaricus bisporus, le champignon de Paris, est un basidiomycète saprophytenaturellement présent dans la litière de cyprès (Cupressus macrocarpa). Il possède différentsmodes de reproduction. Pour étudier leur rôle dans la dynamique spatio-temporelle et l’évolutionde la diversité génétique au cours du temps, des dispositifs expérimentaux ont été mis en place.Dix souches sauvages d’Agaricus bisporus var. bisporus ont été sélectionnées, à partir de deuxpopulations françaises, sur leurs traits phénotypiques et génotypiques. L’étude de l’évolutionmoléculaire de leurs génomes a montré que, pour le génome mitochondrial, la mobilité desintrons de groupe I apparait comme la principale source de polymorphisme. Des taux desubstitution nucléotidique (nt) faibles ont été observés chez tous les types de séquencesmitochondriales (éléments mobiles, séquences géniques et inter géniques). Cette forteconservation, comparée avec les taux élevés de substitution nt des séquences nucléairessimilaires, contraste avec ce qui est généralement décrit dans l’évolution des séquencesfongiques. Des expériences de croisements entre sporocarpes et mycelia de souches sauvages ontété menées sur du compost, dans une chambre de culture, pour simuler l’implantation d’unepopulation naturelle. Pour les sporocarpes récoltés, les données montrent l’existence d’unphénomène parasexuel de Buller conduisant à des souches hybrides d’A. bisporus dans lachambre de culture et potentiellement dans la nature. Parallèlement, les mycelia de souchessauvages ont été introduits dans deux parcelles expérimentales de cyprès. L’analyse génotypiquedes sporocarpes récoltés la première année d’introduction n’a pas permis de mettre en évidencede souche hybride et les conditions climatiques de la seconde année n’ont pas permis d’obtenir defructification. Les dispositifs et outils mis au point doivent permettre un suivi génétique spatiotemporelde la population sur le long terme. / Agaricus bisporus, the button mushroom, is a saprophytic basidiomycete naturallyfound in cypress litter (Cupressus macrocarpa). It possesses different modes of reproduction. Tostudy their role in the spatio-temporal dynamics and in the evolution of the genetic diversity,experimental systems have been set up. Ten wild strains of Agaricus bisporus var. bisporus havebeen selected, from two french populations, on both phenotypic and genotypic traits. Themolecular evolution of their genomes has shown that, for the mitochondrial genome, group Iintron mobility was the main source of gene polymorphism. Low nucleotide (nt) substitutionrates were found in all types of mitochondrial sequences (mobile elements, genic and intergenicones). This stringent conservation of mitochondrial sequences, when compared with the high ntsubstitution rates of their nuclear counterparts, contrasts to what is widely accepted in fungalsequence evolution. Mating experiments between sporocarps and mycelia of wild strains wereconducted on compost in a room culture, to simulate the implantation of a natural population.Among the collected sporocarps, results indicate the occurrence of a parasexual Bullerphenomenon leading to hybrid strains of A. bisporus in room culture and putatively in the wild.In parallel, mycelia of the wild strains have been introduced in two experimental plots of cypress.Genotypic analysis of the sporocarps collected from these plots in the first year of introduction,failed to evidence a hybrid strain. The climatic conditions of the second year did not allowobtaining fruiting-bodies. The developed experimental systems and tools must allow a followingat the genetic level of the spatio-temporal evolution of the population.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The genus Scleroderma Pers.: Fr. belongs to Order Boletales, Phylum Basidiomycota, and involves a group of fungi whose spore production occurs in an enclosed hymenium with passive basidiospores release. The main morphological characteristic of this genus are the size and ornamentation of the spores and mycorrhizal association with various tree species. The aim of this study is to identify Scleroderma species of Pampa biome region by using techniques of morphological analysis and molecular biology following specific methodologies. For an accurate analysis of the basidiospore ornamentation, scanning electron microscopy (SEM) was used. The specimens collected were deposited in the Herbarium SMDB of the Department of Biology, Federal University of Santa Maria - UFSM. For the analysis of phylogenetic relationships of selected species of Scleroderma was sequenced the rDNA (ITS1-5.8S-ITS2) region of some specimens collected or obtained from other herbaria for comparison with sequences deposited in GenBank. Two species were identified: S. albidum; S. citrinum and a specimen to genus level (Scleroderma sp.). We have generate 20 new ITS sequences, clustering most of them, in S. albidum clade, contrasting, however, with basidiome morphological variation initially observed; they represent the first records of this species for the ITS region in GenBank database. Sequences in clade S. albidum showed high similarity with some other nominated as S. bovista, S. aurantium and Sceroderma sp. from GenBank so, the latter sequences are considered as conspecific in the clade. S. citrinum clustered together with homonymous sequences from China, United States and Germany being conspedifics, and its basidiomes were the most identifable in study area by their morphological characteristics. The specimen ICN:154625, from Rio Grande do Sul, State nominated as S. verrucosum, showed high similarity with sequences of S. areolatum and were identified under the latter name. The phylogenetic analysis confirms the literature trend on formation of two major infrageneric clades combining, respectively, echinulate spores plus simple-septate hyphae and reticulate spores plus fibulae. Scleroderma sp. appeared phylogenetically isolated from these two main clusters suggesting a probable formation of a new clade combining echinulate spores and fibulae. / O gênero Scleroderma Pers.: Fr. pertence à Ordem Boletales, Filo Basidiomycota, e envolve um grupo de fungos cuja produção dos esporos ocorre em um himênio fechado, com liberação passiva dos basidiósporos. As principais características morfológicas deste gênero são o tamanho e ornamentação dos esporos e a associação micorrízica com várias espécies arbóreas. Este trabalho tem por objetivo buscar a identificação das espécies do gênero Scleroderma de ocorrência no bioma Pampa, utilizando, para esta finalidade, técnicas morfológicas e de moleculares, segundo metodologias específicas para o estudo do grupo. Para a análise inequívoca da ornamentação dos basidiósporos, foi utilizada a Microscopia Eletrônica de Varredura MEV. Os espécimes coletados foram depositados no Herbário SMDB do Departamento de Biologia da Universidade Federal de Santa Maria - UFSM. Para a análise das relações filogenéticas das espécies selecionadas de Scleroderma, foram sequenciadas as regiões ITS1-5.8S-ITS2 do rDNA de alguns espécimes coletados ou obtidos de outros herbários para comparação com sequências depositadas no GenBank. Duas espécies foram identificadas: S. albidum; S. citrinum e um espécime a nível somente de gênero (Scleroderma sp.). Foram obtidas 20 novas sequencias de ITS, a maioria delas agrupando-se no clado para S. albidum contrapondo a variação morfológica inicialmente verificada nos basidiomas e representando os primeiros registros desta espécie para a região ITS no banco de dados do GenBank; as sequências deste clado também apresentaram alta similaridade com outras do GenBank nominadas como S. bovista, S. aurantium e Scleroderma sp. (da Estônia e Montenegro), permitindo, assim, a retificação nas suas identidades para S. albidum, S. citrinum alinhou com sequências homônimas da China, Estados Unidos e Alemanha mostrando-se conspecíficas, e seus basidiomas podem ser considerados os de mais fácil identificação da área de estudo pelas suas características morfológicas. O espécime ICN:154625, nominado como S. verrucosum, anteriormente citado para o Rio Grande do Sul, mostrou alta similaridade com sequências de S. areolatum do GenBank, formando um clado S. areolatum. A análise filogenética confirma a tendência na literatura para a formação de dois grandes clados infragenéricos combinando, respectivamente, esporos reticulados e hifas fíbuladas ou esporos equinulados e septos simples. Scleroderma sp. isolou-se filogeneticamente dos dois grupos anteriores sugerindo a provável formação de um novo clado combinando com esporos equinulados e hifas fíbuladas.

Made available in DSpace on 2017-07-10T17:40:56Z (GMT). No. of bitstreams: 1 Lindomar_Assi.pdf: 1380412 bytes, checksum: 04c33cb8b847cbf218a9a050fc0ffc83 (MD5) Previous issue date: 2012-06-25 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Due the environmental pollution and damages in non-target organisms, by the indiscriminate use of pesticides in agriculture, it is necessary the search for new plant diseases control technology. The objective of this study was to evaluate the in vitro antimicrobial activity against Alternaria solani and Xanthomonas vesicatoria and the control of early blight and bacterial spot on tomato plants, using stable formulation, as soluble powder, from Rosmarinus officinalis, Curcuma longa and Pycnoporus sanguineus fruiting body. To determine the shelf life, the soluble powder was kept at 40 oC during 0, 60, 120 and 180 days and assayed with five concentrations (0, 50, 100, 150 and 200 mg L-1). When the shelf life and the best concentrations were defined, it was evaluated the timeinterval of spraying of seven, 14 and 21 days. As control treatments were used the fungicides azoxystrobin (200 mg of active ingredient a.i L-1) to early blight and oxychloride (400 mg a.i. L-1) to bacterial spot. There were no in vitro antifungal and antibacterial activities from all extracts. The calculated concentration of P. sanguineus extract for the best control of diseases was 130 mg L-1, with 24 months of shelf live to soluble powder. In vivo, the three extracts, spraying weekly gave the same control of fungicides to early blight and bacterial spot, with values of disease reduction up to 75% for rosemary and turmeric and 79% for P. sanguineus. These results show that the aqueous extract obtained from R. officinalis, C. longa and P. sanguineus soluble powder formulation, can protect tomato plants against early blight and bacterial spot, probably by resistance induction / Face aos problemas de contaminação ambiental e danos em organismos não alvos resultantes do uso indiscriminado de agrotóxicos, têm-se buscado novas medidas de proteção das plantas contra as doenças. O objetivo deste trabalho foi verificar a atividade antimicrobiana in vitro contra Alternaria solani e Xanthomonas vesicatoria e o controle da pinta preta e da mancha bacteriana no tomateiro, utilizando para tanto, produto formulado estável, na forma de pó solúvel, de folhas de alecrim (Rosmarinus officinalis), rizomas de cúrcuma (Curcuma longa) e basidiocarpo de Pycnoporus sanguineus. Para o teste de estabilidade, o pó solúvel foi armazenado a 40 oC por 0, 60, 120 e 180 dias e testado em cinco concentrações (0, 50, 100, 150 e 200 mg L-1). Definidos o tempo de prateleira e a melhor concentração, avaliaram-se os intervalos de aplicação de sete, 14 e 21 dias. Como tratamentos controle foram utilizados os fungicidas azoxystrobin (200 mg do ingrediente ativo i.a. L-1) para a pinta preta e oxicloreto de cobre (400 mg i.a. L-1) para a mancha bacteriana. Não houve in vitro atividade antifúngica a antibacteriana de todos os extratos. A concentração calculada de extrato de P. sanguineus que resultou em menor severidade das doenças foi em média ao redor de 130 mg L-1, com tempo de prateleira do pó solúvel de 24 meses. In vivo, os extratos proporcionaram o mesmo nível de controle da pinta preta e da mancha bacteriana que os fungicidas utilizados, com eficiência de até 75% para alecrim e cúrcuma e 79%, para P. sanguineus, todos em intervalo de aplicação de sete dias. Estes resultados demonstram a eficiência do controle da pinta preta e da mancha bacteriana do tomateiro, provavelmente por indução de resistência, pelo extrato aquoso obtido de formulação pó solúvel de R. officinalis, C. longa e P. sanguineus.

As mudanças na produção de mudas cítricas, com o uso de compostos a base de casca de pínus, vem levando a uma interação antagônica entre os citros e algumas espécies de cogumelos, ditos saprófitas. Este trabalho caracterizou esta nova interação, identificando e isolando os basidiomicetos associados ao problema e outros macrofungos presentes neste sistema de produção. O principal fungo encontrado foi Leucocoprinus birnbaumii que forma basidiomas de cor creme a amarelo brilhante, ocorrendo dois tipos morfológicos (LB e NTA). O micélio promove a impermeabilização no substrato dependendo das condições hídricas, o qual explica a variação nas perdas. Os esporos são produzidos após a abertura do píleo, sendo que a recomendação da coleta dos cogumelos fechados está sendo uma prática adotada nos viveiros. In vitro, as colônias são brancas para creme, tendo uma taxa de crescimento em diâmetro de 0,41 a 0,72 cm/dia em meio cenoura-ágar (CA), variando em função do isolado e da sua procedência. O pH não teve grande influência, mesmo variando de 4,5 a 8,0. O crescimento das colônias foi decrescendo nos meios de cultura CA, aveia-ágar, V8, BDA, malte-ágar e completo de Pontecorvo; enquanto que nos de composição definida, o meio Czapek (0,56 cm/dia) foi superior ao meio mínimo (0,48 cm/dia). Entre as fontes de carbono, o amido foi o melhor, seguido da galactose, maltose, glicerol, glicose, ágar, sacarose, ácido succínico e lactose, não houve crescimento em acetato de amônio nem em oxalato de sódio. Entre as fontes de nitrogênio, os compostos nítricos foram semelhantes aos aminoácidos (glutamato, glutamina e glicina) e ligeiramente preferenciais aos amoniacais, enquanto a uréia inibiu o crescimento e o nitrito de sódio teve efeito fungicida. Nos experimentos realizados em viveiro comercial, o de controle químico teve aumento da produção de cogumelos para a maioria dos fungicidas avaliados, com exceção da iprodiona; o qual é explicado pelo efeito sobre os fungos agentes do controle biológico, como foram associados aos basidiomas Aspergillus niger, Aureobasidium pullulans e Penicillium olsoni. No experimento de controle biológico, as espécies de Trichoderma foram muito variáveis na sua eficiência, alguns colonizando até o substrato, mas não os cogumelos. O experimento com produtos alternativos teve melhores resultados de controle principalmente com o nitrito de sódio, o qual vem sendo adotado no controle deste basidiomiceto via aplicação no substrato e o vinagre pelo uso superficial. Outros macrofungos foram observados como o ninhode- passarinho (Cyathus stercorius); o cogumelo branco (Leucocoprinus brebissonii); a falsa trufa (Agaricaceae incertae sedis) e o cogumelo vermelho (Gymnopilus dilepis). A presença do cogumelo branco também causou preocupação nos viveiristas, mas não apresentou efeito nas plantas, nem impermeabilização. Entretanto, o cogumelo vermelho vem causando problemas na produção com fibra de coco por dimuir a capacidade de retenção de água. A falsa trufa também provoca prejuízos diretos diminuindo o desenvolvimento das mudas cítricas, tendo menor número de radicelas, provavelmente associado à maior temperatura na decomposição do substrato. Assim, há uma necessidade de um contínuo levantamento dos novos macrofungos que possam se adaptar às condições de substratos e de manejo nos viveiros de mudas cítricas. / Changes in the production of citrus nursery, using compounds from pine bark, have led to an antagonistic interaction between the citrus and some species of mushrooms, so-called saprophytes. This study characterized this new interaction, by identifying and isolating basidiomycetes associated to the problem and other macrofungi present in this production system. The main basidiomycete found was Leucocoprinus birnbaumii which has cream-colored to bright yellow, occurring two morphological types (LB and NTA). The mycelium promotes the sealing of the substrate depending on water conditions, for which there is much variation in losses. The spores are produced after the opening of the pileus, and the collection of closed cap has been adopted as a practice in citrus nurseries. In vitro, the colonies vary from cream to yellow, with a growth rate in diameter from 0.41 to 0.72 cm / day in the carrot-agar (CA) medium, depending on the isolate and its origin. The pH was not of great influence, even varying from 4.5 to 8.0. The colony growths was decreasingly in the culture medium CA, oatmeal agar, V8, PDA, malt agar and complete Pontecorvo, whereas the composition set, the Czapek medium (0.56 cm / day) was higher than the minimal medium (0.48 cm / day). Among the carbon sources, starch was the best, followed by galactose, maltose, glycerol, glucose, agar, sucrose, succinic acid and lactose, no growth in ammonium acetate or sodium oxalate. Among the sources of nitrogen, nitric compounds were similar to aminoacids (glicine, glutamate and glutamine) and were slightly preferred to ammonium, whereas urea inhibited the growth and sodium nitrite had fungicidal effect. In the experiments in citrus nursery, the chemical control assay, most fungicides has led to an increased production of mushrooms, except for iprodione, which is explained by the effect on fungal-biological control agents, which have been associated as Aspergillus niger, Aureobasidium pullulans and Penicillium olsoni. In the biological control experiment, Trichoderma species were highly variable in their efficiency, some of them colonized the substrate, but not the mushrooms. The experiment with unconvencional products has led to better results, especially with sodium nitrite, which has been adopted to control the substrate and vinegar in surface application. Other macrofungi were observed as the nest-to-bird (Cyathus stercorius), the white mushroom (Leucocoprinus brebissonii), the false truffle (Agaricaceae incertae sedis) and the Red Mushroom (Gymnopilus dilepis). The presence of the white mushroom has also caused concern in the nursery, but had no effect on plants, or sealing. However, the red mushroom is causing problems in the production of coconut fiber, leaving the fibers, which have no function in water retention. The false truffle also causes direct losses decreasing the development of the citrus trees, and fewer fibrous roots, probably associated to higher temperature in the decomposition of the substrate. Thus, there is a need for a new continuous survey of macrofungi that can adapt to the conditions of substrate and management in citrus nurseries

As mudanças na produção de mudas cítricas, com o uso de compostos a base de casca de pínus, vem levando a uma interação antagônica entre os citros e algumas espécies de cogumelos, ditos saprófitas. Este trabalho caracterizou esta nova interação, identificando e isolando os basidiomicetos associados ao problema e outros macrofungos presentes neste sistema de produção. O principal fungo encontrado foi Leucocoprinus birnbaumii que forma basidiomas de cor creme a amarelo brilhante, ocorrendo dois tipos morfológicos (LB e NTA). O micélio promove a impermeabilização no substrato dependendo das condições hídricas, o qual explica a variação nas perdas. Os esporos são produzidos após a abertura do píleo, sendo que a recomendação da coleta dos cogumelos fechados está sendo uma prática adotada nos viveiros. In vitro, as colônias são brancas para creme, tendo uma taxa de crescimento em diâmetro de 0,41 a 0,72 cm/dia em meio cenoura-ágar (CA), variando em função do isolado e da sua procedência. O pH não teve grande influência, mesmo variando de 4,5 a 8,0. O crescimento das colônias foi decrescendo nos meios de cultura CA, aveia-ágar, V8, BDA, malte-ágar e completo de Pontecorvo; enquanto que nos de composição definida, o meio Czapek (0,56 cm/dia) foi superior ao meio mínimo (0,48 cm/dia). Entre as fontes de carbono, o amido foi o melhor, seguido da galactose, maltose, glicerol, glicose, ágar, sacarose, ácido succínico e lactose, não houve crescimento em acetato de amônio nem em oxalato de sódio. Entre as fontes de nitrogênio, os compostos nítricos foram semelhantes aos aminoácidos (glutamato, glutamina e glicina) e ligeiramente preferenciais aos amoniacais, enquanto a uréia inibiu o crescimento e o nitrito de sódio teve efeito fungicida. Nos experimentos realizados em viveiro comercial, o de controle químico teve aumento da produção de cogumelos para a maioria dos fungicidas avaliados, com exceção da iprodiona; o qual é explicado pelo efeito sobre os fungos agentes do controle biológico, como foram associados aos basidiomas Aspergillus niger, Aureobasidium pullulans e Penicillium olsoni. No experimento de controle biológico, as espécies de Trichoderma foram muito variáveis na sua eficiência, alguns colonizando até o substrato, mas não os cogumelos. O experimento com produtos alternativos teve melhores resultados de controle principalmente com o nitrito de sódio, o qual vem sendo adotado no controle deste basidiomiceto via aplicação no substrato e o vinagre pelo uso superficial. Outros macrofungos foram observados como o ninhode- passarinho (Cyathus stercorius); o cogumelo branco (Leucocoprinus brebissonii); a falsa trufa (Agaricaceae incertae sedis) e o cogumelo vermelho (Gymnopilus dilepis). A presença do cogumelo branco também causou preocupação nos viveiristas, mas não apresentou efeito nas plantas, nem impermeabilização. Entretanto, o cogumelo vermelho vem causando problemas na produção com fibra de coco por dimuir a capacidade de retenção de água. A falsa trufa também provoca prejuízos diretos diminuindo o desenvolvimento das mudas cítricas, tendo menor número de radicelas, provavelmente associado à maior temperatura na decomposição do substrato. Assim, há uma necessidade de um contínuo levantamento dos novos macrofungos que possam se adaptar às condições de substratos e de manejo nos viveiros de mudas cítricas. / Changes in the production of citrus nursery, using compounds from pine bark, have led to an antagonistic interaction between the citrus and some species of mushrooms, so-called saprophytes. This study characterized this new interaction, by identifying and isolating basidiomycetes associated to the problem and other macrofungi present in this production system. The main basidiomycete found was Leucocoprinus birnbaumii which has cream-colored to bright yellow, occurring two morphological types (LB and NTA). The mycelium promotes the sealing of the substrate depending on water conditions, for which there is much variation in losses. The spores are produced after the opening of the pileus, and the collection of closed cap has been adopted as a practice in citrus nurseries. In vitro, the colonies vary from cream to yellow, with a growth rate in diameter from 0.41 to 0.72 cm / day in the carrot-agar (CA) medium, depending on the isolate and its origin. The pH was not of great influence, even varying from 4.5 to 8.0. The colony growths was decreasingly in the culture medium CA, oatmeal agar, V8, PDA, malt agar and complete Pontecorvo, whereas the composition set, the Czapek medium (0.56 cm / day) was higher than the minimal medium (0.48 cm / day). Among the carbon sources, starch was the best, followed by galactose, maltose, glycerol, glucose, agar, sucrose, succinic acid and lactose, no growth in ammonium acetate or sodium oxalate. Among the sources of nitrogen, nitric compounds were similar to aminoacids (glicine, glutamate and glutamine) and were slightly preferred to ammonium, whereas urea inhibited the growth and sodium nitrite had fungicidal effect. In the experiments in citrus nursery, the chemical control assay, most fungicides has led to an increased production of mushrooms, except for iprodione, which is explained by the effect on fungal-biological control agents, which have been associated as Aspergillus niger, Aureobasidium pullulans and Penicillium olsoni. In the biological control experiment, Trichoderma species were highly variable in their efficiency, some of them colonized the substrate, but not the mushrooms. The experiment with unconvencional products has led to better results, especially with sodium nitrite, which has been adopted to control the substrate and vinegar in surface application. Other macrofungi were observed as the nest-to-bird (Cyathus stercorius), the white mushroom (Leucocoprinus brebissonii), the false truffle (Agaricaceae incertae sedis) and the Red Mushroom (Gymnopilus dilepis). The presence of the white mushroom has also caused concern in the nursery, but had no effect on plants, or sealing. However, the red mushroom is causing problems in the production of coconut fiber, leaving the fibers, which have no function in water retention. The false truffle also causes direct losses decreasing the development of the citrus trees, and fewer fibrous roots, probably associated to higher temperature in the decomposition of the substrate. Thus, there is a need for a new continuous survey of macrofungi that can adapt to the conditions of substrate and management in citrus nurseries

Se presenta el estudio taxonómico y ecológico, con respecto a gradientes altitudinales, de los políporos en el Corredor Biológico Marcapata-Camanti (CBM-C), ubicado en la provincia de Quispicanchi, al sureste del departamento del Cusco. Se realizaron colectas intensivas de los basidiomas de los hongos poliporoides en los siguientes gradientes altitudinales: 500-1000 msnm, 1000-1500 msnm, 2000-2500 msnm y 2500-3000 msnm. Para el estudio taxonómico se elaboraron descripciones morfológicas, con información del hábito, distribución y comentarios, de las especies identificadas de Hymenochaetales y Polyporales. Se reportan 45 especies de 102 especímenes colectados. Para el primer orden, se identificaron nueve especies, todas pertenecientes a la familia Hymenochaetaceae. Para el segundo orden, se identificaron 36 especies pertenecientes a las siguientes familias: Fomitopsidaceae (dos especies), Ganodermataceae (siete especies), Meripilaceae (seis especies), Meruliaceae (Lamelloporus americanus), Phanerochaetaceae (dos especies) y Polyporaceae (18 especies). En el estudio ecológico, Hymenochaetaceae, Ganodermataceae, Phanerochataceae y Polyporaceae presentaron ocurrencia en todos los gradientes altitudinales, desde 500-3000 msnm, como en los ecosistemas en el área de estudio, bosque amazónico, pre-montano y montano o nublado, dentro del CBM-C. De la misma manera ocurrió para Fuscoporia gilva, Ganoderma australe y Polyporus leprieurii. En general la micodiversidad entre las gradientes comparadas resultó ser variable, tanto para Hymenochaetales como Polyporales con relación negativa a la altitud. Entre 500-1000 msnm y 1000-1500 msnm el bosque amazónico presentó mayor ocurrencia de especies que el bosque pre-montano. Para el bosque nublado, entre 2000-2500 msnm y 2500-3000 msnm, la micodiversidad fue mayor en la parte baja que la parte alta. Palabras claves: Bosques Andes-Amazónicos, Hymenochaetales, Polyporales, Taxonomía, Ecología. / --- Taxonomic and ecological studies of Aphyllophorales through the altitudinal gradients Corridor Biological Marcapata-Camanti (southeast of Cusco) were studied. The basidiomata of polypore among the altitudinal gradients: 500-1000 msnm, 1000-1500 msnm, 2000-2500 msnm and 2500-3000 msnm was collected intensively. For taxonomic study, keys and descriptions were developed, with information about habit, distribution and comments of Hymenochaetales and Polyporales. Forty six species from 102 specimens collected were indentified. For the first orden, 10 species belonging to Hymenocahtaceae. For the second order, 36 species were identified belonging to the families: Fomitopsidaceae (two species), Ganodermataceae (seven species), Meripilaceae (six species), Meruliaceae (Lamelloporus americanus), Phanerochaetaceae (two species) and Polyporaceae (18 species). In the ecological study, Hymenochaetaceae, Ganodermataceae, Phanerochataceae and Polyporaceae occurred in all altitudinal gradients (500-3000 msnm), as well as in the ecosystems: the Amazon, Pre-Montano and Montano or Cloudy Forest in the CBM-C. Fuscoporia gilva, Ganoderma australe and Polyporus leprieurii as well. The mycodiversity among the gradients tourned out to be variable, both Hymenochaetales and Polyporales. The Amazon Forest (500-1000 msnm) had a higher occurrence of species than Pre-Montane Forest (1000-1500 msnm). For the Montane Forest, 2000-2500 msnm presented higher micodiversity then 2500-3000 msnm. Key words: Andean Amazonian Forest, Hymenochaetales, Polyporales, Taxonomy, Ecology.

O fungo fitopatogênico hemibiotrófico Moniliophthora perniciosa, causador da doença Vassoura-de-Bruxa no cacaueiro (Theobroma cacao), é um dos maiores responsáveis pela contribuição do declínio da produção brasileira de cacau e constitui um problema fitopatológico em todas as regiões cacauicultoras. O gênero Solanum, do qual faz parte o tomate, berinjela, batata entre outros, é bem conhecido na literatura por conterem compostos que exibem atividade biológica tais como antifúngica, antiviral, moluscida, anticancerígena entre outras. O presente trabalho explorou o potencial de extrato bruto de folhas de Solanum lycopersicum como agente inibidor no controle de crescimento e desenvolvimento de três biotipos de M. perniciosa (cacau, solanácea e liana). Através de ensaios biológicos e métodos cromatográficos foi identificado o metabólito secundário -tomatina como responsável pela atividade antifúngica. Este glicoalcalóide, que apresentou potente atividade antifúngica em ensaios in vitro, foi utilizado como agente de controle no desenvolvimento do fungo e manifestação dos sintomas em plantas in vivo, onde foi verificado uma redução significativa no número de plantas com sintomas. / The hemibiotrophic phytopathogenic fungi Moniliophthora perniciosa, agent of witches broom disease of cocoa (Theobroma cacao) is one of the major responsible for the Brazilian cocoa production decay and it is a phytopathological problem in every cocoa yield area. Solanum genera, from each belongs tomatoes, eggplants, potatoes, among others, is well known as producer of biological activity such as antifungal, antiviral, anti mollus, anticancer, etc. This study explored the potential of crude extract leaves from Solanum lycopersicum as an inhibitor agent for the control of growing and development of three M. perniciosa biotypes. By the use of biological assays and chromatography, -tomatine, a secondary metabolite was identified as responsible for antifungal activity. This glycoalcaloid presented great antifungal activity on in vitro assays and it was used on the control and development of witches broom symptoms in vivo, where a significant reduction of symptomatic plants could be observed.

O fungo fitopatogênico hemibiotrófico Moniliophthora perniciosa, causador da doença Vassoura-de-Bruxa no cacaueiro (Theobroma cacao), é um dos maiores responsáveis pela contribuição do declínio da produção brasileira de cacau e constitui um problema fitopatológico em todas as regiões cacauicultoras. O gênero Solanum, do qual faz parte o tomate, berinjela, batata entre outros, é bem conhecido na literatura por conterem compostos que exibem atividade biológica tais como antifúngica, antiviral, moluscida, anticancerígena entre outras. O presente trabalho explorou o potencial de extrato bruto de folhas de Solanum lycopersicum como agente inibidor no controle de crescimento e desenvolvimento de três biotipos de M. perniciosa (cacau, solanácea e liana). Através de ensaios biológicos e métodos cromatográficos foi identificado o metabólito secundário -tomatina como responsável pela atividade antifúngica. Este glicoalcalóide, que apresentou potente atividade antifúngica em ensaios in vitro, foi utilizado como agente de controle no desenvolvimento do fungo e manifestação dos sintomas em plantas in vivo, onde foi verificado uma redução significativa no número de plantas com sintomas. / The hemibiotrophic phytopathogenic fungi Moniliophthora perniciosa, agent of witches broom disease of cocoa (Theobroma cacao) is one of the major responsible for the Brazilian cocoa production decay and it is a phytopathological problem in every cocoa yield area. Solanum genera, from each belongs tomatoes, eggplants, potatoes, among others, is well known as producer of biological activity such as antifungal, antiviral, anti mollus, anticancer, etc. This study explored the potential of crude extract leaves from Solanum lycopersicum as an inhibitor agent for the control of growing and development of three M. perniciosa biotypes. By the use of biological assays and chromatography, -tomatine, a secondary metabolite was identified as responsible for antifungal activity. This glycoalcaloid presented great antifungal activity on in vitro assays and it was used on the control and development of witches broom symptoms in vivo, where a significant reduction of symptomatic plants could be observed.

by Wang He-Xiang. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 161-179). / ACKNOWLEDGEMENTS --- p.i / ABSTRACT --- p.ii / LIST OF CONTENTS --- p.v / LIST OF TABLES --- p.xi / LIST OF FIGURES --- p.xii / LIST OF ABBREVIATIONS --- p.xvi / Chapter CHAPTER 1. --- General Introduction --- p.1 / Chapter CHAPTER 2. --- Literature Review --- p.5 / Chapter 2.1. --- Lectins --- p.5 / Chapter 2.1.1. --- Aspects of lectins --- p.5 / Chapter 2.1.2. --- Isolation and purification of lectins --- p.8 / Chapter 2.1.3. --- Characteristics of lectins --- p.9 / Chapter 2.1.4. --- Effects of lectins on biological activities --- p.10 / Chapter 2.1.4.1. --- The role of lectins in plant defence --- p.11 / Chapter 2.1.4.2. --- The specificity of some legume lectins --- p.13 / Chapter 2.1.4.3. --- Some properties of animal lectins --- p.14 / Chapter 2.1.4.4. --- Hypotensive activity of the lectins --- p.18 / Chapter 2.1.4.5. --- Lectins in immunology --- p.20 / Chapter 2.2. --- Mushroom Lectins and Polysaccharides --- p.24 / Chapter 2.2.1. --- General aspects of mushroom lectins and polysaccharides --- p.24 / Chapter 2.2.2. --- Mushroom lectins --- p.25 / Chapter 2.2.2.1. --- Hericium erinaceum lectin --- p.26 / Chapter 2.2.2.2. --- Lactarius deterrimus lectin --- p.26 / Chapter 2.2.2.3. --- Laetiporus sulfureus lectin --- p.27 / Chapter 2.2.2.4. --- Grifola frondosa lectin --- p.28 / Chapter 2.2.2.5. --- Volvariella volvacea lectin --- p.28 / Chapter 2.2.2.6. --- Flammulina veltipes lectin --- p.29 / Chapter 2.2.2.7. --- Ischnoderma resinosum agglutinin --- p.31 / Chapter 2.2.2.8. --- Lectins from Agaricus spp --- p.31 / Chapter 2.2.3. --- Mushroom polysaccharides --- p.34 / Chapter 2.2.3.1. --- Lentinan --- p.35 / Chapter 2.2.3.2. --- "PSK (trade name, Krestin)" --- p.35 / Chapter 2.2.3.3. --- PSP (Polysaccharopeptide) --- p.37 / Chapter 2.2.3.4. --- PSPC (polysaccharide-peptide complex) --- p.38 / Chapter CHAPTER 3. --- Isolation and Characterization of Two Distinct Lectins from the Cultured Mycelium of the Edible Mushroom Tricholoma mongolicum --- p.44 / Chapter 3.1. --- Introduction --- p.44 / Chapter 3.2. --- Materials and Methods --- p.45 / Chapter 3.2.1. --- Strain and culture condition --- p.45 / Chapter 3.2.2. --- Extraction --- p.46 / Chapter 3.2.3. --- Purification --- p.46 / Chapter 3.2.4. --- Hemagglutination activity --- p.47 / Chapter 3.2.5. --- Test of hemagglutination inhibition by various carbohydrates --- p.47 / Chapter 3.2.6. --- MW estimation by gel filtration and SDS- PAGE --- p.48 / Chapter 3.2.7. --- Glycoprotein staining with PAS reagent --- p.49 / Chapter 3.2.8. --- Carbohydrate content --- p.49 / Chapter 3.2.9. --- Thermal stability --- p.49 / Chapter 3.2.10. --- pH stability --- p.49 / Chapter 3.2.11. --- Effect of cations --- p.50 / Chapter 3.2.12. --- Amino acid analysis --- p.50 / Chapter 3.2.13. --- Antiproliferative activity of lectins --- p.50 / Chapter 3.2.14. --- Statistics --- p.51 / Chapter 3.3. --- Results --- p.51 / Chapter 3.3.1. --- Extraction and purification --- p.51 / Chapter 3.3.2. --- General characteristics of lectins --- p.52 / Chapter 3.3.3. --- Antiproliferative activity of lectins --- p.54 / Chapter 3.4. --- Discussion --- p.55 / Chapter 3.5. --- Summary --- p.58 / Chapter CHAPTER 4. --- The Immunomodulatory and Antitumor Activities of Lectins from the Mushroom Tricholoma mongolicum --- p.79 / Chapter 4.1. --- Introduction --- p.79 / Chapter 4.2. --- Materials and Methods --- p.81 / Chapter 4.2.1. --- Lectins --- p.81 / Chapter 4.2.2. --- Animals --- p.81 / Chapter 4.2.3. --- Assay for antitumor activity --- p.81 / Chapter 4.2.4. --- Assessment of tumor growth and host survival after lectin treatment --- p.82 / Chapter 4.2.5. --- Mitogenic activity of lectins --- p.82 / Chapter 4.2.6. --- Production of nitrite ions in response to lectin treatment --- p.83 / Chapter 4.2.7. --- Preparation of concanavalin A-stimulated lymphokines --- p.84 / Chapter 4.2.8. --- Assay for macrophage activating factor --- p.85 / Chapter 4.2.9. --- Production of tumor necrosis factor (TNF) --- p.86 / Chapter 4.2.10. --- Bioassay for tumor necrosis factor --- p.86 / Chapter 4.2.11. --- Statistics --- p.87 / Chapter 4.3. --- Results --- p.87 / Chapter 4.3.1. --- Antitumor activity --- p.87 / Chapter 4.3.2. --- Assessment of tumor growth and host survival --- p.87 / Chapter 4.3.3. --- Mitogenic activity --- p.88 / Chapter 4.3.4. --- Production of nitrite ions --- p.89 / Chapter 4.3.5. --- Production of macrophage activating factor --- p.89 / Chapter 4.3.6. --- Tumor necrosis factor assay --- p.90 / Chapter 4.4. --- Discussion --- p.90 / Chapter 4.5. --- Summary --- p.94 / Chapter CHAPTER 5. --- Hypotensive and Vasorelaxing Activities of a Lectin (TML-1) from the Edible Mushroom Tricholoma mongolicum --- p.109 / Chapter 5.1. --- Introduction --- p.109 / Chapter 5.2. --- Materials and Methods --- p.111 / Chapter 5.2.1. --- Animals --- p.111 / Chapter 5.2.2. --- In vivo blood pressure measurement in rats --- p.112 / Chapter 5.2.3. --- Study employing blockade of autonomic ganglion transmission --- p.113 / Chapter 5.2.4. --- Study employing alpha-adrenergic blockade --- p.113 / Chapter 5.2.5. --- Study employing beta-adrenergic blockade --- p.114 / Chapter 5.2.6. --- Study employing cholinergic blockade --- p.114 / Chapter 5.2.7. --- Study employing histaminergic blockade --- p.114 / Chapter 5.2.8. --- Study employing inhibitor of the renin- angiotensin system --- p.115 / Chapter 5.2.9. --- Preparation of right atrium for in vitro studies --- p.115 / Chapter 5.2.10. --- Preparation of aorta for in vitro studies --- p.116 / Chapter 5.2.11. --- Adenosine receptor binding assays --- p.116 / Chapter 5.2.12. --- Effect of methylene blue on the hypotensive activity of TML-1 --- p.118 / Chapter 5.2.13. --- Statistics --- p.118 / Chapter 5.3. --- Results --- p.118 / Chapter 5.3.1. --- Blood pressure changes in vivo --- p.118 / Chapter 5.3.2. --- Pharmacological studies using receptor antagonists --- p.119 / Chapter 5.3.3. --- Adenosine receptor binding assay --- p.119 / Chapter 5.3.4. --- Effects on the right atrium in vitro --- p.120 / Chapter 5.3.5. --- Effect of TML-1 on vascular relaxation --- p.120 / Chapter 5.3.6. --- Effect of methylene blue on the hypotensive activity of TML-1 --- p.120 / Chapter 5.4. --- Discussion --- p.120 / Chapter 5.5. --- Summary --- p.123 / Chapter CHAPTER 6. --- A Polysaccharide-Peptide Complex with Immunoenhancing and Antitumor Activities from Cultured Mycelia of the Mushroom Tricholoma mongolicum --- p.134 / Chapter 6.1. --- Introduction --- p.134 / Chapter 6.2. --- Materials and Methods --- p.135 / Chapter 6.2.1. --- Extraction --- p.135 / Chapter 6.2.2. --- Purification --- p.135 / Chapter 6.2.3. --- PSP for purpose of comparison --- p.136 / Chapter 6.2.4. --- Polysaccharide and protein contents --- p.136 / Chapter 6.2.5. --- MW determination of F1 using gel filtration --- p.136 / Chapter 6.2.6. --- Animals --- p.136 / Chapter 6.2.7. --- Antiproliferative activity assay --- p.137 / Chapter 6.2.8. --- Mitogenic activity --- p.137 / Chapter 6.2.9. --- Production of nitrite ions --- p.138 / Chapter 6.2.10. --- Macrophage activating factor assay --- p.138 / Chapter 6.2.11. --- Antitumor activity assay --- p.139 / Chapter 6.2.12. --- Statistics --- p.139 / Chapter 6.3. --- Results --- p.140 / Chapter 6.3.1. --- Purification of polysaccharide-peptide complex --- p.140 / Chapter 6.3.2. --- Antiproliferative activity --- p.140 / Chapter 6.3.3. --- Mitogenic activity in vitro --- p.140 / Chapter 6.3.4. --- Molecular weight of Fl --- p.141 / Chapter 6.3.5. --- Mitogenic activity in vivo --- p.141 / Chapter 6.3.6. --- Production of nitrite ions --- p.141 / Chapter 6.3.7. --- Production of macrophage activating factor --- p.141 / Chapter 6.3.8. --- Antitumor activity in vivo --- p.142 / Chapter 6.4. --- Discussion --- p.142 / Chapter 6.5. --- Summary --- p.144 / GENERAL DISCUSSION --- p.155 / CONCLUSIONS --- p.158 / REFERENCES

by Liu Fang. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 159-178). / ACKNOWLEGEMENTS --- p.i / ABSTRACT --- p.ii / TABLE OF CONTENTS --- p.v / LIST OF TABLES --- p.ix / LIST OF FIGURES --- p.xi / ABBREVIATIONS --- p.xiv / Chapter Chapter 1. --- General Introduction --- p.1 / Chapter Chapter 2. --- Literature Review --- p.7 / Chapter 2.1. --- Biologically active polysaccharides --- p.7 / Chapter 2.2. --- Antitumor activities of polysaccharides --- p.11 / Chapter 2.2.1. --- In vivo studies --- p.11 / Chapter 2.2.2. --- In vitro studies --- p.15 / Chapter 2.3. --- Antitumor mechanisms of polysaccharides --- p.17 / Chapter 2.4. --- Structure and antitumor activities of polysaccharides --- p.25 / Chapter 2.4.1. --- The effect of molecular mass --- p.26 / Chapter 2.4.2. --- The impact of branching configuration --- p.21 / Chapter 2.4.3. --- The relationship between antitumor activity and conformation --- p.28 / Chapter 2.4.4. --- Improvement of antitumor activity by chemical modification --- p.29 / Chapter 2.5. --- Other biological activities --- p.30 / Chapter 2.5.1. --- Antiviral activity --- p.30 / Chapter 2.5.2. --- Antimicrobial activity --- p.31 / Chapter 2.5.3. --- Free radical scavenging activity --- p.32 / Chapter 2.5.4. --- Hepatic protective effect --- p.32 / Chapter Chapter 3. --- Isolation and Characterization of a Polysaccharide-Protein Complex (PSPC) from Tricholoma lobayense --- p.34 / Chapter 3.1. --- Introduction --- p.34 / Chapter 3.2. --- Materials and methods --- p.36 / Chapter 3.2.1. --- Strain --- p.36 / Chapter 3.2.2. --- Culture conditions --- p.36 / Chapter 3.2.3. --- Extraction of T. lobayense --- p.39 / Chapter 3.2.4. --- Purification of polysaccharide-protein complex --- p.40 / Chapter 3.2.5. --- Molecular mass determination --- p.43 / Chapter 3.2.6. --- High performance liquid chromatography --- p.43 / Chapter 3.2.7. --- SDS-polyacrylamide gel electrophoresis --- p.44 / Chapter 3.2.8. --- Ultraviolet scanning --- p.44 / Chapter 3.2.9. --- Chemical analysis --- p.45 / Chapter 3.2.10. --- Experimental animals --- p.47 / Chapter 3.2.11. --- In vivo antitumor assay --- p.48 / Chapter 3.2.12. --- Safety tests --- p.49 / Chapter 3.2.13. --- Statistical analysis --- p.51 / Chapter 3.3. --- Results --- p.51 / Chapter 3.3.1. --- Extraction and purification --- p.51 / Chapter 3.3.2. --- Biochemical analysis --- p.52 / Chapter 3.3.3. --- Chemical analysis --- p.60 / Chapter 3.3.4. --- In vivo antitumor activity --- p.68 / Chapter 3.3.5. --- Safety evaluation --- p.68 / Chapter 3.4. --- Discussion --- p.75 / Chapter 3.5. --- Summary --- p.84 / Chapter Chapter 4. --- "Immunomodulating, Antitumor and other Biological Activities of Polysaccharide-Protein Complex (PSPC) from Tricholoma lobayense" --- p.85 / Chapter 4.1. --- Introduction --- p.85 / Chapter 4.2. --- Materials and methods --- p.87 / Chapter 4.2.1. --- Experimental animals --- p.87 / Chapter 4.2.2. --- Cultivation of tumor cells --- p.87 / Chapter 4.2.3. --- Preparation of peritoneal exudate cells and splenocytes --- p.87 / Chapter 4.2.4. --- Mitogenic response of T cells --- p.89 / Chapter 4.2.5. --- Responses of peritoneal exudate cells --- p.89 / Chapter 4.2.6. --- In vitro antitumor assay --- p.92 / Chapter 4.2.7. --- Transmission electron microscope --- p.93 / Chapter 4.2.8. --- Evaluation of other biological activities --- p.94 / Chapter 4.2.9. --- Statistical analysis --- p.99 / Chapter 4.3. --- Results --- p.99 / Chapter 4.3.1. --- Immunomodulating activity --- p.99 / Chapter 4.3.2. --- In vitro antitumor action --- p.107 / Chapter 4.3.3. --- Observation on tumor regression induced by PSPC --- p.107 / Chapter 4.3.4. --- Other biological actions --- p.112 / Chapter 4.4. --- Discussion --- p.121 / Chapter 4.4.1. --- Immunomodulating activity --- p.121 / Chapter 4.4.2. --- Antitumor activity --- p.125 / Chapter 4.4.3. --- Other biological activities --- p.127 / Chapter 4.5. --- Summary --- p.130 / Chapter Chapter 5. --- Induction of Gene Expression of Immunomodulating Cytokines by Polysaccharide-Protein Complex (PSPC) from Tricholoma lobayense --- p.132 / Chapter 5.1. --- Introduction --- p.132 / Chapter 5.2. --- Materials and methods --- p.135 / Chapter 5.2.1. --- Experimental animals --- p.135 / Chapter 5.2.2. --- Preparation of peritoneal exudate cells and splenocytes --- p.136 / Chapter 5.2.3. --- RNA extraction --- p.137 / Chapter 5.2.4. --- Reverse transcription- polymerase chain reaction --- p.137 / Chapter 5.2.5. --- Dot blot --- p.138 / Chapter 5.2.6. --- Hybridization --- p.141 / Chapter 5.3. --- Results --- p.142 / Chapter 5.3.1. --- mRNA phenotyping of cytokines and cytokine receptors in normal mice --- p.142 / Chapter 5.3.2. --- mRNA phenotyping of cytokines and cytokine receptors in tumor-bearing mice --- p.142 / Chapter 5.4. --- Discussion --- p.150 / Chapter 5.5. --- Summary --- p.153 / Chapter Chapter 6. --- General Discussion and Conclusion --- p.155 / REFERENCES --- p.159