Cell Engineering Breeding And Application Of Agrocybe Aegirita Based By MPMS Mutation System

It is intersection period of poverty alleviation and rural revitalization in 2020,edible fungus industry has played an important role in my country’s poverty alleviation plan and will continue to contribute to rural revitalization.In order to enrich the cultivation of edible fungi in Hebei Province and develop new raw materials for edible fungi cultivation,breeding is carried out with the dominant shrub species Vitex negundo var.heterophylla in Hebei Province as the main material,and the excellent strains of Agrocybe aegerita with strong ability to decompose Vitex negundo var.heterophylla are selected.In this project,taking advantage of the strong ability of A.aegerita to decompose sawdust,a multifunctional plasma mutagenesis technology（MPMS）was used to induce mutation breeding of A.aegerita,and a high-yielding A.aegerita strain suitable for decomposing wattle chips was selected.A.aegerita AS7 was subjected to mutagenesis through the MPMS mutagenesis system,and a total of 362 mutagenic strains suitable for decomposing the Vitex negundo var.heterophylla stalk substrate were obtained.The preliminary screening was carried out based on whether there was an antagonistic line,and 33 mutagenic strains with obvious antagonistic reaction were obtained.Passed a series of re-screening tests（the growth rate and vigor of the mutagenized strain,three extracellular enzyme activities of carboxymethyl cellulose,laccase and polyphenol oxidase,the biological efficiency and agronomy of the excellent mutagenized strain Characters,polysaccharides and dietary fiber content）,the excellent mutagenic strain AM220,which is most suitable for decomposing Vitex negundo var.heterophylla shavings,was obtained.Its biological efficiency under Vitex negundo var.Heterophylla striata cultivation substrate was 81.70%,which was 11.57%higher than that of CK（A.aegerita AS7）,which was significantly better than CK.The ISSR molecular marker technology was used to identify the authenticity of the 5excellent mutant strains and CK.The results showed that the genetic distance between the 5excellent mutant strains and CK was between 0.678-0.883,indicating that these 5 excellent mutagenesis strains.There are different degrees of molecular genetic differences between the strain and CK The excellent strain AM220 of A.aegerita was renamed as Xinheda AS7.In the optimized cultivation experiment of Xinheda AS7,the most suitable stock medium formula for the growth of Xinheda AS7 is:78%wheat grains,20%Vitex negundo var.heterophylla shavings,and 2%lime.Under this formula,Xinheda AS7 mycelium grows faster.It is 3.53 mm/d.The most suitable cultivation formula for the growth of Xinheda AS7 is:65%Vitex,20%corncob,13%bran,2%quicklime.Under this formula,the biological efficiency of Xinheda AS7 is 85.29%.After covering with soil,the biological efficiency of Xinheda AS7increased by 4.78%.After adding the sleeve,a small range of high CO₂can be formed,and the high CO₂helps to stimulate the differentiation of the primordium to obtain high-quality A.aegerita with small caps and long stalks.By comparing the effects of different ratios of crustacean stimulating hormone on the mycelial growth rate of A.aegerita,the most suitable dilution ratio of crustacean stimulating hormone was determined to be 1:1500,and the mycelial growth rate of A.aegerita was 3.67mm/d.Mix different proportions of crustacean stimulating hormone into the cultivation material,and determine the most appropriate mixing ratio of 1:1500.The biological efficiency of A.aegerita at this ratio is 86.34%,which increases the yield by 15.81%compared with CK.Through the spraying of different proportions of crustacean stimulating hormone in the reproductive growth stage of A.aegerita,it was determined that the most suitable spraying ratio of chitin-stimulating hormone was 1:1000,and the biological efficiency of A.aegerita at this ratio was 83.08%,which increased production by 13.19% compared with CK.