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