Studies On The Protoplast Fusion Between Polystictus Versicolor And Ganoderma Lucidum

With the increase of people’s healthy consciousness and the development of science, mushroom is popularly welcomed as the delicious food of fungi with the function of health care. Polystictus versicolor and Ganoderma lucidum are both the traditional food treasure of land with overall nutritive materials and high medical value, with efficacy of resisting cancer, lowing down the blood sugar and enhancing natural immunity effectively, so they are very expensive.Protoplast fusion technology has been one of the heredity and breed means. It enlarged hybridize range between species, made far conditioning cause hybridize become possible, enlarged the memory reservation of mushroom. The research used protoplast fusion technology make Polystictus versicolor and Ganoderma lucidum protoplast come together, and carried the fusant on filtration and identification. The aim was to get the fusant in possession of parents’ merits.Some main factors in the course of Polystictus versicolor and Ganoderma lucidum protoplast preparation and regeneration were studied in this paper. Through orthogonal experiment, we defined that using 5d cultivated mycelia of in liquid static state, which was taken as proper strain age, digested into 2% lywallzyme at 30℃for 3h, with osmotic pressure stabilizer of 0.6mol/L MgSO₄ as optimal preparation condition; using 7d cultivated mycelia of in liquid static state, which was taken as proper strain age, digested into 1.5% lywallzyme at 33℃for 2h, with osmotic pressure stabilizer of 0.6mol/L MgSO₄ as optimal preparation condition. In this condition, the quantity and quality is the best. Both kinds of protoplast regeneration effects will be higher, adding 10g maltose, 4g glucose, 4g yeast extraction to the culture media, 5.5g agar with 0.6mol/L sucrose used as stabilizing osmotic pressure.Using different appearance between Polystictus versicolor and Ganoderma lucidum on hygromycin B and heat inactivated technique, we did fusion experiment between the above two strains under PEG, Ca²⁺ system. Thirteen fusion strains were selected. The frequency of fusion was about 1.76×10^-5. Compared the phenomena that mycelia growth speed, biomass of mycelium in liquid culture media , colony and mycelia microcosmic feature, contents of DNA and antagonistic reaction between randomly selected the fusant F5 and the parents on biological characteristics, we observed that the fusant originated from the two parents fusion and the fusant is close to Polystictus versicolor.