Study On Protoplast Mutagenesis Of Hypsizygus Marmoreus By High Voltage Corona Field

The Hypsizygus marmoreus is a kind of high-quality edible fungus that can be used as both medicine and food.Its fruiting body is rich in vitamins,amino acids,natural antioxidant substances and micronutrient.with high nutritional value and health care value in improving immunity and anti-tumor.Strain selection,improvement and factory cultivation are the key contents of current research.Protoplast mutagenesis is one of the important methods for strain breeding and improvement,and has been widely used in the field of microbial edible fungi.In combination with the sensitivity of protoplasts and the environmental protection of physical mutagenesis,the conditions of protoplasts preparation and regeneration were studied and optimized with the Hypsizygus marmoreus as the starting strain.Then,the protoplasts were mutagenesis by high voltage corona field,the effects of high voltage corona field on the mycelium growth rate and the activities of laccase and protease were studied,and the suitable mutagenic parameters were selected.It is expected to provide new ideas and technical methods for the selection and breeding of Hypsizygus marmoreus and related research.Through experiment,we get the following conclusions:1.Optimization of protoplast preparation conditions of Hypsizygus marmoreus:Age 7 days,with the concentration of 0.6 mol/L mannitol as osmotic pressure stabilizer,the lysozyme concentration of 2 %,enzymolysis for 4 hours.Regeneration conditions: Age of 6 days,with the concentration of 0.6 mol/L mannitol as osmotic pressure stabilizer,the lysozyme concentration of 2 %,enzymolysis for 3 hours.2.The protoplasts of Hypsizygus marmoreus were sensitive to the mutagenic conditions of high voltage corona field.On the whole,the protoplast regeneration rate of low voltage and long time treatment group was higher,and the mutant strains were more easily obtained.3.The growth rate of the mutants treated with high voltage corona field for 12 min was 22.60 % higher than that of the original strain.4.The activity of Laccase was 7.67 times as high as that of the original strain after 14 min of high voltage corona field treatment.On the 7th day,the protease of the mutant strain was 1.56 times that of the original strain.These results provided the experimental basis for obtaining high quality new strains from protoplasts mutagenesis of the Hypsizygus marmoreus by high voltage corona field.