The Key Techniques Of Nutritional Physiologyand Preparation Conditions Of The Protoplast Of Sparassis Crispa

Sparassis crispa with tender meat,delicious taste and rich nutrition,and has anti-tumor,improve human hematopoietic function,promoting wound healing,is developed in recent years rare edible medicinal fungi in the world.For the study of Sparassis crispa,domestic just started,mainly concentrated in nutrition,pharmacology and cultivation research.In this paper,the nutritional physiology,shaking flask liquid culture condition,protoplast preparation and regeneration condition were studied.The main results are as follows:1.Studied the effects of 12 carbon sources on the growth of the mycelium of the Sparassis crispa.The results showed that the best carbon source was galactose and mannitol,the worst of maltose.In 12 kinds of tested nitrogen sources,the optimum nitrogen source for yeast extract and glutamine,ammino acid was the worst,In 9 kinds of inorganic salt,Zinc sulfate,potassium chloride to promote the best effect.Suitable dosage of 16 mg / L and 0.5 g /L,In 6 kinds of vitamins,VB1 concentration(8 mg / L)has the best effect.In 8 kinds of plant growth regulators,IAA concentration(6 mg / L)effect best,Melissyl alcohol in the tested concentration inhibited mycelial growth.2 The optimal conditions of the liquid culture of the flask were determined.The temperature was 25?,the rotational speed was 256 rpm,the liquid volume was 84 mL/(150mL),the inoculation volume 10 mL/ bottle,the glycoprotein content 227.72 g,and the initial pH 6.0.3.Determine the Sparassis crispa protoplast preparation suitable conditions: steady infiltration agents(mannitol)0.2 mol / L,pH 6.0,1% cellulase and 1% snailase mixed enzyme liquid(mycelial wet weight /g:enzyme solution volume / mL= 1:1).Under these conditions,protoplast preparation rate and regeneration rate is the highest.