Dissertations / Theses on the topic 'Mycelial growth and mycelial colonization'

Resumo: Objetivou-se o ajuste de metodologia e seleção de genótipos de guandu para resistência a Macrophomina phaseolina a partir de material obtido pela Embrapa Pecuária Sudeste, e verificar o desenvolvimento micelial e esporulação do fungo em meios de cultura. O trabalho foi conduzido em casa de vegetação na UNESP/Jaboticabal no período de agosto de 2004 a dezembro de 2005. Para o ajuste de metodologia e seleção de genótipos resistentes ao fungo as sementes foram submetidas a escarificação com lixa d'água e inoculação artificial através do método de exposição das mesmas ao patógeno por diferentes períodos, que variaram de O a 72 horas. Foram avaliadas porcentagem de plantas sobreviventes e massa fresca. Já para o crescimento micelial e esporulação do fungo foi utilizado o método de sobreposição de discos de diferentes hospedeiros no meio de cultura. A escarificação das sementes contribuiu para a penetração do fungo nas mesmas o período de 24h de exposição das sementes ao fungo são suficientes para detectar diferenças no grau de resistência dos genótipos. Os genótipos mais resistentes são g167-97, g124-95, g27-94, g40-95, g154-95, g127-97 e g9m-97, e os mais suscetíveis são g48-95, g123-95, g8-95, g168-99 e g1m-95. A sobreposição de discos foliares de guandu em meio BDA e folha de papel de filtro em meio sojinha proporcionam um incremento na esporulação de M. phaseolina.<br>Abstract: This work had the objective of determining the best schedule for artificial inoculation and select pigeon pea genotypes resistant to Macrophomina phaseolina in material obtained by Embrapa Pecuária Sudeste, and verify the mycelial growth and sporulation of the fungi in middle of culture. The work were carried in greenhouse at the UNESP/Jaboticabal, from August 2004 to December 2005. For the methodology and selection adjustment of resistant genotypes to the fungi the seeds were submitted scarified with water sandpaper and artificial inoculation the seeds were the contact method to fungi for different periods, which varied from O to 72 hours. They were evaluated percentage of surviving plants and fresh mass. For the mycelial growth and sporulation of the fungi was used the superposition of disks method of different hosts in the middle of culture. The scarified of the seeds contributed for penetration of the fungi at the seeds; the period of 24h of contact of the seeds to the fungi enough to detect differences in the resistance degree ofthe genotypes. The genotypes g167-97, g124-95, 927-94, g40-95, g154-95, g127-97 and g9m-97 were found to be the most resistant and most susceptible were g48-95, g123-95, g8-95, g168-99 and g1m-95. The treatment with superposition of the leaf disks of pigeon pea in BDA and disks of filter paper in middle of soybean extract were the treatments that provided better sporulation levei in the conditions of that experiment were half.<br>Orientador: Rita de Cássia Panizzi<br>Coorientador: Rodolfo Godoy<br>Banca: Antonio de Goes<br>Banca: Patrícia Menezes Santos<br>Mestre

Submitted by Geyciane Santos (geyciane_thamires@hotmail.com) on 2015-07-03T15:37:47Z No. of bitstreams: 1 Dissertação - Tamiris Rio Branco da Fonseca.pdf: 1102398 bytes, checksum: e70c786229e1b956a358698df9fb090b (MD5)<br>Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-08T18:09:02Z (GMT) No. of bitstreams: 1 Dissertação - Tamiris Rio Branco da Fonseca.pdf: 1102398 bytes, checksum: e70c786229e1b956a358698df9fb090b (MD5)<br>Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-08T18:13:13Z (GMT) No. of bitstreams: 1 Dissertação - Tamiris Rio Branco da Fonseca.pdf: 1102398 bytes, checksum: e70c786229e1b956a358698df9fb090b (MD5)<br>Made available in DSpace on 2015-07-08T18:28:04Z (GMT). No. of bitstreams: 1 Dissertação - Tamiris Rio Branco da Fonseca.pdf: 1102398 bytes, checksum: e70c786229e1b956a358698df9fb090b (MD5) Previous issue date: 2013-05-02<br>CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico<br>Mushrooms arouse interest for presenting high nutritional and medicinal value. The representatives of the genus Pleurotus are able to grow in different agricultural and agro-industrial waste, since they secrete numerous enzymes that hydrolyze the substrates prepared from waste providing nutrient The search for alternative substrates increased in recent years due to the availability of regional waste little or never used in the production of edible mushrooms. The use of waste in solid-state fermentation has emerged also in the production of enzymes. Proteases are a group of enzymes commercially exploited, however the use of Basidiomycetes as producers of proteases is still a relatively unexplored. The aim of this study was to evaluate the mycelial growth as well as to investigate the production and partially characterize extracellular proteases from Pleurotus ostreatoroseus DPUA 1720 in different agro-industrial waste, selecting a mixture for the production of the mushroom and check their microbiological quality and nutritional value. Cultures were prepared in potato dextrose added yeast extract 0.5% (w/v). The vertical mycelial growth and production of proteases from P. ostreatoroseus DPUA 1720 were performed in agro-industrial waste (acai seed, sawdust, cupuassu bark, rice bran, bark and pineapple crown) for 15 days under two growing conditions (presence and absence of light). The production of basidiomata was performed in the substrate which presented satisfactory mycelial growth and vigorous mycelium. From this bioprocess evaluated the formation and development of early basidiomata, total time of cultivation, biological efficiency, productivity, production rate, loss of organic matter, microbiological and nutritional value of the mushroom. P. ostreatoroseus DPUA 1720 grew in all industrial residues also tested serving as sources for the production of proteases. In bark cupuaçu added rice bran mycelium was strongly vigorous and the mycelial growth satisfactory, therefore selected for the production of P. ostreatoroseus DPUA 1720. Under the experimental conditions the proteolytic activity was determined in crude extracts from the substrates of vertical mycelial growth with maximum proteolytic activity (7.89 U/mL) in bark cupuassu added rice bran in the presence of light. This substrate was the most efficient for the production of proteases with activity atpH 6.0 and optimum temperature at 40 ˚C. The basidiomata showed no contaminants, low in fat, high in fiber and protein, macro and micro minerals, essential and non-essential amino acids, may therefore be inserted into the feed as a safe and nutritious food.<br>Os cogumelos despertam interesse por apresentar alto valor nutricional e medicinal. Os representantes do gênero Pleurotus são capazes de crescer em diferentes resíduos agrícolas e agroindustriais, pois secretam inúmeras enzimas que hidrolisam os substratos elaborados a partir dos resíduos disponibilizando nutrientes. A busca por substratos alternativos cresceu nos últimos anos em virtude da disponibilidade de resíduos regionais pouco ou nunca utilizados na produção de cogumelos comestíveis. O uso de resíduos na fermentação semi-sólida vem despontando também na produção de enzimas. As proteases são o grupo de enzimas mais exploradas comercialmente, no entanto o uso de Basidiomicetos como produtores de proteases ainda é um tema pouco explorado. O objetivo deste trabalho foi avaliar o crescimento micelial, bem como investigar a produção e caracterizar parcialmente proteases extracelulares de Pleurotus ostreatoroseus DPUA 1720 em diferentes resíduos agroindustriais, selecionar uma mistura para produção dos basidiomas e verificar sua qualidade microbiológica e valor nutricional. As culturas foram preparadas em ágar batata dextrose acrescido de extrato de levedura 0,5% (p/v). O crescimento micelial vertical e a produção de proteases de P. ostreatoroseus foram realizados em resíduos agroindustriais (semente de açaí, serragem, casca de cupuaçu, farelo de arroz, casca e coroa de abacaxi) durante 15 dias, sob duas condições de cultivo (presença e ausência de luz). A produção do basidioma foi realizada no substrato que apresentou crescimento micelial satisfatório e micélio vigoroso. A partir desse bioprocesso foi avaliada a formação e desenvolvimento dos primórdios, tempo total de cultivo, eficiência biológica, produtividade, taxa de produção, perda de matéria orgânica, qualidade microbiológica e valor nutricional dos basidiomas. P. ostreatoroseus DPUA 1720 cresceu em todos os resíduos agroindustriais testados, também servindo como fontes para a produção de proteases. Em casca de cupuaçu adicionado de farelo de arroz o micélio foi fortemente vigoroso e o crescimento micelial satisfatório, por isso selecionado para produção de P. ostreatoroseus DPUA 1720. Nas condições experimentais a atividade proteolítica foi determinada nos extratos brutos provenientes dos substratos miceliados do crescimento micelial vertical com máxima atividade proteolítica (7,89 U/mL) em casca de cupuaçu adicionado de farelo de arroz na presença de luz. Este substrato foi o mais eficiente para a produção das proteases com atividade em pH 6,0 e temperatura ótima a 40˚C. Os basidiomas apresentaram ausência de contaminantes, baixo teor de lipídios, alto teor de fibras e proteínas, macro e microminerais, aminoácidos essenciais e não essenciais, podendo assim ser inserido na alimentação como um alimento saudável e nutritivo.

Made available in DSpace on 2015-04-11T13:55:47Z (GMT). No. of bitstreams: 1 Marcus Vinicius Lopes de Lima.pdf: 1036854 bytes, checksum: ac6f00a175ea31a35d39d2469140ccaf (MD5) Previous issue date: 2012-08-31<br>Coordenação de Aperfeiçoamento de Pessoal de Nível Superior<br>The dark-brown spot is a disease that occurs in the leaves of the tomato tree and is caused by the fungus Corynespora cassiicola. The fungus is considered cosmopolitan and nonspecific because of the wide host range and geographical distribution. Due to the absence of resilient and grow commercial chemicals registered in Brazil to control the disease the same is done with the use of alternative products so healing. This paper reports the in vitro antifungal effect of the ethanol extract phases and volatile oil from Piper aduncum and prophylactic and curative effect of aqueous extract of P. aduncum in tomato seedlings. The aqueous extract was obtained by macerating 300 g of fresh leaves in 2 L of water, ethanol extract by means of cold maceration of leaves with ethanol, the essential oil obtained by the method of hydrodistillation of the dry leaves, the the hexane, chloroform, N-butanol and hydroalcoholic through the fractionation of the ethanol extract by the process of liquid-liquid partition. For in vitro assays were performed in petri dishes, the antifungal effect of the ethanol extract phases and volatile oil of P. aduncum on mycelial growth, and germination of conidia blades while in vivo via inoculation of conidia suspension of 104 conidia mL-1 in tomato seedlings of the cultivar Santa Cruz Kada, the analyzes were done before and after treatment with aqueous extract, evaluations of prophylactic and curative effect, respectively. The volatile oil of P. aduncum showed no inhibition on the mycelial growth of C. cassiicola concentrations tested. The ethanol extract, the hexane and chloroform showed antifungal effect in 10000 μg.mL-1 concentration on mycelial growth and spore germination. The minimum inhibitory concentration on the mycelial growth was in 2000 μg.mL-1 of the hexane phase, this same concentration was found for the germination of conidia in the ethanol extract. The aqueous extract of P. aduncum the test of curative effect at dosages 1:1 and 1:2, showed significant effect in reducing the severity of the disease. The calculation of AUDPC confirming curative effect at all doses tested. In testing the prophylactic aqueous extract showed no effectiveness in controlling the disease<br>A mancha alvo do tomateiro é uma doença que ocorre nas folhas e é causada pelo fungo Corynespora cassiicola. O fungo é considerado cosmopólita e inespecífico devido à ampla gama de hospedeiros e distribuição geográfica. Devido à inexistência de cultivarem comerciais resistentes e produtos químicos registrados no Brasil para o controle da doença, o mesmo é feito com uso de produtos alternativos de forma curativa. Este trabalho relata o efeito antifúngico in vitro do extrato etanólico, fases e óleo volátil de Piper aduncum e o efeito curativo e profilático do extrato aquoso de P. aduncum em mudas de tomateiro. O extrato aquoso foi obtido por meio da maceração de 300 g de folhas verdes em 2 L de água; O extrato etanólico, por meio da maceração a frio de folhas secas com etanol; o óleo volátil obtido pelo método da hidrodestilação das folhas secas; as fases hexânica, clorofórmica, N-butanólica e hidroalcóolico por meio do fracionamento do extrato etanólico pelo processo de partição líquido-líquido. Para as análises in vitro, foram realizadas em placas de petri o efeito antifúngico do extrato etanólico, fases e óleo volátil de P. aduncum sobre o crescimento micelial, e em lâminas a germinação dos conídios, enquanto que, in vivo, via inoculação de suspensão de inóculo na concentração de 104 conídios.mL-1 em mudas de tomateiro do cultivar Santa Cruz Kada, as análises foram feitas antes e após tratamento com extrato aquoso, nas avaliações do efeito curativo e profilático, respectivamente. O óleo volátil de P. aduncum não apresentou nenhuma inibição sobre o crescimento micelial de C. cassiicola nas concentrações testadas. O extrato etanólico, fases hexânica e clorofórmica apresentaram efeito antifúngico na concentração 10000 µg.mL-1 sobre o crescimento micelial e germinação dos conídios. A concentração inibitória mínima sobre o crescimento micelial foi em 2000 µg.mL-1 da fase hexânica, nesta mesma concentração foi encontrado para a germinação dos conídios no extrato etanólico. O extrato aquoso de P. aduncum no teste de efeito curativo nas dosagens 1:1 e 1:2, apresentaram efeito significativo na redução da severidade da doença. O cálculo da AACPD confirma o efeito curativo em todas as dosagens testadas. No teste profilático o extrato aquoso não apresentou nenhuma efetividade no controle da doença

Made available in DSpace on 2015-04-11T13:55:51Z (GMT). No. of bitstreams: 1 Francy Mary Sousa.pdf: 1356959 bytes, checksum: a7db48bf3163f386db3bca96f187f815 (MD5) Previous issue date: 2010-11-30<br>Coordenação de Aperfeiçoamento de Pessoal de Nível Superior<br>Corynespora cassiicola has been related causing damages in several crops of economic importance in Amazonas State, principally Solanaceae and Cucurbitaceae families affecting production by significative form. This study aimed to characterize C. cassiicola isolates through colony morphology, sporulation, mycelial growth and spore dimension on culture media PDA, PSA, OEA and CA. Five mm-diameter disks taken from colonies grown on PDA medium for 15 days were transferred to Petri dishes containing each medium under room temperature and continuous light. The study was installed using randomized block design in a factorial 4 x 5 x 3 (culture media x isolates x hosts) with five replications. The effects of culture media varied to isolates of same host and among different hosts. Mycelial growth was observed in CA. The most spore production was observed in PDA and PSA media. A high variation of aspect and colonies coloration was observed among same host isolates and isolates of different hosts in all culture media. Variation trough spore dimension of C. cassiicola isolates was observed in all culture media tested.<br>O fungo Corynespora cassiicola já foi relatado causando danos em diversas culturas de interesse comercial no Estado do Amazonas, principalmente das famílias Solanaceae e Cucurbitaceae, afetando de forma significativa a produção. Este estudo teve como objetivo caracterizar isolados de C. cassiicola quanto a morfologia das colônias, esporulação, crescimento micelial e dimensão dos conídios nos meios de cultura BDA, BSA, AvA e CA. Discos de 5mm de diâmetro retirados de colônias crescidas em meio BDA por 15 dias, foram transferidos para o centro de placas de Petri contendo os meios de cultura e mantidas à temperatura ambiente sob luz contínua. O estudo seguiu esquema fatorial 4 x 5 x 3 (meios de cultura x isolados x hospedeiros) com cinco repetições. Os efeitos dos meios de cultura variaram para isolados do mesmo hospedeiro e entre hospedeiros diferentes. Maior crescimento micelial foi observado no meio CA. A maior produção de esporos foi observada nos meios BDA e BSA. Foi verificado uma ampla variação de aspecto e coloração das colônias entre os isolados do mesmo hospedeiro e entre isolados de hospedeiros diferentes em todos os meios de cultura. Também houve variação quanto às dimensões dos conídios dos isolados de C. cassiicola em todos os meios de cultura testados.

Submitted by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2018-03-06T14:24:29Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Reprodução Não Autorizada.pdf: 47716 bytes, checksum: 0353d988c60b584cfc9978721c498a11 (MD5)<br>Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2018-03-06T14:24:43Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Reprodução Não Autorizada.pdf: 47716 bytes, checksum: 0353d988c60b584cfc9978721c498a11 (MD5)<br>Made available in DSpace on 2018-03-06T14:24:43Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Reprodução Não Autorizada.pdf: 47716 bytes, checksum: 0353d988c60b584cfc9978721c498a11 (MD5) Previous issue date: 2009-02-21<br>FAPEAM - Fundação de Amparo à Pesquisa do Estado do Amazonas<br>This work presents the analysis of mycelial growth of Pleurotus ostreatus (Jacq .: Fr.) Kumm. DPUA 1533, Pleurotus ostreatus (Jacq.: Fr.) Quél. cv. Florida DPUA 1534, Lentinus citrinus Walleyn & Rammeloo DPUA 1535 and Neolentinus lepideus (Fr.) Redhead & Ginns DPUA 1536, in natural and complex media under different growing conditions to verify production of biocompounds against fungi and bacteria, to evaluate vertical mycelial growth, to analyze the production of proteases and biomass in agroindustrial residues selected by solid fermentation. Antimicrobial activity of mushrooms against Candida albicans DPUA 1336, Cryptococcus laurentii DPUA 1501, Aspergillus flavus DPUA1836, Escherichia coli DAUPE 224 and Mycobacterium smegmatis ATCC 607 was determined by the Gelose Block Method and Minimum Inhibitory Concentration. For the production of Pleurotus and Lentinus proteases by solid state fermentation, the cultivations were realized on 200 g of cupuaçu shell + rice husk (CC+CA); cupuaçu shell + bran rice (CC+FA), in flask of transparent glass, humidity 60% and pH 6,0 and the sterilization of the substrates was accomplished at 121 °C/90 minutes for two consecutive days. Each flask was inoculated with 12 agar disks (10 mm in diameter) from stock culture and allowed to fermented for 15 days at 25 °C, without light and mycelial growth was determined in centimeters every 24 hours. For determination of proteolityc activity at 440 nm solution of azocasein 1% (p/v), pH 7,2 was used. L. citrinus was the specie selected for biomass production. In this stage for the accomplishment of the fermentation in the solid-state were prepared 5 kg plastic bags containing different combinations and proportions of cupuaçu shell, rice husk and rice bran. Methods were carried out to evaluate the formation, development of the primordia, total culture time, number of production flows and analysis of centesimal composition of basidiomas of L. citrinus in triplicate. The results demonstrate that the composition of culture media, the time of cultivation, the light and the temperature influence significantly in the growth of Lentinus and Pleurotus species, on culture media analyzed. In the experimental conditions, those mushrooms express antimicrobial activity against most fungi and bacteria tested, however L. citrinus and N. lepideus didn't express antimicrobial activity against Aspergillus flavus. The mixture of cupuaçu shell and rice husk (1: 1) stimulated the vertical growth rate of L. citrinus (13 cm), P. ostreatus (12.6 cm) and N. lepideus (12.3 cm). The best averages of the activity and effect of the proteolytic enzyme extraction conditions were determined in the crude extract of P. ostreatus (24.0 U / g) and L. citrinus (22.0 U / g) obtained in the NaCl 2 solution % at 36 °C and with distilled water at 25 °C. In the biomass production, the best averages of biological efficiency (61.0%), productivity (0.86%), production rate (32.0%) and protein (13.44%) were observed in the formulations different agroforestry substrates that depending on the concentration have demonstrated the viability of the process for the growth and production of L. citrinus DPUA 1535.<br>Este trabalho apresenta a análise do crescimento de Jacq.: Fr.) Kumm. DPUA 1533, Pleurotus ostreatus (Jacq.: Fr.) Quél. cv. Florida DPUA 1534, Lentinus citrinus Walleyn & Rammeloo DPUA 1535 e Neolentinus lepideus (Fr.) Redhead & Ginns DPUA 1536, em meios natural e complexo sob diferentes condições de cultivo, para verificar a produção de biocompostos contra fungos e bactérias, avaliar o crescimento micelial vertical, analisar a produção das proteases e biomassa em resíduos agroindustriais selecionados por fermentação sólida. A atividade antimicrobiana dos cogumelos frente à Candida albicans DPUA 1336, Cryptococcus laurentii DPUA 1501, Aspergillus flavus DPUA 1836, Escherichia coli DAUPE 224 e Mycobacterium smegmatis ATCC 607 foi determinada pelo Método do Bloco de Gelose e pela Concentração Mínima Inibitória. Para a produção de proteases de Pleurotus e Lentinus por fermentação no estado sólido, os cultivos foram realizados em 200 g de casca de cupuaçu + casca de arroz (CC+CA); casca de cupuaçu + farelo de arroz (CC+FA), em frasco de vidro transparente, umidade 60% e pH 6,0. A esterilização dos substratos foi realizada a 121 °C/90 minutos/dois dias consecutivos. Em cada frasco foram inoculados 12 discos de 10 mm de diâmetro e a fermentação foi conduzida a 25 oC, sem luminosidade por 15 dias, determinando-se a velocidade de crescimento micelial em centímetros, a cada 24 horas. Para determinação da atividade proteolítica a 440 nm foi utilizado solução de azocaseina 1% (p/v), pH 7,2. L. citrinus foi a espécie selecionada para produção de biomassa. Nesta etapa para a realização da fermentação no estado sólido foram preparados sacos plásticos de 5 kg contendo diferentes combinações e proporções de casca de cupuaçu, casca de arroz e farelo de arroz. Efetuaram-se processos para avaliar a formação, desenvolvimento dos primórdios, tempo total de cultivo, número de fluxos de produção e análises de composição centesimal de basidiomas de L. citrinus em triplicata. Os resultados demonstraram que a composição dos meios de cultura, o tempo de cultivo, a luz e a temperatura influenciam significativamente no crescimento das espécies de Lentinus e Pleurotus, nos meios de cultura analisados. Nas condições experimentais, esses cogumelos expressaram atividade antimicrobiana contra a maioria dos fungos e bactérias testados, contudo L. citrinus e N. lepideus não expressaram atividade antimicrobiana frente Aspergillus flavus. A mistura de casca de cupuaçu com casca de arroz (1:1) estimulou a velocidade de crescimento vertical de L. citrinus (13 cm), P. ostreatus (12,6 cm) e N. lepideus (12,3 cm). As melhores médias da atividade e efeito das condições de extração de enzimas proteolíticas foram determinadas no extrato bruto de P. ostreatus (24,0 U/g) e L. citrinus (22,0 U/g), obtidos na solução de NaCl 2%, a 36 °C e com água destilada, a 25 °C. Na produção de biomassa, as melhores médias de eficiência biológica (61,0 %), produtividade (0,86 %), taxa de produção (32,0 %) e proteína (13,44 %) foram observadas nas formulações realizadas com os diferentes substratos agroflorestais que dependendo da concentração têm demonstrando a viabilidade do processo para o crescimento e produção de L. citrinus DPUA 1535.

Made available in DSpace on 2014-06-11T19:28:30Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-05-15Bitstream added on 2014-06-13T20:58:17Z : No. of bitstreams: 1 rosa_j_me_jabo.pdf: 154567 bytes, checksum: 0d2897a42007e37a9a5f314b75f0f0f0 (MD5)<br>Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)<br>Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)<br>Objetivou-se o ajuste de metodologia e seleção de genótipos de guandu para resistência a Macrophomina phaseolina a partir de material obtido pela Embrapa Pecuária Sudeste, e verificar o desenvolvimento micelial e esporulação do fungo em meios de cultura. O trabalho foi conduzido em casa de vegetação na UNESP/Jaboticabal no período de agosto de 2004 a dezembro de 2005. Para o ajuste de metodologia e seleção de genótipos resistentes ao fungo as sementes foram submetidas a escarificação com lixa d'água e inoculação artificial através do método de exposição das mesmas ao patógeno por diferentes períodos, que variaram de O a 72 horas. Foram avaliadas porcentagem de plantas sobreviventes e massa fresca. Já para o crescimento micelial e esporulação do fungo foi utilizado o método de sobreposição de discos de diferentes hospedeiros no meio de cultura. A escarificação das sementes contribuiu para a penetração do fungo nas mesmas o período de 24h de exposição das sementes ao fungo são suficientes para detectar diferenças no grau de resistência dos genótipos. Os genótipos mais resistentes são g167-97, g124-95, g27-94, g40-95, g154-95, g127-97 e g9m-97, e os mais suscetíveis são g48-95, g123-95, g8-95, g168-99 e g1m-95. A sobreposição de discos foliares de guandu em meio BDA e folha de papel de filtro em meio sojinha proporcionam um incremento na esporulação de M. phaseolina.<br>This work had the objective of determining the best schedule for artificial inoculation and select pigeon pea genotypes resistant to Macrophomina phaseolina in material obtained by Embrapa Pecuária Sudeste, and verify the mycelial growth and sporulation of the fungi in middle of culture. The work were carried in greenhouse at the UNESP/Jaboticabal, from August 2004 to December 2005. For the methodology and selection adjustment of resistant genotypes to the fungi the seeds were submitted scarified with water sandpaper and artificial inoculation the seeds were the contact method to fungi for different periods, which varied from O to 72 hours. They were evaluated percentage of surviving plants and fresh mass. For the mycelial growth and sporulation of the fungi was used the superposition of disks method of different hosts in the middle of culture. The scarified of the seeds contributed for penetration of the fungi at the seeds; the period of 24h of contact of the seeds to the fungi enough to detect differences in the resistance degree ofthe genotypes. The genotypes g167-97, g124-95, 927-94, g40-95, g154-95, g127-97 and g9m-97 were found to be the most resistant and most susceptible were g48-95, g123-95, g8-95, g168-99 and g1m-95. The treatment with superposition of the leaf disks of pigeon pea in BDA and disks of filter paper in middle of soybean extract were the treatments that provided better sporulation levei in the conditions of that experiment were half.

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-03-23T11:38:02Z No. of bitstreams: 1 Paulo Cezar das Merces Santos.pdf: 945938 bytes, checksum: 4dcffae9a84258c9e4e04fc53d685685 (MD5)<br>Made available in DSpace on 2017-03-23T11:38:02Z (GMT). No. of bitstreams: 1 Paulo Cezar das Merces Santos.pdf: 945938 bytes, checksum: 4dcffae9a84258c9e4e04fc53d685685 (MD5) Previous issue date: 2011-02-17<br>Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq<br>From a socioeconomic perspective, the banana, one of the most consumed fruit, has great importance worldwide. The Brazilian production of this fruit presents particular geographic distribution, since it is present in all States of the federation. In the State of Pernambuco, the banana is grown in all the regions, occupying about 80000 permanent jobs in the field, with two distinct poles of banana production: the Zona da Mata and Vale do São Francisco. Among the limiting factors for this crop, there are the post-harvest diseases such as anthracnose, caused by Colletotrichum musae. This disease causes significant losses in production, manifesting itself mainly in the fruit at maturity stage. In order to verify the diversity of this pathogen isolated from C. musae from banana production fields of three municipalities (Vicência, São Vicente Férrer and Machados) of the Pernambuco State were characterized for daily mycelial growth rate (DMGR), colony color, reverse plate, topography, presence of sectors and microsclerotia, conidial morphology, biochemical tests, pathogens and molecular. The 60 isolates, based on morphological characters of shape and size of conidia, were identified as C. musae, followed with morphological measurements from 9.3 to 30.2 x 2.6 to 12.6 μm ( x = 15.0 x 4.9 μm, n = 6000). Most isolates showed conidia straight, oblong, with rounded apices. As for the DMGR, all isolates showed growth averaged more than 1.0 cm.day–1, ranging from 1.21 to 1.63 cm.day–1. It was found three groups of striking color to the colonies: white, cream and salmon, while the presence of sectors ranging from zero to eight per isolate. Most isolates (53) metabolized the ammonium tartrate as the sole source of carbon, while for citric acid, all isolates except the isolate CMM 3219, were not able to metabolize this carbon source. The pathogenic characterization showed significant differences in the epidemiological parameter AUDPC in fruits inoculated with injury, indicating the existence of variation in aggressiveness among isolates. For fruit inoculated and no injuries, the AUDPC was not estimated for 20 isolates because there was no injury. The dendrogram generated by UPGMA (Unweighted Pair Group Method with Arithmetic Mean) cluster analysis of DNA bands obtained with the combination of three primers (GTG5, GACA and 820) by ISSR-PCR allowed the separation of three groups of isolates to 62% by Jaccard’s similarity coefficient. The group I was composed of isolates from Vicência Group II isolates São Vicente Férrer and group III, Machados. The results suggest that isolates of C. musae can be discriminated as to their origin because they had genetic similarity between them. The ISSR markers in this study appear to be useful tools in the study of C. musae.<br>Do ponto de vista socioeconômico, a banana, uma das frutas mais consumidas, possui grande importância mundial. A produção brasileira dessa frutífera apresenta particular distribuição geográfica, já que está presente em todos os Estados da federação. No Estado de Pernambuco, a banana é cultivada em todas as microrregiões, ocupando cerca de 80 mil postos de trabalho permanente no campo, havendo dois pólos distintos de produção de banana: a Zona da Mata e o Vale do São Francisco. Dentre os fatores limitantes para esta cultura, destacam-se as doenças pós-colheita, como a antracnose, causada pelo fungo Colletotrichum musae. Essa doença causa perdas significativas na produção, manifestando-se, principalmente, na fruta em estádio de maturação. Com a finalidade de verificar a diversidade desse fitopatógeno, isolados de C. musae provenientes de campos de produção de banana de três municípios (Vicência, São Vicente Férrer e Machados) do Estado de Pernambuco foram caracterizados quanto à taxa de crescimento micelial diária (TCMD), coloração da colônia, reverso da placa, topografia, presença de setores e de microescleródios, além da morfologia de conídios, testes bioquímicos, patogênicos e moleculares. Os 60 isolados, com base nos caracteres morfológicos de formato e tamanho de conídios, foram identificados como C. musae, com as seguidas medidas morfológicas 9,3-30,2 x 2,6-12,6 μm ( x = 15,0 x 4,9 μm, n = 6000). A maioria dos isolados avaliados apresentaram conídios retos, oblongo, com ápices arredondados. Quanto à TCMD, todos os isolados apresentaram crescimento médio superior a 1,0 cm.dia–1, variando de 1,21 a 1,63 cm.dia–1. Foram encontrados três grupos marcantes de coloração de colônia: branca, creme e salmão, enquanto que a presença de setores variou de zero a oito por isolado. A maioria dos isolados (53) metabolizou o tartarato de amônio como fonte exclusiva de carbono, enquanto que para o ácido cítrico, todos os isolados, com exceção do isolado CMM 3219, não foram capazes de metabolizar essa fonte de carbono. A caracterização patogênica mostrou diferenças significativas para o parâmetro epidemiológico AACPD em frutas inoculadas com ferimento, indicando a existência de variação da agressividade entre os isolados. Para as frutas sem ferimentos e inoculadas, a AACPD não foi estimada para 20 isolados, pois não houve lesões. O dendrograma gerado pela análise de agrupamento UPGMA (Unweighted Pair Group Method with Arithmetic Mean) das bandas de DNA obtidas com a combinação dos três primers (GTG5, GACA e 820) por ISSR-PCR possibilitou a separação de três grupos de isolados a 62% pelo coeficiente de similaridade de Jaccard. O grupo I foi formado pelos isolados provenientes de Vicência; grupo II, isolados de São Vicente Férrer e grupo III, de Machados. Os resultados sugerem que isolados de C. musae podem ser discriminados quanto à sua origem por apresentarem similaridade genética entre si. Os marcadores ISSR, nesse estudo, parecem ser ferramentas bastante úteis no estudo populacional de C. musae.

Made available in DSpace on 2016-12-08T16:44:32Z (GMT). No. of bitstreams: 1 PGPV07MA022.pdf: 293867 bytes, checksum: 7be800f613b76704595bb5911a0eeab8 (MD5) Previous issue date: 2007-02-26<br>The apple culture has great social and economic importance for the South region of Brazil and in the 2003/04 crop season produced approximately 970,000 tons. During the 1995/96 crop season, a diferente symptom from the mosto f symptoms observed in apple fruit and branches were evidenced in Rio Grande do Sul and Santa Catarina orchards. After etiological studies, it was determined the occurrence of fungus Cryptosporiopsis perrennans. This pathogen can cause rota round of 15% on storage apples and for the time been there is few studies about the patogen and the epidemiology of disease. The purpose of this work was to study the development and the production of C. perennans spores over diferente cultures medium with and without fungicides, and to determinethe effect of temperature and humidity periods over the Bull s eye roto n Fuji apple. The experiments carriedout were the following: a) evaluatoon of the effect of potato-dextrose-agar (PDA), malt extract (MEM) and V8 juice (V8) médium on the development and production of spores by 9 isolates of C. perennans; b) evaluation of sensibility in vitro of C. perennans to six diferente concentrations of the fungicides captan, cooper hidroxide, dithianon, mancozev and trifloxistrobin diluted in PDA; c) evaluation of incidence and severity of the Bull eye s rot when Fuji apples inoculated with C. perennans had been submited to diferente combinations of humidity and temperature periods. The results showedthat the medium AEM and V8 promoted better stimulus to mycelium growth and that five C. perennans isolates produced more spores when cultivated in V8 and four isolates in MEM. The diferente fungicides tested were eficiente to control C. perennans micelial growth, however the most promising results were obtained the fungicides dithianon and mancozeb. Higher incidence and severity of the Bull eye s roto n Fuji apples were obteined with 20 hours of incubation at 22°C<br>A cultura da macieira tem grande importância sócio-econômica para a região sul do Brasil tendo atingido no ciclo 2003/04 aproximadamente 970.000 toneladas. Nos ciclos vegetativos de 1995 e 1996, um sintoma diferente daqueles relatados até então no Brasil em frutos e ramos da macieira foi constatado em pomares do Rio Grande do Sul e de Santa Catarina. Após o estudo de sua etiologia, identificou-se a ocorrência do fungo Cryptosporiopsis perennans. Este patógeno pode causa podridões ao redor de 15% das maçãs no armazenamento e, no presente, há poucos estudos que contribuam ao conhecimento do patógeno ou à epidemiologia da doença. Este trabalho visou estudar o crescimento micelial e a produção de conídios de isolados de C. perennans em meios de cultura com e sem fungicidas e determinar o efeito de temperaturas e períodos de umidade na infecção de maçãs Fuji . As ações conduzidas foram: a) determinação os efeitos dos meios de cultura batata-dextrose-ágar (BDA), ágar-extrato-de-malte (AEM) e ágar-sucoV8 (V8) no crescimento micelial e produção de conídios de nove isolados de C. perennans; b) avaliação a sensibilidade in vitro de C. perennans a seis concentrações dos fungicidas captan, hidróxido de cobre, dithianon, mancozeb e trifloxistrobin incorporados ao meio de cultura BDA; c) avaliação da incidência e severidade da podridão olho-de-boi em maçãs Fuji inoculadas com C. perennans e submetidas a diferentes combinações de períodos de umidade e temperatura. Os resultados mostraram que o maior estímulo ao crescimento do micélio do patógeno foi induzido pelos meios de cultura AEM e V8 e que cinco dos nove isolados avaliados produziram maior quantidade de conídios quando cultivados em suco V8 e quatro em AEM. Todos os fungicidas apresentaram-se eficientes para o controle do crescimento micelial de C. perennans quando comparados a testemunha, porém os resultados mais promissores foram obtidos com os fungicidas dithianon e mancozeb. A temperatura e período de incubação que proporcionaram maior índice de incidência e severidade da podridão olho-de-boi em frutos de maçã cv. Fuji, foram de 22°C com 20 horas de incubação

Orientador: Robinson Antonio Pitelli<br>Banca: Rita de Cassia Panizzi<br>Banca: Margarete Camargo<br>Banca: Glaucia de Figueiredo Nachtigal<br>Banca: Marta Cecília Malassen Nemoto<br>Resumo: Sagittaria montevidensis é uma planta aquática emergente, de difícil controle desde que se tornou problemática em arroz irrigado. Infestante em corpos hídricos de usinas hidrelétricas, o biocontrole com o fungo Cylindrocarpon sp. empregado na estratégia inundativa, tem sido sugerido para manejo dessa macrófita onde o uso do controle químico tem se mostrado ineficiente devido ao desenvolvimento de resistência aos herbicidas inibidores da enzima ALS (acetolactato sintase). O presente trabalho foi conduzido em casa de vegetação e no laboratório de Controle Biológico de Plantas Daninhas Prof. Giorgio de Marinis, da FCAV - UNESP, Jaboticabal, visando avaliar a especificidade do agente de biocontrole em plantas não alvo de diferentes famílias botânicas e determinar a severidade da doença em função do número e intervalos de aplicações do bioherbicida, do período de orvalho e idade da planta além de estimar os efeitos de adjuvantes sobre o desenvolvimento de Cylindrocarpon sp. no controle de S. montevidensis. O fungo mostrou-se específico a S. montevidensis, não sendo constatado sintomas da infecção fúngica em plantas de interesse econômico e nas plantas aquáticas Pistia stratiotes, Eichhornia crassipes, E. azurea, Salvinia sp., Azolla caroliniana, Lemna minor, Heteranthera reniformis, Hydrocotyle verticillata, Pontederia rotundifolia, P. parviflora, P. cordata, Polygonum lapathifolium, Ludwigia sedoides, L. elegans, L. repens, L. helminthorrhiza, Potamogeton pectinatus, Cerathophylum demersum, Myriophyllum aquaticum, Egeria densa, E. najas, Najas guadalupensis, Hydrilla verticillata, Alternanthera phyloxeroides, Nimphoides indica. A doença foi mais severa quando o bioherbicida foi aplicado em plantas com 3 a 4 folhas e na fase de inflorescência, submetidas as condições de período de orvalho superior a 6 horas ...(Resumo completo, clicar acesso eletrônico abaixo)<br>Abstract: Sagittaria montevidensis is a very important aquatic in shallow water bodies in wetland rice, mainly recentely because tolerant biotipes is raising in areas with repetitive spraying with ALS inhibitor herbicide. The fungus Cylindrocarpon sp. has a good potential as biocontrol agent for this control action of this fungus against S. montevidensis under different conditions of spore concentration, dew period, number and intervals of spraying and surfactants adittion in the spraying solution. The fungus specificity also was evaluated in aquatic and crop plants. The fungus showed high specificity to S. montevidensis. The disease severity was more strong when the plants were inoculated a 3-4 leaf stage and during blooming. A dew period longer than 6 hours was enough for to reach the maximum disease severity. Two a three spraying with the fungus enhanced the disease severity comparing with a single application and the intervals of 21 days between spraying produced better control than intervals of 7 and 14 days. Among the surfactants, Silwet L-77AG and Iharaguen-S improved the disease severity while Energic and Herbitensil reduced the Cylindrocarpon sp. disease severity.<br>Doutor

Made available in DSpace on 2014-06-11T19:33:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-07-11Bitstream added on 2014-06-13T20:25:33Z : No. of bitstreams: 1 fiorillo_cmt_dr_jabo.pdf: 399101 bytes, checksum: 0abac3d781532f7b3d6e83aad7d31df7 (MD5)<br>Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)<br>Funep/Light<br>Sagittaria montevidensis é uma planta aquática emergente, de difícil controle desde que se tornou problemática em arroz irrigado. Infestante em corpos hídricos de usinas hidrelétricas, o biocontrole com o fungo Cylindrocarpon sp. empregado na estratégia inundativa, tem sido sugerido para manejo dessa macrófita onde o uso do controle químico tem se mostrado ineficiente devido ao desenvolvimento de resistência aos herbicidas inibidores da enzima ALS (acetolactato sintase). O presente trabalho foi conduzido em casa de vegetação e no laboratório de Controle Biológico de Plantas Daninhas Prof. Giorgio de Marinis, da FCAV – UNESP, Jaboticabal, visando avaliar a especificidade do agente de biocontrole em plantas não alvo de diferentes famílias botânicas e determinar a severidade da doença em função do número e intervalos de aplicações do bioherbicida, do período de orvalho e idade da planta além de estimar os efeitos de adjuvantes sobre o desenvolvimento de Cylindrocarpon sp. no controle de S. montevidensis. O fungo mostrou-se específico a S. montevidensis, não sendo constatado sintomas da infecção fúngica em plantas de interesse econômico e nas plantas aquáticas Pistia stratiotes, Eichhornia crassipes, E. azurea, Salvinia sp., Azolla caroliniana, Lemna minor, Heteranthera reniformis, Hydrocotyle verticillata, Pontederia rotundifolia, P. parviflora, P. cordata, Polygonum lapathifolium, Ludwigia sedoides, L. elegans, L. repens, L. helminthorrhiza, Potamogeton pectinatus, Cerathophylum demersum, Myriophyllum aquaticum, Egeria densa, E. najas, Najas guadalupensis, Hydrilla verticillata, Alternanthera phyloxeroides, Nimphoides indica. A doença foi mais severa quando o bioherbicida foi aplicado em plantas com 3 a 4 folhas e na fase de inflorescência, submetidas as condições de período de orvalho superior a 6 horas...<br>Sagittaria montevidensis is a very important aquatic in shallow water bodies in wetland rice, mainly recentely because tolerant biotipes is raising in areas with repetitive spraying with ALS inhibitor herbicide. The fungus Cylindrocarpon sp. has a good potential as biocontrol agent for this control action of this fungus against S. montevidensis under different conditions of spore concentration, dew period, number and intervals of spraying and surfactants adittion in the spraying solution. The fungus specificity also was evaluated in aquatic and crop plants. The fungus showed high specificity to S. montevidensis. The disease severity was more strong when the plants were inoculated a 3-4 leaf stage and during blooming. A dew period longer than 6 hours was enough for to reach the maximum disease severity. Two a three spraying with the fungus enhanced the disease severity comparing with a single application and the intervals of 21 days between spraying produced better control than intervals of 7 and 14 days. Among the surfactants, Silwet L-77AG and Iharaguen-S improved the disease severity while Energic and Herbitensil reduced the Cylindrocarpon sp. disease severity.

A citricultura brasileira apresenta números bastante significativos e expressivos, refletindo a grande importância social e econômica que essa atividade agrícola e industrial tem para a economia do país. O Brasil se destaca como o maior produtor de laranja do mundo. As plantas cítricas podem ser produzidas e multiplicadas de várias formas e a enxertia é a forma de propagação vegetativa mais utilizada comercialmente no Brasil. Dentre os porta-enxertos mais empregados, sobressaem-se o citrumeleiro Swingle e a tangerineira Sunki. Apesar das vantagens presentes nesses dois genótipos, a produção deles está sendo limitada por doenças causadas por fungos e estraminipilas. Em citros, as diversas espécies de Phytophthora são responsáveis pelas doenças gomose e podridões, principalmente de radicelas. P. nicotianae é a espécie que mais afeta as plantas cítricas no Brasil. Não obstante a grande importância de P. nicotianae para os porta-enxertos, alguns mecanismos de resistência ou susceptibilidade ainda requerem mais investigações cientificas. O presente trabalho visa a elucidação de alguns mecanismos de resistência dos citros a P. nicotianae, por meio de estudos fisiológicos e bioquímicos dos porta-enxertos, além da ação de meios de cultura e da qualidade da luz no estudo da fisiologia desse estraminipila. Para isso realizou-se a detecção de alguns equivalentes de compostos fenólicos através da técnica HPLC e o comportamento do genótipo resistente frente à possível inibição da síntese de compostos fenólicos através do tratamento das raízes do porta-enxerto Swingle com Pro-Ca; avaliou-se a atração de zoósporos de P. nicotianae por exsudatos radiculares provenientes dos dois porta-enxertos com o uso de uma armadilha adaptada; verificou-se o comportamento do zoósporo, a motilidade e a zoosporogênese frente à ação de vários compostos fenólicos e testou-se a ação de diferentes meios de culturas e da qualidade da luz no crescimento micelial e na esporulação de P. nicotianae. Os resultados revelaram que a tangerineira Sunki possui maiores quantidades de equivalentes em apigenina que o citrumeleiro Swingle nos dois períodos de avaliação, para o HPLC. O porta-enxerto resistente apresentou mais equivalentes de ácido clorogênico que o genótipo susceptível. Quanto ao Pro-Ca, os equivalentes de compostos fenólicos em raízes de plântulas do citrumeleiro Swingle não diferiram daquelas plantas tratadas com água. O Pro-Ca não inibiu e não desativou a síntese de compostos fenólicos. O uso do anticorpo contra a elicitina \"&alpha;-plurivorina\" teve resultados intermediários no consumo de água por plântulas de tangerineira Sunki e a atividade zoosporicida dos compostos fenólicos apresentou resultados variáveis no comportamento da motilidade de zoósporos. Os compostos fenólicos escopoletina e tricetinpentametoxi conseguiram paralisar a motilidade e proporcionaram as menores porcentagens de zoosporogênese. O meio de cultura AA permitiu a maior TCMD, seguido de CA e V8-CaCO3-Ágar. Os maiores valores de esporulação foram encontrados nesses dois últimos meios, respectivamente. Todas as faixas de luzes (e a ausência de luz) estudadas no presente trabalho podem ser avaliadas para o crescimento micelial, porém o tratamento claro foi aquele que mais induziu a esporulação de zoósporos de P. nicotianae.<br>The Brazilian citrus industry has very significant and expressive numbers, reflecting the great social and economic importance of this industrial and agricultural activity has for the economy. Brazil stands out as the world\'s largest orange producer. Citrus plants can be produced and multiplied in various ways and the bud is the form of vegetative propagation more commercially used in Brazil. Among the most commonly used rootstocks, stand up the Swingle citrumelo and Sunki mandarin. Despite the advantages these two genotypes present, their production is being limited by diseases caused by fungi and estraminipilas. In citrus, all species of Phytophthora are responsible for diseases gummosis and fibrous root rot, especially. P. nicotianae is the kind that affects citrus plants in Brazil. Regardless of the great importance of P. nicotianae for rootstocks, some mechanisms of resistance or susceptibility still require further scientific investigation. This study aims to elucidate some mechanisms of resistance of citrus to P. nicotianae, through physiological and biochemical studies of rootstocks, beyond of action culture media and quality of light in the physiological study of this estraminipila. To this was carried out to detect certain equivalent of phenolic compounds by HPLC and genotype resistant behavior against the possible inhibition of the synthesis of phenolic compounds by treatment of the roots of Swingle rootstock with Pro-Ca; evaluated the zoospores attraction P. nicotianae by exudates from the two rootstocks using an adapted trap; it was the zoospore behavior, motility and zoosporogenesis to the action of various phenolic compounds and tested the action of different culture media and quality of light on mycelial growth and sporulation of P. nicotianae. The results for HPLC revealed that Sunki mandarin has larger amounts of apigenin equivalents Swingle citrumelo in that the two periods. The sturdy rootstock had more chlorogenic acid equivalents of the susceptible genotype. As for the Pro-Ca equivalent of phenolic compounds in Swingle seedling roots not differ from those plants treated with water. The Pro-Ca not inhibited and not disabled the synthesis of phenolic compounds. The use of the antibody against elicitin \"&alpha;-plurivorin\" had intermediate results in water consumption by Sunki mandarin seedlings and zoosporicide activity of phenolic compounds showed variable results in the behavior of zoospore motility. The scopoletin and tricetinpentametoxi phenolic compounds managed to paralyze the motility and provided the smallest percentages of zoosporogenesis. The culture medium AA allowed greater DMGR, followed by CA and V8-CaCO3-Agar. The highest sporulation values were found in the last two media, respectively. All lights tracks (and the absence of light) studied in this work can be evaluated for mycelial growth, but the treatment light was that more induced sporulation of zoospores of P. nicotianae.

La maîtrise du développement des moisissures retrouvées dans le contexte agro-alimentaire parmi les flores microbiennes d'altération ou technologiques répond à des enjeux économiques et sanitaires importants. Le développement des moisissures peut être affecté par des facteurs abiotiques comme la température, l'activité de l'eau (aw) ou la composition gazeuse. L'évaluation de l'effet de ces facteurs via des outils de mycologie prévisionnelle vise à prévoir l'altération fongique des aliments. Ce travail a pour objectif d'explorer l'impact des conditions environnementales sur la physiologie de moisissures d'intérêt pour l'industrie agro-alimentaire.L'effet de la température, de l'aw ajustée par du glycérol ou du chlorure de sodium (NaCl) du pH et de la composition gazeuse a été évalué sur la germination des spores et/ou la croissance radiale de cinq moisissures: Paecilomyces niveus, Mucor lanceolatus, Penicillium brevicompactum, Penicillium expansum et Penicillium roqueforti. Au niveau appliqué, ces travaux ont montré que l'effet du NaCl ou de la composition gazeuse peuvent être inclus dans une approche de mycologie prévisionnelle. Le choix des souches représentatives d'une espèce fongique et l'état physiologique des spores utilisées comme inoculum ont un impact significatif sur les modèles prédictifs.Au niveau fondamental, des marqueurs ont été recherchés pour évaluer l'effet des facteurs abiotiques sur la physiologie des spores. La température et l'aw ont un effet significatif sur l'état physiologique des spores et leur germination. La recherche de marqueurs moléculaire, contribuera aux connaissances de l'effet des facteurs abiotiques sur la physiologie des moisissures<br>In the food processing industry, controlling the development of filamentous fungi encountered as spoilers or technological cultures address significant economic and sanitary issues. Fungal development in foods is mainly determined by abiotic factors including temperature, water activity (aw) or the headspace gas composition. The quantification of these respective effects through a predictive mycology approach aims at preventing fungal food spoilage. The present work aims at exploring the effect of environmental conditions on the physiology of filamentous fungi of interest in the food processing industry.The effect of temperature, aw (adjusted with glycerol of sodium chloride), pH and headspace gas composition was evaluated on conidial germination and/or radial growth of five fungi isolated from dairy products: Paecilomyces niveus, Mucor lanceolatus, Penicillium brevicompactum, Penicillium expansum and Penicillium roqueforti. The present work suggests that the specific effects of sodium chloride or gas composition could be included in predictive mycology approaches. It was also demonstrated that the selection of strains representative of a fungal species and the physiological state of conidia utilized as inoculum have a significant effect on the final predictive models.At the fundamental level, markers were investigated to study the effect of abitoic factors on the physiological state of spores. The temperature and aw significantly affected the physiological state of spores and their germination kinetics. The investigation of markers at the molecular level could provide better knowledges on the effect of abiotic factors on the physiology of filamentous fungi

Submitted by Johnny Rodrigues (johnnyrodrigues@ufcg.edu.br) on 2018-05-03T18:31:57Z No. of bitstreams: 1 ANDRESSA LACERDA NÓBREGA UGULINO - DISSERTAÇÃO PPGSA ACADÊMICO 2017..pdf: 676544 bytes, checksum: 0dd2e41c1011a60929ca0702449f6287 (MD5)<br>Made available in DSpace on 2018-05-03T18:31:57Z (GMT). No. of bitstreams: 1 ANDRESSA LACERDA NÓBREGA UGULINO - DISSERTAÇÃO PPGSA ACADÊMICO 2017..pdf: 676544 bytes, checksum: 0dd2e41c1011a60929ca0702449f6287 (MD5) Previous issue date: 2017<br>O fungo Macrophomina phaseolina (Tassi.). Goid é o agente etiológico de uma das principais doenças na cultura do feijão-caupi, vulgarmente conhecida como podridão cinzenta do caule. Em decorrência da falta de produtos fitossanitários registrados para o controle desta doença um aumento na busca por métodos de controle alternativo é cada vez mais comum. O objetivo do presente trabalho foi avaliar o efeito de óleos essenciais sobre o crescimento micelial do fungo M. phaseolina. Os óleos testados foram de hortelã (Mentha sp.), eucalipto (Eucalyptus spp.), copaíba ( Copaifera sp.) e alecrim-da-chapada (Lippia gracilis), nas concentrações 0,4; 0,6; 0,8 e 1,0%. Os óleos foram incorporados ao meio de cultura do tipo BDA e posteriormente vertidos em placas de Petri de 90x15mm, após a solidificação do meio, discos de 8mm contendo o micélio do patógeno foram repicados para as placas com os respectivos tratamentos. O experimento foi conduzido no delineamento experimental inteiramente casualizado (DIC) em arranjo fatorial 4 x 4 + 1 +1, sendo dezesseis tratamentos mais uma testemunha negativa e uma testemunha positiva que constou da aplicação suplementar no meio (BDA) do fungicida Sportak 450 EC (Procloraz), e cinco repetições, totalizando 90 parcelas experimentais. Posteriormente todas as placas foram transferidas para incubadora do tipo B.O.D. (biochemical oxygen demand) a 27 ± 2 ºC. Foram realizadas medições diárias do diâmetro da colônia em dois sentidos perpendiculares até que tomasse toda a superfície do meio de cultura de uma das placas. Os dados obtidos foram interpretados por meio de análise de variância não paramétrica e as médias foram comparadas pelo teste de Kruskal-wallis, a 5% de probabilidade de erro e Scott Knott a 5% de probabilidade nos casos em que existiram diferenças significativas e normalidade dos dados. Os óleos essenciais de hortelã e alecrimda-chapada apresentaram melhores resultados quanto a inibição do crescimento micelial, enquanto que os óleos de eucalipto e copaíba apesar de potencialmente promissores, apresentaram inibição intermediária do crescimento micelial do fungo.<br>Macrophomina phaseolina (Tassi.). Goid is the etiological agent of one of the major diseases in cowpea bean crop, commonly known as gray rot stem. Due to the lack of registered phytosanitary products to control this disease, the search for alternative control methods is increasingly common. The objective of this study was to evaluate the effect of essential oils on the mycelial growth of M. phaseolina. The oils tested were Mentha sp.,Eucalyptus spp., Copaifera sp., and Lippia gracilis at concentrations of 0.4; 0.6; 0.8 and 1.0%. The work was conducted in Phytopathology Laboratory of the Federal University of Campina Grande. Daily measurements of the colony diameter were performed in two perpendicular directions until it filled the entire surface of the culture medium of one of the plates. The experiment was conducted in a completely randomized experimental design in a factorial arrangement 4 x 4 + 1+1, with sixteen treatments plus one negative control and one positive control, which consisted of the supplemental application in the medium (BDA) of the fungicide Sportak 450 EC (Procloraz), and five replications, totaling 90 experimental plots. The data were interpreted through non-parametric analysis of variance and the means were compared by the Kruskal-wallis test, with a 5% probability of error and Scott Knott at 5% probability in cases where there were significant differences and data normality. The Mentha sp. essential oil and Lippia gracilis showed better results in inhibiting mycelial growth, while the Eucalyptus essential oil and Copaifera, although potentially promising, showed intermediate inhibition of fungal mycelial growth.

香菇是一種重要的藥用蘑菇。數千年來，香菇一直被人們作為食物和藥物來使用。許多研究表明，香菇的提取物具有抗氧化活性，而且他們的抗氧化活性與他們的酚類化合物的含量相關。然而，到目前為止該研究大多集中在對香菇子實體的研究，對香菇菌絲體分泌物的研究就少見報導。<br>在本課題研究中，使用不同的體外抗氧化測定方法和酚類化合物含量測定方法來研究兩種香菇菌絲體分泌物（1358DE 和L5458DE）。實驗結果表明， 在不同的體外抗氧化實驗中1358DE 和L5458DE均具有明顯不同的抗氧化活性。在清除DPPH自由基，清除氫氧根離子，清除超氧陽離子，清除過氧化氫離子，螯合亞鐵離子，還原能力，抑制老鼠紅細胞溶血和抑制脂質過氧化的實驗中，1358DE 和L5458DE的IC50 分別為3.3和132.6; 44.5和 > 1000; 26.9和53.7; 153.6和 >175.0; 176.0和521.0; 26.7和746.4; 47.8和736.9; 3.1和 > 1000 μg/ml。他們的多酚化合物的含量分別為237.33 and 24.08 mg (GAE)/g of DE。實驗資料表明，1358DE的抗氧化活性高於L5458DE，其原因可能是1358DE的酚類化合物含量較高。<br>由於1358DE具有較好的抗氧化活性，採用有機溶劑萃取的方法將其分成水溶性部位和乙酸乙酯部位。體外抗氧化實驗表明，水溶性部位的抗氧化活性與1358DE相近，而乙酸乙酯部位則沒有表現出抗氧化活性。因此，使用聚醯胺柱色譜（可以將多酚類化合物從其他成分中分離出來）對水溶性部位進行進一步的分離，可以得到兩個聚醯胺洗脫部位（P-1和P-2）。與原來的水溶性部位比較，P-1的糖的含量明顯增加，而多酚化合物含量明顯減少，抗氧化活性也明顯降低；相反，P-2的糖的含量明顯減少，而多酚化合物含量明顯增加，抗氧化活性也明顯增加。該實驗結果表明，糖對抗氧化活性的貢獻遠不及多酚化合物。因此，多酚化合物是1358DE的抗氧化活性成分。基質輔助鐳射解吸電離飛行時間質譜和三氯化鐵試劑測定結果表明，P-2是一類水溶性多酚低聚物（WSP），它的分子量在600~1200Da之間。<br>水溶性多酚低聚物（WSP）是1358DE的主要抗氧化活性成分。採用過氧化氫引導細胞毒性的細胞(V79-4)模型來進一步研究WSP的抗氧化活性。在細胞毒性試驗中，在所有測試濃度，WSP在濃度6.25~50 μg/ml均能明顯抑制過氧化氫引致的細胞毒性。此外，WSP還能明顯抑制由過氧化氫引起的丙二醛（MDA）增加和抗氧化酶（SOD，CAT，GSH-Px）的減少。<br>許多抗氧化劑被報導具有抗血管增生活性，該活性與其抗氧化活性相關。由於WSP具有非常好的抗氧化活性，因此，採用斑馬魚模型來研究WSP的抗血管增生活性。在內源性鹼性磷酸酶測定實驗結果表明，WSP在濃度50，100，150，200，250 μg/ml，斑馬魚（野生型）的血管生成明顯分別減低為87.2, 85.6, 74.8, 69.4, and 62.8%（與空白對照組相比）。此外，在螢光顯微鏡下可觀察到WSP在濃度為250μg/ml能明顯抑制螢光斑馬魚（fli1a:EGFP）的節間血管形成。<br>本研究表明，水溶性多酚低聚物（WSP）是香菇菌絲體分泌物的抗氧化成分，WSP不僅具有抗氧化活性，同時還具有抗血管增生活性。此外，本研究結果表明香菇菌絲體分泌物是很好的天然抗氧化劑的來源。<br>Shiitake mushroom (Lentinus edodes), known in China as Xiang-gu, is one of the most valuable medicinal mushrooms, and has been used for thousands of years both as food and medicine. Shiitake mushroom extracts have also been found to have antioxidant properties and their antioxidant ability is positively correlated with their phenolic content. However, thus far, investigation of the antioxidant ability of shiitake mushroom has mainly focused on the fruiting body, and the antioxidant properties of its mycelial exudates are rarely reported.<br>In this study, exudates (DE) secreted from two shiitake mushroom mycelia (strains 1358 and L5458) were evaluated for their antioxidative properties and phenolic content. 1358DE and L5458DE showed distinct antioxidant activity in different in vitro assays, including scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, hydroxyl radical, superoxide anions and hydrogen peroxide; the ability to chelate ferrous ions; reducing power; hemolysis inhibition activity in rat erythrocyte; and lipid peroxidation inhibition (IC₅₀ values of 1358DE and L5458DE were 3.3 and 132.6; 44.5 and > 1000; 26.9 and 53.7; 153.6 and >175.0; 176.0 and 521.0; 26.7 and 746.4; 47.8 and 736.9; and 3.1 and > 1000 μg/mL, respectively). Their total phenolic content was 237.33 and 24.08 mg gallic acid equivalent (GAE)/g of dry DE, respectively. Overall, these results show that 1358DE generally possesses better antioxidant properties than L5458DE, possibly due to its larger total phenolic content.<br>1358DE were selected to further investigate for its better antioxidant effect. 1358DE was fractionated using water-solvent partition and two fractions [water soluble fraction (WF) and ethyl acetate fraction (EF)] were obtained. The antioxidant effects of WF were similar to those of the original 1358DE, while EF did not possess any antioxidant activities. The WF was further isolated with polyamide column, which can separated the polyphenols from other components, and two sub-fraction (P-1 and P-2) were obtained. After the WF passing through the polyamide column, carbohydrate content in the sub-fraction 1 (P-1) was significantly increased, while its total phenolic content reduced dramatically, and its antioxidant activity decreased. However, the sub-fraction 2 (P-2) was the opposite. Carbohydrate content in P-2 was significantly reduced, while its total phenolic content increased dramatically, and its antioxidant activity increased. Apparently, carbohydrate contributed little to the antioxidant effect than that of the phenolic compounds as shown from this investigation. These results suggest that the antioxidant effect in 1358DE was contributed by the presence of polyphenols. Besides, results from Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and ferric trichloride reaction suggested that P-2 was oligomers of water soluble polyphenols (WSP) with the molecular weight of about 600~1200 Da.<br>The water soluble polyphones (WSP) were the potent antioxidant components in 1358DE and further study its protective effect against the hydrogen peroxide which induced cytotoxicity in V79-4 cells. In the cell viability experiments, pretreatment of WSP at the concentrations of 6.25~50 μg/ml increased the cell viability significantly more than at the presence of H₂O₂ only. Besides, the pretreatment of cells with WSP significantly inhibited the increase of Malondialdehyde (MDA, which is a by-product of lipid peroxidation) production and the decrease of antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase) activities induced by H₂O₂.<br>Quite a few antioxidant compounds have been reported that a causative relationship may exist between the anti-angiogenic activity and antioxidant effect. Therefore, a zaebrafish model was using to investigate the anti-angiogenic activity of the WSP because of its excellent antioxidant activity. In quantitative of endogenous alkaline phosphatase (EAP) assay, after the embryos treated with WSP at final concentrations of 50, 100, 150, 200, 250 μg/ml, and the vessel formation were significantly (p < 0.05) reduced to 87.2, 85.6, 74.8, 69.4, and 62.8% of the control value, respectively. Moreover, from the microscope, compare to the control, WSP at the concentration of 250 μg/ml also showed potent inhibition on the intersegmental blood vessels (ISVs) formation in Tg(fli1a:EGFP)y1 zebrafish embryos. Thus, the finding indicated that WSP could inhibit vessel formation in zeabrafish.<br>Overall, this study revealed that water soluble polyphenols (WSP) was the active components of 1358DE. Besides of the antioxidant effect, the WSP could inhibit vessel formation significantly in zebrafish. The findings indicate that exudates of shiitake mushroom mycelia have good potential as a source of natural antioxidants.<br>Detailed summary in vernacular field only.<br>Detailed summary in vernacular field only.<br>Detailed summary in vernacular field only.<br>Detailed summary in vernacular field only.<br>Detailed summary in vernacular field only.<br>Detailed summary in vernacular field only.<br>Huang, Weihuan.<br>Thesis (Ph.D.)--Chinese University of Hong Kong, 2012.<br>Includes bibliographical references (leaves 90-103).<br>Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.<br>Abstract also in Chinese.<br>Acknowledgements --- p.i<br>Abstract --- p.ii<br>摘要 --- p.v<br>List of Abbreviations --- p.vii<br>List of Figures --- p.ix<br>List of Tables --- p.xi<br>Chapter Chapter 1 --- Introduction --- p.1<br>Chapter 1.1 --- Oxidation and antioxidant --- p.1<br>Chapter 1.1.1 --- Reactive oxygen species (ROS) --- p.1<br>Chapter 1.1.2 --- Sources of ROS --- p.1<br>Chapter 1.1.3 --- The role of ROS in normal physiology --- p.2<br>Chapter 1.1.4 --- Oxidative damage to DNA, lipids and proteins --- p.2<br>Chapter 1.1.5 --- Antioxidant defense systems in vivo --- p.4<br>Chapter 1.1.6 --- Sources of antioxidants --- p.6<br>Chapter 1.2 --- Assessment of antioxidant capacity in vitro and in vivo (Antioxidant methodology) --- p.9<br>Chapter 1.2.1 --- Assessment of Free Radical Scavenging Capacity in vitro --- p.9<br>Chapter 1.2.2 --- Antioxidant capacity in cultured Cells --- p.10<br>Chapter 1.2.3 --- Antioxidant capacity in vivo --- p.11<br>Chapter 1.3 --- Mushrooms --- p.12<br>Chapter 1.3.1 --- Mushroom life cycle --- p.12<br>Chapter 1.3.2 --- Nutritional and medicinal values of mushroom --- p.14<br>Chapter 1.4 --- Shiitake mushroom (Lentinus edodes) --- p.15<br>Chapter 1.5 --- Objectives --- p.17<br>Chapter Chapter 2 --- Antioxidant activity and total phenolic content in Shiitake mycelial exudates --- p.18<br>Chapter 2.1 --- Introduction --- p.18<br>Chapter 2.2 --- Materials and methods --- p.19<br>Chapter 2.2.1 --- Materials --- p.19<br>Chapter 2.2.2 --- Sample preparation --- p.20<br>Chapter 2.2.3 --- In vitro antioxidant activity assays --- p.20<br>Chapter 2.2.4 --- Determination of total phenolic content --- p.25<br>Chapter 2.2.5 --- Statistical analysis --- p.25<br>Chapter 2.3 --- Results and discussion --- p.25<br>Chapter 2.3.1 --- Antioxidant activity --- p.25<br>Chapter 2.3.2 --- Total phenolic content --- p.37<br>Chapter 2.3.3 --- Antioxidant activity (IC₅₀ values) and phenolic content --- p.37<br>Chapter 2.4 --- Conclusion --- p.39<br>Chapter Chapter 3 --- The antioxidant components of Shiitake mycelial exudates (1358DE) --- p.42<br>Chapter 3.1 --- Introduction --- p.42<br>Chapter 3.2 --- Materials and methods --- p.42<br>Chapter 3.2.1 --- Materials --- p.42<br>Chapter 3.2.2 --- Sample preparation --- p.43<br>Chapter 3.2.3 --- HPLC analytical condition --- p.43<br>Chapter 3.2.4 --- Sample fractionated using solvent-water partition and polyamide column chromatographic method guided by in vitro antioxidant assays --- p.43<br>Chapter 3.2.5 --- Determination of total phenolic content --- p.44<br>Chapter 3.2.6 --- Determination of the contents of carbohydrate --- p.45<br>Chapter 3.2.7 --- MALDI-TOF MS analysis --- p.45<br>Chapter 3.2.8 --- Statistical analysis --- p.46<br>Chapter 3.3 --- Results and discussions --- p.46<br>Chapter 3.3.1 --- HPLC analytical results --- p.46<br>Chapter 3.3.2 --- Samples fractionation using solvent-water partition --- p.47<br>Chapter 3.3.3 --- Water soluble fraction (WF) was further isolated using polyamide column chromatographic --- p.50<br>Chapter 3.3.4 --- Molecular weight determination of P-2 --- p.54<br>Chapter 3.4 --- Conclusion --- p.56<br>Chapter Chapter 4 --- Antioxidative effect of water soluble polyphenols (WSP) in Shiitake mycelial exudates (1358DE) against H2O2-induced cytotoxicity in V79-4 cells --- p.57<br>Chapter 4.1 --- Introduction --- p.57<br>Chapter 4.2 --- Sample preparation, materials and methods --- p.58<br>Chapter 4.2.1 --- Preparation of the water soluble polyphenols (WSP) --- p.58<br>Chapter 4.2.2 --- Materials --- p.58<br>Chapter 4.2.3 --- Cell culture and treatment --- p.59<br>Chapter 4.2.4 --- MTT assay --- p.60<br>Chapter 4.2.5 --- Lactate dehydrogenase (LDH) release assay --- p.60<br>Chapter 4.2.6 --- Assay for lipid peroxidation measuring the malondialdehyde (MDA) --- p.61<br>Chapter 4.2.7 --- Assay for antioxidant enzymes --- p.62<br>Chapter 4.2.8 --- Protein determination --- p.62<br>Chapter 4.2.9 --- Statistical analysis --- p.63<br>Chapter 4.3 --- Results and discussion --- p.63<br>Chapter 4.3.1 --- Cytotoxicty of WSP in V79-4 cells --- p.63<br>Chapter 4.3.2 --- Determined the time of WSP pretreatment in V79-4 cell against H₂O₂-induced cytotoxicity --- p.63<br>Chapter 4.3.3 --- Protective effect of WSP treated cell against H₂O₂-induced cytotoxicity --- p.64<br>Chapter 4.3.3 --- Inhibition of WSP on lipid peroxidation --- p.66<br>Chapter 4.3.4 --- Effects of WSP on antioxidant enzyme activities --- p.66<br>Chapter 4.4 --- Conclusion --- p.72<br>Chapter Chapter 5 --- Anti-angiogenic property of water soluble polyphenols (WSP) in Shiitake mycelial exudates (1358DE) --- p.73<br>Chapter 5.1 --- Introduction --- p.73<br>Chapter 5.1.1 --- Angiogenesis --- p.73<br>Chapter 5.1.2 --- Angiogenesis as a therapeutic target --- p.73<br>Chapter 5.1.3 --- Tumors angiogenesis --- p.75<br>Chapter 5.1.4 --- Reactive oxygen species (ROS) and tumor angiogenesis --- p.75<br>Chapter 5.1.5 --- Anti-angiogenic effects of polyphenols --- p.76<br>Chapter 5.1.6 --- Experimental model for studying anti-angiogenic agents --- p.76<br>Chapter 5.2 --- Sample preparation, materials and methods --- p.78<br>Chapter 5.2.1 --- Preparation of the water soluble polyphenols (WSP) --- p.78<br>Chapter 5.2.2 --- Materials --- p.78<br>Chapter 5.2.3 --- Methods --- p.79<br>Chapter 5.2.4 --- Statistical analysis --- p.80<br>Chapter 5.3 --- Results --- p.81<br>Chapter 5.3.1 --- Anti-angiogenic effect of WSP on zebrafish model --- p.81<br>Chapter 5.3.2 --- Microscopic imaging --- p.82<br>Chapter 5.4 --- Discussion and conclusion --- p.82<br>Chapter Chapter6 --- Conclusions --- p.84<br>Chapter 6.1 --- Conclusion --- p.86<br>Chapter 6.2 --- Future works --- p.88<br>References --- p.89<br>Chapter Appendix 1 --- Publication --- p.104

碩士<br>亞洲大學<br>生物科技學系碩士班<br>100<br>Grifola frondosa, also named Maitake, is a polypore mushroom recognized by clusters of rosettes arising from a single, branches stem structure. Grifola frondosa has been used as a traditional medicine in China and Japan. White Grifola frondosa is the particular species in Japan. Medical research confirmed that Grifola frondosa efficacy, including anti-cancer,enhance the immune system, lower blood sugar, blood pressure and blood cholesterol, protect the liver, prevent diabetes, and weight-loss. Although its edible and medicinal value was extremely high, however the yield produced in Taiwan is not so stable, mostly rely on imported from Japan and China. This study was to investigate the Grifola frondosa mycelial growth and fruiting body development on agar plate medium and sawdust substrate and tried to find out the most optimum methods in cultivation. Results showed the best environment for mycelial growth on PDA medium were 24 ℃ and pH6.5 medium environment mycelial growth faster and mycelium dense, PDA and FPDA were most suitable media for mycelial growth. Light (500 lux/24hrs) also cause mycelium grow sparsely and slowly. Test of water content, sawdust ratio, nutrient composition and peat moss supplement in sawdust substrate revealed that 65% of water content and 7:3 of sawdust and nutrients composition enhanced mycelial growth faster and primordium development better; 25% of wheat bran, and 5% of rice bran in sawdust substrate were the best proportion for mycelial growth and primordium development, addition of 5% to 20% peat moss accelerated mycelial growth. Results also showed that rice straw, Pleurotus eryngii waste sawdust substrate, sweet sorgum and corn cob were the available alternative substrates for cultivation of Grifola frondosa, 100% of Pleurotus eryngii waste sawdust substrate could completely replaced of sawdust substrate, sawdust 70% + wheat bran 25% + rice bran 5% and 65% of the water content in culture substrate produced highest biological efficiency of Grifola frondosa. Fourth-generation cultivation bottle developed by Dr. Chien-Yih Lin was the best container for culture of Grifola frondosa, because of its high biological efficiency (57.3%) and low contamination in cultivation. Fourth-generation cultivation bottle also used in automatic culture for Grifola frondosa.

碩士<br>亞洲大學<br>生物科技學系碩士班<br>99<br>One of the foremost medicinal and culinary mushrooms of China and Taiwan, Tremella fuciformis has the most unusual strategies for cultivation by utilizing the nutrients gained from a common wood-decomposing Ascomycetes fungus, Hypoxylon archeri. Goals of this study are trying to understand factors affecting mycelial growth and fruiting of T. fuciformis. The results revealed that T. fuciformis mycelium was white or beige, slow growth, had upright, inclined up or flat aerial hyphae on the surface of the medium, the mycelium formed gel-like colony on PDA medium. H. archeri mycelium was white, fast growth, had slightly feathery and flat branching hyphae on the PDA medium , old mycelium became pale yellow to light brown and secreted melanin in medium. T. fuciformis mycelium could grow among 23℃-27℃ and 25℃was the most optimum temperature. H. archeri mycelium could grow among 25℃-31℃ and 29℃was the best temperature. It was also found that FPDA was the best medium for both growth of T. fuciformis and H. archeri mycelium among eight media. Test of T. fuciformis confront culture with H. archeri , the result revealed T. fuciformis formed white and dense mycelium mat on PDA medium. Liquid filtrates of T. fuciformis or H. archeri mycelium were added to PDA separately, the growth of mycelium of H. archeri or T. fuciformis was stimulated individually. Although T. fuciformis mycelium growing in sawdust alone was slow and no primordium developed , however, fruiting body of T. fuciformis was successfully produced by mixed culture with H. archeri in sawdust medium due to T. fuciformis could utilize the nutrition decomposed from sawdust by H. archeri. The formation procedures of T. fuciformis fruiting body included (1) secretion of yellow stick material, (2) formation of primordium, (3) development of fruiting body. 25℃ was found the best temperature for mycelium growth of T. fuciformis in sawdust substrate and 23℃ was the best temperature for development of fruiting body. The result revealed that rice bran was the best supplementary nutrient added in sawdust substrate. 24% of high fruiting body yields were increased by adding 30% of rice bran in sawdust compared to those adding 20% of rice bran. The result also indicated that 98.2% of fruiting body yields were increased if 50% sawdust were replaced by corn cob.

碩士<br>國立屏東科技大學<br>植物保護系<br>94<br>For studying the positive effects on mycelial growth and mushroom production of Agaricus blazei Murill, the affecting factors of various compost materials, casing materials, thickness of casing, nutrient supplements and microbes were experimented. The optimum temperature for mycelial growth of A. blazei on PDA was 28℃；optimum pH was pH 6. A. blazei cultivated on various composts made by pangola grass, rice straw, sawdust, or sugarcane bagasse and with various pH values of 4 to 8, the most suitable pH for mycelial growth was at pH 6 with 35.4 mm in colony diameter in petri dish plate of pangola grass compost, whereas the highest biological efficiency with 40.54﹪was at pH 8 requiring 63 days after peat moss casing in bag culture to harvast the first mushroom. However, in rice straw compost, the biggest colony diameter with 60.6 mm was at pH 8, and the best biological efficiency 38.01﹪was at same pH 8 for 50 days for beginning harvesting. In sawdust, the 37.4 mm diameter was the biggest at pH 7.0, but the biological efficieny 32.24﹪was the best and harvesting on 54th day after casing at pH 8. In sugancane bagasse compost, pH 7 was good for mycelial growth, and pH 8 also was the best for biological efficiency, but only 7.86﹪, whereas there is no mushroom production at pH 5. Using peat moss, peat mixed with sand, red soil or sand as the casing materials for casing on sawdust compost in beaker culture, the peat and sand mixture had the highest biological efficiency 40.2﹪, requiring 44 days for first mushroom production. The 3 cm thickness of peat moss obtained the best biogical efficiency 49.21﹪with 51 days for mushroom harvesting. In experiment of casing peat sprayed with soil microbes, the LS33-2 isolate treatment had gotten a biggest biological efficiency in 36.3﹪and required 37 days for beginning harvesting. In simple culture house, fruiting body extract sprayed to casing soil of ground culture bed had obtained the best biological efficiency 16.5﹪with 33 days requiring for mushroom harvesting.

碩士<br>國立臺灣大學<br>森林學研究所<br>90<br>Cinnamomum kanehirae is an endemic broadleaf tree species in Taiwan. Heartwood of C. kanehirae in natural forest was decayed by one brown rot fungus, which is Chang-chih (Antrodia camphorata). However, Chang-chih is a precious fungus species that has been used for traditional medical purpose as an antidote and hepatic medicine in Taiwan. Unfortunately, any attempt to reproduce the fruiting body of Chang-chih by artificial cultivation was unsuccessful, therefore it has to be collected from natural forest by damaging the C. kanehirae tree. In order to protect this unique endemic species, this study is to determine the effects of the extracts of C. kanehirae on the mycelial growth of A. camphorata. Hopefully, it is expected to find the active constituents which might promote the mycelial growth, and generate the fruiting body of A. camphorata. Results revealed that, due to its specific extracts, C. kanehirae inhibited the growth of several rot fungi (Laetiporus sulphureus, Gloeophyllum trabeum, Lenzites betulina and Trametes versicolor), whereas promoted A. camphorata to grow better on MEA medium plate. Besides, A. camphorata was proven to excrete some antibiotics which inhibit the growth of fungi based on the results from dual culture. These results demonstrated clearly that A. camphorata is the only kind fungus on C. kanehirae. Results obtained from the chemical analyses of C. kanehirae decayed by A. camphorata revealed that the amounts of its extracts increased, whereas the contents of total phenolics decreased and a significant reduction of pH-value was also observed. Comparing the variations in ethyl acetate soluble fraction of extracts between sound wood and decayed wood, it is obvious that changes in the contents of total phenolics and pH-value were due to the degradation of ethyl acetate soluble extract caused by A. camphorata. According to the results of agar plate test, the decreasing order of growth index of four subfractions that was partitioned from extract of sound wood was Hex. sol.> EA sol.> BuOH sol.= H2O sol.. Therefore, hexane soluble fraction was identified to be the most significant one to promote the mycelial growth of A. camphorata. Furthermore, some constituents of essential oil from C. kanehirae, such as α-Terpineol, Geraniol, Citronellol, l-Linalool, d-Camphor and Eugenol, were proven to promote effectively the mycelial growth of A. camphorata.

碩士<br>國立屏東科技大學<br>熱帶農業暨國際合作系所<br>96<br>This study was to elucidate the rates of mycelial growth, under different temperatures in vitro culture, from tissues excised from the top, middle and bottom sections of a fruiting body stem of Agaricus blazei Murrill. Also determined were, after the mycelia had been transferred to a jar containing wheat grains as medium, the effects of different water contents in wheat grains and different additives to the medium on the growth of mycelia. Based on these results, an optimal procedure for producing wheat grain spawn in the laboratory was developed. It was the intent of this study to utilize wheat grains to minimize the reliance of wood or sawdust as commonly used materials for the production of a fungal spawn. Results of this study showed that mycelia from the tissue, excised from the bottom section and cultured in potato dextrose agar (PDA), developed significantly faster at 28 and 30oC than the mycelia from other sections of a fresh fruiting body (p<0.05). A comparison of the mean diameters of mycelia on the 10th day after the inoculation of the fungus on the PDA medium showed that the diameters decrease with the decrease of the temperature. The sequence of the decrease, in descending order, was 30oC (7.27±0.33cm) = 28oC (6.80±0.63cm) > 26oC (5.71±0.51cm) > 24oC (4.87±0.68cm). After transferring from the PDA medium to the wheat grain medium containing different water contents and being subjected to the second shaking, the mycelia grew the fastest in covering the entire jar in a medium containing 150 ml wheat grains with 90 ml water. A comparison of the growth rates showed, in descending order, 90mL (5.5±0.52 days) was significantly differed from 100mL (7.9±2.23 days); however, both of them are not significantly differed from 110mL and 120mL. After the first shaking, the mycelial growth in the medium containing 150 ml wheat grains with 90 ml water fortified with the additive of 2.7g citrus juice was significantly faster, in descending order, orange juice（6.2±0.83days）＞sucrose（7.3±0.87days）=fructose（7.8±0.87days）＞control（10.1±0.45days）. A comparison of the enhancement of mycelial growth by the different additives showed, in descending order, orange juice (6.42±1.31 days) > control (9.10±0.67 days); however, both of them were not significantly differed from sucrose and fructose after the second shaking.

碩士<br>亞洲大學<br>生物科技學系<br>103<br>Antrodia cinnamomea belonging to Fungi, Basidiomycota, Basidiomycetes, Polyporales, Polyporaceae, Antrodia, is a special fungus only found in Taiwan . A. cinnamomea normally used as the healthy food is rarely and difficultly cultivated, therefore, price of A.cinnamomea fruiting body is much higher than those of other mushrooms. This studies were trying to understand the effect of environment and culture substrates on the mycelial growth of A.cinnamomea AC 11 strain .The results revealed that the best temperature was 28℃, best medium was PDA, and best pH value was 5.5 . The higher proportion of antrodia sawdust extract replaced water of PDA resulted in better mycelial growth of A. cinnamomea 100% of antrodia sawdust extract was the best replacement of water in PDA . Different proportion of brown rice , buckwheat or barley powder was added to PDA for studying the effect of mycelial growth of A.cinnamomea AC 11 revealed that 40% of buckwheat , brown rice or barley powder was the best treatment .Studies on imported camphor (from Laos country) sawdust was added with different nutrients revealed that 21% of wheat middlings was the test supplement and proportion , where as the imported camphor mixed with Taiwan camphor sawdust showed the test added nutrient was 21% of rice bran；best nutrient for single Taiwan camphor sawdust was 21% of corn meal. A.cinnamomea AC 11 cultured in test tube of sawdust substrates studies revealed that the non-camphor sawdust could inhibit the mycelial growth of A.cinnamomea AC 11.Imported camphor wood filled with different media studies showed that the best formula was the imported camphor 78% , PDB 21% , CaCO3 1%. The best formula to camphor wood filled with different media was Taiwan camphor sawdust 78% , PDB 21% , CaCO3 1%. The best temperature studies for mycelial growth of A. cinnamomea AC 11 on substrates of Taiwan camphor 78% , water 21%, CaCO3 1% was 28℃.

碩士<br>國立虎尾科技大學<br>生物科技系碩士班<br>107<br>[Cordyceps cicadae (Miq.) Massee] is an sexual stage of Isaris cicadae Miq. [Syn. Paecilomyces cicadae (Mique) Samson] has been used as a valuable Chinese medicine, which was known with its immunomodulatory, antipyretic, anti-fatigue ability. The artificial cultivation of C. cicadae is the best way to solve the difficulty of obtaining natural C. cicadae, the high price, the less production area and the long interval of harvesting fruiting bodies (FB). My thesis detected that using sucrose as carbon source and cicada shell meal as nitrogen source at 24 °C, pH 5.5, 24 hours light resulted the longest of the C. cicadae mycelium. Solid culture 1 % maltose+ 5 % fish meal condition increased 58.5% average of fruiting bodies (FB) height (4.58 cm) as compare to 1 % glucose+ 5 % fish meal (2.89 cm). Moreover, the optimal water content for culture FB is 50%. Incubated C. cicadae for 3 days had the suitable biomass of mycelia and spore density (1×107 spores/ml) for pre-activated spore for farther rice-solid culture. In addition, it will be discussed whether C. cicadae FB can be dyed in the medium by adding the natural pigment.

碩士<br>國立虎尾科技大學<br>生物科技研究所<br>102<br>Medicinal mushrooms have attracted much attention recently owing to their potent therapeutic activity, especially as chemotherapy and immunomodulatory agents. Antrodia cinnamomea is an endemic medicinal fungus that grows mainly inside the rotten trunk of Cinnamomum kanehirae in natural conditions in Taiwan. Due to the limited distribution of the host plant and the slow growth rate of the fungus, the mass production of the fungus through in vitro culture systems including on the artificial cultural camphor tree for pharmaceutical usage has been attempted. Asarum heterotropoides F. Schmidt var. mandshuricumon that was assigned to birthwort family Aristolochiaceae, have been reported to promote growth of A. cinnamomea, also named Taiwanofungus camphorates. Therefore, the purpose of this study is to determine the effect of A. heterotropoides (Ashe) on sporulation and mycelial growth of A. cinnamomea in vitro and to detect the shorten growth time and formation of fruiting body quality compared with A. cinnamomea in natural. For sporulation and mycelial growth tests, two different strains of A. cinnamomea: AC1CK and AC-N were used and cultured with the extract from A. heterotropoides (Ashe). The malt extract agar (MEA) and a series of dilution of Ashe were made to culture for A. cinnamomea spores. Five fractions of Ashe, including Ashe E, Ashe W, Ashe C, Ashe CH, Ashe CM with different concentration 500 ppm, 1000 ppm and 2000 ppm were tested in this study. The results indicated that strain AC1CK and strain AC-N grew fastest in 2000 ppm among tested concentration as compared with other dosage. Ashe CH was the best among the partial fraction of Ashe to promote mycelial growth of A. cinnamomea at 2000 ppm. For mycelial growth tests, the results revealed that the Ashe CH obviously promoted the mycelial growth of A. cinnamomea to 99.28 -150.36 % in dose dependent manner with control. For pH tests, the maximum yield of 908 mg/100 ml, and 929 mg/100 ml, occurred at pH 5.0 in PDB with 2000 ppm Ashe CH of both strains AC1CK and AC-N. Furthermore, pure compounds from Ashe CH, asarinin and sesamin were used at concentrate 500 ppm to detect the effect on spore germination. The results indicated that spore germination rate of A. cinnamomea could be promoted to 120% as compared with untreated control. Based on the above results, the reliable production of the medicinal fungus can promote by Ashe treatment under laboratory conditions.

碩士<br>實踐大學<br>食品營養與保健生技研究所<br>95<br>Tip growth is a model of cellular expansion of hyphae, especially at extreme tip. It involves many cytoplasmic activities, such as the synthesis of the cell wall, cytoplasmic streaming, motility and positioning of vesicle. Vesicles are present at the extreme tip of growing hyphae, which contain enzymes and precursors for the new cell membrane and cell wall. Filamentous or F-actin plays important roles in tip growth of hyphae such as the maintenance of tip shape, the transport of wall vesicles and organelles, and cytoplasmic streaming. Latrunculin B (LB) or cytochalasin D (CD), inhibitor of actin polymerization, was used to investigate the roles of F-actin on growth and morphology of hyphae and mycelium of Achlya bisexualis. LB reduced growth rate of the hyphae in a concentration-dependent manner and also induced vacuolation and swelling, and swelling was time and concentration-dependent. At 50μg/ml CD reduced hyphae growth rate, without changing morphology. After removing LB, the growth rate and swelling were fully reversible when LB concentration is less than 1μg/ml, and the hyphae resumed grow in a time-dependent manner and some were induced hyperbranching. When LB concentration more than 1μg/ml, the hyphae were still vacuolation and swelling at the tips and even ceased to grow, and some were hyperbranching at subapical regions. Sub-apical swelling indicated vesicles failed forward to the tip, thus accumulated in this region owing to LB disrupted the formation of F-actin. Fractal analysis was used to investigate actin effects on growth and morphology of mycelium. LB affected extension area, diameter, and border fractal of mycelia in dose-dependent manner. Actin organizations were disrupted by LB, reflected in Alexa 488 phalloidin staining patterns. LB significantly affected actin organizations of hyphae such as actin caps and fibril cables in dose-dependent manner, but plaques were still the same as without treatment. In conclusion, F-actin plays an important role in maintaining growth and morphology of hyphae and mycelia of Achlya bisexualis. Fractal analysis is more convenient to quantify changes in growth and morphology of mycelia.

碩士<br>國立虎尾科技大學<br>生物科技研究所<br>103<br>Trichoderma spp. were known as good biological agents; they not only inhibit pathogens but also promote the growth of plants. Although Trichoderma can be grown in a variety of carbohydrates, with different supply matrix, the biological quality of the product will be significantly different. To effectively increase the biomass production of Trichoderma and shorten its incubation time for reducing the development and production costs of biological agents, this study intends to screen the crude extracts from 20 kinds of Chinese herbs for promoting the mycelial growth and spore germination of Trichoderma strains. The results show, gardenia (Gardenia jasminoides) crude extracts with 1000 to 2000 ppm can significantly promote spore germination of Trichoderma strains T2 (Trichoderma harzianum), T3 (T. virens) and T1-1 (T. harzianum) as compared to the control group cultured in 1% glucose + 1% L-Asparagine solution for 10 hours. The spore germination of T1-1 strain , T2 strain, and T3 strain treated with 1000 ppm of gardenia crude extract was 90.33%, 90.17% and 93.67%, respectively, and treated with control group was 48.67%, 50% and 51.83%, respectively. Similarily, cultured in sterile water for 14 hours, treatment with 1000 ppm gardenia crude extract had, was higher germination rate of three strains of Trichoderma above than that of untreated control groups. To understand the mechanism of gardenia crude extract for promoting the mycelial growth of Trichoderma spp. The hypothesis that it may play a role in carbon or nitrogen source, and complex medium was tested. The carbon sources, and nitrogen sources were compared with 1000 ppm gardenia for mycelial growth of different Trichoderma strains in water agar. The carbon sources included Glucose, Sucrose, Arabinose, Lactose, Maltose, Mannitol, Xylose, and Fructose. The nitrogen sources were Sodium nitrate, Ammonium chloride, Calcium nitrate, Ammonium acetate, and Ammonium nitrate. The complex media were Czpek''s agar, Potato dextrose agar (PDA), and Malt extract agar (MEA). The results showed that the mycelial growth of three strains of Trichoderma spp. in water agar containing 1000 ppm gardenia crude extract were growth faster than that of with different tested carbon source, and nitrogen source or complex medium alone. In shortening the incubation time tests, rice straw, sugarcane bagasse were soaked in 1000 ppm gardenia crude extract for 24 hours. The medium were sterilized, and then inoculated with 105 spores / ml of Trichoderma strain. The results indicated that sporulation of various Trichoderma strains in rice straw and sugarcane bagasse with 1000 ppm gardenia crude extract was faster than that of with untreated control. The incubation time at the same spore concentration per dry gram was shorten 2-4 days.

碩士<br>國立中興大學<br>植物病理學系<br>88<br>英文摘要1 Effect of nutrient supply and cultural conditions on the growth and sporulation of Metarhizium anisopliae MA850 isolate Li-Te Tseng Abstract The effect of nutrient supply and cultural condition on the growth and sporulation of the entomopathogenic Metarhizium anisopliae MA850 isolate was investigated. The main objective of this work was to provide physiological data base for constructing an optimized mass production system for the microbial pesticide application. The fungus tested was found to have better mycelial growth at pH ranging from 4 to 6, temperature ranging from 25 to 30 ℃, and water potential ranging from —6.8 to —17.5 bars. For sporulation, it had optimum pH at 5 to 7, optimum temperature at 25 ℃, optimum water potential at —6.8 to —11.3 bars, and was greatly favored by the provision of light illumination. Mannose, sucrose, maltose and molasses were among 15 carbon sources the best for supporting the mycelial growth; whereas for the sporulation, sucrose, raffinose, mannose and molasses showed best performance. Inorganic nitrogens including KNO3, (NH4)2SO4 and urea, all supported the mycelial growth well. For the sporulation, urea and KNO3 were the best. Asparagine, tyrosine and histidine, when applied as the sole nitrogen source, supported the growth and sporulation well. However, when these amino acids were applied as additives in a KNO3 containing medium, only histidine was effective in increasing the sporulation. In Czapek’s solution, in which mannose and KNO3 were used as the carbon and nitrogen source’s respectively, the fungus appeared to grow best at C/N ratio around 60 to 80; whereas for the sporulation, the optimum C/N ratio was around 20 to 50. The addition of yeast extract and malt extract enhanced greatly the myceial growth and sporulation; the optimum concentration was 0.5 and 2 %, respectively. Peptone supplemetation, on the contrary, did not enhance the sporulation although significant enhancement of mycelial growth was also observed. Yeast powder was also shown to be an effective additive for supporting the fungal growth and sporulation. In a 2 % yeast powder amended medium, a more than 2 order increase of sporulation was observed. Among 7 animal protein additives screened, pig liver, beef, fish and locust powders appeared to be the superior ones in terms of sporulation. A total of 8 grain decoctions were screened for the efficacy as supporting growth substrate. Among them, peanut decoction performed best in supporting the growth and sporulation. The addition of peanut meal and sesame pomace was also shown to enhance the growth and sporulation of test fungus. For the sporulation, 2 complex media MKYS [M (3 % mannose)+K(0.2 % KNO3)+Y (1 % yeast powder)+S (0.2 % sesame pomace)] and MOUYS [MO (3 % molasses)+U (0.2 % urea)+Y (1 % yeast powder)+S (0.2 % sesame pomace)] were formulated and their superior efficacy were both demonstrated. The value of the constructed formulation was further supported by the evidence that the use of mannose, molasses, KNO3 and urea greatly favored the virulence expression of the test fungus against Spodoptera litura. 英文摘要2 Improvement in conidial mass production of Metarhizium anisopliae MA850 for practical application Li-Te Tseng Abstract The main objective of this study was to establish an improved 2-step (semi-solid) fermentation system for the mass production of Metarhizium anisopliae MA850 for practical application. Using mycelial homogenate as the seed inoculum, an optimization of the solid fermentation system for sporulation was attempted. Eight kinds of grain substrates were screened; pearl barley, oat, corn and rice grains appeared to be among them ideal culture substrate for the attempted purpose. On corn grain, maximum sporulation, (approximately 1.2x1010 conidia/g substrate) was generally detected 8-14 days after inoculation. The sporulation on corn grains appeared to perform best with the addition of water at 50-75 % and the seed inoculum at 105~106 CFU/ml. The addition of wheat bran significantly reduced the agglutination of corn grain and thus facilitate a better- reproducible and enhanced spore yield. The addition of urea, molasses, pig liver powder, sesame pomace and peanut powder, each respectively, also increased the sporulation on corn. On corn/wheat bran (CW, 1/3) -mix, the sporulation was significantly enhanced by the addition of molasses, urea, sesame pomace and yeast powder. The maximum yield of conidia reached approximately 4.62x1010 per gram of the combined supplements-amended CW-mix, which appeared to be 3x and 4.9x of that obtained from corn and rice grains respectively. The established two-step fermentation conidia production system was further valued by the evidence that the total yield and the virulence of the conidia were all comparable to that produced by using conidia as the seed inoculum.

碩士<br>國立交通大學<br>生物科技研究所<br>89<br>Ganoderma lucidum and related species are fungi used in traditional Chinese medicine. Recent studies on this fungus have demonstrated many interesting biological activities, including increasing non-specific immunoactivity. Therefore, this study is carried out to determine the effect of cultivating conditions on the mycelial growth and polysaccharide formation of Ganoderma lucidum in submerged cultures. This research is carried out by “ One factor at a time technique ” and “ Response surface methodology, RSM “. The results of “ One factor at a time technique “ show that olive oil added, medium C/N ratio and medium initial pH have significant effect on Ganoderma lucidum submerged cultures. We study on olive oil added, medium C/N ratio and medium initial pH for optimization of polysaccharide formation of Ganoderma lucidum by RSM technique. The optimal olive oil added, medium C/N ratio and medium initial pH are 0.35~0.55mL, 45~55 and 4~5.

碩士<br>國立屏東科技大學<br>農園生產系所<br>98<br>Clitocybe maxima Gaertn. et G. Mey.: Fr. is an innovative mushroom belong to the group of Basidiomycetes. It has great potential to cultivate in southern Taiwan for the high temperatures weather. The purpose of this study was to evaluate the fine conditions for mycelium growth and mushroom production. The results showed that the mycelium grew the best at the temperature of 25~30℃. In medium pH trials, pH 6 present the best mycelium growth followed in order by the pH of 6.8, 7, 5, 8, 9, 10, and 4. The best media for spawn was made of sorghum grains followed in order by the grains of wheat, oat, and sawdust, coffee residue. The mycelium would completely spread over the cultivation bag when media in bags inoculated with spawn after 45~60 days, their mean fresh weight of Glitocybe maxima was 450 g per bag. In the trials of appropriate temperature for fruiting body initiation, the day-requirement at 30℃ were shorter than those of 20, 24, 28, 32 and 36℃, and the highest yield (around 500 gram per bag) were obtained as well. In casing substrate trials, mushroom extrusion in medium of peat moss mixed with recycled casing substrate need shorter time and obtained the highest yield. The thickness of casing substrate would be better at 8cm for mushroom production than those of 4, 10, 12, 14cm. In the trials of antioxidant capacity, fruiting body of Clitocybe maxima were dried with different method. The results showed that the antioxidant capacity is increased with the methanol extract concentration increasing. Treatment with freeze drying, 121℃ 15 min and 30 min have the higher antioxidant capacity than 40 and 80℃ treatment. Keywords: Clitocybe maxima Gaertn. et G. Mey.: Fr., cultivation bag, casing , antioxidant

碩士<br>東海大學<br>食品科學系<br>96<br>Polyporus umbellatus is a widely used traditional Chinese medicinal fungus. Its sclerotium can be used for treating edema and diuresis. The water-soluble polysaccarides isolated from P. umbellatus possess anti-bacteria, anti-virus, immunomodulatory and anti-tumor activities. P. umbellatus in the wild is growing slowly and has been depleted. Research on the methods of artificial cultivation for production of the biomass and the bioactive metabolites by P. umbellatus is necessary. This study was conducted to optimize the growth conditions in submerged culture. Effect of different aeration rate in the fermentor was also investigated. The effects of differential combination with carbon and nitrogen sources of culture media on mycelial growth and polysaccharide biosynthesis were evaluated in shake flask culture. Glucose and corn steep powder（CSP） combination was the favorable carbon and nitrogen source for the culture media of P. umbellatus. The hightest mycelial dry weight（CDW） and intra-polysaccharides（IPS） production, 5.65 mg/ml and 0.0457 mg/ml, were obtained respectively. The maximum production of exopolysaccharides（EPS） was 0.356 mg/ml, with a relative high molecular weight of 432 kDa was obtained. The hightest production of CDW（4.65 mg/ml）, IPS（0.036 mg/ml） and EPS（0.315 mg/ml） were achieved with concentration of 30 g/l in the glucose medium. The enhancement of CDW was obtained when the glucose concentration was increasing to 40 g/l, but the EPS production was not significant as compared to 30 g/l. CSP in 5 g/l was the suitable nitrogen source concentration. The optimal initial pH of growth medium was pH 7, the highest CDW（7.55 mg/ml） was obtained. The mycelial yield was diminished by decreasing pH, initial pH 4 was unsuitable for cultivaton. The productions of IPS were not significant among unadjest pH, pH 5, pH 6 and pH 7 of the medium, the maximum IPS content was reached to 0.097 mg/ml. The highest production of EPS（0.588 mg/ml） was obtained in unadjest pH culture. The optimal initial pH for mycelial growth and polysaccharides production was the unadjest pH of the culture medium. In fermentor studies, the high aeration rate was favorable for mycelial growth. After 18 days of cultivation, the highest CDW（7.25 mg/ml）and IPS（0.100 mg/ml）production were abtained at 2 vvm with a molecular weight of 206 kDa and intra-β-1,3-glucan content（1.372 μg/ml LE）. The maximum production of EPS（0.600 mg/ml）and exo-β-1,3-glucan（2.773 μg/ml LE） were obtained after 16 days in 1.5 vvm culture. A two-stage aeration rate process was evaluated. The highest production of CDW, IPS and EPS obtained from two-stage fermentation was two days earlier than any other single-control. The maximum CDW（7.14 mg/ml）, IPS（0.076 mg/ml）and intra-β-1,3-glucan content（1.221 μg/ml LE）were abtained after 16 days of cultivation. The molecular weight of IPS was distributed on 331 to 398 kDa. The highest production of EPS（0.583 mg/ml） with the molecular weight of 20.2 kDa and exo-β-1,3-glucan（1.193 μg/ml LE） were obtained after 14 days of cultivation.

碩士<br>國立屏東科技大學<br>熱帶農業暨國際合作系所<br>97<br>This study investigated effects of different culture media and conditions on the growth of mycelia of a specific fungus in Taiwan, Antrodia camphorata T. T. Chang & W. N. Chou. The main purpose of this study is to find out the best conditions and procedures for the growth of A. camphorata mycelia , then setup a complete model for culturing A. camphorata and even commercial production. This study focused on the effects of different culture media and temperature conditions on the growth of Atrodia camphorate. The mycelia of A. camphorata had same growing rate in PDA (potato dextrose agar) at 26 and 28 oC and both are better than at 30 oC. The descending order of mean diameters of mycelia on the 21st day after inoculating A. camphorata on PDA, PDA + malt ectract, PDA + yeast extract, and PDA + camphorata broth were PDA + malt extract (3.45 cm) > PDA (2.95 cm) > PDA + yeast ectract (0.1 cm) and there were significant different among the three data sets (p &amp;lt; 0.05); however, PDA + camphoratebroth (3.20 cm) was not significant different from PDA and PDA+malt extract (p > 0.05). PDA with malt extract and camphorate broth can definitely improve the growing rate of A. camphorata mycelia. When the dosages of malt extract that added to PDA varies from 20 g/L to 30 g/L to 40 g/L, the mycelia growing rates are not significantly different from each other (p > 0.05); however, the the higher is the dosage of camphorata broth, the higher is the growing rate of the mycelia . Wheat grain was not a good substrate for A. camphorata mycelia growing no matter whether the grain were added the camphorata broth or not. The 40th days records of average diameters of the mycelia growing on 100 mL (136 g) brown rice added by 60 mL camphorata broth (5.52 cm) was higher than 100 mL (90 g) wheat grain added by 60 mL camphorata broth (1.75 cm), and there was significant difference between them. Brown rice is a good substrate for the camphorata mycelia growth. Wild C. kaneherai wood is better than cultured C. kaneherai wood for A. camphorata mycelia growing.

Wax-based edible coatings for strawberry fruit were formulated and optimized. The weight loss was the preliminary quality parameter for determining the appropriate composition of the waxbased coating. Different combinations of wax to coconut oil ratio and amount of ethanol were performed to create a coating solution that would result in reduction of the weight loss while giving a desirable appearance. After conducting the preliminary study, it was concluded that the combination of 4 wax: 1 coconut oil in 70 mL ethanol was the best combination in order to significantly reduce the weight loss. Beewax and candelilla wax were used as the wax-basis, with coconut oil in the ratio 4:1. Both types of coating were applied onto strawberry fruits and these were evaluated for weight loss, surface color, ascorbic acid content, decay incidence, yeasts and mold count. A sensory evaluation was also performed. Strawberries were stored for 10 days at 10ºC. Both wax-based coatings produced significant reduction in weight loss in comparison to the uncoated fruits. Furthermore, a significant decrease of 20% and 15% in the reduction of ascorbic acid was noted on the 7th day of storage, for beewax - coconut oil coating and candelilla wax - coconut oil coating, respectively. The presence of mycelial growth often dictates the shelf-life of the strawberry fruit. In this study, a significant reduction in the decay incidence was observed both in beewax-coconut oil and candelilla wax-coconut oil coating. This was further supported by the lower yeast and mold count obtained for the coatings used. The changes in color between coated and uncoated fruits were not significantly different except for the L*value with higher values for coated fruits. In the sensory evaluation of the uncoated and coated fruits, judges also distinguished the color between the control and the treated fruits, after two days of storage at 4ºC. Significant differences were also noticed between fruits coated with candelilla wax- coconut oil and the control, regarding other sensory parameters. However, there were not significant differences between beewax-coconut oil based coating and the uncoated fruits. Edible wax-based coating of beewax-coconut oil and candelilla wax-coconut oil could help to extend the shelf life of strawberry since after 10 days of storage the yeasts and molds count was 4.0 × 105 cfu/g and 5.3 × 105 cfu/g for beewax-based and candelilla wax-based coated samples, respectively, lower than 9.4 × 107 cfu/g for the uncoated ones. However, those counts are still high, indicating a shelf life of beewax-based and candelilla wax-based coated strawberries shorter than 10 days.