| Ganoderma is not Mesona chinensis, but it is often treated as Mesona chinensis to bring the dying back to life in numerous tales, which shows the strong love of people for Ganoderma. The mystery of Ganoderma has been unveiled through the long-term effort of many researchers. From artificial cultivation to the separation of active ingredients, from pharmacological effect experiments to finishing full genome sequencing, each step marked that the study of Ganoderma boarded a new level. With the expanding of Ganoderma health care market, its cultivation scale increased sharply, which put forward higher requirements on replacement of new species of Ganoderma Lucidum. Therefore, it has a very vital significance to research the genetic breeding of Ganoderma Lucidum. This study mainly contains the following parts:1. Collected and separated 18 varieties from parts of Ganoderma Lucidum’s main producing areas, and made a classification and identification for them by using morphology, ITS and ITS-RFLP methods. The results showed they could be divided into 3 broad categories:13 strains of the common Ganoderma Lucidum,1 strain of the common Ganoderma Sinense, and 4 strains of the rare Ganoderma Resinaceum. And we got Ganoderma Lucidum of different specific agronomic traits, such as non-spore Ganoderma Lucidum and so on, which could provide rich germplasm resources for hybrid breeding.2. we found there existed big differences in the germination characteristics of three kinds of Ganoderma Lucidum. Ganoderma Resinaceum germinated germ tubes in 24h in the sterile water, and after 48h the germination rate was 6.77% while the basidiospores of Ganoderma Lucidum and Ganoderma Sinense couldn’t germinate. Further tests showed that the fresh sprouting basidiospores could germinate in malt extrat medium, and after 48h the germination rate of Ganoderma Resinaceum was 23.39%, but the basidiospores of Ganoderma Lucidum and Ganoderma Sinense germination rate was extremely low. Through the observation of the germination spores, we found the germination was irrelevant to the size of the germination spores while the germination of Ganoderma Lucidum and Ganoderma Sinense was relevant to the width of the spores. The average length and width of Ganoderma Lucidum germination spores were 7.38μm and 4.46μm, respectively, and they were 10.92μm and 4.95μm respectively for Ganoderma Sinense, all smaller than those of their mature basidiospores. The germination process of Ganoderma Resinaceum is featured with a germ tube on the top or umbilical bottom part. Only a few germinate two germ tubes. Get 92 mononuclear strains of Ganoderma Lucidum,166 mononuclear strains of Ganoderma Resinaceum,35 mononuclear strains of Ganoderma Sinense.3. We obtained the mononuclear strains from xue-zhi and wu-bao and non-spore Ganoderma Resinaceum, through protoplast nucleation. We also obtained mononuclear strains from Gutian based on the research of the chlamydospore of Ganoderma Resinaceum. It reflected that the life circle of Ganoderma Resinaceum possibly had some asexual minor-circle like Flammmulina velutipes:mononuclear mycelia chlamydospore germinated into monocaryon hypha; dicaryohypha chlamydospore germinated into dicaryohypha; and dicaryohypha chlamydospore germinated into monocaryon hypha.4. We made an accurate determination of mating factor for the mononuclear strains using conventional mating type determining methods and nuclear transfer experiments combining OWE-SOJ experiment. The result showed that there were four kinds of mating type in mononuclear strains germinated by basidiospore of Ganoderma Lucidum, Ganoderma Sinense, and Ganoderma Resinaceum. The chi-square result showed that mononuclear strains mating type of Ganoderma Lucidum existed segregation distortion phenomenon. Several kinds of mating type of Ganoderma Lucidum were determined. We presumably got 8 A factors and 8 B factors in 7 strains of Ganoderma Lucidum, and 4 A factors and 4 B factors in 3 strains of Ganoderma Resinaceum. There were no compatibility between mononuclear strains of Ganoderma Resinaceum and Ganoderma Lucidum, and of Ganoderma Lucidum and Ganoderma Sinense; they couldn’t pair and formed clamp connection.5. We determined the growth rate for part of the mononuclear trains. We obtained 40 hybrids of Ganoderma Resinaceum and 95 hybrids of Ganoderma Lucidum by preliminary identification of hybrids based on observing parent antagonism experiment and clamp connection. The result of growth rare experiment indicated that hybridization can obtain heterobeltiosis hybrid which accounted for 22.50% in the hybrids of Ganoderma Resinaceum,10.87% of Ganoderma Lucidum. According to the initial screening result, We choosed 9 hybrids of Ganoderma Resinaceum and 51 hybrids of Ganoderma Lucidum to be used in fruiting experiment. It showed that 5 hybrids of Ganoderma Resinaceum can fruit normally and the production of 2 hybrids increased significantly. In order to make a comprehension aseessments, we applied grey relational degree method and determined the agronomic trait including single bag weight, drying rate, growth speed, stem length, lid thickness, et al. Then we got 5 hybrids whose comprehension assessments are above all the parents. And we made a preliminary analysis on basidiospore ejection of xue zhi hybrids. |
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