Study On The Optimization Of Fir Culture Medium And The New Cultivation Matrix Of Oudemansiella Raphanipes

The Oudemansiella raphanipes is crisp,tender,delicious,nutritious,with a variety of effective nutrients and flavor substances,and it is a top-grade fungus loved by consumers.With the increasing demand of people,its planting scale has been increasing year by year,which has become a new type with better economic benefits in the production of edible fungi.In this study,the O.raphanipes fruiting body was used as material,and the strain was obtained by tissue separation.Response surface methodology was used to optimize the composition of solid and liquid media which affected the growth O.raphanipes mycelia,and the optimal media formula was determined;combined with solid plate culture and liquid culture methods,the effects of four growth regulators（NAA,GA,Triacontanol,IBA）on the growth and enzyme activity of O.raphanipes mycelia were studied;three kinds of agriculture and forestry were used The waste material（Eucommia ulmoides Oliv,grape branch and cassava stem）was used as the culture medium,and the new culture medium of O.raphanipes was studied.The results are as follows:（1）Study on the optimization of solid culture medium of O.raphanipes the biological characteristics of mycelia were studied by the methods of plate culture and single factor test.Taking the daily growth rate of mycelia as an index,it was found that the addition of the residue extract of Pleurotus eryngii had a significant promoting effect on the growth of mycelia,and the effect of carbon and nitrogen sources on the growth of mycelia was explored.The results showed that glucose was the best carbon source and peptone was the best nitrogen source.Through single factor screening,the optimal range of carbon and nitrogen sources and mushroom dregs was obtained,and the optimal level was selected.Using box Behnken response surface design,the best solid medium formula was obtained:the amount of mushroom dregs was 90.53g/l,glucose was 1.96%,yeast powder was 0.48%.After verification,the daily growth rate of mycelia increased to 0.608cm/d.（2）The single factor test method,Plackett Burman test were used to select the significant factors,the steepest climbing path method and box Behnken response surface optimization.The most suitable liquid medium formula of O.raphanipes was determined.Using Plackett Burman design,three significant factors were screened out from eight factors affecting the biomass of O.raphanipes mycelium:Pleurotus eryngii residue,peptone and VB1.Then,the steepest climbing experiment method was used to approach the maximum response area,and the box Behnken response surface was used to design the optimal medium combination:Pleurotus eryngii residue 44.2g/L,peptone1.26%,VB1 0.015%,glucose2%,potato starch 1.5%,KH₂PO₄ 0.15%,Mg SO₄ 0.05%,rotational speed 140r/min.The average biomass of mycelium was 1.581±0.032g/100m L,which was increased by52.3%compared with the liquid medium（1.038g/L±0.175）.（3）Study on the effect of growth regulators on the growth of O.raphanipes mycelium by single factor screening test,liquid culture method,mycelium enzyme activity test,to explore whether the growth regulators can be applied in the actual production.The results showed that the growth regulators could promote the growth of O.raphanipes mycelium,and the optimum concentration was 5 mg/L^-1GA.NAA is 10 mg/L^-1,IBA is 5 mg/L^-1 and Triacontanol is 5 mg/L^-1.The order of mycelium dry weight was:Triacontanol>GA>IBA>NAA>CK.The results showed that GA>Triacontanol>IBA>NAA>CK.The results showed that IBA>Triacontanol>NAA≥CK>GA.The activity of HMC was determined as follows:Triacontanol>IBA>CK>NAA>GA.The mycelial growth of black skin chicken fir in the cultivation bag was Triacontanol>IBA>NAA>CK>GA.The results show that Triacontanol and IBA can be used in the actual production,which can improve the growth rate of mycelium and reduce the production cost.（4）Study on the new culture medium of O.raphanipes the effect of mushroom production of grape branch and cassava stem is worse than that of the control group,so it is not suitable for the production of O.raphanipes Eucommia ulmoides can be used as a new substrate to replace part of the cottonseed shell for the cultivation of O.raphanipes.The formula is:Eucommia ulmoides 30%+cottonseed shell 10%+sawdust 40%+bran 15%+soybean meal 3%+rapeseed cake 2%+gypsum powder 1%.The results showed that the biotransformation rate of Eucommia group was 53.87%and that of the control group was55.20%.There was no significant difference between the Eucommia group and the control group.However,Eucommia ulmoides belongs to the waste of agriculture and forestry,the cost of raw materials is low,and the market price of cottonseed shell is quite high.According to the calculation of the total output of mushroom and the prices of mushroom classification,Eucommia ulmoides ulmoides ulmoides can effectively reduce the cost and improve the efficiency as the cultivation medium to replace part of cottonseed shell.