| The protoplast fusion technology has been a new fast developing breeding technology in cell engineering recent years. It has many exceptional advantages that many other normal hybridization methods cant compare with. The concentrations of lywallzyme, the time of lywallzyme treatment and the culture age regeneration medium were studied to find out the effects on preparation and yield of and Tricholoma mongolicam Imai and Agaricits bispopus Sing in this paper. The optimum conditions for regeneration the yield of Tricholoma mongolicam Imai were obtained by using 5-day-cultured mycelia with the treatment of 2.5% (w/w) lywallzyme for 4 hours and the osmotic-pressure stabilizer 0.6MKCL,were regeneration MB. The rate of formation 7.53 l07 /ml, the rate of regeneration 8.4x10 -4.The optimum conditions for regeneration the yield of Agaricus bispopus Sing were obtained by using 5-day-cultured mycelia with the treatment of 2% (w/w) lywallzyme for 3 hours and the osmotic-pressure stabilizer 0.6MKCL, were regeneration PQA. The rate of formation 6.96 107 /ml, the rate of regeneration7.4 10-4. The releasing process of two protoplasts were also observed morphologically. Tricholoma mongolicam Imai and Agaricus bispopus Sing protoplasts fusion were studied by using the inactivated parents protoplasts fusion technique. Used lethality method, Tricholoma mongolicam Imai was handled in 65 C 30min.Lefhality rate of Tricholoma mongolicam Imai were 100%; Used UV method, Agaricus bispopus Sing was handled in 15W 20min. Lethality rate of Agaricus bispopus Sing were 100%. The ratio of Tricholoma mongolicam Imai and Agaricus bispopus Sing 1 to l,they were mixed and fused in 10 min. The fusion rate was 5.6 10-5.The fusion strain heredity stability was tested to find out its stability 80%. The zygote was identified from its colony character, morphological state, growth rate, is dynamic enzyme of EST, soluble free holophote, LDH, ADH, POD and so on. A strain containing the characters of both parents was picked out. |
