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Intergeneric hybridization of schizophyllum commune and pleurotus florida by protoplast fusion.

by To Siu-wing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references (leaves 182-195). / ACKNOWLEDGEMENTS --- p.VI / ABSTRACT --- p.VII / LIST OF TABLES --- p.IX / LIST OF FIGURES --- p.XI / ABBREVIATIONS --- p.XVII / Chapter PART I --- GENERAL ASPECTS / Chapter CHAPTER 1 --- GENERAL INTRODUCTION --- p.1 / Chapter CHAPTER 2 --- LITERATURE REVIEW / Chapter 2.1. --- History of fungal protoplast fusion / Chapter 2.1.1. --- Fungal protoplast preparation technique --- p.4 / Chapter 2.1.2. --- Application of fungal protoplasts --- p.5 / Chapter 2.2. --- Protoplast fusion by polyethene glycol (PEG) --- p.9 / Chapter 2.3. --- Incompatibility system in fungi --- p.10 / Chapter 2.4. --- Characterization of fusion products by genetic markers --- p.12 / Chapter PART II --- OPTIMIZATION OF PROTOPLAST RELEASE AND PROTOPLAST FUSION STUDIES / Chapter CHAPTER 3 --- PROTOPLAST ISOLATION OF Pleurotus florida AND Schizophyllum commune / Chapter 3.1. --- Introduction --- p.14 / Chapter 3.2. --- Materials and methods / Chapter 3.2.1. --- Strains and culture media --- p.14 / Chapter 3.2.2. --- Protoplast isolation in different types and concentrations of lytic enzyme --- p.15 / Chapter 3.2.3. --- Protoplast isolation using mycelium with different culture ages --- p.17 / Chapter 3.2.4. --- Protoplast isolation in different types and concentrations of osmotic stabilizers --- p.17 / Chapter 3.2.5. --- Collection of protoplast by centrifugation --- p.18 / Chapter 3.3. --- Results / Chapter 3.3.1. --- Effect of type and concentration of lytic enzyme --- p.19 / Chapter 3.3.2. --- Efficiency of protoplast isolation from mycelia with different culture ages --- p.25 / Chapter 3.3.3. --- Effect of types and concentrations of osmotic stabilizers --- p.28 / Chapter 3.3.4. --- Collecting efficiency of protoplast by centrifugation --- p.31 / Chapter 3.4. --- Discussion / Chapter 3.4.1. --- Choice of lytic enzyme system and time for enzyme digestion --- p.33 / Chapter 3.4.2. --- Culture age for maximum protoplast yield --- p.34 / Chapter 3.4.3. --- Choice of concentration and type of osmotic stabilizers --- p.35 / Chapter CHAPTER 4 --- PROTOPLAST FUSION OF Pleurotus florida AND Schizophyllum commune / Chapter 4.1. --- Introduction --- p.38 / Chapter 4.2. --- Materials and methods / Chapter 4.2.1. --- Protoplast formation and size of protoplasts --- p.39 / Chapter 4.2.2. --- Fluorescent staining of protoplasts' nuclei --- p.39 / Chapter 4.2.3. --- Stability of the genetics markers / Chapter 4.2.3.1. --- Preparation of media for checking the presence of genetics markers --- p.40 / Chapter 4.2.3.2. --- Determining the presence of auxotrophic as well as drug resistance markers --- p.42 / Chapter 4.2.4. --- Regeneration of mycelium from protoplast --- p.42 / Chapter 4.2.5. --- Protoplast fusion and screening of fusion products --- p.45 / Chapter 4.3. --- Results / Chapter 4.3.1. --- Size of protoplasts ofPf67 and Scl7 --- p.48 / Chapter 4.3.2. --- Proportion of protoplasts bearing nucleus --- p.48 / Chapter 4.3.3. --- Protoplast regeneration in regeneration medium / Chapter 4.3.3.1. --- Protoplasts regeneration morphologies --- p.52 / Chapter 4.3.3.2. --- Regeneration frequencies and back mutation frequencies of Pf67 and Scl7 protoplasts --- p.58 / Chapter 4.3.4. --- Effect of PEG fusion treatment on auxotrophic and drug resistance markers of Pf67 and Scl7 --- p.60 / Chapter 4.3.5. --- Fusion products obtained from screening process --- p.61 / Chapter 4.4. --- Discussion / Chapter 4.4.1. --- Effect of protoplast isolation and PEG treatment on the two fusion parents --- p.63 / Chapter 4.4.2. --- Structural heterogeneity of protoplasts --- p.64 / Chapter 4.4.3. --- Polymorphic nature of protoplast regeneration --- p.67 / Chapter 4.4.4. --- Protoplast fusion frequence --- p.67 / Chapter PART III --- ANALYSIS OF FUSION PARENTS AND FUSION PRODUCTS / Chapter CHAPTER 5 --- MORPHOLOGICAL AND CYTOLOGICAL STUDIES / Chapter 5.1. --- Introduction --- p.69 / Chapter 5.2. --- Materials and methods / Chapter 5.2.1. --- Strains --- p.69 / Chapter 5.2.2. --- Study on colonial and mycelial morphology --- p.70 / Chapter 5.2.3. --- Fluorescent staining of mycelial nuclei with DAPI --- p.70 / Chapter 5.2.4. --- Study on fruit body and basidial morphology / Chapter 5.2.4.1. --- Fruiting on agar plate --- p.71 / Chapter 5.2.4.2. --- Scanning electron microscopic examination --- p.73 / Chapter 5.3. --- Results / Chapter 5.3.1. --- Variation of colonial morphology --- p.74 / Chapter 5.3.2. --- Morphologies and the number of nuclei in the mycelial cells of fusion parents and fusion products --- p.76 / Chapter 5.3.3. --- Fruit body morphology --- p.82 / Chapter 5.3.4. --- Basidial morphology --- p.84 / Chapter 5.4. --- Discussion --- p.87 / Chapter CHAPTER 6 --- PHYSIOLOGICAL STUDIES OF FUSION PARENTS AS WELL AS FUSION PRODUCTS BY INVESTIGATING THE GROWTH RESPONSES TO DRUGS / Chapter 6.1. --- Introduction --- p.90 / Chapter 6.2. --- Materials and methods / Chapter 6.2.1. --- Strains and media --- p.96 / Chapter 6.2.2. --- Growth responses of the strains to different concentrations of drugs --- p.97 / Chapter 6.3. --- Results / Chapter 6.3.1. --- Comparison of growth pattern as well as growth rate between fusion parents and fusion regenerants --- p.98 / Chapter 6.3.2. --- Growth responses of fusion parents and fusion products on complete medium --- p.105 / Chapter 6.3.3. --- Growth responses of fusion parents and fusion regenerants on complete medium with acriflavin --- p.108 / Chapter 6.3.4. --- Growth responses of fusion parents and fusion products on complete medium with guaiacol --- p.111 / Chapter 6.4. --- Discussion / Chapter 6.4.1. --- General considerations on experimental design --- p.115 / Chapter 6.4.2. --- Growth responses of protoplast regenerants of either fusion parents --- p.116 / Chapter 6.4.3. --- Growth responses on complete medium without fungitoxic drug --- p.117 / Chapter 6.4.4. --- Growth responses on the acriflavin agar medium --- p.118 / Chapter 6.4.5. --- Growth responses on guaiacol agar medium --- p.119 / Chapter 6.4.6. --- Summary --- p.120 / Chapter CHAPTER 7 --- GENETICAL STUDIES / Chapter 7.1. --- Introduction --- p.121 / Chapter 7.2. --- Materials and methods / Chapter 7.2.1. --- Segregation tests of auxotrophic and drug resistance markers in progeny of dikaryotic fusion product --- p.127 / Chapter 7.2.2. --- Complementation test of fusion products as well as the spore germinants of dikaryotic fusion product PS1 --- p.129 / Chapter 7.2.3. --- Recovery of the individual nuclear type of dikaryotic fusion product PS1 --- p.130 / Chapter 7.2.4. --- Genomic fingerprinting / Chapter 7.2.4.1. --- Strains and culture medium --- p.133 / Chapter 7.2.4.2. --- Genomic DNA preparation by cesium chloride (CsCl) method --- p.135 / Chapter 7.2.4.3. --- Genomic DNA preparation by chloroform :TE saturated phenol method --- p.136 / Chapter 7.2.4.4. --- Qualitative analysis of genomic DNA --- p.137 / Chapter 7.2.4.5. --- Quantitative analysis of genomic DNA --- p.137 / Chapter 7.2.4.6. --- DNA amplification by arbitrarily primed -polymerase chain reaction --- p.138 / Chapter 7.3. --- Results / Chapter 7.3.1. --- Progeny analysis and determination of auxotrophic as well as drug resistance markers --- p.140 / Chapter 7.3.2. --- Complementation tests of the fusion products as well as the spore germinants of dikaryotic fusion product PS1 --- p.143 / Chapter 7.3.3. --- Monokaryotic protoplast regenerants of dikaryotic fusion product PS1 --- p.147 / Chapter 7.3.4. --- Studies on extraction of undigested genomic DNA --- p.148 / Chapter 7.3.5. --- Genomic fingerprinting by AP-PCR --- p.155 / Chapter 7.4. --- Discussion / Chapter 7.4.1. --- Genomic DNA extraction --- p.161 / Chapter 7.4.2. --- Recovery of the individual nuclear type of dikaryotic fusion product PS1 --- p.165 / Chapter 7.4.3. --- Genomic changes in fusion products --- p.167 / Chapter 7.4.4. --- Progeny analysis and determination of auxotrophic as well as drug resistance markers --- p.171 / Chapter PART IV --- SUMMING-UP / Chapter CHAPTER 8 --- GENERAL SUMMARY AND CONCLUSION REMARKS / Chapter 8.1. --- General summary --- p.176 / Chapter 8.2. --- Conclusion remarks and future studies --- p.179 / REFERENCES --- p.182 / APPENDIX A SOLUTIONS

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_319110
Date January 1993
ContributorsTo, Siu-wing., Chinese University of Hong Kong Graduate School. Division of Biology.
PublisherChinese University of Hong Kong
Source SetsThe Chinese University of Hong Kong
LanguageEnglish
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xviii, 195 leaves : ill. (some mounted col.) ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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