Using the dilution plating method,bacteria in casing soil of Oudemansiella raphanipes were isolated,and then those with mycelium growth promotion activity on O.raphanipes were screened by confrontation culture analysis.The beneficial bacterial strains were determined for biological activities,including phosphorus solubilization,nitrogen fixation,potassium releasing,cellulose production,siderophore production,and indoleacetic acid production.Four of the beneficial bacterial strains were then identified based on morphological features,physio-biochemical characteristics,and molecular phylogeny.The results showed that a total of 256 bacterial strains were isolated from the casing soil samples,among which,54 strains belonging to 8 genus showed mycelium-promoting activity.The dominant genera were Bacillus and Pseudomonas,accounting for 42.59％and 25.93％,respectively.Among the 54 bacterial strains,14,8 and 6 strains showed phosphorus solubilization,nitrogen fixation and potassium releasing activity,respectively.There were 10,9,and 7 strains capable of producing cellulose,indoleacetic acid and siderophore,respectively.The beneficial strains 54-14,41-60,57-71 and 27-17 were identified as P.fluorescens,P.putida,Serratia marcescens and B.subtilis,respectively.This study provided a theoretical basis for exploring the biological function of bacteria in casing soil of O.raphanipes.
Wild Naematelia aurantialba fruiting bodies with bright colors,strong growth and thick flesh were collected from fallen oak trees along Degang Highway between the Lisu Autonomous Prefecture of Nujiang and Diqing Tibetan Autonomous Prefecture of Yunnan Province.After tissue separation,preliminary screening and secondary screening,an excellent strain named JE-4 was obtained.Using N.aurantialba strain J-5 isolated from mushrooms sold in the market as the control,antagonistic reaction,mycelial characteristics,agronomic traits,and SSR molecular markers were compared between the two strains.JE-4 and J-5 were then compared for biological efficiency and yield in production experiments.The results showed that JE-4 had better mycelial growth than J-5,and the mycelial growth temperature,fruiting temperature,optimal fruiting temperature for JE-4 were 20-30 ℃,10-35 ℃ and 18-20 ℃,respectively.The cultivation period(from inoculation to harvest)for JE-4 was 48 d.JE-4 produced golden yellow fruiting bodies with the first flush biological efficiency at(71.26±0.55)％.During 2021-2022,multiple demonstration experiments were carried out,and JE-4 showed a significantly higher first flush biological efficiency in all production sites as compared with J-5.JE-4 also showed a shorter production cycle.Fruiting bodies of JE-4 were highly uniform,large(9.7 cm in diameter and 3.9 cm in fleshy thickness),fully flowered,uniformly changed in color,golden yellow on both front and back of dried fruiting bodies(white root),and spot-free white.In contrast,fruiting bodies of J-5 were small sized(7.7 cm in diameter and 3.5 cm in fleshy thickness,and insufficiently flowered.Dried fruiting bodies of J-5 were dark yellow on the front and off-white on the back.Using six SSR primers,JESSR003,JESSR010,JESSR075,JESSR090,JESSR098,JESSR100,JE-4 was distinguished from J-5 in DNA polymorphism analysis.JE-4 was found rich in nutrients,and the total amount of polysaccharides and amino acids were higher than those of J-5.In 2023,JE-4 was recognized as a non-major crop variety in Yunnan Province,and was named'Zhongjun N.aurantialba No.4'.
By measuring bleeding time,coagulation time,coagulation function index,platelet parameters and red blood cell parameters of mice,different kinds of Inonotus hispidus fruiting body samples,including fruiting body powder(high dose 1.2 g·kg-1 and low dose 0.6 g·kg-1),petroleum ether extract(high dose 12 g·kg-1 and low dose 6 g·kg-1),ethyl acetate extract(high dose 17 g·kg-1 and low dose 8.5 g·kg-1),n-butanol extract group(high dose 45 g·kg-1 and low dose 22.5 g·kg-1)and water extract(high dose 620 g·kg-1 and low dose 310 g·kg-1)were studied for hemostatic activity in mice.The results showed that high doses of I.hispidus powder,ethyl acetate extract and water extract extremely significantly reduced mouse bleeding time as compared with the blank control.Coagulation time was extremely significantly decreased in I.hispidus powder,high dose petroleum ether extract,high dose ethyl acetate extract,and high dose water extract groups as compared with that in the blank control.Prothrombin time was extremely significantly decreased in I.hispidus powder groups,high dose petroleum ether extract group,high dose ethyl acetate extract group,high dose n-butanol extract group and high dose water extract group.Activated partial thromboplastin time was significantly reduced in I.hispidus powder groups,high dose ethyl acetate extract group,high dose n-butanol extract group,and high dose water extract group.Plasma thrombin time was significantly reduced in I.hispidus powder groups,high dose ethyl acetate extract group,and water extract groups.High dose I.hispidus powder and high dose petroleum ether extract significantly reduced fibrinogen content.High dose petroleum ether extract and the low dose water extract resulted in a significant increase in mean platelet volume.High dose petroleum ether extract also significantly increased platelet distribution width and average red blood cell volume.Platelet count was significantly increased in I.hispidus powder and water extract groups.Plateletcrit was significantly increased in I.hispidus powder and high dose water extract groups.On the other hand,high dose petroleum ether extract extremely significantly decreased red blood cell number and hemoglobin content,and extremely significantly increased corpuscular volume.The results of this study provided a reference for the application of the hemostatic activity of I.hispidus.
The effects of Sparassis latifolia polysaccharides(SLP)on oxidative stress in liver of high-fat diet-fed mice were studied.C5BL/6J mice were randomly divided into six groups:normal control group(NC),high-fat model group(HF),low dose SLP group(LD,100 mg·kg-1),medium dose SLP group(MD,200 mg·kg-1),high dose SLP group(HD,400 mg·kg1),and positive control group(PC).After 12 weeks,mice in different groups were sacrificed and measured for liver index,liver lipid metabolism indices including total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C)and antioxidant capacity related indices including malondialdehyde(MDA),superoxide dismutase(SOD),glutathione(GSH),glutathione peroxidase(GSH-Px).HE staining was used to observe liver tissue morphology.Real-time fluorescence quantitative PCR was used to detect expression of genes associated with antioxidant activity and liver inflammation.The results showed that medium and high doses of SLP significantly reduced liver index and MDA content as compared with the model.Contents of TC and TG were significantly decreased in HD,and LDL-C was extremely significantly decreased in all SLP groups.High dose SLP also significantly promoted GSH-Px activity and increased GSH level.For antioxidant-related genes,the expression of HO-1 and GST was significantly increased in all SLP groups.The expression of NQO1 was significantly increased in HD,and the expression of γ-GCL was significantly increased in both MD and HD.For genes associated with liver inflammation,the expression of CRP,IL-6,and TNF-α was extremely significantly decreased in HD.The expression of VCAM-1 was extremely significantly decreased in MD and HD,and the expression of ICAM-1 was extremely significantly decreased in all SLP groups.In summary,SLP ameliorated high-fat diet induced liver injury in mice by improving antioxidant capacity of liver and by regulating the expression of inflammation-related genes.These results provided a reference for the development of S.latifolia polysaccharides.
Fourteen HMG-box protein coding genes,PoHMG1-PoHMG14,were identified in the whole genome of Pleurotus ostreatus,and then analyzed for their characteristics,structure and phylogeny.The results showed that the tertiary structures of these HMG-box transcription factors were highly similar.Their relative molecular weight ranged from 6 893.79 to 60 397.29 Da.PoHMG6,PoHMG11,PoHMG12 and PoHMG13 were phylogenetically closely related.Based on the transcriptome data of P.ostreatus in mycelium,primordium,young fruiting body and mature fruiting body,PoHMG11 was cloned and analyzed for its function and expression at different growth stages.The full-length of PoHMG11 was found to be 1 517 bp,which encodes 489 amino acid residues and contains a HMG-box domain.PoHMG11 was heterologously expressed in E.coli to yield a protein product of 5.5× 104,and the highest expression level was achieved when the recombinant protein was induced by 1 mol L-1 IPTG at 25 ℃ for 6 h.Real-time quantitative PCR analysis showed that the expression level of PoHMG11 was significantly increased by more than five times in mature fruiting bodies compared with other stages,suggesting that the gene may be involved in regulating fruiting body maturation of P.ostreatus.
