Studies On Protoplast Of Pholiota Nameko Induced By High Voltage Corona Electric Field

Pholiota nameko is a rare edible fungus,rich in dietary fiber,crude protein,vitamins and various amino acids,with the advantages of improving immunity,anti-cancer,relieving muscle pain,and delicious taste,smooth taste,is strongly welcomed by consumers,has high economic value and practical value.In order to expand its breeding methods and create high-quality resources.Based on the protoplast of pholiota nameko,this paper intends to explore the new technology and ideas of pholiota nameko mutagenesis breeding by optimizing the method of corona electric field mutagenesis,as well as the growth rate of the mycelium of pholiota nameko mutagenesis and the changes of the activities of laccase,protease and cellulase.The experimental results are as follows:1.The optimal conditions for the preparation of the pleulococcus mushroom protoplasm system were as follows: bacterial age of 7 days,infiltration stabilizing agent mannitol,concentration of infiltration stabilizing agent 0.6mol/L,enzymatic hydrolysis time of 4h,and concentration of lysozyme 1.5%.2.When the discharge voltage was 6kv and 7kv,and the time was 10 s,30s,1min,3min,5min and 10 min,respectively,the regeneration rate of pholiota nameko protoplast decreased gradually with the increase of discharge voltage and time.3.The growth rate of protoplast regenerated mycelium was significantly different after mutagenesis,and the growth rate of mycelium on the fifth day in MS+solid medium had the greatest difference,which was increased by 70.81% compared with CK group.The mycelium of 6kv 30 s group grew best in wheat grain medium,and the growth length to 14 days was 1.47 times that of CK group.4.The activity of laccase was different in different media,among which,the activity of laccase in the 7k V and 30 s treatment groups was the strongest on the seventh days of culture in CMC-NA liquid medium,which was 12.5 times that of the control group.5.The protease activity in MS+,6kv and 1min treatment group,was the strongest on the 5days culture,which increased by 23.25% compared with the control group.It’s the most varied group of all Cellulase activity was different in different media,Cellulase activity in CMC-Na liquid medium was the strongest,which was9.06 times of CK group on the fifth days culture.6.The experiment on the emergence of mycelium regeneration from the protoplast of the mutagenized pholiota nameko showed that the mycelium treated with 6kv 5min had the fastest growth rate,with mycelium and bright color fruit body at 6kv 30 s treat while the shape of 7kv 10 s and 7kv 1min was poor and the surface was rough.These results provide a new idea for corona electric field mutagenesis of pholiota nameko,and provide experimental basis for improving strains.