A Study On The Biological Characteristics And Cultivation Of One Strain Of Wild Coprinus Comatus

China is lack of good Coprinus comatus isolates resources,however the development of industry requires the strains to have high yield,stronger resistance to stress and disease.Domesticating wild isolates resources,especially,native wild isolates resources,are one of the reliable ways to solve this problem.In July 31^st,2015,the author collected a large wild mushroom up to 25cm in the grassland of Luyashan Nature Reserve,Xinzhou City,and named as ZMM-3 strain.In this work,the biological characteristics of ZMM-3 were studied based on the identification of this strain,and then the artificial cultivation of this strain was carried out,the nutritional components and antioxidant properties of fruit bodies was analyzed and food safety was evaluated.The main results were as follows:The pure culture of strain ZMM-3 was obtained by tissue isolation method.Its Internal transcribed space?ITS?was amplified after the genomic DNA was extracted and then sequenced.According to results of ITS sequence and morphological characteristics,ZMM-3 strain was identified as Coprinus comatus.The newly derived ITS sequence was deposited in GenBank under accession number KX015766.The results showed that the optimum culture conditions for ZMM-3 strain were the following:temperature of 25?,sucrose and yeast powder as carbon and nitrogen sources,ratio of carbon to nitrogen of 10:1-20:1,pH 7-8.The cultivation of ZMM-3 strain was carried out.The effects of different media?i.e.original spawn media and spawn media?and substrates on ZMM-3 were studied.The fruit bodies of ZMM-3 were successfully cultivated,and the formation time of primordia,the mature time and number of fruit bodies produced in each cultivating bag,and all the indexes about the fresh weight,dry weight,cap diameter,stalk length and the biological efficiency of mushrooms were counted.The results showed that the optimum original spawn medium for ZMM-3 strain was the mixture of glucose,malt and yeast extract.The optimum spawn medium was wheat medium.There was no significant difference in the formation time of primorda,maturation time and the stipe length of fruit bodies on different substrates?P<0.05?.In addition,the number of fruit bodies?9.66±0.58?and the biological efficiency?63.87±12.09?of mushrooms on straw substrate were significantly higher than that on other four substrates?P<0.05?.So straw substrate was the optimal substrate to cultivate ZMM-3.In order to evaluate the safety of ZMM-3 fruit bodies,icr mice were used to study the sub-acute toxicity of ZMM-3 fruit bodies.The results showed that all the mice of the experimental group lived well after feeding with dry ZMM-3 fruit bodies of high dose?5000 mg/kg·d?,medium dose?500 mg/kg·d?and low dose?50 mg/kg·d?.The physical signs,body weight,behavior,appearance,organ coefficient,the physiological and biochemical indexes of blood and organ structures all were normal.No obvious sub-acute toxicity was seen in tested animals,the fruit bodies of ZMM-3 proved to be non-toxic.The nutritional components and antioxidant properties of ZMM-3 fruit bodies were analyzed and compared with of Tebai-8 strain.The results showed that the ash content and crude protein content in the two samples had no significant difference?P<0.05?,The crude fat content of ZMM-3 was significantly lower than that of Tebai-8?P<0.05?.The reducing sugar,crude fiber,total phenol contents of ZMM-3 were significantly higher than of that Tebai-8?P<0.05?.The Ca and Fe content between these two samples had no significant difference?P<0.05?.Tebai-8 fruit bodies was significantly higher than ZMM-3 in the Zn content?P<0.05?.The harmful heavy metals Cd and Pb were not detected.The fruit bodies of both ZMM-3 and Tebai-8 strains contained eight kinds of essential amino acids,The ratio of the total essential amino acids to the total amino acids of the two protein sources was nearly40%.The ratio of essential amino acids to non-essential amino acids was close to 0.6,conforms to the United Nations Food and Agriculture Organization?the Food and Agriculture Organization,FAO?/World Health Organization?World Health Organization,WHO?put forward the ideal condition of protein.By evaluating and analyzing the amino acids of the two protein sources,the fruit bodies of the two strains were as available protein sources.Antioxidant properties research results showed that only the EC₅₀value of Fe²⁺chelating ability of ZMM-3 strain was higher than that of Tebai-8 strain,the other antioxidant properties indexes of alcohol extract of ZMM-3 fruit bodies were stronger than that Tebai-8 strain.To sum up,ZMM-3 strain has strong resistance to stress and disease resistance,and its fruiting body has high nutritional value.It is a high quality edible fungus germplasm resource with great potential for development.