Protoplast Fusion Breeding Of Auricularia Cornea CV.Yumuer And Auricularia Heimuer CV.Baimuer

In this paper,albino strains of Auricularia heimuer were collected、self-crossed and systematic bred to obtain excellent strain of Auricularia heimuer CV.Baimuer with stable genetic characters.The protoplast preparation system of Auricularia heimuer CV.Baimuer was optimized.The critical lethal conditions for thermal inactivation and UV inactivation of protoplasts were screened.The inactivated Auricularia heimuer CV.Baimuer and Auricularia cornea CV.yumuer protoplasts were fused with PEG.Antagonistic test,colony morphology observation,fruiting test and ISSR analysis method were used to identify fusants.The main findings are as follows:1.The albino Auricularia heimuer strain were isolated,purified and cultivated.The white Auricularia heimuer strain JAUH-W-591 with stable genetic characters was obtained by collecting spores and self-crossed.The strain was identified as a white variety of Auricularia heimuer by morphological observation and molecular biological analysis.The experiments showed that the most suitable carbon source for mycelial growth of JAUH-W-591 strain is lactose,the most suitable nitrogen source is yeast extract,the most suitable cultivation temperature is 25-28℃,and the most suitable p H is 6.5.The results of the production experiment showed that the average yield of JAUH-W-591 was 53.5 g per bag,which increased by 7.1% compared with the control.It had obvious commercial advantages and cultivation value.2.The protoplast preparation conditions of JAUH-W-591 were optimized from four aspects: mycelial age,concentration of enzymatic hydrolysis solution,enzymatic hydrolysis time and enzymatic hydrolysis temperature.The results showed that the suitable mycelial age is 10 days,the enzyme hydrolysis liquid concentration is 2%,the enzymatic hydrolysis time is 7 h,and the enzymatic hydrolysis temperature is31℃.3.The critical lethal conditions for UV inactivation of Auricularia cornea CV.yumuer and thermal inactivation of Auricularia heimuer CV.baimuer wrere screened.The results showed that the protoplasts can be completely inactivated under the conditions of UV inactivation of Auricularia cornea CV.yumuer for 3 minutes and thermal inactivation of Auricularia heimuer CV.yumuer at 60℃ for 20 minutes.Using the parental inactivation method as the primary screening marker,the fusion agent PEG was used for fusion,and 10 fusants formed obvious antagonistic lines with the parent strains.The colony morphology of fusants on PDA medium had the characteristics of both parent strains,the yellow-brown primordium was obtained by the fruiting test.Cluster analysis was performed on the parents and 10 fusants using ISSR molecular markers,and the parents and fusants were divided into two groups.