Study On The Breeding Of Resistant Ganoderma Lucidum With Rich Bioactive Macromolecular Polysaccharide

Ganoderma lucidum belonging to Eumyeota, Basidiomycotina, Hymenomycetes,Aphyllophorales, Ganodermataeeae, Ganoderma, is a kind of Chinese herbal medicine contained in Chinese Pharmacopoeia and is also a kind of edible and medicinal fungi.The main active components of Ganoderma lucidum are polysaccharides and triterpenes. Modern pharmacological studies have shown that Ganoderma lucidum has anti-tumor, immune regulation, liver protection, anti-aging, improve the body’s ability to resist hypoxia activities and so on. Ganoderma lucidum and its spore powder has been understood and acknowledged by the public, and its products have been favored by consumers.In this study, "Hunong No. 1" and "Longzhi No. 2" were used as the parents by using the protoplast hybridization technique to breed new strains of Ganoderma lucidum. The conditions of protoplast preparation and regeneration were optimized and 35 new strains of Ganoderma lucidum were obtained. Then, we collected the data of their fruiting bodies yields, spore powder yields and the rate of final products, at the same time HPLC analysis was carried out on the water crude extracts from fruit bodies.The results are as follows:(1) The optimal conditions of protoplast preparation and regeneration are as follws: concentration of the enzyme 2%, pH 5.0, enzymolysis for 3 hours at temperature of 30 ℃. In this condition, the number of protoplasts was about 3.2 × 107cell/mL.Regeneration medium formula is as follows: 10g Glucose, 5g maltose, 5g yeast extract,11% 0.6M mannitol, 20g agar, add distilled water to 1000mL. Regeneration condition:26℃ for 6-7d. Regeneration rate reached 3%.(2 ) Through the microscopic and the antagonistic test, we obtained 14 mononuclear protoplast strains of "Hunong No.1" and 58 mononuclear protoplast strains of "Longzhi No.2",and the mating types of of "Hunong No. 1" strains were determined. Finally, 35 new strains of Ganoderma lucidum were obtained by hybridization. According to the results of the antagonistic tests with their parents, the 35 strains were determined to be different from their parents.(3) Through the bag cultivation, the fruiting bodies of 35 hybrid strains were obtained. At the same time, the data of their fruiting bodies yields, spore powder yields and the rate of final products were collected (The data of spore powder yields are not accurate, and can only be used for the simple comparison between the strains in the same cultivation batch.), and HPLC analysis was carried out on the water crude extracts from fruit bodies. The result shows that Hi-174×L2-56 and Hi-80×L2-56 fruiting body yield are higher, Hi-174×L2-53 and Hi-80×L2-56 fruiting bodies macromolecular polysaccharide are higher, Closing to the data of "Longzhi No.2n" Hi-100×L2-56 spore powder yield are higher, 28 hybrid strains rate of final products are over 90%. Among them, Hi-80×L2-56 combined favorable trait of parental pair, have a better performance in cultivation than other strains.