Inaug.-diss.--Göttingen.

Thesis (M.A.)--Guo li Zhengzhi da xue. / Cover title. Mimeo. copy. Includes bibliography.

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by Li Shuxian. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1991. / Bibliography: leaves 169-192. / ACKNOWLEDGEMENTS --- p.ii / ABSTRACT --- p.iii / LIST OF TABLES --- p.viii / LIST OF FIGURES --- p.x / ABBREVIATIONS --- p.xiv / Chapter CHAPTER ONE: --- GENERAL INTRODUCTION --- p.1 / Chapter CHAPTER TWO: --- LITERATURE REVIEW / Chapter 2.1. --- Introduction --- p.6 / Chapter 2.2. --- Sexuality --- p.6 / Chapter 2.2.1. --- Heterothallism --- p.9 / Chapter 2.2.2. --- Homothallism --- p.11 / Chapter 2.2.3. --- Sexuality of Volvariella volvacea --- p.14 / Chapter 2.3. --- "Induction and selection of genetic ., markers in higher fungi" --- p.18 / Chapter 2.3.1. --- Physical mutagenesis --- p.19 / Chapter 2.3.2. --- Chemical mutagenesis --- p.22 / Chapter 2.3.3. --- Biological mutagenesis --- p.24 / Chapter 2.4. --- Nuclear behaviour in edible fungi --- p.27 / Chapter 2.4.1. --- Somatic division --- p.27 / Chapter 2.4.2. --- Meiotic events --- p.28 / Chapter 2.4.3. --- Electrophoretic karyotype --- p.31 / Chapter CHAPTER THREE : --- VARIATIONS IN MONOSPOROUS ISOLATES OF VOLVARIELLA VOLVACEA / Chapter 3.1. --- Introduction --- p.32 / Chapter 3.2. --- Materials and methods --- p.33 / Chapter 3.2.1. --- Strains and culture media --- p.33 / Chapter 3.2.2. --- Monosporous isolation --- p.34 / Chapter 3.2.3. --- Linear and radial growth --- p.34 / Chapter 3.2.4. --- Fertility test --- p.35 / Chapter 3.2.5. --- Determination of extracellular cellulase activity --- p.33 / Chapter 3.3. --- Results --- p.41 / Chapter 3.3.1. --- Variations in colonial morphology --- p.41 / Chapter 3.3.2. --- Variations in linear growth rate --- p.44 / Chapter 3.3.3. --- Variations in fertility --- p.46 / Chapter 3.3.4. --- A comparison of extracellular cellulase activity between fertile and sterile monosporous isolates --- p.52 / Chapter 3.4. --- Discussion --- p.61 / Chapter 3.4.1. --- Study on the colonial morphology of monosporous isolates --- p.61 / Chapter 3.4.2. --- Study on the growth rate of monosporous isolates --- p.62 / Chapter 3.4.3. --- Study on the extracellular cellulase activity --- p.63 / Chapter 3.4.4. --- Study on the fertility of monosporous isolates --- p.64 / Chapter 3.5. --- Summary --- p.66 / Chapter CHAPTER FOUR: --- GENETICAL STUDIES ON VARIATIONS OF V. VOLVACEA / Chapter 4.1. --- Introduction --- p.67 / Chapter 4.2. --- Materials and methods --- p.68 / Chapter 4.2.1. --- Strains and cultural condition --- p.68 / Chapter 4.2.2. --- Chemical compounds --- p.70 / Chapter 4.2.3. --- Determination of sensitivities to growth inhibitor and chemical --- p.72 / Chapter 4.2.4. --- UV mutagenesis --- p.72 / Chapter 4.2.5. --- Chemical mutagenesis --- p.73 / Chapter 4.2.6. --- Enrichment methods for auxotrophs --- p.74 / Chapter 4.2.7. --- Isolation and establishment of resistance markers --- p.74 / Chapter 4.2.8. --- Isolation and characterization of auxotrophic mutants --- p.75 / Chapter 4.2.9. --- Fruiting and marker segregation test --- p.75 / Chapter 4.3. --- Results --- p.75 / Chapter 4.3.1. --- Effects of UV irradiation on mycelium and spore germination rate --- p.76 / Chapter 4.3.2. --- Median lethal concentrations (LC50) of chemical mutagens --- p.79 / Chapter 4.3.3. --- Sensitivity of various strains to growth inhibitors and tested compounds --- p.82 / Chapter 4.3.4. --- Identification and characterization of resistant mutants --- p.94 / Chapter 4.3.5. --- Identification and characterization of auxotrophic mutants --- p.115 / Chapter 4.3.6. --- Analysis of genetic markers in progenies of mutants --- p.116 / Chapter 4.4. --- Discussion --- p.118 / Chapter 4.4.1. --- Significance of mutant induction in V. volvacea --- p.119 / Chapter 4.4.2. --- Methods for mutant induction --- p.120 / Chapter 4.4.3. --- Resistant mutants of V. volvacea --- p.121 / Chapter 4.4.4. --- Auxotrophic mutants of V. volvacea --- p.124 / Chapter 4.4.5. --- Study on the sexuality pattern of V.volvacea by analyzing mutant progeny --- p.125 / Chapter 4.5. --- Summary --- p.128 / Chapter CHAPTER FIVE: --- CYTOLOGICAL STUDIES ON VARIATIONS OF V. VOLVACEA / Chapter 5.1. --- Introduction --- p.129 / Chapter 5.2. --- Materials and methods --- p.130 / Chapter 5.2.1. --- Strains --- p.130 / Chapter 5.2.2. --- Feulgen staining method --- p.130 / Chapter 5.2.3. --- Fluorescent staining of nuclei with DAPI --- p.131 / Chapter 5.2.4. --- Microscopic autoradiography --- p.131 / Chapter 5.2.5. --- Scanning electron microscopic examination --- p.132 / Chapter 5.2.6. --- DNA measurement --- p.132 / Chapter 5.3. --- Results --- p.133 / Chapter 5.3.1. --- Number of nuclei in V. volvacea --- p.133 / Chapter 5.3.2. --- Comparison of cytological differences between self-fertile and self-sterile monosporous isolates --- p.143 / Chapter 5.3.3. --- Spore patterns on basidia --- p.145 / Chapter 5.3.4. --- Nuclear DNA content of V.volvacea --- p.148 / Chapter 5.4. --- Discussion --- p.154 / Chapter 5.4.1. --- Staining method --- p.154 / Chapter 5.4.2. --- Analysis of the possible sources of variation in V. volvacea --- p.155 / Chapter 5.5. --- Summary --- p.162 / Chapter CHAPTER SIX: --- GENERAL SUMMARY AND CONCLUSION --- p.164 / REFERENCES: --- p.169

The classical taxonomic methodologies for determining taxa of mushrooms are based primarily on morphological traits. If the mushroom specimens are dried or pulverized, the morphological characteristics are frequently disguised or no longer apparent. Included among these fungi are known to produce psilocybin, which are hard to identify by their macroscopic features. Molecular techniques could help to overcome these problems. In this study, twenty samples of two genera, Psilocybe and Panaeolus, of fungi from genera known to produce psilocybin were analyzed. The profiles of random amplified polymorphic DNA (RAPD) produced by two primer sets were analyzed. Fingerprints of amplified fragment length polymorphism (AFLP) produced by five selective primer pairs were also analyzed. By the analysis of genetic similarity, RAPD and AFLP method were capable of providing genus and species information. AFLP band patterns can provide reliable species test and can be separated in a simple way using PAGE electrophoresis and stained by silver nitrate. The sequencing data of twenty psilocybin-producing mushrooms were established by fluorescent sequencing method on the nuclear small subunit ribosomal DNA (nuc-ssu-rDNA) and the internal transcribed spacer 1 (ITS-1) DNA regions. The sequences of 877 bp DNA fragment of nuc-ssu-rDNA exhibited genus specific DNA sequences. The size (307-344 bp) and sequences variation of ITS-1 showed not only genus-specific but also species-specific DNA motifs. In the results of RNA gene analysis, the internal transcribed spacer DNA has much faster evolving sequences than the small subunit rDNA. A simple method was developed to perform the genus identification using a primer that is specific to either members of the Psilocybe or Panaeolus genus. A second primer was used to amplify a common product. This multiplex PCR was successfully developed for the quick screening of a large number of samples for identification of the genus. The SSCP pattern of the common product is sufficient to reveal the species that was present. If dye-labeled primer was used, the accurate size of common product is also a valuable evidence of species. If an automatic sequencer is available in the lab, the best method of species and genus identification is sequencing the RNA gene. The multiallele DNA fragments of ITS-1 DNA in Panaeolus subalteatus and Psilocybe semilanceata gave strong evidence in species identification. By the phylogenetic analysis of sequence in nuclear small subunit rDNA and the variable internal transcribed spacer 1, even the other species (not included in this study) of the same genus can be easily determined their relationship to the fungal genera known to produce psilocybin. This study developed DNA profiling methods for the identification of members of the genera Psilocybe and Panaeolus, which can provide information not only in taxon determination but also in forensic identification.

Thesis (Ph. D.)--University of Washington. / Vita. Bibliography: L. [103]-105.

Thesis--University of Wisconsin--Madison. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

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Thesis (M.Sc. (Microbiology)) -- University of Limpopo, 2011 / Lectins are among a large number of proteins produced by mushrooms. Mushroom lectins with important biological functions have been isolated and studied. However, none of the studies were reported on lectins isolated from mushrooms indigenous to southern Africa. A galactose-specific lectin from one of the common mushroom species in southern Africa, Schizophyllum commune, was isolated and characterized. Initially, twenty mushroom samples were collected and their crude extracts screened for lectin activities. Assays involved in the screening procedures included heamagglutination, carbohydrate inhibition, enzyme linked glycoprotein (ELGA) and various stability assays. Four different mushroom samples exhibited positive lectin activities with varying stabilities towards thermal treatment and susceptibility to proteolytic degradation. Further screening assays resulted in ZHR1 being selected for identification and purification of the lectin. This was due to its ability to agglutinate rabbit erythrocytes. In addition to its activity being destroyed after 3 hours of treatment with trypsin-NIPAAM conjugate, the activity of this lectin was also completely destroyed after an hour incubation in boiling water. In contrast to other mushroom extracts assayed, heamagglutination activity of the crude extract of ZHR1 was not inhibited by glycoproteins only but also by the sugars such as galactose, lactose and mannose. ZHR1 was identified as S. commune. S. commune lectin (ScL) was purified using affinity chromatography on a fetuin-agarose column and further purified using gel-filtration chromatography on Biogel P-100 column. ScL was characterized as a glycosylated, galactose-specific dimeric lectin with a molecular weight of approximately 32 and 33 kDa. ScL is a thermolabile lectin which loses its activity as early as 5 minutes after being incubated at 60°C. Anti-ScL antibodies, to be employed in screening for the presence of ScL in the protein extracts, were developed in the rabbits and their interaction with ScL was detected using the double immunodiffusion assays whereas their specificity towards the lectin was detected using Western blot. ScL is one of the first mushroom lectins to be isolated and studied in southern African region. If the lectin is found to be exhibiting important biological functions, ScL can be of commercial importance in the region.