Cheng, Chi Keung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 144-160). / Abstracts in English and Chinese. / English Abstract --- p.ii / Chinese Abstract --- p.iv / Acknowledgements --- p.v / Abbreviations --- p.vi / Table of Contents --- p.vii / List of Figures --- p.x / List of Tables --- p.xii / Chapter Chapter 1 --- Literature Review / Chapter 1.1 --- Introduction & Taxonomy --- p.1 / Chapter 1.2 --- Life cycle and morphology --- p.1 / Chapter 1.3 --- Growth requirements --- p.4 / Chapter 1.3.1 --- Nutritional requirements --- p.4 / Chapter 1.3.2 --- Environment factors --- p.5 / Chapter 1.4 --- Fruiting body development in Coprinopsis cinerea --- p.6 / Chapter 1.4.1 --- Physiology of the fruiting process --- p.6 / Chapter 1.4.2 --- Other studies related to the fruiting process --- p.7 / Chapter 1.5 --- Other biological studies in Coprinopsis cinerea --- p.8 / Chapter 1.5.1 --- Meiosis studies --- p.9 / Chapter 1.5.2 --- Mating analyses --- p.10 / Chapter 1.5.3 --- Peroxidase production --- p.11 / Chapter 1.5.4 --- Transformation and gene silencing --- p.12 / Chapter 1.5.5 --- Other studies --- p.13 / Chapter 1.6 --- C. cinerea genome project --- p.13 / Chapter 1.7 --- Transcriptome analyses --- p.14 / Chapter 1.7.1 --- Serial Analysis of Gene Expression (SAGE) --- p.14 / Chapter 1.7.2 --- Analyzing the 5´ةend of transcripts --- p.16 / Chapter 1.7.3 --- Mapping of SAGE tags to the genome --- p.19 / Chapter 1.8 --- High throughput sequencing --- p.20 / Chapter 1.8.1 --- Pyrophosphate sequencing --- p.20 / Chapter 1.8.2 --- Application of pyrosequencing --- p.21 / Chapter 1.9 --- Aims of project --- p.22 / Chapter Chapter 2 --- 5' Serial Analysis of Gene Expression (5' SAGE) from mycelial and primordial stages of C. cinerea / Chapter 2.1 --- Introduction --- p.24 / Chapter 2.2 --- Materials and Methods --- p.29 / Chapter 2.2.1 --- 5' SAGE libraries construction --- p.29 / Chapter 2.2.1.1 --- Mushroom mycelium and primordium cultivation --- p.29 / Chapter 2.2.1.2 --- RNA extraction --- p.29 / Chapter 2.2.1.3 --- Isolation of mRNA --- p.30 / Chapter 2.2.1.4 --- cDNA synthesis --- p.31 / Chapter 2.2.1.5 --- Mmel digestion and Polyacrylamide gel electrophoresis --- p.32 / Chapter 2.2.1.6 --- Formation and amplification of ditag --- p.33 / Chapter 2.2.2 --- Identification of lOObp ditag --- p.34 / Chapter 2.2.3 --- High throughput pyrosequencing --- p.35 / Chapter 2.2.4 --- Tags extraction from ditags --- p.35 / Chapter 2.2.5 --- Genome mapping and annotation --- p.36 / Chapter 2.3 --- Results --- p.37 / Chapter 2.3.1 --- 5´ةSAGE libraries construction --- p.37 / Chapter 2.3.1.1 --- cDNA synthesis --- p.37 / Chapter 2.3.1.2 --- Mmel digestion and ditag formation --- p.38 / Chapter 2.3.2 --- Identification of lOObp ditags --- p.39 / Chapter 2.3.3 --- High throughput pyrosequencing --- p.40 / Chapter 2.3.4 --- Tags extraction from ditags --- p.41 / Chapter 2.3.5 --- Genome mapping and annotation --- p.42 / Chapter 2.4 --- Discussion --- p.46 / Chapter 2.4.1 --- 5´ةSAGE libraries construction --- p.46 / Chapter 2.4.2 --- Tags extraction and genome mapping --- p.46 / Chapter 2.4.3 --- Observations based on the genome mapping data --- p.48 / Chapter Chapter 3 --- Validation of expression patterns of 5' SAGE libraries and analysis of differentially expressed genes / Chapter 3.1 --- Introduction --- p.55 / Chapter 3.2 --- Materials and Methods --- p.58 / Chapter 3.2.1 --- Identification of housekeeping gene by Northern Blot analysis --- p.58 / Chapter 3.2.1.1 --- RNA fractionation by formaldehyde gel electrophoresis --- p.58 / Chapter 3.2.1.2 --- Transfer of RNAs --- p.58 / Chapter 3.2.1.3 --- Probe preparation --- p.59 / Chapter 3.2.1.4 --- "Hybridization, Stringency washes and signal detection" --- p.60 / Chapter 3.2.2 --- Quantitative real-time PCR --- p.61 / Chapter 3.2.2.1 --- cDNA synthesis from 2 developmental stages --- p.61 / Chapter 3.2.2.2 --- Primer design and verification --- p.62 / Chapter 3.2.2.3 --- Real time PCR reaction and data analysis --- p.65 / Chapter 3.2.3 --- Gene expression level comparison --- p.65 / Chapter 3.3 --- Results --- p.67 / Chapter 3.3.1 --- Identification of housekeeping gene by Northern Blot analysis --- p.67 / Chapter 3.3.2 --- Quantitative real-time PCR analysis --- p.71 / Chapter 3.3.3 --- Gene expression level comparison --- p.78 / Chapter 3.4 --- Discussion --- p.126 / Chapter 3.4.1 --- Validation of 5´ة SAGE libraries --- p.126 / Chapter 3.4.2 --- Analysis of highly and differentially expressed genes --- p.127 / Chapter Chapter 4 --- General discussion --- p.135 / References --- p.144 / Appendix --- p.161

Submitted by Gizele Lima (gizele.lima@inpa.gov.br) on 2017-04-11T13:27:50Z No. of bitstreams: 2 2016_TESE_PPGBOT_INPADLKOMURA.pdf: 76912377 bytes, checksum: b240865b92577d7a79887db3c4b61e24 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-04-11T13:27:50Z (GMT). No. of bitstreams: 2 2016_TESE_PPGBOT_INPADLKOMURA.pdf: 76912377 bytes, checksum: b240865b92577d7a79887db3c4b61e24 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-03-04 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / This thesis is an ecological and taxonomic account of Agaricales macrofungi in a terra firme forest at the Estação Experimental de Manejo Florestal do INPA (ZF-2). Over a two year period 669 basidiomes (fruiting bodies) were collected in eight transects (5 × 50 m each), located in primary and secondary forests. A total of 290 species/ mophospecies were identified during the study. The macrofungal composition was different among primary and secondary forests, and this difference was also observed among seasons and collection year. In the dry season, the secondary forest presented the lowest macrofungal richness and abundance when compared with primary forest. In relation to substrate type (trunk, branches, leaves and soil), basidiome richness of leaf litter guilds was higher in secondary forest plots, whereas, in contrast, soil guild richness was greater in primary forest. Marasmioid and gymnopoid fungi were the most representative group in this study, and taxonomic descriptions using macro and microscopic characters complemented with molecular data, ITS (internal trancribed spacer), were carried out in these groups. The taxonomic stage resulted in description of six new species for Tetrapyrgos (provisional names: T. albonigripes, T. brevipileocystidiata, T. brunneolucida, T. cystidiacrassa, T. pileobrunnea, and T. pseudonigripes) and T. longicystidiata as newly registered new recording to Amazonia. Marasmius calvocystidiatus (provisional name) is described as new species and its sister species M. horridulus, which just its type specimen was known is recollected. Nine species with rhizomorph are described, among them, M. cupressiformis, M. populiformis e M. microdendron. In addition, 37 taxa of Marasmius are presented, from which 21 are describe at the species level. For the species included in Omphalotaceae (Gymnopus, Marasmiellus and Rhodocollybia), we restrict to the presentation of molecular data and synopses about them. Overall, this work intends to open up paths for future studies of macrofungal taxonomy and ecology in the Amazon basin. / Esta tese refere-se ao estudo de macrofungos Agaricales em floresta de terra firme na Estação Experimental de Manejo Florestal do INPA (ZF-2). Durante o período de dois anos, 669 basidiomas (corpos de frutificação) foram coletados nos oito transectos (5 × 50 m cada) localizados em áreas de floresta primária e secundária. Um total de 287 espécies/morfotipos foram identificados durante o estudo. A composição de macrofungos foi diferente entre floresta primária e secundária, diferindo também de acordo com a estação e o ano da coleta. A detecção de macrofungos foi muito baixa na estação seca, principalmente na floresta secundária. Observou-se que, em relação aos substratos (troncos, galhos, folhas e solo), a floresta primária apresentou maior índice de riqueza e diversidade para “solo”. Por outro lado, no substrato “folhas”, estes índices foram maiores para floresta secundária. Os fungos marasmioides e gymnopoides apresentaram um número expressivo de espécies/morfotipos. Em virtude disso, a descrição morfológica usando caracteres macro e microscópicos aliada aos dados moleculares com sequenciamento da região ITS (internal trancribed spacer) foi realizada para estas espécies. Esta etapa do trabalho resultou na descrição de seis espécies novas de Tetrapyrgos (nomes provisórios: T. albonigripes, T. brevipileocystidiata, T. brunneilucida, T. cystidiacrassa, T. pileobrunnea, e T. pseudonigripes) e no registro de nova ocorrência para a Amazônia de T. longicystidiata. Marasmius calvocystidiatus (nome provisório) é descrito como nova espécie, cuja espécie irmã, Marasmius horridulus, é recoletada pela primeira vez após sua descrição. Nove espécies de Marasmius que apresentam a formação de rizomorfos foram descritas, dentre estas, M. cupressiformis, M. populiformis e M. microdendron. Além disso, foram também descritos 37 táxons de Marasmius da seção Marasmius, dos quais 21 em nível de espécie. Em relação às espécies compreendidas em Omphalotaceae (Gymnopus, Marasmiellus e Rhodocollybia), restringiu-se à apresentação de dados moleculares e sinopses. De modo geral, o presente trabalho abre veredas que, espera-se que, possam auxiliar novos estudos interessados na compreensão de macrofungos da Amazônia.

Li, Longfei. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 221-238). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Coprinopsis cinerea, 亦稀灰蓋鬼傘，是研究擔子菌發育過程的模範生物。它的生命週期短，容易培養，亦有已建立的遺傳和分子生物研究技術。最近，它的完整基因組序列亦被發表。C. cinerea 的子實體萌生和發展是快速而複雜的過程。它受多種因素影響，如交配基因，營養消耗，光照和溫度。然而，我們對於C. cinerea 出菇的基本機制和涉及的分子途徑仍不清楚。在這項研究中，我採用NimbleGen 微陣列，以了解13,320 個C. cinerea 基因模型的表達。此微陣列覆蓋不同發育階段，包括雙核菌絲體，子實體初體，第2階段原基體，年青和成熟子實體。11,815個預測，在至少一個發育階段表達。707個基因模型在出菇的萌生過程有差異表達。我發現一些可能參與子實體的萌生和發展的份子。它們可能在檢測養分、形態、信號轉導和應激反應方面，擔當重要角色。此外，我亦分析了轉錄因子 、蛋白激酶組和細胞色素P450的基因表達模式。 / C. cinerea 的子實體發展是與光暗週期同步。當子實體初體在沒有光的環境培植，使會形成dark stipe。101個基因在dark stipe 表達差異。它們可能參與原基體成熟的過程。此微陣列基因表達數據，對了解菇機制有價格的信息。 / 胞吞作用是真核細胞透過質膜內陷將外來物質攝取的過程。Rab5 和Rab7 分別控制早期和晚期的胞吞作用。C. cinerea 的胞吞作用是組織由110個基因組成。FM4-64 螢光顯示在C. cinerea 菌絲體的胞吞作用是依賴肌動蛋白和能源。從菌絲體到年青子實體，Cc. Rab5的表達維持相同水平，而Cc.Rab7 的表達則不斷增加，兩者在成熟子實體的表達都是最高，原位雜交體技術顯示 Cc.Rab5 和Cc.Rab7 的 mRNA在年青子實體的子實層以及整個子實層上菌摺表達。我在第2階段原基體進行RNA 乾擾，致使Cc.Rab5 和Cc.Rab7的基因表達敲落。這導致原基體的生長遲緩。及後形成的成熟子實體亦有異常形態。因此，我推測Cc. Rab5 和Cc.Rab7參與出菇過程，並影響擔孢子的形成。這些結果表明，胞吞作用在C.Cinerea 子實體發育過程中發揮定一定的作用。 / Coprinopsis cinerea, is a model organism for studying developmental processes in basidiomycetous fungi. It has a short life cycle, easy to be cultivated in laboratory and can be accessed by various genetic and molecular techniques. Recently, its complete genome sequence was released. The fruiting body development in C. cinerea is a rapid yet complicated process. It is under the regulation of various factors such as mating type genes, nutrients depletion, light and temperature. However, the underlying mechanism and molecular events involved during fruiting body initiation and development remains unclear. / In this study, fruiting body developmental stages including mycelium, fruiting initials, stage 2 primordium, young and mature fruiting body, were analyzed with a comprehensive NimbleGen microarray. 11,815 out of 13,320 predicted gene models were expressed in at least one of the stages. 707 genes were differentially expressed during fruiting body initiation. Potential players involved in nutrients sensing, morphogenesis, signaling pathways and stress response were identified. In particular, expression patterns of all transcription factors, kinome and cytochrome P450s were analyzed. / The fruiting body development of C. cinerea is synchronized with the light/dark cycle. Differentially expressed genes were found in dark stipe produced by keeping fruiting initials in complete darkness. 101 genes, which are likely to be involved in maturation of primordium were identified. / Endocytosis is an essential process in eukaryotes through which cells take up extracellular substrates by membrane invaginations. Rab5 and Rab7 control the early and late stage of endocytosis respectively. The C. cinerea endocytic machinery composed of 110 genes models. The endocytic pathway was traced by FM4-64 and was found to be actin- and energy-dependent. Temporal and spatial expressions of Cc.Rab5 and Cc.Rab7 during fruiting body development were studied. Cc.Rab5 expressed constitutively from mycelium to young fruiting body stage, and reached the highest in the mature fruiting body. The expression of Cc.Rab7 increased continually from mycelium to mature fruiting body stage. From the in situ RNA-RNA hybridization results, both transcripts were localized at the hymenium layer in the young fruiting body and throughout the gill tissue of the mature cap. Knock-down of Cc.Rab5 and Cc.Rab7 by siRNA resulted in retarded growth of the stage 2 primordium and abnormal mature fruiting body. Cc.Rab5 and Cc.Rab7 may be involved in the formation of basidiospores. Endocytosis may play some roles during fruiting body development in C. cinerea. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Lee, Yung Yung. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 156-179). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract --- p.i / 論文摘要 --- p.iii / Abbreviations --- p.iv / Acknowledgements --- p.v / Table of Contents --- p.vi / List of Figures --- p.x / List of Tables --- p.xiii / Chapter Chapter 1 --- Literature Review --- p.1 / Chapter 1.1 --- Importance of fruiting body (mushrooms) production --- p.1 / Chapter 1.2 --- Introduction on Coprinopsis cinerea --- p.1 / Chapter 1.2.1 --- General introduction --- p.1 / Chapter 1.2.2 --- Life cycle and morphology --- p.2 / Chapter 1.2.3 --- Growth conditions for C. cinerea --- p.4 / Chapter 1.3 --- Regulation of fruiting development in C. cinerea --- p.5 / Chapter 1.3.1 --- Regulation by mating types --- p.5 / Chapter 1.3.2 --- Regulation by light-dark cycle --- p.6 / Chapter 1.3.3 --- Regulation by physiological factors --- p.9 / Chapter 1.4 --- Fruiting-specific genes --- p.9 / Chapter 1.5 --- Genome project of C. cinerea --- p.10 / Chapter 1.6 --- Transformation and gene silencing in C. cinerea --- p.11 / Chapter 1.7 --- DNA microarray --- p.12 / Chapter 1.8 --- Endocytosis --- p.13 / Chapter 1.8.1 --- The endocytic pathway --- p.14 / Chapter 1.8.2 --- Rab GTPase --- p.17 / Chapter 1.8.2.1 --- Control of the active and inactive state of Rab proteins --- p.18 / Chapter 1.8.2.2 --- Functions of Rab GTPases in vesicular transport --- p.19 / Chapter 1.8.2.3 --- Rab5 and Rab7 GTPase --- p.20 / Chapter 1.8.3 --- Endocytosis in fungi --- p.21 / Chapter 1.9 --- Aims of project --- p.22 / Chapter Chapter 2 --- Whole genome expression analysis during fruiting body development --- p.25 / Chapter 2.1 --- Introduction --- p.25 / Chapter 2.2 --- Materials and Methods --- p.26 / Chapter 2.2.1 --- NimbleGen 12x135K gene expression microarray --- p.26 / Chapter 2.2.1.1 --- Strains and culture conditions --- p.26 / Chapter 2.2.1.2 --- RNA extraction --- p.27 / Chapter 2.2.1.3 --- Overall design of the NimbleGen custom Microarray --- p.28 / Chapter 2.2.1.4 --- Microarray hybridization, data extraction and normalization --- p.28 / Chapter 2.2.2 --- Microarray data analysis, clustering and GO assignment --- p.29 / Chapter 2.2.3 --- Validation of expression patterns of NimbleGen microarray and analysis of differentially expressed genes by quantitative real-time PCR --- p.29 / Chapter 2.2.3.1 --- cDNA synthesis --- p.29 / Chapter 2.2.3.2 --- Primer design and verification --- p.30 / Chapter 2.2.3.3 --- Real time PCR and data analysis --- p.32 / Chapter 2.3 --- Results --- p.33 / Chapter 2.3.1 --- Whole-genome expression during fruiting body development --- p.33 / Chapter 2.3.2 --- Differentially expressed genes during fruiting body initiation --- p.48 / Chapter 2.3.2.1 --- Fruiting body initiation-specific genes --- p.52 / Chapter 2.3.3 --- Gene expression analysis during fruiting body development --- p.53 / Chapter 2.3.4 --- The C. cinerea kinome --- p.55 / Chapter 2.3.5 --- Transcription factors in C. cinerea --- p.60 / Chapter 2.3.6 --- The cytochrome P450 family in C. cinerea --- p.65 / Chapter 2.3.7 --- Validation of NimbleGen microarray data by quantitative real-time PCR --- p.68 / Chapter 2.4 --- Discussion --- p.77 / Chapter Chapter 3 --- Effect of light on gene expression of fruiting body development --- p.92 / Chapter 3.1 --- Introduction --- p.92 / Chapter 3.2 --- Materials and Methods --- p.93 / Chapter 3.2.1 --- NimbleGen 12x135K gene expression microarray --- p.93 / Chapter 3.2.2 --- Validation of expression patterns of NimbleGen microarray and analysis of differentially expressed genes by quantitative real-time PCR --- p.93 / Chapter 3.2.2.1 --- cDNA synthesis --- p.94 / Chapter 3.2.2.2 --- Primer design and verification --- p.94 / Chapter 3.3 --- Results --- p.95 / Chapter 3.3.1 --- Differentially expressed genes in dark stipes --- p.95 / Chapter 3.3.2 --- Validation of expression patterns of NimbleGen microarray by real-time PCR --- p.102 / Chapter 3.4 --- Discussion --- p.107 / Chapter Chapter 4 --- Endocytosis in C. cinerea and its role in fruiting body development --- p.111 / Chapter 4.1 --- Introduction --- p.112 / Chapter 4.2 --- Materials and Methods --- p.112 / Chapter 4.2.1 --- The endosomal machinery of C. cinerea --- p.112 / Chapter 4.2.2 --- Tracing the endocytic pathway sing FM-64 --- p.113 / Chapter 4.2.2.1 --- Strains and culture conditions --- p.113 / Chapter 4.2.2.2 --- FM4-64 internalization in mycelium of C. cinerea --- p.113 / Chapter 4.2.2.3 --- Drug treatment effect on the internalization of FM4-64 dye --- p.114 / Chapter 4.2.3 --- Temporal and spatial expression of Cc.Rab5 and Cc.Rab7 --- p.114 / Chapter 4.2.3.1 --- Cloning of Cc.Rab5 and Cc.Rab7 --- p.114 / Chapter 4.2.3.1.1 --- RNA extraction and cDNA synthesis --- p.114 / Chapter 4.2.3.1.2 --- TA cloning of amplification products and bacterial transformation --- p.115 / Chapter 4.2.3.1.3 --- PCR screening for positive transformants and sequencing --- p.115 / Chapter 4.2.3.2 --- Quantitative real-time PCR --- p.116 / Chapter 4.2.3.2.1 --- RNA extraction and cDNA synthesis --- p.116 / Chapter 4.2.3.2.2 --- Primer design and verification --- p.116 / Chapter 4.2.3.2.3 --- Real time PCR and data analysis --- p.117 / Chapter 4.2.3.3 --- In situ RNA-RNA hybridization --- p.117 / Chapter 4.2.3.3.1 --- Tissue preparation --- p.117 / Chapter 4.2.3.3.2 --- RNA probe synthesis --- p.117 / Chapter 4.2.3.3.3 --- Hybridization, signal development and image viewing --- p.118 / Chapter 4.2.4 --- Knock-down of endogenous Cc.Rab5 and Cc.Rab7 by siRNA --- p.119 / Chapter 4.2.4.1 --- Strains and culture conditions --- p.119 / Chapter 4.2.4.2 --- Production of dsRNA of Cc.Rab5 and Cc.Rab7 --- p.119 / Chapter 4.2.4.3 --- Digestion of dsRNA to give siRNA --- p.120 / Chapter 4.2.4.4 --- Effects of Cc.Rab5 and Cc.Rab7 siRNA on fruiting body development --- p.120 / Chapter 4.2.4.4.1 --- Application of siRNA to C. cinerea culture --- p.120 / Chapter 4.2.4.4.2 --- Validation of the knock-down efficacy by real-time PCR --- p.121 / Chapter 4.3 --- Results --- p.122 / Chapter 4.3.1 --- The endosomal machinery of C. cinerea --- p.122 / Chapter 4.3.2 --- The endocytic pathway of C. cinerea --- p.127 / Chapter 4.3.2.1 --- Time-course of FM4-64 internalization --- p.127 / Chapter 4.3.2.2 --- Validation of active transport of FM4-64 --- p.129 / Chapter 4.3.3 --- Cloning of Cc.Rab5 and Cc.Rab7 --- p.131 / Chapter 4.3.4 --- Temporal expression of Cc.Rab5 and Cc.Rab7 --- p.133 / Chapter 4.3.5 --- Spatial expression of Cc.Rab5 and Cc.Rab7 --- p.136 / Chapter 4.3.6 --- Effects of Cc.Rab5 and Cc.Rab7 knock-down by siRNA --- p.140 / Chapter 4.3.6.1 --- Observation of effect of siRNA on fruiting body development --- p.140 / Chapter 4.3.6.2 --- Validation of the efficacy of external application of siRNA --- p.143 / Chapter 4.4 --- Discussion --- p.146 / Chapter Chapter 5 --- Concluding remarks --- p.152 / References --- p.156 / Appendix --- p.180

Submitted by Pedro Barros (pedro.silvabarros@ufpe.br) on 2018-07-11T21:06:50Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) DISSERTAÇÃO Ana Emília de Medeiros Roberto.pdf: 2427887 bytes, checksum: 98fa3502db60df7b1feef1a3e2887aa1 (MD5) / Rejected by Alice Araujo (alice.caraujo@ufpe.br), reason: Ajustes para Pedro realizar. on 2018-07-11T21:10:32Z (GMT) / Submitted by Pedro Barros (pedro.silvabarros@ufpe.br) on 2018-07-11T21:13:25Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) TESE Carla Rejane Sousa de Lira.pdf: 8856315 bytes, checksum: 49b2bcdfabf0b25b09c24662646ef705 (MD5) / Approved for entry into archive by Alice Araujo (alice.caraujo@ufpe.br) on 2018-07-13T22:41:22Z (GMT) No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) TESE Carla Rejane Sousa de Lira.pdf: 8856315 bytes, checksum: 49b2bcdfabf0b25b09c24662646ef705 (MD5) / Made available in DSpace on 2018-07-13T22:41:22Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) TESE Carla Rejane Sousa de Lira.pdf: 8856315 bytes, checksum: 49b2bcdfabf0b25b09c24662646ef705 (MD5) Previous issue date: 2016-05-12 / CAPES / Os Agaricomycetes são caracterizados por desenvolverem basidiomas onde produzem basídios e basidiosporos. Grande parte dos representantes deste grupo degrada componentes da madeira, sendo assim chamados de lignolíticos ou lignocelulolíticos. Para ampliar o conhecimento sobre a diversidade taxonômica e ecológica desse grupo de fungos em áreas de Caatinga, foram realizadas coletas em seis áreas de caatinga xerófila e em seis de brejos de altitude, totalizando 72 transectos percorridos. Para avaliar a diversidade, foram utilizados testes de χ² e ANOSIM. No presente estudo, foram coletados 2249 espécimes correspondendo a 191 espécies de Agaricomycetes lignocelulolíticos. Dentre estas espécies, há 132 novos registros para estados, região, bioma, país, continente ou ciência. Após 102 visitas a campo, as curvas cumulativas de espécies não se estabilizaram, indicando que mais coletas são necessárias. Apesar disso, os esforços amostrais, de modo geral, variaram entre 58 e 85% da riqueza estimada e foram considerados suficientes no presente trabalho. De acordo com a frequência relativa das espécies, em todas as áreas analisadas a maioria dos táxons foram classificados como raros, sendo a minoria classificada como abundante. Os resultados dos testes mostraram que há diferença significativa na composição, riqueza e a abundância de espécies entre as fitofisionomias, sendo coletados um maior número de espécies e espécimes nas áreas de brejo de altitude. Entre as áreas preservadas e antropizadas do Parque Nacional do Catimbau, não houve diferença significativa na riqueza e na abundância de espécies, indicando que não há influencia das atividades antrópicas na ocorrência de Agaricomycetes poróides no local. Foram coletados mais espécimes durante o período chuvoso, porém não houve diferenciação na riqueza e na composição de espécies entre as estações seca e chuvosa nas áreas de Caatinga estudadas. Também foram empregadas ferramentas moleculares para elucidação de complexos de espécies. A partir delas, foi possível encontrar as novas espécies Datroniella minuta, Megasporoporiella variabilicolor, Perenniporia brasiliensis e P. paraguyanensis e ainda propor as duas novas combinações Megasporoporiella amazonica e M. anoectopora. / The Agaricomycetes are characterized by developing basidiomata in which basidia and basidiospores are produced. Most of the representatives of this group degrades components of wood, so called lignolitics or lignocellulolytic fungi. To increase knowledge about the taxonomic and ecological diversity of this group of fungi in areas of Caatinga, collections were made in six xerophytic areas and six montane areas, totaling 72 visited transects. To assess the diversity of these fungi, χ² and ANOSIM tests were used. In this study, were collected 2249 specimens representing 191 species lignocellulolytic Agaricomycetes. Among these species, 132 are new records for states, region, biome, country, continent or science. After 102 field visits, the cumulative curves of species did not stabilized, indicating that more collections are required. Despite this, effort sampling, generally ranged between 58 and 85% of the estimated richness and was considered sufficient in present work. According to the relative frequency of the species in all analyzed areas, most taxa were classified as rare, being the minority classified as abundant. The test results showed significant differences in composition, richness and abundance of species between vegetation types, and more species and specimens being collected in the areas of altitude. Among the native and disturbed areas of Catimbau National Park, there was no significant difference in the richness and abundance of species, indicating no influence of human activities on the occurrence of poroid Agaricomycetes on site. More specimens were collected during the rainy season, but there was no difference in the richness and species composition between the dry and rainy seasons in the studied areas of Caatinga. Also used were molecular tools to elucidate complex species. It was possible to find the new species Datroniella minuta, Megasporoporiella variabilicolor, Perenniporia brasiliensis and P. paraguyanensis and also to propose two new combinations Megasporoporiella amazonica and M. anoectopora.

Submitted by Pedro Barros (pedro.silvabarros@ufpe.br) on 2018-09-10T21:21:56Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) DISSERTAÇÃO Fernando Cezar Sebastião Silva Júnior.pdf: 1756808 bytes, checksum: b289bb06ecff8d7fced8a0598a5d9354 (MD5) / Approved for entry into archive by Alice Araujo (alice.caraujo@ufpe.br) on 2018-09-17T18:14:04Z (GMT) No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) DISSERTAÇÃO Fernando Cezar Sebastião Silva Júnior.pdf: 1756808 bytes, checksum: b289bb06ecff8d7fced8a0598a5d9354 (MD5) / Made available in DSpace on 2018-09-17T18:14:04Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) DISSERTAÇÃO Fernando Cezar Sebastião Silva Júnior.pdf: 1756808 bytes, checksum: b289bb06ecff8d7fced8a0598a5d9354 (MD5) Previous issue date: 2015-04-15 / CNPq / O gênero Gymnopilus foi descrito por P. Karsten (1980), posteriormente foi incluído na família Strophariaceae por suas espécies serem lignícolas e por apresentar estirilpironas, mais recentemente esse pigmento também foi encontrado em Pholiota P. Kumm em estudos de Biologia molecular. Singer (1986) incluiu este gênero em Cortinariaceae por conta da ornamentação do esporo. As espécies do gênero possuem como características píleos coloridos, himenóforo lamelado e cor marrom–ferrugínea, estipe amarelo, os esporos são ferrugíneo–méleos sob microscópio, com parede dupla e ornamentados, trama da lamela regular e geralmente apresentando queilocistídios. A ‘Lista de Espécies da Flora do Brasil’ cita a ocorrência de apenas 11 espécies do gênero no país. O objetivo deste trabalho é ampliar o conhecimento do gênero Gymnopilus para região Nordeste do Brasil, sua distribuição e riqueza. As coletas foram realizadas em seis áreas da região Nordeste do Brasil. O checklist das espécies encontradas no Brasil foi feito a partir de artigos e livros contendo registros de coleta do gênero. Foi encontrada uma nova espécie para o gênero, Gymnopilus purpureograminicola, em João Pessoa–PB. Novos registros de Gymnopilus sp.1 são apresentados na RPPN Fazenda Almas, e Gymnopilus aff. dilepis na Área de Proteção Ambiental da Serra de Ibiapaba–Ceará e Gymnopilus sp.2 para Recife–PE. O trabalho traz ainda um checklist contendo as 24 espécies do gênero encontradas no Brasil, com breve descrição e estados onde foram coletadas. / The Gymnopilus genus was described by P. Karsten (1980). Which was included in the Strophariaceae family, their species are lignicolous and presents styrylpyrones, more recently this pigment was also found in Pholiota P. Kumm in molecular biology studies. Singer (1986) included this genus under Cortinariaceae due to the spore ornamentation. The Cortinariaceae belonging to the family was later proposed its inclusion in Strophariaceae to present styrylpyrones and proximity to Pholiota in molecular biology studies. The species of the genus have as characteristics colored pileus, lamellar hymenium in rusty-brown and stipe yellow color, the spores are rufous-meleos under microscope, double and ornate wall, plot regular slide and generally presenting cheilocystidia. The 'Flora Species List of Brazil' mentions the occurrence of only 11 species of the genus in the country. The objective of this study is to increase knowledge of Gymnopilus genus to northeastern Brazil, its distribution and wealth. Samples were collected in six areas of the Northeast region of Brazil. The checklist of the species found was made from articles and books containing the genus collection records. A new species have been found for the genus, Gymnopilus purpureograminicola in João Pessoa-PB. New records of Gymnopilus sp.1 are presented in RPPN Fazenda Almas, and Gymnopilus aff. dilepis in the Área de Proteção Ambiental of Serra de Ibiapaba-Ceará and Gymnopilus sp.2 to Recife-PE. The work also contains a checklist incorporate 24 species of the genus found in Brazil, with a brief description and states where they were collected.

Made available in DSpace on 2014-06-12T15:04:52Z (GMT). No. of bitstreams: 2 arquivo545_1.pdf: 3871753 bytes, checksum: 29520b91f30821a5d7d20428e1c61753 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2010 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Amanita é um gênero morfologicamente e molecularmente bem definido, dividido em dois subgêneros, Amanita (com as seções Amanita, Vaginatae e Caesarea) e Lepidella (com as seções Lepidella, Amidella, Phalloideae e Validae). Amanita é importante ecologicamente nos ecossistemas devido à associação ectomicorrízica com gimnospermas e angiospermas. Algumas espécies são venenosas e outras são usadas para fins recreativos. O objetivo deste estudo foi contribuir com o conhecimento da diversidade de Amanita no Brasil. Estudos de materiais correspondendo a espécies de Amanita depositados em Herbários e coletas foram realizados, e 32 táxons são registrados. Destes, 12 ocorreram nas regiões Norte e Nordeste, enquanto nove táxons foram registrados para a região Sul do Brasil e dois táxons para a Sudeste. Os táxons de distribuição geográfica mais ampla no país são A. muscaria ssp. flavivolvata, que ocorre nas regiões Sul e Sudeste, A. coacta, que ocorre nas regiões Norte e Sudeste, e A. crebresulcata, com ocorrência no Norte e Nordeste. Os espécimes de A. campinaranae, A. coacta, A. craseoderma, A. crebresulcata, A. lanivolva, A. sulcatissima, A. viscidolutea e A. xerocybe foram revisados e efetuou-se a lectotipificação dos isótipos de A. craseoderma, A. coacta, A. crebresulcata e A. sulcatissima. Amanita lippiae foi descrita para a região semi-árida do Nordeste e A. petalinivolva sp. ined. mais 14 espécimes representam espécies novas para a ciência. As ocorrências de A. ameghinoi, A. crysoleuca, A. multisquamosa, A. spissa e A. strobiliformis não foi confirmada para o Brasil. Materiais depositados em herbários brasileiros como Amanita sp. , Amanitopsis regalis e Amanitopsis bresadolae pertencem a outros gêneros de Agaricales

Five novel lectin isoforms, Volvariella volvacea (VV) lectins, designated VVA, VVB, VVC, VVD & VVE, were isolated and purified from the fruiting bodies of an edible mushroom, Volvariella volvacea , by ion-exchange chromatographies in a FPLC system. Their molecular masses are very close, as measured by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS); they are 12740, 12737, 12709, 12708 & 12707 Da for VVA, VVB, VVC, VVD & VVE, respectively, but the pI values between VVA and the others show distinct differences; the pI value of VVA is around 6.7 and the others are much closer to each other (higher than pI 9.3). Their sugar-binding specificities are thyroglobulin, N-acetylneuramic acid and galacturonic acid. The mitogenic activities of VVA and VVE with distinct pI values were measured using [methyl -3H]thymidine (3H-TdR) incorporation assays, nucleic acid sequence by rapid amplification of cDNA ends analysis, amino acid sequencing and molecular masses by MALDI-TOF/MS and gel electrophoresis, respectively. / VVA and VVE share 98.2% amino acid sequence similarities. Both VVA and VVE are potent mitogens toward mouse CD3+ & CD4 + T-cells, which were mediated through a calcium-dependent activation signaling pathway (Sze et al., 2004). VVA is slightly more effective than VVE in the induction of T cell activation and proliferation, as demonstrated by 3H-TdR incorporation assays, cell flow cytometry for calcium ion mobilization, immunoblotting blot analysis for tyrosine phosphorylation of Lck proteins and Lck shift (p60lck protein), and two dimensional gel electrophoresis for up-regulated proteins. The gene encoding VV lectin was cloned and characterized. The recombinant protein possessed hemagglutinating activity and mitogenic activity, as demonstrated by hemagglutination assays and 3H-TdR incorporation assays respectively. The endoproteinase Arg-C-digested VVA retained the mitogenic activity but lost the hemagglutinating activity, indicating that the mitogenic activity of VVA is not only dependent on the dimerization and tertiary structure of the protein (Paaventhan et al., 2003; Lin et al., 1997), but also on the primary structure of unique amino acid sequences. These endoproteinase fragments have also been used for study of structure-function relationship. / Sze Cho Wing. / "July 2004." / Adviser: Ken W. K. Liu. / Source: Dissertation Abstracts International, Volume: 67-01, Section: B, page: 0171. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 156-189). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

by Lee Man Yi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 125-137). / Abstract also in Chinese. / THESIS COMMITTEE --- p.i / ACKNOWLEDGEMENTS --- p.ii / ABSTRACT --- p.iii / ABSTRACT (Chinese version) --- p.v / TABLE OF CONTENTS --- p.vi / LIST OF TABLES --- p.xi / LIST OF FIGURES --- p.xv / LIST OF ABBREVIATIONS --- p.xvii / Chapter CHAPTER ONE: --- INTRODUCTION --- p.1 / Chapter 1.1 --- Background of mushrooms --- p.1 / Chapter 1.1.1 --- Cultivated mushrooms --- p.1 / Chapter 1.1.1.1 --- Volvariella volvacea --- p.3 / Chapter 1.1.1.2 --- Pleurotus sajor-caju --- p.4 / Chapter 1.1.1.3 --- Pleurotus tuber-regium --- p.5 / Chapter 1.1.2 --- Chemical composition and nutritional value --- p.6 / Chapter 1.1.3 --- Composition and structure of fungal cell wall --- p.9 / Chapter 1.1.4 --- Medicinal attributes of β-glucan in mushrooms --- p.10 / Chapter 1.1.5 --- Structure and antitumor activity β-glucan --- p.11 / Chapter 1.2 --- Dietary fiber --- p.14 / Chapter 1.2.1 --- Composition of dietary fiber --- p.14 / Chapter 1.2.2 --- Preparation of dietary fiber --- p.18 / Chapter 1.3 --- Structural analysis of polysaccharides --- p.19 / Chapter 1.3.1 --- Isolation of polysaccharides --- p.19 / Chapter 1.3.2 --- Methylation analysis --- p.20 / Chapter CHAPTER TWO : --- MATERIALS AND METHODS --- p.22 / Chapter 2.1 --- Sources and preparation of mushroom samples --- p.22 / Chapter 2.1.1 --- V. volvacea --- p.22 / Chapter 2.1.2 --- P. sajor-caju --- p.22 / Chapter 2.1.2.1 --- Fungal strain --- p.22 / Chapter 2.1.2.2 --- Production of spawn --- p.22 / Chapter 2.1.2.3 --- Production of fruiting bodies --- p.23 / Chapter 2.1.3 --- p. tuber-regium --- p.23 / Chapter 2.2 --- Analysis of mushroom composition --- p.24 / Chapter 2.2.1 --- Moisture content --- p.24 / Chapter 2.2.2 --- Starch content --- p.24 / Chapter 2.2.2.1 --- Total glucose --- p.24 / Chapter 2.2.2.2 --- Free glucose --- p.25 / Chapter 2.2.2.3 --- Measurement of glucose content --- p.25 / Chapter 2.2.2.4 --- Total starch content --- p.25 / Chapter 2.2.3 --- Crude protein content --- p.26 / Chapter 2.2.4 --- Amino acid analysis --- p.27 / Chapter 2.3 --- Preparation of mushroom fiber material --- p.28 / Chapter 2.3.1 --- Enzymatic method --- p.28 / Chapter 2.3.1.1 --- Total dietary fiber (TDF) --- p.28 / Chapter 2.3.1.2 --- Insoluble dietary fiber (IDF) and soluble dietary fiber (SDF) --- p.29 / Chapter 2.3.2 --- Chemical method --- p.29 / Chapter 2.3.2.1 --- Cell wall material --- p.29 / Chapter 2.4 --- Chemical composition of mushroom fiber material --- p.31 / Chapter 2.4.1 --- Monosaccharide composition of non-starch polysaccharides (NSP) --- p.31 / Chapter 2.4.1.1 --- Acid depolymerisation --- p.31 / Chapter 2.4.1.2 --- Neutral sugar derivatization --- p.31 / Chapter 2.4.1.3 --- Determination of neutral sugars by gas chromatography (GC) --- p.32 / Chapter 2.4.1.4 --- Uronic acid content --- p.32 / Chapter 2.4.2 --- Resistant starch content --- p.33 / Chapter 2.4.3 --- Residual protein content --- p.34 / Chapter 2.5 --- Fractionation of mushroom fiber material --- p.34 / Chapter 2.5.1 --- Solvent extraction --- p.34 / Chapter 2.5.2 --- Anion-exchange chromatography --- p.35 / Chapter 2.5.3 --- Gel permeation chromatography --- p.36 / Chapter 2.6 --- Structural analysis of mushroom fiber material --- p.37 / Chapter 2.6.1 --- Linkage analysis by methylation --- p.37 / Chapter 2.6.1.1 --- Preparation of methylsufinyl carbanion (Dimsyl) --- p.37 / Chapter 2.6.1.2 --- Preparation and dissolution of sample --- p.37 / Chapter 2.6.1.3 --- Methylation --- p.38 / Chapter 2.6.1.4 --- Hydrolysis --- p.38 / Chapter 2.6.1.5 --- Reduction and acetylation --- p.39 / Chapter 2.6.1.6 --- Determination of partially methylated alditol acetate (PMAA) by gas chromatograph-mass spectrometry (GC-MS) --- p.39 / Chapter 2.6.2 --- Fourier-transform infrared (FTIR) spectroscopy --- p.40 / Chapter CHAPTER THREE : --- RESULTS AND DISCUSSION --- p.41 / Chapter 3.1 --- Chemical composition of mushrooms --- p.41 / Chapter 3.1.1 --- Moisture content --- p.41 / Chapter 3.1.2 --- Carbohydrate content --- p.41 / Chapter 3.1.3 --- Protein content --- p.44 / Chapter 3.1.4 --- Amino acid profile --- p.44 / Chapter 3.1.5 --- Dietary fiber content --- p.48 / Chapter 3.1.6 --- Cell wall material --- p.53 / Chapter 3.1.7 --- Comparison of the yield and composition of TDF and CWM --- p.55 / Chapter 3.1.8 --- "Monosaccharide composition of the dietary fiber (TDF, IDF and SDF) and cell wall material (CWM)" --- p.57 / Chapter 3.2 --- Fractionation of TDF and CWM --- p.69 / Chapter 3.2.1 --- Solvent extraction --- p.69 / Chapter 3.2.2 --- Monosaccharide composition of solvent fractionated TDF and CWM --- p.71 / Chapter 3.2.3 --- Anion-exchange chromatography --- p.78 / Chapter 3.2.4 --- Gel permeation chromatography --- p.82 / Chapter 3.2.5 --- Monosaccharide composition of fractionated fiber material by anion-exchange chromatography --- p.84 / Chapter 3.3 --- Structural analysis --- p.86 / Chapter 3.3.1 --- Partially methylated alditol acetate (PMAA) --- p.86 / Chapter 3.3.1.1 --- Alkali-extracted water-soluble fractions of V. volvacea fiber material --- p.95 / Chapter 3.3.1.2 --- Alkali-extracted water-soluble fractions of P. sajor-caju fiber material --- p.99 / Chapter 3.3.1.3 --- Alkali-extracted water-soluble fractions of P. tuber-regium fiber material --- p.102 / Chapter 3.3.1.4 --- Alkali-extracted water-insoluble fractions of the mushroom fiber material --- p.106 / Chapter 3.3.1.5 --- Alkali- and acid- resistant fractions of the mushroom fiber material --- p.109 / Chapter 3.3.2 --- Infrared spectroscopy --- p.112 / Chapter 3.4 --- "β (l→3), (→4) glucan" --- p.119 / Chapter CHAPTER FOUR : --- CONCLUSION --- p.121 / REFERENCES --- p.125 / RELATED PUBLICATIONS --- p.137

Lo Kit Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 193-208). / Abstracts in English and Chinese. / THESIS COMMITTEE --- p.i / ACKNOWLEDGEMENTS --- p.ii / ABSTRACT --- p.iii / ABSTRACT (Chinese version) --- p.v / CONTENT --- p.vii / LIST OF TABLES --- p.xiii / LIST OF FIGURES --- p.xviii / LIST OF ABBREVIATIONS --- p.xx / Chapter CHAPTER 1: --- INTRODUCTION --- p.1 / Chapter 1.1 --- An introduction of natural antioxidants --- p.1 / Chapter 1.1.1 --- Definition of antioxidants --- p.1 / Chapter 1.1.2 --- Application of natural antioxidants in foods --- p.3 / Chapter 1.1.2.1 --- Oxidation of foods --- p.3 / Chapter 1.1.2.1.1 --- Autoxidation of food --- p.3 / Chapter 1.1.2.1.2 --- Photo-oxidation of food --- p.4 / Chapter 1.1.3 --- Free radicals and antioxidants --- p.6 / Chapter 1.1.3.1 --- Free radicals and reactive oxygen species --- p.6 / Chapter 1.1.3.1.1 --- Superoxide anion radical --- p.8 / Chapter 1.1.3.1.2 --- Hydrogen peroxide --- p.9 / Chapter 1.1.3.1.3 --- Hydroxyl radical --- p.10 / Chapter 1.1.3.1.4 --- Peroxyl radical --- p.12 / Chapter 1.1.3.1.5 --- Lipid peroxidation of cell membranes --- p.13 / Chapter 1.1.3.1.6 --- Oxidation of LDL and atherosclerosis --- p.16 / Chapter 1.1.4 --- Natural antioxidants and their mechanisms --- p.18 / Chapter 1.1.4.1 --- Carotenoids --- p.18 / Chapter 1.1.4.2 --- Phenolic compounds --- p.20 / Chapter 1.1.4.2.1 --- Flavonoids --- p.21 / Chapter 1.1.4.2.2 --- Phenolic acids --- p.22 / Chapter 1.1.4.3 --- Sterols --- p.24 / Chapter 1.1.4.4 --- Vitamins --- p.25 / Chapter 1.2 --- Antioxidant in mushrooms --- p.27 / Chapter 1.2.1 --- Antioxidant properties of mushrooms --- p.27 / Chapter 1.2.2 --- Characterization of mushroom phenolic antioxidants --- p.30 / Chapter 1.2.3 --- Biosynthesis of phenolic compounds in mushrooms or fungi --- p.33 / Chapter 1.3 --- Assays for evaluation of antioxidants --- p.35 / Chapter 1.3.1 --- Beta-carotene bleaching method --- p.35 / Chapter 1.3.2 --- Scavenging activity of DPPH radical --- p.36 / Chapter 1.3.3 --- Erythrocyte hemolysis --- p.36 / Chapter 1.3.4 --- Scavenging activity of ABTS ´Ø + radical cation --- p.37 / Chapter 1.3.5 --- Scavenging activity of hydroxyl radical --- p.37 / Chapter 1.3.6 --- Assay for lipid peroxidation of rat brain homogenate --- p.38 / Chapter 1.3.7 --- Inhibition of low-density lipoproteins (LDLs) oxidation --- p.39 / Chapter 1.4 --- Analysis of phenolic antioxidants --- p.41 / Chapter 1.4.1 --- Extraction of phenolic compounds --- p.41 / Chapter 1.4.2 --- Determination of total phenolic content --- p.43 / Chapter 1.4.3 --- Chromatographic fractionation of phenolic compounds --- p.44 / Chapter 1.4.4 --- Characterization of phenolic compounds --- p.45 / Chapter 1.4.4.1 --- Thin-layer chromatography (TLC) --- p.45 / Chapter 1.4.4.2 --- High performance liquid chromatography (HPLC) --- p.46 / Chapter 1.4.4.3 --- Liquid chromatography-Mass spectrometry (LC-MS) --- p.47 / Chapter 1.5 --- Objectives --- p.49 / Chapter CHAPTER 2: --- MATERIALS AND METHODS --- p.50 / Chapter 2.1 --- Sample preparation --- p.50 / Chapter 2.2 --- Sample extraction. --- p.51 / Chapter 2.2.1 --- Small-scale methanol and water extraction --- p.51 / Chapter 2.2.2 --- Large-scale methanol and water extraction and fractionation --- p.54 / Chapter 2.2.2.1 --- Large-scale methanol and water extraction --- p.54 / Chapter 2.2.2.2 --- Fractionation of crude extracts --- p.55 / Chapter 2.2.2.2.1 --- Fractionation of methanol crude extract --- p.55 / Chapter 2.2.2.2.2 --- Fractionation of water crude extract --- p.55 / Chapter 2.3 --- Fractionation by column chromatography --- p.58 / Chapter 2.4 --- Assays for measuring antioxidant activity --- p.60 / Chapter 2.4.1 --- Beta-carotene bleaching method --- p.60 / Chapter 2.4.2 --- Scavenging activity of DPPH radical --- p.62 / Chapter 2.4.3 --- Erythrocyte hemolysis --- p.63 / Chapter 2.4.4 --- Scavenging activity of ABTS ´Ø + radical cation --- p.64 / Chapter 2.4.5 --- Scavenging activity of hydroxyl radical --- p.65 / Chapter 2.4.6 --- Assay for lipid peroxidation of rat brain homogenate --- p.66 / Chapter 2.4.7 --- Inhibition of human low-density lipoproteins (LDLs) oxidation --- p.67 / Chapter 2.4.7.1 --- Isolation of human LDLs --- p.67 / Chapter 2.4.7.2 --- Calculation of density --- p.68 / Chapter 2.4.7.3 --- Lowry's method for determination of protein content --- p.69 / Chapter 2.4.7.4 --- Preparation of reagents --- p.69 / Chapter 2.4.7.5 --- Determination of thiobarbituric acid reactive substance (TBARS) --- p.70 / Chapter 2.5 --- Total phenolic content --- p.70 / Chapter 2.6 --- Total carbohydrate content --- p.71 / Chapter 2.7 --- Determination of protein content- the Biuret method --- p.71 / Chapter 2.8 --- Thin-layer chromatography (TLC) --- p.72 / Chapter 2.9 --- High performance liquid chromatography (HPLC) --- p.73 / Chapter 2.9.1 --- Analysis of subfractions of methanol crude extract --- p.73 / Chapter 2.9.2 --- Analysis of fractionated subfractions and subfractions of Pevf and Aa mushrooms --- p.74 / Chapter 2.10 --- Liquid chromatography- Mass spectrometry (LC-MS) --- p.74 / Chapter 2.10.1 --- Liquid chromatography --- p.74 / Chapter 2.10.2 --- Mass spectrometry --- p.75 / Chapter 2.11 --- Data analysis --- p.75 / Chapter CHAPTER 3: --- RESULTS AND DISCUSSION --- p.77 / Chapter 3.1 --- Small-scale extraction scheme --- p.77 / Chapter 3.1.1 --- Extraction yield --- p.77 / Chapter 3.1.2 --- Assays for measuring antioxidant activity --- p.80 / Chapter 3.1.2.1 --- Beta-carotene bleaching method --- p.80 / Chapter 3.1.2.2 --- Scavenging activity of DPPH radical --- p.91 / Chapter 3.1.2.3 --- Erythrocyte hemolysis --- p.98 / Chapter 3.1.2.4 --- Summary for small-scale extraction --- p.105 / Chapter 3.2 --- Large-scale extraction and fractionation scheme --- p.107 / Chapter 3.2.1 --- Extraction yield for crude extracts and subfractions --- p.107 / Chapter 3.2.2 --- Antioxidant activity of subfractions and crude extracts of Aa and Pevf mushrooms --- p.111 / Chapter 3.2.2.1 --- Scavenging activity of ABTS ´Ø + radical cation --- p.111 / Chapter 3.2.2.2 --- Scavenging activity of hydroxyl radical --- p.114 / Chapter 3.2.2.3 --- Assay for lipid peroxidation of rat brain homogenate --- p.125 / Chapter 3.2.2.4 --- Summary for large-scale extraction --- p.135 / Chapter 3.2.3 --- Chemical characterization of the crude extracts and their sub fractions of Aa and Pevf mushrooms --- p.13 8 / Chapter 3.2.3.1 --- Total phenolic content --- p.139 / Chapter 3.2.3.1.1 --- Total phenolic content of crude extract and their sub fractions of Aa and Pevf mushrooms --- p.139 / Chapter 3.2.3.1.2 --- Correlation between total phenolic content and antioxidant activity --- p.140 / Chapter 3.2.3.2 --- Total carbohydrate content --- p.144 / Chapter 3.2.3.2.1 --- Total carbohydrate content of water crude extract and their sub fractions of Aa and Pevf mushrooms --- p.144 / Chapter 3.2.3.2.2 --- Correlation between total carbohydrate content and antioxidant activity --- p.144 / Chapter 3.2.3.3 --- Determination of protein content- the Biuret method --- p.146 / Chapter 3.2.3.3.1 --- Protein content of water crude extract and their sub fractions of Aa and Pevf mushrooms --- p.146 / Chapter 3.2.3.3.2 --- Correlation between protein content and antioxidant activity --- p.147 / Chapter 3.2.3.4 --- Summary of correlation between chemical components and antioxidant activity --- p.148 / Chapter 3.3 --- Column fractionation of ethyl acetate and butanol subfractions of Aa mushroom --- p.151 / Chapter 3.3.1 --- Rf value in TLC and yield of fractionated subfractions --- p.151 / Chapter 3.3.2 --- Total phenolic content of fractionated subfractions --- p.153 / Chapter 3.3.3 --- Antioxidant activity of fractionated subfractions --- p.155 / Chapter 3.3.3.1 --- Scavenging activity of ABTS ´Ø + radical cation --- p.155 / Chapter 3.3.3.2 --- Scavenging activity of DPPH radical --- p.158 / Chapter 3.3.3.3 --- Inhibition of human low-density lipoprotein (LDL) oxidation --- p.163 / Chapter 3.4 --- Chromatographic characterization of the subfractions of methanol crude extract of Aa and Pevf mushrooms --- p.167 / Chapter 3.4.1 --- Thin-layer chromatography (TLC) --- p.167 / Chapter 3.4.2 --- High performance liquid chromatography (HPLC) --- p.172 / Chapter 3.5 --- Chromatographic and spectrometric characterization of the fractionated subfractions of the ethyl acetate and butanol subfractions of Aa --- p.177 / Chapter 3.5.1 --- High performance liquid chromatography (HPLC) --- p.177 / Chapter 3.5.2 --- Liquid chromatography- Mass spectrometry (LC-MS) --- p.186 / Chapter CHAPTER 4: --- CONCLUSION --- p.189 / REFERENCES --- p.193 / RELATED PUBLICATION --- p.209