Study On Genetic Relationship Of Macrofungi In Bijie Of Guizhou Province And Analysis Of Biological Properties Of Coprinellus Disseminatus

Macrofungal is a class of fungi existing in nature with large fruiting.Bijie is located in the northwest of Guizhou Province,which is suitable for the growth of large fungi due to its remarkable topography and diverse climate.However,there are few reports on systematic investigation and resource utilization of large fungi in Bijie.Therefore,this paper through random sampling method of Guizhou bijie parts fungal sampling,investigation,and based on morphological markers and molecular markers technology for strains of genetic diversity analysis,the widely distributed strains white false ghost umbrella(Coprinellus disseminatus)(more than five places have this fungi)biological characteristics,mainly obtain the following results:1)Large fungal collection and classificationSurveying,photographing and sampling of large fungi in some areas of Bijie,Guizhou province,a total of 102 large fungal strains were collected,which were mainly distributed in 6 forest types.According to the primary color map of the sequfungi of ITS(or LSU),the morphology of the collected strains belonged to 2phyla,3 classes,11 orders,25 families and 39 genera.Among them,Gymnopus accounted for the most,accounting for 15.6% of the total,followed by Russula and Lactarius,accounting for 13.7% of the total,respectively.Pleurotus,Entoloma and Hymenochaete accounted for the lowest proportion 0.9%.2)Phylogenetic tree construction based on ITS conserved regionThe phylogenetic tree was constructed with ITS conserved region.Strains0914YY13,0527ZSF9,0527SNJ14 and 0527SNJ13 were associated with Russula rosea(MT555414.1),R.grisea(KX963793.1),R.vesca(MW722739.1)and R.compacta(MK167417.1)The ITS sequence homology ofwere 100%,so they were considered to be the same fungi.The sequence homology of strains 1019YY4 and0917LB26 with Pleurotus pulmonarius(MN17941.1)and Lepiota clypeolaria(OL527693.1)was respectively 95% and 98%,The results indicated that strains1019YY4 and 0917LB26 were different species strains of P.pulmonarius and L.clypeolaria,respectively.The similarity of ITS sequence between strain 0527YY6 and strain Coprinellus disseminatus(KC771477.1)was as high as 99.53%.Furthermore,phylogenetic tree was constructed based on ITS conserved sequences,and the homology between strain 0527YY6 and C.disseminatus(KY243924.1)was100%,which suggested that the strain was C.disseminatus.3)Phylogenetic analysis based on ISSR and RAPD markersFrom 40 ISSR universal primers,10 had good polymorphisms and relatively stable amplification.Ten ISSR primers coamplified 96 sites in 72 fungi,Of the polymorphic sites were 69,On average,we amplified 9.6 sites per primer,The average number of polymorphic loci was 6.9;A total of two of the 10 primers reached100%,The average polymorphism ratio of the 10 primers was as high as 71.8%,The stering of ISSR markers showed an overall 75% similarity between strains,When the genetic coefficient was 0.79,72 strains were clustered into four groups,Among them,the maximum genetic similarity coefficient of strains 0914 ZSF 13 and 0914 WXH 9was 90%;Seven polymorphisms from 20 RAPD universal primers,Amplification results in more stable primers.Seven RAPD primers coamplified 64 sites in 72 large fungi,Of the polymorphic sites were 41,On average,we amplified 9.1 sites per primer,The average number of polymorphic loci was 5.9;The average polymorphism ratio of the 10 primers was 65%,The RAPD marker clustering results showed an overall strain similarity of 62%,The overall similarity was 62%,When the genetic coefficient was 0.65,72 strains were clustered into three major groups,Among them,the maximum genetic similarity coefficient between strain 0917 LB24 and0917ZSF21 and strain 0917ZSF19 and 0919YY5 was 84%;Among the two molecular markers,different primers showed some differences in the DNA genetic map of the same strain,strain 0527YY6(C.disseminatus)amplified 84 bands in 10 ISSR primers,53 bands in 7 RAPD primers,amplified 9 and 7 sites respectively,polymorphic bands were between 7 and 11;and when the genetic coefficient was 0.89 and 0.83,respectively,indicating that there is rich genetic diversity among the strains.4)Study on biological characteristics of C.disseminatusTo study the biological characteristics of C.disseminatus,The mycelium was obtained by tissue isolation and purification culture;Observation and comparison of hyphae under different culture conditions found in different media(no carbon source media,Medium of glucose carbon source,Xylan carbon source medium)in the mycelium will increase with the culture time,There is a trend of growth,And basically full culture dishes for 7 days of culture,But there will be some differences in growth,color,and density,Growth on the carbon-free medium is relatively sparse,Mphelium in a loose state from the center to the edge,Light and white in color,Mycelium margin is not flat;Hyphe cultured on the medium where glucose was the carbon source are near-round,Center density,Loose and close to the flat edge,The hyphae is white;Among them,the best growth was observed on the medium with xylan as the carbon source,Mcelium is white and dense,In a regular circle,Edge is flat.Further determination of xylanase(XLE),cellulase(CLE),acetyl-xylanesterase(AXE)and α-L-arabinofuranosidase(α-L-AF)enzyme activities during the fermentation of Eucommia ulmoides leaves by C.disseminatus showed that AXE andα-L-AF activities in the xylanan carbon source group were the highest at 5 days after inoculation.Up to 4.657 ± 0.036 U/m L and 3.497 ± 0.140 U/m L,respectively.The results showed that the extraction rate of E.ulmoides could be significantly improved after the leaves were treated with C.disseminatus.