| Ganoderma, as one kind of mushrooms, belongs to Basidiomycota, and used by Chinese people for thousands of years. In present study, scientists found that Ganoderma could improve immune, cure cardiovascular and cerebrovascular diseases, and so on.Because of its high value, the study of Ganoderma was focus on its cultivation, and the chemical components and medicine efficacy of Ganoderma were researched little in past years. Meanwhile, the registration system of edible fungus in China is not perfect at present. So the Ganoderma was varietal complexity, and the genetic relationship was not clear. All of these restricted the application and exploitation of Ganoderma in China.In this paper, I compared the genetic specificity and medicine effect of different Ganoderma strains. Here, 30 strains of Ganoderma were studied on their antagonistic ability. And 18 stains that growed potential exuberantly were studied on their genetic specificity from mycelium antagonistic, strains phenotype, isozyme, genome to rDNA. At the same time, the differences in chemical components and medicine efficacy of 18 strains were compared, and strain that had better medicine efficacy was selected. This paper carried out the following studies :1. The genetic specificity of different Ganoderma strains was investigated by antagonistic analysis. Two of 30 Ganoderma strain, were inoculated together on culture plate in a petri dish to test their antagonistic relationship. Then analyzed then-genetic relationships by the antagonistic relationships. There were antagonistic relationships among 30 Ganoderma strains, and there were stronger antagonistic relationships between American Ganoderma strain and the other strains.2. Phenotype transmissibility shapes such as different morphology yield, the growth cycle, sporulation, Spore external morphology phenotype genetic traits were compared by cultivation with wood in sheds. Assured that ZiZhi and ZiLingZhi, JingDa and MeiGuoDaLingZhi had closer genetic relationship, respectively. Meanwhile, the JingDa, MeiGuoDaLingZhi and ChiZhi were suitable for mass production.3. 18 Ganoderma strains were used to study further genetic relationships by esterase isozyme zymogram. In esterase isozyme zymogram, the electrophoresis bands were scored as present (1) or absent (0) and used as a raw data matrix. A square symmetric matrix of genetic distance was obtained using Nei & Li's coefficient. Then the dendrogram was generated by UPGMA. The genetic similarity coefficient was 0.45 among 18 Ganoderma strains, which indicated that all of those strains had further genetic relationships and genetic distance in the whole; No.3 and No.5 strains had the same isozyme zymogram, and their similarity coefficient came to 0.95, which indicated that they had closest genetic relationship; No.7 and No. 8 strains had farthest genetic relationship, and their similarity coefficient was only 0.45. Esterase isozyme zymogram can be used as a valid tool to study the genetic relationships.4.18 Ganoderma strains were used to study their further genetic relationships and genetic diversity by AFLP technique. The Ganoderma strains' genome DNA was extracted by CTAB method. After inactivation of the restriction endonucleases with EcoRI and MseI, the genomic DNA fragments were ligated by EcoRI and MseI adapters. And the template DNA generated was pre-amplified and selective amplified with AFLP primers. After gel electrophoresis, coloration and statistics, the dendrogram was generated by UPGMA. The AFLP analysis of 14 E3/M3 primer combinations on 18 strains yielded a total of 177 polymorphic bands; 18 Ganoderma strains' similarity coefficient was only 0.21, which indicated that they had farther genetic relationships. AFLP has high resolution, and test Ganoderma strains' genetic relationships efficiently.5. By ITS PCR-RFLP technique, 18 Ganoderma strains were used to study their ITS sequences, and test their phylogenetic relationships and genetic relationships. After amplification of ITS region of rDNA, PCR-amplified products were digested by four restriction endonucleases HinfI, HaeIII, TaqI,MspI respectively. Restriction fragments were electrophoresed on agarose. Then the dendrogram was generated by UPGMA. 18 Ganoderma strains were grouped into three groups: American Ganoderma strain was fall into one group and had further genetic relationships from the other strains. ITS PCR-RFLP is able to test Ganoderma strains' genetic relationships efficiently.6. Assured the relationship of the medical component and genetic relationship of Ganoderma stranins according to Ganoderma Polysaccharide and Triterpene contents and compounds. The result showed that the strains would effect Ganoderma polysaccharide, Triterpene contents and compounds directly, and there were no necessity relation between the same strain Ganoderma polysaccharide and Triterpene contents. The Ganoderma polysaccharide and Triterpene contents of hypha and fruit-body had obvious distinguish . These experiment explain that the main medical Ganoderma component may decided by Ganoderma resource diversity. 7. To investigate the effect of Ganoderma on immune fuction. Ip red blood cells of chicken and cyclophosphamide(CY) in mice. After 7 days, all mice showed different levels of humoral immunity. And the lower immune function caused by Cyclophosphamide on mice were observed. The result shows that every kind of Ganoderma has effects on prompting immunity in mice, but only Zilingzhi, Zizhi, Ribenpingzhi and Hongzhi have remarkable effect.8. To observe the effect of Ganoderma on immune function. After injecting CY in mice 7 days, all mice showed different levels of humoral immunity. And the lower immune function caused by Cyclophosphamide on mice were observed. The result shows that every kind of Ganoderma has effects on prompting immunity in mice, but only Zilingzhi, Zizhi, Ribenpingzhi and Hongzhi have remarkable effect.9. Ganoderma can improve lymphocyte proliferation responses induced by ConA of immunosuppress mice and increase IL-2 production by T lymphocyte. Excited by antigen or mitosin like ConA, T lymphocyte begins to produce IL-2. Moreover IL-2 improves proliferation and differentiation of T lymphocyte. Theses results indicated that Ganoderma could enhance cell immunity. This result showed that these kinds of Ganoderma have different effects, and Zilingzhi, Ribenpingzhi, Zizhi have obviously effects.10. To investigate the effects of Ganoderma on anti-wearyin mice. Mice were divided into proups randomly. After giving the Ganoderma fruit body decoction orally to mice for 15 days, each mouse was burdened clog which weight is 8% of its own avoirdupois, measured time of exhausting swimming. The results show that Zilingzhi, Mingxing1~#, Ribenchizhi, Xinzhoulingzhi, Jingda, Meiguodalingzhi, Chizhi, Tianzhi, Zizhi, Lingzhi, Dechanglingzhi4~#, Hongzhi and Dechanglingzhi1~# in this experiment can obviously prolong exhausting swimming time. Although other kinds of Ganoderma have effects too, were not remarkable. 11. The study observed different effects of Ganoderma on improved the effectiveness of sleep in mice. Divided mice into groups randomly. After giving the Ganoderma fruit body decoction orally to mice for 3 days, we did sleep-promoting test in combination with pentobarbital sodium. The result indicated that Ganoderma can reduce the autonomic activity, and promoting sleep in combination with pentobarbital sodium. But many kinds have bit effect on mice, only Zilingzhi, Ribenchizhi, Jingda, Meiguodalingzhi, Tianzhi and Zizhi have the remarkable effects.12. To investigate the anti-anoxia effects of different kinds of Ganoderma in mice, mice were randomly divided into different groups, given the Ganoderma fruit body decoction orally for 15 days. Acute anoxia on mice was produced by decapitation, observing the time of gasping. The result is the Ganoderma of Xinzhou and Chizhi can prolong the gasping time in mice subject to acte anoxia which compared with control group. And the other kinds of Ganoderma can also prolong the survival time of mice but don't have the remarkable effect,13. We used an in vitro model system, PC12 cells, to observe the effects of Ganoderma on protecting P12 cells. Incubating with Ganoderma fruit body extract for 2 day, we compared effect among different kinds of Ganoderma on cell proliferation of PC12 by MTT assay. And we observed that Ganoderma could induce neuronal differentiation of PC12 cells. Moreover, different kinds of Ganoderma have different effects. Compared with control cells, the Zilingzhi, Jingdalingzhi, Jingda, Meiguodalingzhi, Ribenpingzhi, Lingzhi10~# , Hongzhi and Dechanglingzhi1~# can obviously promote the proliferation of PC12 cells. And Zilingzhi, Lingzhi, Yuanyi6~#, Hongzhi, Dechang1~# can induce PC12 cells differentiation remarkably. Comprehensive evaluation revealed : Lingzhi and Dechang1~# on PC12 cells with better protection. |
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