| Panax ginseng belongs to Araliaceaae Panax perennial herb which has great medicinal value. Ginsenoside is one of the important components.Ganoderma lucidum belongs to basidiomycotina, polyporaceae, Ganoderma, a kind of large filamentous fungus,which is a treasure of edible and medicinal Fungi in China.Ganoderma Triterpene and Ganoderma lysaceharides are the most important components.The BIB AC of the Panax ginseng was used in this experiment which had been constructed. Ganoderma lucidum was used as receptor when the large DNA fragment C07D9was transferred to the protoplast of Ganoderma lucidum by electroporation. We establish a large fragment of DNA genetic transformation system, which lay a theoretical and technological foundation for creation of Lucid Ganoderma resource.1. The optimal conditions of Lucidum Ganoderma protoplast preparation and regeneration: the concentration of Lywallzyme is2%, fungus age is5days, temperature of enzymolysis is31℃, enzymatic hydrolysis time is2.5hours, the solution pH is5.5, the osmotic pressure stabilizer selects mannitol which concentration is0.6M. MYG solid medium is regeneration medium.2. The optimal conditions of transformation are described as follows:electrode gap is2mm, the voltage is1200V, addition amount of protoplasts is80μL, dielectric is mannitol, addition amount of large DNA fragment C07D9islOμL.3. PCR test and Southern Blot detection showed that large DNA fragment C07D9was integrated into the genome of Ganoderma lucidum.4. The determination of growth status and observation of growth curve showed that the growth of transformations was less slowly.5. Through the comparision of content of Ganoderma Triterpene, showed that the transformants is lower.6. Through the detection of content of Ganoderma lysaceharides, showed that the content of the transformants is as much as Ganoderma lucidum.7. Through the detection of ganoderma lucidum cellulose activity, showed that the cellulose activity of the transformants is lower. |
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