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Isolation, identification and application of protoplast fusion products in edible mushrooms.

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

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_318198
Date January 1994
ContributorsZhao, Jiong., 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, xi, 217 leaves : mounted ill. ; 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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