Tesis sobre el tema "Edible fungus"

Aflatoxins are highly carcinogenic secondary metabolites that can contaminate approximately 25% of crops and that cause or exacerbate multiple adverse health conditions, especially in Sub-Saharan Africa and South and Southeast Asia. Regulation and decontamination of aflatoxins in high exposure areas is lacking. Biological detoxification methods are promising because they are assumed to be cheaper and more environmentally friendly compared to chemical alternatives. White-rot fungi produce non-specific enzymes that are known to degrade aflatoxin in in situ and ex situ experiments. The aims of this study were to (1) decontaminate aflatoxin-B-1-(AFB(1)) in naturally contaminated maize with the edible, white-rot fungus Pleurotus ostreatus (oyster mushroom) using a solid-state fermentation system that followed standard cultivation techniques, and to (2) and to assess the risk of mutagenicity in the resulting breakdown products and mushrooms. Vegetative growth and yield characteristics of P. ostreatus were not inhibited by the presence of-AFB(1).-AFB(1) was degraded by up to 94% by the Blue strain. No aflatoxin could be detected in P. ostreatus mushrooms produced from-AFB(1)-contaminated maize. Moreover, the mutagenicity of breakdown products from the maize substrate, and reversion of breakdown products to the parent compound, were minimal. These results suggest that P. ostreatus significantly degrades-AFB(1) in naturally contaminated maize under standard cultivation techniques to levels that are acceptable for some livestock fodder, and that using P. ostreatus to bioconvert crops into mushrooms can reduce-AFB(1)-related losses.

Establishing a commercial, lignocellulose-based, second-generation ethanol process has received several decades of attention by both researchers and industry. However, a fully economically viable process still remains a long-term goal. The main bottleneck to this achievement is the recalcitrance of lignocellulosic feedstocks, although there are several other factors, such as the huge investment required for second-generation ethanol facilities. An intelligent alternative solution discussed in this thesis is an integrated approach using firstgeneration ethanol plants for second-generation processes. Wheat is the major feedstock for first-generation ethanol in Europe; therefore, wheat-based lignocellulose waste, such as wheat straw, bran, and whole stillage fiber (a waste stream from first-generation wheat-based ethanol plants) was the primary focus of the integration model in this thesis. Since the major share of first-generation ethanol plant economics focuses on the animal feed DDGS (Distillers’ dried gains with solubles), the integration of lignocellulose should be designed in order to maintain DDGS quality. An ethanol-producing edible filamentous fungus, Neurospora intermedia, a potential protein source in DDGS, was considered for use as the fermenting microbe. The morphological and physiological aspects of this fungus were studied in the thesis, leading to the first report of fungal pellet development. An alternative approach of using dilute phosphoric acid to pretreat lignocellulose, as it does not negatively affect fungal growth or DDGS quality, was demonstrated in both the laboratory and on a 1m3 pilot scale. Furthermore, the process of hydrolysis of pretreated lignocelluloses and subsequent N. intermedia fermentation on lignocellulose hydrolysate was also optimized in the laboratory and scaled up to 1 m3 using an in-house pilot-scale airlift bioreactor. Fungal fermentation on acid-pretreated and enzyme-hydrolyzed wheat bran, straw and whole stillage fiber resulted in a final ethanol yield of 95%, 94% and 91% of the theoretical maximum based on the glucan content of the substrate, respectively. Integrating the first- and second-generation processes using thin stillage (a waste stream from first-generation wheat-based ethanol plants) enhanced the fungal growth on straw hydrolysate, avoiding the need for supplementing with extra nutrients. Based on the results obtained from this thesis work, a new model for integrated first- and second-generation ethanol using edible filamentous fungi processes that also adds value to animal feed (DDGS) was developed.

This thesis focuses on the edible fungus Lepista (Pied Bleu or Wood Blewit). Factors affecting its potential commercial cultivation were explored and a contribution to knowledge of the morphology and cultivation of Australian species of Lepista has been made. Australian collections of Lepista were made within a 200 km zone of Sydney. A study of the morphology and taxonomic species of these collections was undertaken. Intra- and inter-fertility crosses were completed with French L. nuda and L. sordida to determine genetic relationships and biological species. Suitable substrates for agar medium, spawn production and cultivation were explored. The response to temperature of French and Australian Lepista in vitro, and Australian Lepista under cultivation, using cold shock, was observed. The effect of modified atmosphere exchanges per hour, CO2 levels, and cold shock during the cultivation cycle and sporophore production were investigated. A genebank of Australian Lepista was established. Three species of Lepista were found in Australia : L. nuda, L. sordida and L. saeva. Two other groups of Lepista were identified. The use of A. bisporus compost appeared to be optimal for experimental and commercial applications. Australian isolates of Lepista tolerate higher temperatures than French isolates, and grew at double the rate of the French at all temperatures except 5 degrees centigrade. The length of the spawn run was reduced from 43-58 days to 12-16 days with introduced CO2 of 9,000-11,000 ppm, but an erratic cyclic pattern of net CO2 production occurred which could only be stabilised by increasing ventilation. This initial cyclic pattern appeared to inhibit subsequent sporophore formation.

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CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
Ganoderma lucidum it is a fungus that arouses considerable interest worldwide because on their numerous properties, it is reported mainly for their medicinal power, and it may be used in the prevention and treatment of various diseases including cancer and AIDS. It is a mushroom that has the ability to thrive in a multitude of lignocellulosic wastes, since they produce lignocellulolytic enzymes, specialized to degrade this type of raw material. The G. lucidum species has several distinct lineages, and also nutritional requirements that vary in relation to the collection site and the type of substrate. Therefore, it is necessary to know the substrates, and the situation of the most suitable growing to the different strains of G. lucidum. The aim of this study was to evaluate the production of two strains of Ganoderma lucidum on agricultural waste as well as to perform chemical analyzes of basidiomata obtained in cultivation. The experiment was conducted at the premises of the module Mushrooms FCA / UNESP-Botucatu, SP, in which two strains were used: GLM-09/01 GLM and 10/02 both grown in waste, oat straw, bean straw, straw brachiaria grass Tifton straw and sawdust in two situations: with (20%) and without supplementation (0%) of wheat bran. All the waste came from dumps of agricultural activity resulting from Botucatu-SP. Both treatments were performed in 10 replications, totaling 200 packages. The cultivation of mushrooms totaled 95 days, later we assessed the biological efficiency of the treatments and it was performed and their chemical analysis of initial and residual substrates and basidiomata obtained. EB values (%) ranging from 0.0% to 6.7%. In the chemical analysis of the mushrooms, in the parameters, total protein, ether extract, crude fiber and ash, the results ranged from 8.7% to 13.7%, from 2.0% to 6.7%, 0.83% 1.79% to 38.8% and 54.5% respectively. Thus, it is concluded that from the substrates analyzed, those with the highest income were the base of straw brachiaria 20% in both strains tested (GLM 10/02 and GLM 09/01 and bean straw to 20% GLM on 10/02 line. The mushrooms showed high levels of ether extract, ash and fiber and low protein content.
Ganoderma lucidum trata-se de um fungo que desperta bastante interesse mundial devido suas inúmeras propriedades medicinais, são relatados principalmente pelo seu poder medicinal, podendo ser utilizados na prevenção e tratamento de diversas doenças incluindo o câncer e AIDS. É um cogumelo que apresenta a capacidade de desenvolver-se em diversos resíduos lignocelulósicos, pelo fato de produzirem enzimas lignocelulolíticas, especializadas em degradar esse tipo de matéria-prima. A espécie G. lucidum, apresenta diversas linhagens distintas, cujas exigências nutricionais que variam em relação ao local de coleta e ao tipo de substrato. Assim, torna-se necessário conhecer os substratos e a situação de cultivo mais adequados as diferentes linhagens de G. lucidum. O objetivo deste estudo foi avaliar a produção de duas linhagens de Ganoderma lucidum em resíduos agrícolas, bem como realizar a caracterização físico-química dos basidiomas obtidos no cultivo. O experimento foi conduzido nas instalações do Módulo de Cogumelos da FCA/UNESP-Botucatu, SP. As linhagens utilizadas foram GLM-09/01 e GLM 10/02. Os resíduos utilizados na cultura foram palha de aveia, palha de feijão, palha de capim braquiária, palha de capim tifton e serragem de eucalipto, todos em duas situações: com (20%) e sem suplementação (0%) de farelo de trigo. Todos os resíduos foram provenientes de despejos da atividade agrícola decorrente do município de Botucatu-SP. Os tratamentos foram realizados em 10 repetições, totalizando 200 pacotes. O cultivo totalizou 95 dias. Avaliou-se a eficiência biológica dos tratamentos e realizou-se a caracterização físico-química dos substratos inicial e residual e também dos basidiomas obtidos. Os valores de EB (%) variaram de 0,0% a 6,7%. Nas analises da composição centesimal dos cogumelos, os parâmetros, proteína bruta, extrato etéreo, cinzas e fibra bruta, os resultados variaram de 8,7% a 13,7%, de 2,0% a 6,7%, de 0,83% a 1,79% e de 38,8% a 54,5% respectivamente. Dos substratos analisados, os que apresentaram maior rendimento foram os formulados à base de palha de braquiária (20%) em ambas as linhagens testadas (GLM-09/01 e GLM 10/02) e palha de feijão (20%) na linhagem GLM 10/02. Os cogumelos apresentaram teores elevados de lipídios, fibras e cinzas e baixo teor de proteínas.

This thesis focuses on the edible fungus Lepista (Pied Bleu or Wood Blewit). Factors affecting its potential commercial cultivation were explored and a contribution to knowledge of the morphology and cultivation of Australian species of Lepista has been made. Australian collections of Lepista were made within a 200 km zone of Sydney. A study of the morphology and taxonomic species of these collections was undertaken. Intra- and inter-fertility crosses were completed with French L. nuda and L. sordida to determine genetic relationships and biological species. Suitable substrates for agar medium, spawn production and cultivation were explored. The response to temperature of French and Australian Lepista in vitro, and Australian Lepista under cultivation, using cold shock, was observed. The effect of modified atmosphere exchanges per hour, CO2 levels, and cold shock during the cultivation cycle and sporophore production were investigated. A genebank of Australian Lepista was established. Three species of Lepista were found in Australia : L. nuda, L. sordida and L. saeva. Two other groups of Lepista were identified. The use of A. bisporus compost appeared to be optimal for experimental and commercial applications. Australian isolates of Lepista tolerate higher temperatures than French isolates, and grew at double the rate of the French at all temperatures except 5 degrees centigrade. The length of the spawn run was reduced from 43-58 days to 12-16 days with introduced CO2 of 9,000-11,000 ppm, but an erratic cyclic pattern of net CO2 production occurred which could only be stabilised by increasing ventilation. This initial cyclic pattern appeared to inhibit subsequent sporophore formation.
Doctor of Philosophy (PhD)

Terfezia pfeilii is an edible mycorrhizal fungus that thrives in the Kalahari Desert of southern Africa. It is best known by desert dwellers for its flavour and as a source of nutrition. Although the genus Terfezia is generally regarded as being an ectomycorrhizal mycobiont, the exact mycorrhizal type formed by T. pfeilli and its' associated host plants remains uncertain. Discovery of the host plants for T. pfeilii would first be required in order to further investigate the life cycle and cultivation of this truffle. This study focussed on the isolation of mycelia from the ascocarp, optimising the growth conditions of the mycelial cultures, rapid molecular identification of T. pfeilii, investigation of potential helper bacteria and mycorrhizal synthesis experiments. T. pfeilii ascocarps were harvested from the Spitskop Nature Reserve in Upington, South Africa. Ascocarps were successfully identified using both morphological and molecular methods. Despite the delayed growth mostly caused by contaminating microorganisms, the isolation of T. pfeilii mycelia culture was successful. Molecular techniques were used to confirm the identity of the pure culture. Further studies were conducted on ways to improve the growth conditions of the mycelial culture on Fontana medium. An optimum temperature of 32°C, the addition of Bovine Serum Albumin as a nitrogen source and a pH of 7.5 significantly improved the growth of T. pfeilii in vitro. A rapid PeR-based molecular method was developed to speed up the identification of T. pfeilii. Specific primers that can exclusively amplify the ITS region of T. pfeilii were designed and used to identify both the ascocarps and the mycelial culture. The specificity of these primers was confirmed by their inability to amplify DNA from the isolates of contamining fungi obtained during the isolation process. Molecular comparison was made to confirm the reclassification of South African samples of T. pfeilii as Kalaharituber pfeilii as proposed by Ferdman et al.,(2005). However, in this study, the name T. pfeilii has been retained. A total of 17 bacterial isolates were obtained from the fruiting bodies of T. pfeaii and these were tested for stimulation of mycelial growth in vitro, indole production and phosphate solubilising capabilities. Bacterial isolates that showed potential to be Mycorrhization Helper Bacteria (MHB) were identified as Paenibacillus sp., Bacillus sp. and Rhizobium tropici. Selected plant seedlings were inoculated with T. pfeilii cultures or ascocarp slurry in order to re-establish the mycorrhizal association. After 8 months, light microscopy observations revealed an endomycorrhizal type association between Cynodon dactylon and T. pfeilii. This was confirmed with molecular analysis using specific T. pfeilii ITS primers. After 15 months, molecular methods confirmed Acacia erioloba as another host plant. These results have provided essential information paving the way for further investigation into the life cycle and biology of the Kalahari truffle.
KMBT_363
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O fungo Pleurotus ostreatus pertence ao grupo de basidiomicetos que degradam madeira. Este cogumelo cultivado em todo mundo apresenta grande rusticidade e produtividade, e pode ainda ser usado em processos de biorremediação e biopolpação. Devido a seu potencial biotecnológico, torna-se interessante a compreensão da interação deste com outros microrganismos. Neste sentido, recentemente foi observada a presença de bactérias associadas a P. ostreatus em culturas in vitro, que apresentavam grande pleomorfismo. A partir desta observação foram elaborados ensaios que visaram a confirmação da presença de bactérias. Para tanto, foi utilizada a estratégia do “Ciclo Completo de Análise do rRNA” (full-cycle rRNA analysis) empregada em microrganismos não cultiváveis ou de crescimento fastidioso, além do emprego de técnicas de microbiologia básica, e de estudos de ultraestrutura. Os estudos de microbiologia básica indicaram que se tratava de um microrganismo fastidioso e que se desenvolvia melhor na presença do fungo em sistema de co-cultivo em meios contendo Tween 80 ou Tween 20. Por sua vez, a análise de ultraestrutura demonstrou a presença de estruturas pleomórficas, tanto internamente como externamente à hifa. Em relação ao “Ciclo completo de Análise do rRNA” este se iniciou pela amplificação e seqüenciamento de parte do rDNA bacteriano, que revelou a proximidade desta bactéria com o Complexo Burkholderia cepacia (CBC). A partir desta seqüência, foi realizado um estudo de bioinformática que indicou sondas específicas para este grupo de bactérias. Completando o Ciclo completo de Análise do rRNA, foram realizados ensaios de hibridização in situ fluorescente (FISH) para a confirmar a relação entre as estruturas bacterianas e a seqüência obtida. Este método comprovou a presença das bactérias no interior das hifas de P. ostreatus. Este trabalho constitui o primeiro relato de bactérias pleomórficas pertencente ao complexo B. cepacia associados a P. ostreatus.
The fungus Pleurotus ostreatus, which belongs to white rot basidiomycete group, is a widely cultivated mushroom; this species has high productivity and rusticity, besides its use in biobleaching and bioremediation processes. This biotechnological potential justifies microbial interaction studies between this fungi and others microorganisms. In P. ostreatus mycelia, it has been observed pleomorphic bacteria growing on agar media. This research describes several assays to confirm bacterial presence in this sample. Therefore, the full-cycle rRNA analysis (described for unculturable or fastidious microorganism), ultrastructure and basic microbiology approaches were employed. Basic microbiology approaches indicated slow growing bacteria, which grown faster near to fungi colonies in solid media amended with Tween 80 or Tween 20 (co-culture system). Ultrastructure studies confirm the presence of intracellular and extracellular pleomorphic bacteria. The full-cycle rRNA analysis started with 16S rDNA amplification and sequencing. This approach demonstrated a relation between these bacteria with Burkholderia cepacia complex. By bioinformatics analysis was determinate which DNA probes can be use to identified this bacterial group. The last step for full-cycle rRNA analysis was applying fluorescent in situ hybridization (FISH). This technique confirmed the relationship between 16S rDNA bacterial sequence and bacterial forms. This is the first time that a pleomorphic bacteria from B. cepacia complex is found associated with P. ostreatus.

Thesis (Ph.D.) -- University of Western Sydney, Hawkesbury, 1998.
Thesis submitted for the degree of Doctor of Philosophy. Photocopies of articles by Karen Stott ... [et al.] in back. Includes bibliographical references (leaves 137-148).

Volatile fatty acids (VFAs) have become attractive and gained high research interest due to its significance for the chemical industry and economical advantage. These acids can be produced by utilizing organic waste such as food waste as substrate through anaerobic digestion. Anaerobic digestion is an environmental process that occurs naturally and produces biogas as the main product. VFAs are intermediate products formed during anaerobic digestion where acetic acid, a type of VFA, is the primary product. The main objective of this study was to utilize acetic acid as carbon and energy source for production of edible fungi, Rhizopus ologisporus, Mucor indicus and Volvariella volvacea. The first step was to evaluate if acetic acid could be used as carbon and energy source for edible fungi production. The results showed, that acetic acid is suitable as carbon and energy source for fungal biomass production. The second step was to optimize growth in liquid media. The cultivations were carried out by using five different conditions, where the liquid media contained different combinations of acetic acid, yeast extract and minerals as well as comparing orbital and linear oscillations. Fungal cultivation was possible regardless of the medium composition and type of water shaking baths. However, a linear water shaking bath with a combination of acetic acid yeast extract and/or minerals seems to be the best. Finally, as step three, acetic acid concentrations, 0.2 g/l and 2.0 g/l were used under similar conditions as in step two to see whether a higher concentration of acetic acid would be beneficial. Although the cultivation containing 2.0 g/l gave a higher value of dry weight, the value of yield is questionable. Further studies are needed to confirm if a higher concentration is beneficial or if it might act as an inhibitor for fungal cultivation.
Flyktiga fettsyror (VFAs) har ekonomiska fördelar och kan användas inom kemiska industrier i olika sammanhang, detta har lett till ett stort forskningsintresse för att kunna nyttja VFAs. Organiskt avfall, såsom matavfall, kan användas som substrat för att producera fettsyror genom anaerob rötning. Anaerob rötning är en miljövänlig process och VFAs bildas som intermediära produkter under den anaeroba nedbrytningen där annars bildas biogas som slutprodukt. Syftet med denna studie var att använda ättiksyra, (den vanligaste typen av VFAs), som kol- och energikälla vid odling av tre olika ätbara svampar, som Rhizopus oligosporus, Mucor indicus, och Volvariella volvacea. Först odlades dessa ätbara svampar i odlingsmedium innehållande ättiksyra. Resultatet visade att ättiksyra kan användas som kol- och energikälla vid produktion av svampbiomassa. Målet i de nästkommande stegen var att optimera tillväxtförhållande för svampodlingen. Fem olika odlingsmedier som innehöll olika kombinationer av ättiksyra, jästextrakt och mineraler användes. Det undersöktes dessutom hur två olika skakmetoder, orbitalt, eller linjärt, skakbad påverkar odlingen. Svamptillväxt var möjligt vid alla olika förhållanden oavsett sammansättningen av medium och typ av skakbad, däremot verkar odlingsmedium som innehåller ättiksyra, jästextrakt och/eller mineraler i kombination med linjär skakning vara de bästa förutsättningar för tillväxt av biomassa. I det sista steget kultiverades svamp med olika koncentrationer av ättiksyra, 0,2 g/l och 2,0 g/l, under liknande optimerade förhållanden som ovan, för att undersöka om en högre koncentration av ättiksyra skulle vara fördelaktig. Det producerades mer svampbiomassa (som torrvikt) vid koncentration av 2,0 g/l ättiksyra jämfört med när 0,2 g/l ättiksyra användes, dock var det svårt att säkerställa utbytet. Det behövs därför ytterligare fortsatta studier för att kunna bevisa om en högre koncentration av ättiksyra är fördelaktig för odlingen, eller om en högre koncentration skulle verka inhiberande för tillväxten.

Establishment of a new industry in plantations of Pinus spp. in south-western Australia, based on edible ectomycorrhizal fungi, will be dependant on the ability of new fungal introductions to survive and proliferate under limitations imposed by biotic and abiotic factors. This thesis investigated factors that would influence establishment of two most valuable fungi, Matsutake (Tricholoma matsutake) and the Pine Mushroom {T. magnivelare). Factors that were investigated included, extant ectomycorrhizal fungi, plant host - fungus compatibility, climate and soils. A survey of ectomycorrhizal (ECM) fimgi associated with introduced Pinus spp. in south-western Australia increased the number of identified species to 15-16 species, from seven previously reported in the literature. Most species form a sub-set of species that have also been introduced to Eastern Australia, and are early colonizers of pines, forming ECM associations with pine seedlings, but are also found with plantationgrown pine trees older than sixty years. Four species of basidiomycetes with resupinate fruitbodies, are new records for Australia, and probably for the Southern Flemisphere. One resupinate fungus (Amphinema byssoides) has been introduced. In contrast, three Tomentella spp., that are part of a suite of at least 25 tomentelloid fungi found in native forests and woodlands in south-western Australia, appear to have made a transition from native to introduced Pinus hosts. Species within other genera of resupinate fungi, that are known to form ECM associations with pines in the Northern Hemisphere, were also identified but not confirmed as ECM fungi of pines in Western Australia. rDNA sequence data strongly suggested that some previous records of ECM fungi have been mis-identified. In two field experiments, Thelephoraceae {Thelephora terrestris and Tomentella stuposa) formed 50-60% of ectomycorrhizas but sporocarps of both Ilmgi were found infrequently or not at all in surveys. The apparent below-ground dominance by representatives of the Thelephoraceae, as ECM fungi of pines in south-western Australia, is similar to some results from natural conifer forests in Europe and North America. Sporocarp production, by naturally occurring pine ECM fungi, was measured over a season in experimental plots of five year-old P. radiata trees, that comprised trees planted with a standard treatment and trees planted for agro-forestry, with additional P fertilization and a cover crop of the legume Trifolium subterraneum. Sporocarp production in agro-forestry plots was estimated to be at least ca. 73 kg'1 ha'1 yr'1 d.w., that was significantly greater than that of the standard treatment, and up to an order of magnitude greater than productivity in natural conifer forests. There were also changes in phenology of sporocarp production by different species of fungi between treatments. Comparison of climate between selected locations within the ranges of T. matsutake and T. magnivelare, and locations within south-western Australia strongly suggests that T. magnivelare, and T. nauseosum, syn. T. matsutake from the Western Mediterranean, are more suited to the climate of south-western Australia than Asian T. matsutake. Soils within south-western Australia that are potentially suitable for Matsutake, that are derived from similar parent materials and with similar physical and chemical properties, occupy an undefined proportion of about 3250 km'2 of soil sub-systems within the area that may be climatically suitable. In vitro synthesis experiments between isolates of Tricholoma matsutake and T.magnivelare and host plants, Pinus dens [flora, P. radiata and P. pinaster, showed that internal structures comparable with natural Pinus densiflora-T. matsutake ECM were formed between P. radiata and T. matsutake. Macro- and micro-morphological features of infected short roots suggested that P. radiata and P. pinaster are suitable hosts for both T. matsutake and T. magnivelare. In contrast to in vitro synthesis experiments, ECM associations failed to form between T. matsutake and mature P. radiata trees after inoculation with two forms of vegetative mycelium, and ECM also failed to form between T. matsutake and Pinus spp. inoculated with two forms of vegetative mycelium under more controlled conditions in a glasshouse experiment.

Mycelial pellet formation by filamentous fungi is one of the most researched topics in fungal biotechnology research. Pellets are generally formed as a result of a complex interaction process through the influence of many cultivation factors such as inoculum size, pH, dissolved oxygen level, agitation system, nucleating agents, additives, trace metals, CO2, temperature, reactor types, carbon substrate, rheology, culture modes, fermenter geometry, nitrogen and phosphate levels etc. Each factor has varying effects on the growth morphologies of different fungal species. Fungal growths in the form of pellets have several advantages and pose a potential solution to overcome the problems associated with the filamentous fungal growth in large scale industrial bioreactors. The aim of the present work was to study pellet formation of edible filamentous fungus Neurospora intermedia, focusing on the molecular aspects of the fungal pellets with special interest to investigate the role of cell signaling second messenger cyclic 3', 5’-adenosine mono- phosphate (cAMP). It was found that Neurospora intermedia stimulate cAMP in the pellet form than filamentous form. The industrial applications of fungal pellets for generating value added products were also studied and observed fermentation in individual and co fermented first and second-generation ethanol substrate, showed an ethanol yield maximum of 0.25 ± 0.05 g/g dry substrate. The growth of fungal pellets in presence of inhibitors (such as acetic acid, HMF and furfural) resulted in about 11% to 45% increase in ethanol production as compared to filamentous forms, at similar growth conditions in the liquid straw hydrolysate.

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CETENE
Frutas são ricas em vitaminas, minerais, fibras e outros compostos que trazem benefícios à saúde. O conhecimento desses benefícios tem proporcionado aumento no consumo nos últimos anos, pois, além do interesse em fitoconstituintes os consumidores têm buscado produtos de elevada qualidade e seguros para o consumo. Devido à alta perecibilidade da matéria-prima e às falhas ocorridas nas diferentes fases da cadeia pós-colheita, grande parte dos vegetais produzidos no Brasil são desperdiçados. Dentre os diversos fatores envolvidos as doenças ocasionadas por fungos fitopatógenos se destacam. O controle de doenças fúngicas pós-colheita é comumente realizado através da aplicação de fungicidas, de elevado custo de produção e apresentados como ameaça à saúde pública e ao meio ambiente. Tendo em vista a problemática apresentada, no presente estudo foi avaliada a eficácia de revestimentos elaborados com quitosana (QUI) e óleos essenciais de Mentha piperita L. (MPOE) ou Mentha x villosa Huds (MVOE) como alternativa para o controle de infecções causadas pelos fungos Aspergillus niger, Botrytis cinerea, Penicillium expansum e Rhizopus stolonifer em tomates cereja e uvas de mesa durante o armazenamento em temperatura ambiente e em baixas temperaturas. A quitosana foi obtida da carapaça do camarão Litopenaus vannamei, em meio alcalino. Os óleos essenciais foram obtidos por arraste de vapor. Para o preparo do revestimento, o polímero de quitosana foi diluído em ácido acético sob agitação por 6 horas, seguido da adição do óleo e agitação por mais 18 horas em presença de glicerol como agente dispersante. As Concentrações Inibitórias Mínima da quitosana e de cada óleo foram determinadas por meio da macrodiluição em caldo. Os efeitos dos revestimentos sobre as características fúngicas (crescimento micelial radial e germinação espórica), físico-químicas (perda de peso, firmeza, cor, acidez e sólidos solúveis) e sensoriais (aceitação e intenção de compra) dos frutos durante a armazenagem também foram avaliados. A concentração inibitória mínima (CIM) de QUI contra todos os fungos-teste foi de 8 mg / mL, ao passo que a CIM para ambos MPOE e MVOE foi de 5 μL / mL. Combinações de QUI (8 e 4 mg/mL) e MPOE (QUI-MPOE) ou (QUI-MVOE) (5, 2,5 e 1,25 μL/mL) inibiu fortemente a germinação de esporos e o crescimento micelial dos fungos estudados. Os revestimentos compostos de QUI-MPOE ou QUI-MVOE retardaram o crescimento dos fungos causadores de mofos ensaiados em frutos artificialmente infectados durante a armazenagem em temperatura ambiente (12 dias) ou em baixa temperatura (24 dias). Os revestimentos ensaiados também preservaram a qualidade de frutos de tomate cereja e uvas de mesa durante a armazenagem, em termos de características físico-químicas e sensoriais. Nas uvas de mesa, houve uma melhoria nos valores de firmeza, cor L* e cor h* indicando frutos mais brilhosos e um possível retardo no desenvolvimento do "browning" das uvas revestidas com QUI-MPOE ou QUI-MVOE em relação aos frutos controle. Estes resultados indicam que os revestimentos que compreendem CHI-MPOE ou CHI-MVOE, apresentam-se como alternativa promissora para inibir a infecção de fungos pós-colheita em tomate cereja e uvas de mesa durante o armazenamento sem afetar a qualidade desses frutos.
Fruits are rich in vitamins, minerals, fiber and other compounds that provide health benefits. Knowledge of these benefits has provided an increase in vegetable consumption in recent years because in addition to the interest in phytochemicals that benefit the health and nutritional value, consumers have sought high quality products and safe for consumption. Due to the high perishability of the raw material and the failures occurred in different stages of post-harvest chain, most of the vegetables produced in Brazil are wasted. Among the many factors involved diseases caused by fungi pathogens in fruits stand out, they result in significant economic losses. Control of post-harvest fungal diseases commonly is achieved through the application of fungicides, increase the cost of production and presented to public health and the environment. In view of the problems presented, in the present study we evaluated the efficacy comprising shrimp chitosan (CHI) and Mentha piperita L. (MPEO) or Mentha x villosa Huds (MVEO) essential oils as an alternative for the control mold infections caused by Aspergillus niger, Botrytis cinerea, Penicillium expansum and Rhizopus stolonifer of cherry tomatoes and table grapes during storage at room temperature and low temperatures. Chitosan was obtained from the shell of the shrimp Litopenaeus vannamei in alkaline medium. The essential oils were obtained by vapor dragging. To prepare the coating, the polymer chitosan was diluted acetic acid with stirring for 6 hours, followed by addition of the oil and shaken for 18 more hours in the presence of glycerol as a dispersing agent. The Minimum Inhibitory Concentrations in chitosan and each oil were determined by the broth macrodilution. The effects of the coatings on the fungal characteristics (radial mycelial growth and esporic germination), physico-chemical (weight loss, firmness, color, acidity and soluble solids) and sensory (acceptance and purchase intention) of the fruit during storage were also evaluated. The minimum inhibitory concentration (MIC) of CHI against all the test fungi were 8 mg / mL, whereas the MIC for both MPOE and MVOE was 5 uL / mL. Combinations CHI (8 and 4 mg / mL) and MPOE (CHI-MPOE) or (CHI-MVOE) (5, 2.5 and 1.25 uL / mL) strongly inhibited spore germination and mycelial growth of of target fungi. The coatings comprising CHI-MPEO or CHI-MVEO delayed the growth of mold-causing fungi in artificially infected fruits during storage at either room (12 days) or low temperatures (24 days). The assayed coatings preserved the quality of cherry tomato fruits and table grapes during storage, in terms of physical, physicochemical and sensory attributes. In table grapes, there was an improvement in the firmness values, color L * and h * color indicating more glossy fruit and a possible delay in the development of "browning" of grapes coated with CHI-CHI-MPOE or MVOE compared to control fruits. These results indicate that coatings comprising CHI-MPEO or CHI-MVEO represent promising post-harvest treatments to inhibit common postharvest mold infections in cherry tomato fruits and in table grapes during storage without affecting the quality of these fruits.

Food waste and food poisoning derived from food contamination are huge issues in a globalized world like ours, where the distances between producer and consumer are ever greater. Biofilm-forming fungi are among the microorganisms more common in this sector. In the present work, antimicrobial and antibiofilm efficacy of Ethyl Lauroyl Arginate (LAE) was evaluated on some strains of fungi, specifically, Cladosporium cladosporioides, Aspergillus ochraceus, Penicillium italicum, Botryotinia fuckeliana and Fusarium oxysporum. LAE is a surfactant derived from lauric acid and arginine, approved and generally recognized as safe (GRAS) for certain food applications, due to the rapidity with which it is hydrolyzed by the human body. The MIC of the LAE was identified and its activity as an antibiofilm was evaluated for the cited fungal strains. Subsequently, the antimicrobial was used as an active part of a varnish-based antibiofilm coating that could be used for surfaces and packaging. Finally, an edible pectin-based film with LAE was tested in vivo against grapes inoculated with B. fuckeliana.

Kalaharituber pfeilii (Henn.) Trappe & Kagan-Zur commonly known as the “Kalahari truffle” is a desert truffle species identified from the Kalahari region of southern Africa. Two other species, Eremiomyces echinulatus (Trappe & Marasas) Trappe & Kagan-Zur and Mattirolomyces austroafricanus (Trappe & Marasas) Trappe & Kovacs are also known to occur in other parts of southern Africa. Truffles are hypogeous fruiting bodies of Ascomycetes, important to humans for their nutritional value and medicinal characteristics. These truffles are known as desert truffles as they prefer to occur under arid or semi-arid conditions characteristic of deserts. Truffle development depends on the presence of a mycorrhizal host, associated microorganisms as well as soil and climatic characteristics. It has been suggested that K. pfeilii has a suspected broad plant host range which includes herbaceous to woody trees and shrubs. However, these relationships have not been verified. Indigenous people of the Kalahari believe that truffles are found under grasses. In the Kalahari, truffle fruiting bodies are often found entangled in Stipagrostis ciliata (Desf.) De Winter var. capensis (Trin. & Rupr.) De Winter roots. S. ciliata, also known as the tall bushman-grass, is the most common grass found in the Kalahari. The objective of this study was to provide conclusive evidence that S. ciliata var. capensis is a host of the Kalahari truffle. Truffle fruiting bodies and grass roots from where the truffles were found were collected from Upington, South Africa. The fruiting bodies were identified by observing their morphological characteristics using the ‘Keys of Truffle genera’. All observed physical properties were similar to those of K. pfeilii and further identification was done using molecular techniques. DNA was extracted from the fruiting bodies, mycelial cultures, rhizosheaths and from the S. ciliata var. capensis grass roots, which were then amplified using the specific K. pfeilii specific primers TPF3 and TPR1 and sequenced. The obtained sequence results confirmed that the collected fruiting bodies were those of the K. pfeilii and the molecular techniques also confirmed that K. pfeilii DNA was present in the S. ciliata var. capensis rhizosheath and root cells. Microscopy showed an ectendomycorrhizal association between K. pfeilii and S. ciliata var. capensis. Mycorrhizal resynthesis experiments were conducted to establish this mycorrhizal association in-vitro. They were unsuccessful because of the structure of the grass and the availability of contaminants. And more...

Les populations du nord du Gabon consomment 39 taxons de champignons. Pour mieux connaître les taxons consommés et collecter des informations sur leur écologie, une étude fondée sur les connaissances mycologiques traditionnelles de ces populations et des observations de terrain a été entreprise dans les provinces de l’Ogooué-Ivindo et du Woleu-Ntem situées dans le nord du pays. Au cours de cette étude basée sur une enquête ethnomycologique menée sur les axes routiers Makokou-Mékambo et Oyem-Minvoul, ainsi que dans les villages pygmées des environs, deux cents personnes dont les Pygmées Baka et Bakoya, et les Bantu Fang, Kota et Kwélé ont été interrogées (100 personnes par province visitée).

Cette étude a permis non seulement d'établir la correspondance entre les noms scientifiques et les noms vernaculaires attribués aux champignons dans les cinq langues locales étudiées, mais aussi de recueillir d'autres informations liées aux connaissances mycologiques traditionnelles des populations enquêtées. Des descriptions macroscopiques et microscopiques détaillées ont été faites pour tous les taxons inventoriés.

L’étude a également révélé qu’il existe des différences significatives tant en ce qui concerne le nombre de taxons que les quantités de champignons consommés par les différents groupes ethniques: les Pygmées vivant uniquement de la chasse et de la cueillette consomment 96% des taxons inventoriés et des quantités élevées de champignons (environ 3 kg / jour / famille). Les Bantu vivant à l’écart de ces derniers consomment également des quantités de champignons assez élevées (environ 2 kg / jour / famille) mais un nombre réduit de taxons (56% des taxons inventoriés pour les Fang; 69% pour les Kota; 39% pour les Kwélé). Par contre, les Bantu vivant à proximité des Pygmées connaissent et consomment un grand nombre de taxons (environ 90% des taxons inventoriés) mais mangent de plus faibles quantités de champignons que leurs congénères éloignés des Pygmées (environ 800 g / jour / famille).

Plus généralement, l’étude a montré que les connaissances mycologiques traditionnelles de ces populations varient en fonction de l’activité pratiquée, de l’âge, de l’ethnie et du sexe. Les meilleures connaissances mycologiques sont détenues par les chasseurs et les pêcheurs qui identifient environ 80% des taxons. Chez les Pygmées, les connaissances mycologiques des hommes et des femmes sont très diversifiées et identiques, alors que chez les Bantu, les femmes connaissent mieux les champignons (plus de 50% des taxons identifiés) que les hommes (à peine 30% des taxons identifiés). Cependant, quel que soit le groupe ethnique, les représentants de la population active connaissent mieux les champignons (85% des taxons identifiés) que les jeunes et les personnes du troisième âge (environ 30% des taxons identifiés).

Les champignons les plus appréciés par ces populations appartiennent au genre Termitomyces dont les espèces les plus recherchées sont T. fuliginosus, T. robustus et T. microcarpus.

Une étude comparative des champignons consommés au Gabon et dans d’autres pays d’Afrique tropicale a montré que les champignons consommés au Gabon le sont également au Bénin, au Burundi, au Cameroun, en République centrafricaine, en RD Congo, au Malawi, en Tanzanie… et que, après la RD Congo (21 taxons inventoriés), le Gabon présente la plus grande diversité de taxons consommés dans le genre Cantharellus (14 taxons inventoriés). Par contre, sur une trentaine de taxons de Termitomyces signalés en Afrique tropicale, le Gabon est le pays qui présente la plus faible diversité (7 taxons inventoriés).

Une compilation des données bibliographiques a révélé que le nombre de champignons symbiontes comestibles signalés en Afrique tropicale est de loin plus élevé en forêt claire qu’en forêt dense (12 taxons de chanterelles sur les 28 inventoriés en Afrique tropicale sont propres à la forêt claire contre 2 taxons à la forêt dense; 15 taxons de Termitomyces sur 30 sont propres à la forêt claire contre 5 taxons à la forêt dense).

Mots-clés: champignons comestibles, Pygmées, Bantu, ethnomycologie, Gabon

Doctorat en Sciences
info:eu-repo/semantics/nonPublished

Chan Kam Che.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2006.
Includes bibliographical references (leaves 199-219).
Abstracts in English and Chinese.
Acknowledgements --- p.i
Abstracts --- p.iii
摘要 --- p.v
Contents --- p.vii
List of figures --- p.xiv
List of tables --- p.xix
Abbreviations --- p.xxii
Chapter Chapter I --- Introduction --- p.1
Chapter 1.1 --- Persistent organic pollutants --- p.1
Chapter 1.2 --- DDT and DDE --- p.2
Chapter 1.2.1 --- Background --- p.2
Chapter 1.2.2 --- Health effects --- p.4
Chapter 1.2.3 --- Environmental exposure of DDE --- p.4
Chapter 1.2.4 --- Level of DDE in human --- p.9
Chapter 1.2.5 --- Biodegradation of DDE --- p.10
Chapter 1.3 --- Remediation methods --- p.11
Chapter 1.3.1 --- Physical/ chemical treatment --- p.11
Chapter 1.3.2 --- Bioremediation --- p.13
Chapter 1.4 --- Fungal Bioremediation --- p.14
Chapter 1.5 --- Ligninolytic enzymes --- p.15
Chapter 1.5.1 --- Laccase --- p.15
Chapter 1.5.2 --- Peroxidases --- p.20
Chapter 1.5.2.1 --- Manganese Peroxidase (MnP) --- p.20
Chapter 1.5.2.1 --- Lignin Peroxidase (LiP) --- p.24
Chapter 1.6 --- Cultivation of Pleurotus pulmonarius --- p.27
Chapter 1.7 --- Enzyme technology on environmental cleanup and its limitation --- p.28
Chapter 1.8 --- Aims and objectives of this study --- p.29
Chapter Chapter II --- Materials and Methods --- p.30
Chapter 2.1 --- Organism and growth conditions --- p.30
Chapter 2.2 --- Cultivation and the expression of the ligninolytic enzyme-coding genes during solid-state-fermentation of edible mushroom Pleurotus pulmonarius --- p.30
Chapter 2.3 --- Treatment of DDE by living P. pulmonarius --- p.31
Chapter 2.3.1 --- Optimization of DDE removal in broth system --- p.31
Chapter 2.3.1.1 --- Effects of initial DDE concentration on the removal of DDE --- p.32
Chapter 2.3.1.2 --- Effects of inoculum size on the removal of DDE --- p.33
Chapter 2.3.1.3 --- Effects of incubation time on the removal of DDE and transcriptional profiles of the ligninolytic enzyme-coding genes --- p.33
Chapter 2.3.2 --- Optimization of DDE removal in soil system --- p.34
Chapter 2.3.2.1 --- Effects of initial DDE concentration on the removal of DDE --- p.34
Chapter 2.3.2.2 --- Effects of inoculum size on the removal of DDE --- p.35
Chapter 2.3.2.3 --- Effects of incubation time on the removal of DDE --- p.35
Chapter 2.3.2.4 --- Transcription of the ligninolytic enzyme-coding genes --- p.35
Chapter 2.4 --- Treatment of DDE by 1st SMC of p. pulmonarius grown on straw-based compost --- p.36
Chapter 2.4.1 --- Optimization of DDE removal in soil system --- p.36
Chapter 2.5 --- Treatment of DDE by crude enzyme preparations of P. pulmonarius grown on straw-based compost --- p.36
Chapter 2.5.1 --- Optimization of DDE removal in broth system --- p.36
Chapter 2.5.1.1 --- Effects of initial DDE concentration on the removal of DDE --- p.37
Chapter 2.5.1.2 --- Effects of amounts of crude enzyme preparations on the removal of DDE --- p.37
Chapter 2.5.1.3 --- Effects of incubation time on the removal of DDE --- p.37
Chapter 2.5.2 --- Optimization of DDE removal in soil system --- p.37
Chapter 2.5.2.1 --- Effects of initial DDE concentration on the removal of DDE --- p.38
Chapter 2.5.2.2 --- Effects of amount of crude enzyme preparations on the removal of DDE --- p.38
Chapter 2.5.2.3 --- Effects of incubation time on the removal of DDE --- p.38
Chapter 2.6 --- Soil characterization --- p.39
Chapter 2.6.1 --- Identification of organic contaminants in soil sample from Gene Garden using Gas Chromatography/Mass Spectrometry (GC/MS) --- p.39
Chapter 2.6.2 --- Determination of soil texture --- p.42
Chapter 2.6.3 --- Fresh soil/air-dried sample moisture --- p.44
Chapter 2.6.4 --- "Soil pH, electrical conductivity & salinity" --- p.44
Chapter 2.6.5 --- Total organic carbon contents --- p.44
Chapter 2.6.6 --- Total nitrogen and total phosphorus --- p.44
Chapter 2.6.7 --- Available nitrogen --- p.45
Chapter 2.6.8 --- Available phosphorus --- p.45
Chapter 2.6.9 --- Potassium value --- p.46
Chapter 2.7 --- Quantification of residual DDE level --- p.47
Chapter 2.7.1 --- Preparation of DDE stock solution --- p.47
Chapter 2.7.2 --- Extraction and quantification of DDE using Gas Chromatography with Electron Capture Detector (GC/μECD) --- p.47
Chapter 2.7.3 --- Identification of DDE breakdown products by GC/MS --- p.50
Chapter 2.8 --- Extraction of protein and ligninolytic enzymes --- p.53
Chapter 2.8.1 --- Protein assay --- p.53
Chapter 2.8.2 --- Laccase assay --- p.53
Chapter 2.8.3 --- Manganese peroxidase assay --- p.54
Chapter 2.8.4 --- Calculation of activity and specific activity of laccase and manganese peroxidase --- p.54
Chapter 2.9 --- Estimation of fungal biomass --- p.55
Chapter 2.9.1 --- Preparation of ergosterol standard solution --- p.56
Chapter 2.9.2 --- Analysis of ergosterol content --- p.56
Chapter 2.10 --- Expression of the ligninolytic enzyme-coding genes --- p.58
Chapter 2.10.1 --- Preparation of ribonuclease free reagents and apparatus --- p.58
Chapter 2.10.2 --- RNA isolation and purification --- p.58
Chapter 2.10.3 --- cDNA synthesis --- p.59
Chapter 2.10.4 --- Semi-quantification of ligninolytic enzyme-coding gene expression by RT-PCR --- p.59
Chapter 2.11 --- Preparation of crude enzyme preparations from P. pulmonarius compost --- p.63
Chapter 2.12 --- "Assessment criteria: removal efficiency, RE, and removal capacity, RC" --- p.63
Chapter 2.13 --- Statistical analysis “ --- p.64
Chapter Chapter III --- Results --- p.65
Chapter 3.1 --- Soil characterization --- p.65
Chapter 3.2 --- Cultivation and the expression of the ligninolytic enzyme-coding genes during solid-state-fermentation of edible mushroom Pleurotus pulmonarius --- p.66
Chapter 3.2.1 --- Mushroom yield --- p.66
Chapter 3.2.2 --- Protein content --- p.66
Chapter 3.2.3 --- Specific ligninolytic enzymes activities --- p.66
Chapter 3.2.4 --- Ergosterol content --- p.69
Chapter 3.2.5 --- Ligninolytic enzymes productivities --- p.69
Chapter 3.2.6 --- Expression of the ligninolytic enzyme-coding genes during solid-state-fermentation --- p.72
Chapter 3.3 --- Treatment of DDE by living P. pulmonaruis --- p.78
Chapter 3.3.1 --- Optimization of DDE removal in broth system --- p.78
Chapter 3.3.1.1 --- Effects of initial DDE concentration on the removal of DDE --- p.78
Chapter 3.3.1.1.1 --- Effects of DDE on biomass development --- p.78
Chapter 3.3.1.1.2 --- Protein content --- p.78
Chapter 3.3.1.1.3 --- Specific ligninolytic enzyme activities --- p.78
Chapter 3.3.1.1.4 --- Ligninolytic enzyme productivities --- p.79
Chapter 3.3.1.1.5 --- DDE removal and removal capacity --- p.79
Chapter 3.3.1.2 --- Effects of inoculum sizes on the removal of DDE --- p.84
Chapter 3.3.1.2.1 --- Effects of DDE on biomass development --- p.84
Chapter 3.3.1.2.2 --- Protein content --- p.84
Chapter 3.3.1.2.3 --- Specific ligninolytic enzyme activities --- p.85
Chapter 3.3.1.2.4 --- Ligninolytic enzyme productivities --- p.85
Chapter 3.3.1.2.5 --- DDE removal and removal capacity --- p.85
Chapter 3.3.1.3 --- Effects of incubation time on the removal of 4.0 mM DDE/g biomass --- p.89
Chapter 3.3.1.3.1 --- Effects of DDE on biomass development --- p.89
Chapter 3.3.1.3.2 --- Protein content --- p.89
Chapter 3.3.1.3.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.89
Chapter 3.3.1.3.4 --- DDE removal and removal capacity --- p.90
Chapter 3.3.1.3.5 --- Putative degradation derivatives --- p.90
Chapter 3.3.1.3.6 --- Expression of the ligninolytic enzyme-coding genes during the removal of 4.0 mM DDE/g biomass --- p.94
Chapter 3.3.1.4 --- Effects of incubation time on the removal of 10.0 mM DDE/g biomass --- p.100
Chapter 3.3.1.4.1 --- Effects of DDE on biomass development --- p.100
Chapter 3.3.1.4.2 --- Protein content --- p.100
Chapter 3.3.1.4.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.100
Chapter 3.3.1.4.4 --- Expression of the ligninolytic enzyme-coding genes during the removal of 10.0 mM DDE/g biomass --- p.102
Chapter 3.3.2 --- Optimization of DDE removal in soil system --- p.107
Chapter 3.3.2.1 --- Effects of initial DDE concentration on the removal of DDE --- p.107
Chapter 3.3.2.1.1 --- Ergosterol content --- p.107
Chapter 3.3.2.1.2 --- Protein content --- p.107
Chapter 3.3.2.1.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.107
Chapter 3.3.2.1.4 --- DDE removal and removal capacity --- p.108
Chapter 3.3.2.2 --- Effects of inoculum sizes on the removal of DDE --- p.111
Chapter 3.3.2.2.1 --- Ergosterol content --- p.111
Chapter 3.3.2.2.2 --- Protein content --- p.111
Chapter 3.3.2.2.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.111
Chapter 3.3.2.2.4 --- DDE removal and removal capacity --- p.112
Chapter 3.3.2.3 --- Effects of incubation time on the removal of DDE --- p.115
Chapter 3.3.2.3.1 --- Ergosterol content --- p.115
Chapter 3.3.2.3.2 --- Protein content --- p.115
Chapter 3.3.2.3.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.115
Chapter 3.3.2.3.4 --- DDE removal and removal capacity --- p.116
Chapter 3.3.2.3.5 --- Putative degradation derivatives --- p.116
Chapter 3.3.2.4 --- Transcription of the ligninolytic enzyme-coding genes --- p.121
Chapter 3.4 --- Treatment of DDE by 1st SMC of p. pulmonarius grown on straw-based compost --- p.127
Chapter 3.4.1 --- Optimization of DDE removal in soil system --- p.127
Chapter 3.4.1.1 --- Effects of initial DDE concentration on the removal of DDE --- p.127
Chapter 3.4.1.1.1 --- Ergosterol content --- p.127
Chapter 3.4.1.1.2 --- Protein content --- p.127
Chapter 3.4.1.1.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.127
Chapter 3.4.1.1.4 --- DDE removal and removal capacity --- p.128
Chapter 3.4.1.2 --- Effects of inoculum sizes on the removal of DDE --- p.132
Chapter 3.4.1.2.1 --- Ergosterol content --- p.132
Chapter 3.4.1.2.2 --- Protein content --- p.132
Chapter 3.4.1.2.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.132
Chapter 3.4.1.2.4 --- DDE removal and removal capacity --- p.133
Chapter 3.4.1.3 --- Effects of incubation time on the removal of DDE --- p.136
Chapter 3.4.1.3.1 --- Ergosterol content --- p.136
Chapter 3.4.1.3.2 --- Protein content --- p.136
Chapter 3.4.1.3.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.136
Chapter 3.4.1.3.4 --- DDE removal and removal capacity --- p.137
Chapter 3.4.1.3.5 --- Putative degradation derivatives --- p.137
Chapter 3.5 --- Treatment of DDE by crude enzyme preparations of P. pulmonarius grown on straw-based compost --- p.142
Chapter 3.5.1 --- The crude enzyme preparations of P. pulmonarius grown on straw-based compost --- p.142
Chapter 3.5.2 --- Optimization of DDE removal in broth system --- p.143
Chapter 3.5.2.1 --- Effects of initial DDE concentration on the removal of DDE --- p.143
Chapter 3.5.2.2 --- Effects of amounts of crude enzyme preparations on the removal of DDE --- p.145
Chapter 3.5.2.3 --- Effects of incubation time on the removal of DDE --- p.147
Chapter 3.5.2.4 --- Putative degradation derivatives --- p.147
Chapter 3.5.3 --- Optimization of DDE removal in soil system --- p.151
Chapter 3.5.3.1 --- Effects of initial DDE concentration on the removal of DDE --- p.151
Chapter 3.5.3.2 --- Effects of amounts of crude enzyme preparations on the removal of DDE --- p.151
Chapter 3.5.3.3 --- Effects of incubation time on the removal of DDE --- p.154
Chapter 3.5.3.4 --- Putative degradation derivatives --- p.154
Chapter Chapter IV --- Discussions --- p.158
Chapter 4.1 --- Quantification of the expression of the ligninolytic enzyme-coding genes --- p.158
Chapter 4.2 --- Artificial cultivation and the expression of the ligninolytic enzyme-coding genes during solid-state-fermentation of edible mushroom Pleurotus pulmonarius --- p.164
Chapter 4.3 --- Treatment of DDE by living P. pulmonarius --- p.166
Chapter 4.3.1 --- Optimization of DDE removal in broth system --- p.166
Chapter 4.3.2 --- Optimization of DDE removal in soil system --- p.169
Chapter 4.3.3 --- Phylogeny of the ligninolytic enzyme-coding genes --- p.170
Chapter 4.3.3.1 --- Laccase coding genes --- p.170
Chapter 4.3.3.2 --- MnP coding genes --- p.175
Chapter 4.3.4 --- Transcription of the ligninolytic enzyme-coding genes --- p.178
Chapter 4.4 --- Treatment of DDE by 1st SMC of P. pulmonarius grown on straw-based compost --- p.183
Chapter 4.4.1 --- Optimization of DDE removal in soil system --- p.183
Chapter 4.5 --- Treatment of DDE by crude enzyme preparations of P. pulmonarius grown on straw-based compost --- p.184
Chapter 4.6 --- Cost-effectiveness of the bioremediation method --- p.185
Chapter 4.7 --- Further investigations --- p.194
Chapter Chapter V --- Conclusions --- p.197
References --- p.199

碩士
大葉大學
食品工程學系碩士班
91
ABSTRACT Grifola frondosa in Japan as maitake （dancing mushroom）, in China as gray tree flower, it is a Basidiomycete fungus belonging to the order Aphyllopherales, and family Polyporaceae, as a white-rot and acreobe fungus. It is polysaccharides have been reported in many research articles include antitumor, immunological enhanc- ement, antidiabetic and anti-HIV, etc. A β-(1-3)-linked glucan with branches of β-(1-6)-D-glucose showing the main of pharma- cological activity has been isolated from fruit bodies and mycelium. By synthetic-log cultivation, when the young mycelium grown to fruit body need of three months.according to the related studies, employing mycelium of the submerged culture to grow the fungus has the advantages of the shorter growth time, better product quality, and lower cost. This study investigates the process of the growth of Grifola frondosa in terms of the following issues：（1）to screen different strains producing polysaccharides and analysis of free amino acid、total amino acid、main element (C、N及O)、enzyme activity；（2）studies biomass、extracellular polysaccharide and intracellular polysaccharide under shaking and static bottles；（3）to compare of the free amino acid and polysaccharides by the chemical and semi-chemical medium；（4） effect of submerged fermentation in shaking bottles, and to expand of 5 and 20L fermentor. The study shows that, in PDA culture, the CCRC 36434 have the best growing speed of the colony, in PDB and base medium culture, the CCRC 36355 have best yields of intracellular poly- saccharides, CCRC 36357 which yields the highest content of extracellular polysaccharide；in content of free amino acid, shows the higher of CCRC 36355；in content of total amino acid by mycelium, the higher of CCRC 36434；in api-ZYM system, intracellular enzyme have higher activity；in shaking culture by chemical medium , the highest intracellular polysaccharides is achieved under the condition of 4﹪glucose, 0.1﹪ammonium oxalate, 0.15﹪potassium phosphate, the highest the free amino acid is achieved under the condition of 3﹪sucrose, 0.2﹪ammonium oxalate, 0.45﹪potassium phosphate；in shaking culture by semi- chemical medium , the highest intracellular polysaccharides is achieved under the condition of 0.2﹪peptone, the highest extra- cellular polysaccharides is 0.4﹪yeast extract, the highest the mycelium biomass is 0.8﹪tryptone；in static culture, the highest intracellular polysaccharides is achieved under the condition of 3﹪fructose, 0.2﹪ammonium nitrate, the highest the free amino acid is achieved under the condition of 4﹪mannose, 0.4﹪ammonium oxalate；studies submerged fermentation, the higher extracellular polysaccharides on day 11 by shaking culture and 5、20L fermentor on day 5；in 5L fermentor, free amino acid have best yield on day 2, the free amino acid follow time to decreased. extracellular polysaccharides and intracellular polysaccharides contain acidic glucan by ion exchange chromatography and intracellular polysaccharides contain less basic glucan.

Food insecurity can contribute to the advancement of diseases such as growth stunting and HIV/AIDS. A holistic approach to addressing food insecurity includes reviewing local resources; including indigenous food stuffs. Six studies investigate the potential of insect nutrition to meet dietary needs in rural South Africa. A novel trapping method for Trinervitermes sp. is examined by parameters of time, sustainability and bait used. Local grass (Themeda triandra Forssk.) seemed to be the most effective bait, being significantly more attractive than loose mound soil (p=0.01), wet maize stalks (p=0.01) or cardboard (p=0.05). The trapping device was demonstrated as an effective tool in assessing the feeding preferences of Trinervitermes sp., which compete directly with cattle for grazing food resources. The chemical composition of Macrotermes natalensis alates (winged, wingless and fried), soldiers, and Odontotermes sp. alates (wingless) was determined. Alates were rich in fat, ranging between 49.2-60.6% (dry matter basis). The protein content ofM natalensis and Odontotermes sp. alates compared favourably to pork and chicken. Alates were high in glutamic, aspartic and alanine amino acids and low in methionine, serine and threonine. Amino acid digestion for broiler chickens was high, ranging between 87.6-96.1%. In an era where rural and urban cultures are rapidly merging, entomophagy may be discarded as an embarrassment or nonsensical practice. The high nutritional content of M natalensis and Odontotermes sp. should be publicised both to increase the awareness of their high quality as a food source for both poultry and human consumption and to avoid the abandonment of cultural practices that make sense.
Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2004.

Thesis (M.S.) -- University of Tennessee, Knoxville, 2005.
Title from title page screen (viewed on Sept. 1, 2005). Thesis advisor: Karen Hughes. Document formatted into pages (viii, 68 p. : ill. (some col.)). Vita. Includes bibliographical references (p. 56-63).

Submitted in fulfilment for the requirements for the degree of Doctor of Technology: Biotechnology, Durban University of Technology, 2010.
South African edible oil processing plants produce approximately 3 x 105 tonnes of oil annually with up to 3 tonnes of water for every tonne of oil produced. Wastewater that contains oil extracts varies in organic loading from 30,000 to 60,000 mg.l-1 COD. This wastewater can be used to grow oleophilic fungi to produce valuable industrial products. The global vitamin B market is approximately R25.5 billion with 4500 metric tonnes being produced. A large proportion of this is produced using the fungus Eremothecium gossypii using oil substrates. The aim of this study was to to develop a novel method to produce riboflavin with the aid of fungi, using edible oil effluent (EOE) as substrate, and to optimize the production thereof by statistical experimental design. Four fungi were surveyed for their growth potential on EOE and two, E. gossypii (CBS109.51) and C. famata (ATCC 208.50) were found to produce sufficient riboflavin for further study. Mutation of these organisms using ethylmethane sulphonate (EMS) increased riboflavin production from 3.52 mg.l-1 to 38.98 mg.l-1, an 11-fold increase. An enzyme pathway responsible for this was found to involve isocitrate lyase and comparison of this enzyme’s activity in the mutant against the wild-type using Michaelis-Menten kinetics showed a higher reaction velocity (Vmax) with a reduced substrate affinity (Km) indicating that the mutation was associated with this enzyme. Biomass comparisons were fitted to the sigmoid Gompertz model which was used to compare the wild-type to the mutant and increased specific growth rates and doubling times were observed in mutated cultures of E. gossypi. A strategy of statistical experimental design was pursued to optimize media components and iterative fractional factorial experiments culminating in a central composite optimization experiment were conducted. Statistically verified mathematical models were developed at each stage to identify important media components, predict media interactions, show directions for improvement and finally, predict maximum riboflavin production. An eight-factor resolution IV fractional factorial increased riboflavin production to 112 mg.l-1 followed by a four-factor resolution V experimental design which increased riboflavin production to 123 mg.l-1. A two-factor (yeast extract and NaCl) central composite experimental design predicted a maximum riboflavin production of 136 mg.l-1 which was a 3.5-fold increase from the mutant, and 38.6-fold higher than the E. gossypii wild-type. The optimized value was achieved within predicted confidence intervals in confirmatory experiments. Cost implications for production of riboflavin on EOE were calculated and a 10% technology uptake by the edible oil industry could yield a riboflavin industry with a 63.65 million rand turnover and a potential 24.96 million rand gross profit margin.

by Jiong Zhao.
Thesis (Ph.D.)--Chinese University of Hong Kong, 1994.
Includes bibliographical references (leaves 197-217).
Acknowledgments --- p.III
Abstract --- p.IX
Abbreviations --- p.XI
Chapter Chapter 1. --- General Introduction --- p.1
Chapter 1.1 --- What is a mushroom? --- p.1
Chapter 1.2 --- Mushroom Genetics: its development and prospective --- p.1
Chapter 1.2.1 --- Genome karyotype by pulsed field gel electrophoresis analysis --- p.2
Chapter 1.2.2 --- Mitochondrial Genetics --- p.4
Chapter 1.2.3 --- Mating type genes --- p.5
Chapter 1.2.4 --- Transformation --- p.7
Chapter 1.2.5 --- Parasexual processes --- p.8
Chapter 1.2.6 --- Mushroom breeding --- p.11
Chapter Chapter 2. --- Literature review: Protoplast fusion in fungi --- p.14
Chapter 2.1 --- Introduction --- p.14
Chapter 2.2 --- Protoplast fusion in yeasts --- p.14
Chapter 2.2.1 --- Intraspecific fusion --- p.14
Chapter 2.2.2 --- Interspecific fusion --- p.15
Chapter 2.2.3 --- Intergeneric fusion --- p.16
Chapter 2.3 --- Protoplast fusion in some Filamentous fungi --- p.17
Chapter 2.3.1 --- Aspergillus --- p.17
Chapter 2.3.2 --- Fusarium --- p.18
Chapter 2.3.3 --- Tricoderma --- p.19
Chapter 2.4 --- Protoplast fusion in strains --- p.21
Chapter 2.4.1 --- Protoplast isolation and regeneration --- p.21
Chapter 2.4.2 --- Intraspecific fusion in mushroom species --- p.24
Chapter 2.4.3 --- Interspecific fusion in mushroom species --- p.24
Chapter 2.4.4 --- Intergeneric fusion in mushroom species --- p.26
Chapter 2.4.5 --- Transfer of nuclei in mushroom species --- p.27
Chapter 2.5 --- General conclusions about literatures --- p.27
Chapter 2.5.1 --- Brief points about fungal protoplast fusion --- p.27
Chapter 2.5.2 --- Some arguements about fusion works in mushrooms strains --- p.31
Chapter 2.5.2.1 --- Classification of parental strains --- p.31
Chapter 2.5.2.2 --- Control experiments --- p.31
Chapter 2.5.2.3 --- Indentification methods of hybrids --- p.32
Chapter 2.6 --- General research ideas about experiments --- p.33
Chapter Chapter 3 --- Protoplast isolation and regeneration in some mushroom species --- p.37
Chapter 3.1 --- Introduction --- p.37
Chapter 3.2 --- Materials and Methods --- p.38
Chapter 3.2.1 --- Strains --- p.38
Chapter 3.2.2 --- Media --- p.38
Chapter 3.2.3 --- Protoplast release --- p.40
Chapter 3.2.4 --- Protoplast regeneration --- p.41
Chapter 3.3 --- Results and Discussion --- p.41
Chapter 3.3.1 --- Effect of culture age --- p.41
Chapter 3.3.2 --- Effect of lytic enzyme --- p.42
Chapter 3.3.3 --- Effect of concentration of mycelium --- p.45
Chapter 3.3.4 --- Effect of filter system --- p.46
Chapter 3.3.5 --- Effect of different regeneration protocols --- p.48
Chapter 3.3.6 --- Effect of soluable starch --- p.49
Chapter 3.3.7 --- Effect of PEG on the regeneration frequency --- p.50
Chapter 3.4 --- Conclusions --- p.53
Chapter Chapter 4 --- Monokaryotization by protoplasting technique in some heterothallic mushroom species --- p.54
Chapter 4.1 --- Introduction --- p.54
Chapter 4.2 --- Materials and Methods --- p.55
Chapter 4.2.1 --- Strains and media --- p.55
Chapter 4.2.2 --- Production of neo-monokaryons by protoplast technique --- p.55
Chapter 4.2.3 --- Identification of mating types in protoplasted monokaryons --- p.57
Chapter 4.3 --- Results
Chapter 4.3.1 --- Formation of neo-monokaryons --- p.57
Chapter 4.3.2 --- Monokaryotization in different strains --- p.60
Chapter 4.3.3 --- Comparison of parental and protoplasted monokaryons --- p.60
Chapter 4.3.4 --- Comparison of regeneration rate of parental monokaryons --- p.62
Chapter 4.4 --- Discussion
Chapter 4.4.1 --- Differences of regeneration time in monokaryons and dikaryons --- p.64
Chapter 4.4.2 --- Genetic differences between parental and neo-monokaryons --- p.64
Chapter 4.4.3 --- Mechanism for the production of neo-monokaryons --- p.65
Chapter 4.4.4 --- Advantages of protoplasting technique in mushroom breeding --- p.65
Chapter 4.4.5 --- Protoplasting technique in the identification of fusion hybrids --- p.67
Chapter 4.5 --- Couclusions --- p.68
Chapter Chapter 5 --- Intraspecific hybridization in Coprinus cinereus and Schizophyllum commune by PEG-induced protoplast fusion and electrofusion --- p.69
Chapter 5.1 --- Introduction --- p.69
Chapter 5.2 --- Materials and Methods
Chapter 5.2.1 --- Strains and Media --- p.70
Chapter 5.2.2 --- Fusogen --- p.70
Chapter 5.2.3 --- Inactivation chemicals --- p.71
Chapter 5.2.4 --- Inactivation of protoplasts --- p.71
Chapter 5.2.5 --- PEG induced protoplast fusion --- p.72
Chapter 5.2.6 --- Electrofusion --- p.72
Chapter 5.2.7 --- Investigation of protoplast fusion yield and fusion frequency --- p.73
Chapter 5.2.8 --- Comparison of mycelium growth rate --- p.73
Chapter 5.2.9 --- Fruiting test --- p.74
Chapter 5.3 --- Results
Chapter 5.3.1 --- Inactivation by IA and DP --- p.76
Chapter 5.3.2 --- Effect of different fusogens on fusion frequency --- p.79
Chapter 5.3.3 --- Effect of different fusion protocols on fusion frequency --- p.79
Chapter 5.3.4 --- Optimization of electrofusion --- p.80
Chapter 5.3.5 --- Fusion frequency resulted by PEG and electrofusion --- p.83
Chapter 5.3.6 --- Comparison of colony diameters and fruiting time --- p.84
Chapter 5.4 --- Discussion
Chapter 5.4.1 --- Inactivation of protoplasts by biochemical inhibitors --- p.85
Chapter 5.4.2 --- Optimization of PEG induced fusion --- p.86
Chapter 5.4.3 --- Optimization of electrofusion --- p.86
Chapter 5.4.4 --- Identification of fusion heterokaryons --- p.87
Chapter 5.4.5 --- Comparison of PEG and electrofusion --- p.89
Chapter 5.4.2 --- Effect of mitochondria --- p.90
Chapter 5.5 --- Couclusions --- p.91
Chapter Chapter 6 --- Interspecific hybridization between Volvariella volvacea and Volvariella bomhycina by protoplast fusion --- p.92
Chapter 6.1 --- Introduction --- p.92
Chapter 6.2 --- Materials and Methods
Chapter 6.2.1 --- Strains and Media --- p.93
Chapter 6.2.2 --- Protoplast production and regeneration --- p.94
Chapter 6.2.3 --- Inactivation of protoplasts --- p.94
Chapter 6.2.4 --- Protoplast fusion --- p.94
Chapter 6.2.5 --- Selection of fusion products --- p.95
Chapter 6.2.6 --- Analyses of progeny --- p.95
Chapter 6.2.7 --- Identification of fusants by protoplasting technique --- p.96
Chapter 6.2.8 --- Nuclear DNA contents in parents and hybrids --- p.96
Chapter 6.2.9 --- Genomic DNA amplification by arbitraly primers --- p.96
Chapter 6.2.10 --- Amplification by nuclear and mitochondrial rDNA --- p.97
Chapter 6.2.11 --- Fruiting test --- p.97
Chapter 6.3 --- Results
Chapter 6.3.1 --- Inactivation of Vb10 protoplasts --- p.98
Chapter 6.3.2 --- Low temperature effect on Vv34 --- p.100
Chapter 6.3.3 --- Selection of fusants --- p.100
Chapter 6.3.4 --- Analyses of progeny --- p.106
Chapter 6.3.5 --- Identification by protoplasting technique --- p.108
Chapter 6.3.6 --- Nuclear DNA contents in parents and hybrids --- p.110
Chapter 6.3.7 --- Arbitraly primer amplified PCR fingerprinting --- p.113
Chapter 6.3.8 --- rDNA PCR results --- p.119
Chapter 6.3.9 --- Interspecific variations
Chapter 6.3.10 --- Genome analysis of hybrids by pulse field gel electrophoresis
Chapter 6.3.11 --- Fruiting test
Chapter 6.4 --- Discussion
Chapter 6.4.1 --- Strain choice --- p.125
Chapter 6.4.2 --- Low temperature strains --- p.125
Chapter 6.4.3 --- Nuclear DNA content --- p.125
Chapter 6.4.4 --- AP-PCR and RAPDs markers --- p.126
Chapter 6.4.5 --- Interspecific fusion in Volvariella --- p.126
Chapter 6.5 --- Couclusions --- p.130
Chapter Chapter 7 --- Intergeneric hybridization between Schizophyllum commune and Pleurotus florida by protoplast fusion --- p.131
Chapter 7.1 --- Introduction --- p.131
Chapter 7.2 --- Materials and Methods
Chapter 7.2.1 --- Strains and Media --- p.132
Chapter 7.2.2 --- Protoplast fusion --- p.133
Chapter 7.2.3 --- Analyses of progeny --- p.134
Chapter 7.2.4 --- Phylogenetic analysis --- p.135
Chapter 7.2.5 --- Fruiting test --- p.135
Chapter 7.3 --- Results
Chapter 7.3.1 --- Selection of fusion products --- p.135
Chapter 7.3.2 --- Analyses of fusion progeny --- p.139
Chapter 7.3.3 --- Identification by protoplasting technique --- p.143
Chapter 7.3.4 --- Determination of nuclear DNA contents --- p.145
Chapter 7.3.5 --- rDNA PCR analysis in fusion --- p.148
Chapter 7.3.6 --- Identification of hybrids by AP-PCR and RAPDs markers --- p.151
Chapter 7.3.7 --- Phylogenetic analysis --- p.162
Chapter 7.3.8 --- Fruiting test --- p.164
Chapter 7.4 --- Discussion --- p.165
Chapter 7.5 --- Couclusions --- p.169
Chapter Chapter 8 --- Protoplast fusion in shiitake and other species --- p.171
Chapter 8.1 --- Introduction --- p.172
Chapter 8.2 --- Materials and Methods --- p.172
Chapter 8.3 --- Results and Discussion --- p.173
Chapter 8.4 --- Couclusion --- p.179
Chapter Chapter 9. --- General discussion and conclusions --- p.180
Appendix 1. Determination of ploidy in some mushrooms --- p.187
Appendix 2. Genomic DNA Isolation --- p.188
Appendix 3. Arbitrary primer polymerase chain reaction --- p.190
Appendix 4. rDNA PCR Amplification conditions --- p.193
Appendix 5. Pulsed Field Gel Electrophoresis --- p.195
Appendix 6. Genetic distance analysis in hybrids and their parents --- p.196
References --- p.197

Dissertação de Mestrado em Biodiversidade e Biotecnologia Vegetal apresentada à Faculdade de Ciências e Tecnologia
São Tomé e Príncipe é arquipélago localizado no Golfo da Guiné, África Ocidental. A ilha de São Tomé é um hotspot de biodiversidade, apresentando elevadas taxas de endemismo. Estas taxas de endemismo são maioritariamente reportadas para plantas vasculares, aves, repteis, anfíbios e organismos marinhos. A diversidade fúngica da ilha de São Tomé tem vindo a ser estudada desde 1851, somando já cerca de 300 espécies de macrofungos reportados até 2019. No entanto, o conhecimento micológico é escasso e disperso, pelo que a necessidade de um estudo aprofundado é premente. O objetivo deste trabalho foi compilar todas as informações acerca da diversidade macrofúngica da ilha de São Tomé e adicionar novas informações obtidas em inventários micológicos in situ. Através de análise bibliográfica e missões de campo, foi possível construir uma lista de espécies de macrofungos e avaliar o seu potencial comestível e medicinal. Foi também possível avaliar a etnomicologia local, que se resume a quatro nomes comuns: Utu, Utu-sandjá, Cloçon-son e Ntuda renda. Tentou-se fazer a correspondência entre estes nomes comuns e os nomes científicos. O conhecimento dos cogumelos comestíveis e medicinais de São Tomé será uma ferramenta extremamente útil para o futuro das populações desta ilha, uma vez que o projeto Tesouros d’Obô pretende utilizar este conhecimento para estabelecer pequenas unidades de produção de cogumelos em comunidades-alvo.Palavras-chave: São Tomé; Fungos; Cogumelos; África; Funga; Micologia; Cogumelos comestíveis; Cogumelos medicinais; África Ocidental; Diversidade fúngica
São Tomé e Príncipe is an archipelago situated on the Gulf of Guinea, West Africa. São Tomé’s island is a biodiversity hotspot, with reported high endemism rates. These endemism rates are mainly reported for vascular plants, birds, reptiles, amphibians, and marine organims. São Tomé’s island’s fungal diversity has been studied since 1851, summing up to around 300 macrofungi species reported until 2019. However, the mycological knowledge is scarce and dispersed, therefore the need for a deeper study is pressing. The objective of this work was to compile all of the information related to the macrofungal diversity of São Tomé’s Island and add new information obtained in in situ mycological inventories. Through the bibliographic analysis and the field missions, it was possible to build a species list and evaluate the edibility and medicinal potential of those species. It was also possible to evaluate the local ethnomycology, that sums up to four common names: Utu, Utu-sandjá, Cloçon-son e Ntuda renda. It was attempted to establish the link between these common names and the respective scientific names. The knowledge of São Tomé’s edible and medicinal mushrooms will be an extremely useful tool for the future of this island’s populations because the Tesouros d’Obô project is aiming to use this knowledge to establish small mushroom production units in the target communities.Keywords: São Tomé; Fungi; Mushrooms; Africa; Funga; Mycology; Edible mushrooms; Medicinal mushrooms; West Africa; Fungal diversity
Outro - This dissertation was funded by the African Union under the Project AURG II-1-254-2016 – “Implementation of Agroforestry Systems in S. Tomé and Principe and development of non-wood forest products (NWFP) in Angola and S. Tomé and Principe to improve income-generation and food security” (Acronym: Tesouros d’Obô).

Dissertação de Mestrado em Biodiversidade e Biotecnologia Vegetal apresentada à Faculdade de Ciências e Tecnologia da Universidade de Coimbra.
Edible mushrooms are consumed globally whether as a delicacy or due to their medicinal and nutritional proprieties. As a seasonal resource, wild edible mushrooms are not available naturally during all year, existing, therefore, a need on their cultivation. Some ectomyc- orrhizal (ECM) fungi produce the most sought and valuable sporocarps worldwide. Unlike saprotrophic species, a pro table cultivation of edible mycorrhizal mushrooms is very diffi- cult due to several issues, being its association with ne roots of plants, dependency of plant partner's biomass and growth rates some of them. Several species of the Mediterranean genus Cistus are common shrubs in the understory of Pinus pinaster dominated forests in Portugal and are ectomycorrhizal species. Besides their ability of establishing ectomycorrhizas, Cistus are also shrubs with relative high growth rates, capable of reach their highest biomass in less time than trees, being suitable candidates for cultivation of ectomycorrhizal mushrooms. One of the main objectives, of the present work, is the production of edible ECM mush- rooms, thus were performed assays with mycelial cultures of ECM species as well as synthesis of ectomycorrhizas. Fungal mycelial cultures were newly isolated from sporocarps of edible ectomycorrhizal species - Boletus fragans and Tricholoma equestre- and compared with the saprobic species Pleurotus ostreatus and Agaricus bisporus. The mycelial cultures were grown in several cul- ture media - Potato Dextrose agar (PDA), Malt extract agar (MEA), Biotin-Aneurin-Folic acid (BAF), Merlin-Norkrans modi ed medium (MNM) and Murashige & Skoog medium (MS) - and different temperatures, 4oC/24oC/30oC. Growth responses of ECM and sapro- trophs fungi were compared. The experiment allowed also the establishment of optimal culture conditions for mycelial growth of each species. Seedlings of three Portuguese native species - Cistus psilosepalus, Cistus salviifolius and Cistus ladanifer - were used as plant partners to axenically induce the establishment of mycor- rhizas with the edible ECM fungal species Boletus fragans, Lactarius deliciosus, Tricholoma equestre and Tricholoma portentosum. Different methods for mycorrhizal synthesis were tested, including containers with different shapes and several substrates and, to our knowl- edge, a new method of ectomycorrhizal synthesis, adapted to these shrubs growth habit, was developed. In order to get some insights about the ECM fungal partners of Cistus plants in natural conditions, a preliminary sampling was performed in a maritime pine forest. Ectomycorrhizal root tips from Cistus psilosepalus, Cistus salviifolius and Pinus pinaster were collected ina natural Portuguese pinewood to identify natural fungal symbionts associated with these plants and sharing of fungal partners between P. pinaster and Cistus spp. ECM root tips were sorted into morphological groups, photographed and symbiotic fungi were molecularly identi ed by ampli cation and sequencing of fungal ITS region (Internal Transcribed Spacer). ECM fungi showed having in general lower growth rates than saprobic fungi, as expected due to their life styles. The optimal temperature for almost all species was 24oC being sub- sequently the best temperature when producing spawn for cultivation of saprobic fungi and fungal inoculum for mycorrhizal synthesis. The only exception was P. ostreatus that grew better at 30oC. The culture medium PDA demonstrated to be the most suitable medium for axenic growth of all fungi, being a nutritional comprehensive medium ful lling the require- ments of fungi of different lifestyles. It was also proved the growth of almost all fungi in MS medium, mainly used to grow plant tissues. Ectomycorrhizas were formed, using the novel technique, in almost all mycorrhizal fungus- plant combinations. The novel technique showed in many different ways to be very useful in testing partners compatibility as well as in monitoring mycorrhizal development. Moreover, results suggest a clear plasticity of these shrubs concerning their fungal partners diversity. Several species were found associated naturally with Cistus spp. as well as sharing of fungal symbionts between Cistus salviifolius and P. pinaster. Moreover, were also found species that produce edible mushrooms associated with Cistus sp., showing these shrubs' potential to produce edible sporocarps. Results are discussed in relation to optimize methods for culture and synthesis of ecto- mycorrhizas, to understand the future application of these relationships in using mycorrhized shrubs with edible fungi as inoculum source to naturally guide pinewoods towards cultivation of a speci c edible ectomycorrhizal mushroom as well as in using Cistus shrubs to produce edible mushrooms.
Os cogumelos comest veis s~ao consumidos por todo o mundo ora como uma iguaria, ora devido as suas propriedades nutricionais e medicinais. Alguns dos mais apreciados e valoriza- dos cogumelos silvestres comest veis s~ao os carp oforos de fungos ectomicorr zicos (ECM) pelo que o interesse econ omico no seu cultivo enorme. No entanto, o cultivo economicamente rent avel de cogumelos micorr zicos comest veis e muito dif cil, contrariamente as esp ecies de fungos sapr o tas, devido, nomeadamente a necessidade de associa c~ao com ra zes de plantas e a depend^encia das taxas de crescimento e biomassa do seu simbionte vegetal. Um dos objectivos deste trabalho e a produ c~ao de cogumelos comest veis de esp ecies ECM, pelo que se realizaram ensaios com culturas de mic elio das esp ecies f ungicas e de s ntese de ectomicorrizas. Foram isoladas culturas de mic elio a partir de carp oforos das esp ecies ectomicorr zicas comest veis - Boletus fragans e Tricholoma equestre - e, para compara c~ao, das esp ecies sapr o tas Pleurotus ostreatus e Agaricus bisporus. As culturas de mic elio foram colocadas a crescer em diferentes meios de cultura - Potato Dextrose agar (PDA), Malt extract agar (MEA), Biotin-Aneurin-Folic acid (BAF), Merlin-Norkrans modi ed (MNM) e Murashige & Skoog (MS) - e em diferentes temperaturas, 4oC/24oC/30oC. As respostas de crescimento dos fungos ECM e dos sapr o tas foram comparadas. Foi tamb em poss vel determinar as condi c~oes optimas de cultura para o crescimento de mic elio para cada esp ecie testada. As esp ecies vegetais utilizadas como simbionte dos fungos ectomicorr zicos pertencem ao g enero mediterr^anico Cistus (famlia Cistaceae). E conhecido serem esp ecies ectomicorr zicas e s~ao comuns no sub-bosque dos pinhais dominados por pinheiro-bravo (Pinus pinaster) em Portugal. Para al em da capacidade de estabelecerem ectomicorrizas, os Cistus s~ao arbustos e como tal, capazes de atingir a sua biomassa m axima em menos tempo que as arvores, sendo, por isso, candidatos adequados para o cultivo de cogumelos ectomicorr zicos. Para a s ntese de ectomicorrizas foram usadas pl^antulas de tr^es esp ecies espont^aneas de Portugal - Cistus psilosepalus, Cistus salviifolius e Cistus ladanifer - e as de fungos ectomi- corr zicos comest veis Boletus fragans, Lactarius deliciosus, Tricholoma equestre e Tricholoma portentosum. Foram testados diferentes m etodos para a s ntese de micorrizas incluindo re- cipientes com diferentes formas e diferentes substratos e foi desenvolvido um novo m etodo, tanto quanto sabemos, para a s ntese de ectomicorrizas adaptado ao h abito de crescimento destes arbustos. Para conhecer a comunidade de fungos ectomicorr zicos simbiontes de Cistus em condi c~oesnaturais, foi feita uma amostragem preliminar num pinhal de pinheiro-bravo. Foram colhi- das ra zes de Cistus psilosepalus, Cistus salviifolius e de Pinus pinaster num pinhal da zona costeira. A inclus~ao de P. pinaster neste estudo visou identi car os simbiontes f ungicos naturais associados, simultaneamente, com Cistus sp. e P. pinaster. No laborat orio as pon- tas ectomicorr zicas foram separadas em grupos morfol ogicos e fotografadas. Os simbiontes f ungicos foram identi cados molecularmente atrav ez da ampli ca c~ao e sequencia c~ao da regi~ao ITS (internal transcribed spacer). Os fungos ECM mostraram ter, em m edia, taxas de crescimento mais baixas compar- ativamente aos sapr o tas, como seria de esperar devido aos seus modos de nutri c~ao. A temperatura optima para quase todas as esp ecies f ungicas foi de 24oC, sendo, desta forma, a temperatura mais adequada para a produ c~ao de in oculo para a s ntese de micorrizas, assim como de "semente" para o cultivo de fungos sapr o tas. A unica excep c~ao foi o P. ostrea- tus que cresceu melhor a 30oC. O meio de cultura PDA demonstrou ser o mais adequado para o crescimento ax enico de todos os fungos, sendo, portanto, um meio nutricionalmente abrangente que preenche os requisitos de grupos de fungos com modos de nutri c~ao difer- entes. Foi tamb em comprovado o crescimento de quase todos os fungos testados no meio MS - desenvolvido e usado maioritariamente para o crescimento de tecidos vegetais. Registou-se a forma c~ao de ectomicorrizas, em quase todas as combina c~oes fungo-planta, com o uso da nova t ecnica. A nova t ecnica mostrou ser util, de diferentes formas, tanto na monitoriza c~ao do desenvolvimento de micorrizas, como para testar a compatibilidade entre parceiros. Para al em disso, os resultados obtidos sugerem uma evidente plasticidade das esp ecies de Cistus no que respeita a diversidade de parceiros f ungicos. Relativamente a comunidade de simbiontes f ungicos ECM foi poss vel identi car um el- evado n umero de esp ecies encontradas naturalmente associadas com Cistus spp., apesar da reduzida amostragem realizada. Foi tamb em encontrada partilha de esp ecies f ungicas entre Cistus salviifolius e P. pinaster. A identi ca c~ao (at e ao momento) de uma esp ecie produtora de cogumelos comest veis naturalmente associada com Cistus sp. evidencia o potencial destes arbustos para a produ c~ao de carp oforos comest veis. Os resultados s~ao discutidos no sentido da optimiza c~ao dos m etodos para a cultura e s ntese de ectomicorrizas e tamb em com vista a aplica c~ao futura destas rela c~oes no uso dosarbustos micorrizados como fonte de in oculo para guiar, de forma natural, pinhais no sentido do cultivo de um determinado cogumelo ectomicorr zico, assim como usar os Cistus para produzir cogumelos comest veis.

by Joyce Chui Kwan Ho.
Thesis (M.Phil.)--Chinese University of Hong Kong, 1999.
Includes bibliographical references (leaves 61-75).
Abstracts in English and Chinese.
Acknowledgments --- p.i
List of Abbreviations --- p.iii
Abstract
English --- p.1
Chinese --- p.2
Chapter Chapter 1 --- General Introduction
Chapter 1.1 --- Tumor Formation --- p.3
Chapter 1.2 --- Anti-tumor Pathways --- p.4
Chapter 1.3 --- Aim of Project --- p.13
Chapter Chapter 2 --- The In Vivo effect of Polysaccharopeptide
Chapter 2.1 --- Introduction --- p.15
Chapter 2.2 --- Materials and Methods --- p.17
Chapter 2.3 --- Results --- p.18
Chapter 2.4 --- Discussion --- p.19
Chapter Chapter 3 --- Cytotoxicity
Chapter 3.1 --- Introduction --- p.23
Chapter 3.2 --- Materials and Methods --- p.26
Chapter 3.3 --- Results --- p.28
Chapter 3.4 --- Discussion --- p.28
Chapter Chapter 4 --- Anti-angiogenic Effect
Chapter 4.1 --- Introduction --- p.30
Chapter 4.2 --- Materials and Methods --- p.35
Chapter 4.3 --- Results --- p.39
Chapter 4.4 --- Discussion --- p.42
Chapter Chapter 5 --- Immunomodulation
Chapter 5.1 --- Introduction --- p.45
Chapter 5.2 --- Materials and Methods --- p.47
Chapter 5.3 --- Results --- p.50
Chapter 5.4 --- Discussion --- p.52
Chapter Chapter 6 --- General Discussion --- p.57
References --- p.61

碩士
明道大學
精緻農業學系碩士班
106
The tolerance and detoxification mechanisms of plants for heavy metals (HMs) are associated with chemical forms. Inoculation of arbuscular mycorrhizal fungi (AMF) could improve growth of water spinach, increase phosphorus (P) in plants which decrease accumulation of HMs in edible part and also decrease bioaccessibility of HMs by blanching. Experimental result showed that inoculation of AMF on water spinach could improve plant growth. It also decreased accumulation of nickel (Ni) but was opposite for cadmium (Cd). Inorganic form (FE) of cadmium and nickel are the major chemical form in water spinach. Blanching could decrease concentration of cadmium and nickel of the chemical form with high mobility in comparison to fresh tissues. In conclusion, AMF could improve plant growth in cadmium and nickel contaminated soil, decrease nickel accumulation in edible parts and bioaccessibility of Cd, chromium (Cr) and Ni by blanching.