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.
Through bag cultivation experiments,wild type(type Ⅰ),Pofst3 overexpression(type Ⅱ),and Pofst3 antisense silencing(type Ⅲ)Pleurotus ostreatus were compared for agronomic traits,including mycelial growth rate,number of primordia and fruiting bodies,stipe diameter and pileus diameter.These strains were also measured for activities of laccase,xylanase,and cellulase.The results showed that there was no significant difference in mycelial growth rate between the three strains.Compared with type Ⅰ,type Ⅲ had more primordia,more fruiting bodies,greater stipe diameter,smaller pileus diameter and expressed a lower level of Pofst3 in primordium and fruiting body.Compared with type Ⅰ and type Ⅱ,type Ⅲ showed higher activities of laccase,xylanase,and cellulase,suggesting that the effects of Pofst3 on the development of P.ostreatus may be associated with its extracellular enzyme activities.
Sixteen batches of Ganoderma lucidum spore powder samples(S1-S16)from different sources were used to establish the chromatographic fingerprint of lipid components in G.lucidum spores by HPLC-ELSD.The HPLC-ELSD fingerprint data of S1-S16 were imported into the similarity evaluation system for chromatographic fingerprints of traditional Chinese medicine(2012 edition)to identify common peaks and evaluate similarity.Then the contents of common peaks in S1-S16 were determined by HPLC-ELSD.The results showed that the common peaks were identified as linoleic acid,palmitic acid,oleic acid and ergosterol.Except S14,the HPLC-ELSD fingerprints of the rest 15 batches of G.lucidum spore powder were highly similar.There were variations in the contents of linoleic acid,palmitic acid and oleic acid in S1-S1 6,whereas the ergosterol content was stable across the samples.
Current problems in China's edible fungi processing industry,such as lack,lag and limitation of regulations for edible fungi used as raw food materials were reviewed.These problems have a negative impact on the high-quality development of edible fungi processing industry and bring difficulties to industry supervision.Countermeasures and recommendations were raised to address these problems,such as to formulate a list of edible fungi that can be used for ordinary food as soon as possible,to formulate incentive policies to promote the application of new food ingredients,to establish standards for edible fungi extracts,to accelerate safety analysis of novel food resources(edible fungal spores,mycelia,and fermentation products).
The effects of exogenous signal substances H2O2,Ca2+,NO(sodium nitroprusside as the donor)and putrescine(Put)on the adaptability of Pleurotus ostreatus strain Xin831 to heat stress were studied.Using sterile water as the control,different concentrations of H2O2(0,0.01,0.05,0.25,1.25,6.25 mmol·L-1),Ca2+(0,2.5,5,10,20,40 mmol·L-1),sodium nitroprusside(0,0.1,0.2,0.4,0.6,0.8 mmol·L1)and Put(0,25,50,100,200,400 μmmol L-1)were added to the medium,respectively.After incubation at 25 ℃ for five days,mycelia of different treatments were further incubated for 24 h at 25 ℃(room temperature)and 40 ℃(heat stress),respectively.Then mycelia of different treatments were measured for mycelial growth rate respectively.The results showed that P.ostreatus mycelial growth was promoted under 0.01-1.25 mmol L-1 H2O2,2.5,10,20 mmol L-1 Ca2+,0.2 mmol·L-1 sodium nitroprusside and 25-200 μmol·L-1 Put at room temperature,but it was inhibited under 6.25 mmol L-1 H2O2,40 mmol L-1 Ca2+and 0.4-0.8 mmol L-1 sodium nitroprusside at room temperature.In the experimental range,H2O2,Ca2+,sodium nitroprusside and Put promoted recovery growth of P.ostreatus mycelia under heat stress.
Transcriptome analysis was used to investigate the effects of light quality on coloration of Naematelia aurantialba fruiting bodies.N.aurantialba fruiting bodies were cultivated under dark(D_2),white light(W_2),and blue light(B_2)conditions,respectively.Differential gene analysis revealed that 1797 genes were differentially expressed between W_2 and D_2.Among these genes,1308 were up-regulated and 489 were down-regulated.There were 1617 differentially expressed genes(DEGs)between B_2 and D_2,among which 1056 were up-regulated and 561 were down-regulated.In comparison with white light induction,blue light induction resulted in differential expression of 152 genes,with 74 up-regulated and 78 down-regulated.GO enrichment analysis showed that most DEGs as compared with D_2 were up-regulated and mainly enriched in translation,peptide metabolism,ribosome structural components,structural molecules activity and other processes induced by blue or white light.Compared with W_2,the DEGs of B_2 were mainly enriched in respiratory regulation,transmembrane transport,and membrane components.KEGG enrichment analysis showed that genes involved in ribosome,amino acid biosynthesis,and 2-oxoacid metabolism pathways were up-regulated during the coloration process.According to KEGG metabolic pathway analysis,nine DEGs were identified to be associated with pigment metabolism,e.g.carotenoid biosynthesis and riboflavin metabolism.The qRT-PCR results of the nine genes were consistent with the transcriptome data.These results provided a reference for understanding the underlying mechanism of N.aurantialba coloration,identification of key functional genes,and light condition regulation during cultivation.
The effects of different concentrations(0.1,1,10,50 μg·mL-1)of Sanghuangporus vaninii water extract on average life span,median-lethal time,maximum life span,number of pupae and number of F1 adults of Drosophila melanogaster were studied.The expression levels of 13 genes pertaining to longevity,immunity and energy metabolism were determined by qPCR at 1,3,5,10 and 20 d upon feeding D.melanogaster with 1 μg·mL-1 S.vaninii water extract.The results showed that in the experimental range,1.0 μg·mL1 water extract prolonged median-lethal time,average life span,and increasesd F1 adult number as compared with the control.Median-lethal time was also prolonged under 0.1 μg·mL-1 S.vaninii water extract.The relative expression levels of TEP,BrC,E74B and JHEH were higher at 3 d than other time.The expression levels of INDY,EcRB,Srl and TPI were higher at 10 d than other time.The expression levels of TotA,Dros and TotX were higher at 20 d than other time.
Five kinds of sawdust,Haining mulberry sawdust(HN),Yulin jujube sawdust(SZ),Yulin mulberry sawdust(SS),Yuan'an mulberry sawdust(YS),and Yuan'an oak sawdust(YL),were analyzed for chemical composition and used to cultivate Sanghuangporus vaninii.The resultant fruiting bodies were analyzed for contents of polysaccharides,triterpenoids,flavonoids,total phenols,and monosaccharide composition.Alcohol extracts of the five kinds of S.vaninii fruiting bodies were determined for their antioxidant activity(ABTS and DPPH)and inhibitory activity on tumor cells in vitro.The results showed that fruiting bodies cultivated with SZ had the highest polysaccharide content(3.06％),and those cultivated with HN had the highest contents of total triterpenoids(0.63％),total phenols(16.09％)and total flavonoids(15.59％).The alcohol extract of fruiting bodies cultivated by HN showed the highest scavenging rate on DPPH and ABTS.It also showed the highest inhibition rate on HepG-2,MCF-7 and HeLa cells.These results provided a reference for enhancing bioactive components in S.vaninii through optimizing cultivation substrate.
采用CCK-8法检测粗毛纤孔菌(Inonotus hispidus)子实体乙酸乙酯萃取物(CM1)和乙醇分离物(CM2)对5种癌细胞(SGC-7901、MDA-MB-231、HeLa、NCI-H1299、HepG2)增殖的抑制作用.结果表明:在实验质量浓度范围内,与癌细胞作用48 h后,CM1的抑制作用弱于CM2,CM1抑制SGC-7901、HeLa、MDA-MB-231 细胞增殖的 IC50分别为 338.64、224.13、542.07 μg·mL-1;CM2 抑制 SGC-7901、HeLa、MDA-MB-231、HepG2、NCI-H1299 细胞增殖的 IC50分别为 104.28、100.51、122.32、196.35、277.66 μg·mL-1.利用超高效液相色谱-串联四极杆飞行时间高分辨率质谱(UPLC-Q-TOF/MS)技术在正负离子模式下表征CM1和CM2的化学成分.从CM 1中鉴定出26种化合物,从CM2中鉴定出30种化合物,CM 1和CM2中存在24种相同的化合物.CM 1和CM2的主要化学成分是多酚,其结构主要由hispidin为骨架聚合或缩合而成,其中相对含量较高的化合物有 osmundacetone、phellinin B、phaeolschidin A、phaeolschidin C、pinillidine、3,3'-亚甲基双｛6-[2-(3,4-二羟苯基)乙烯基]-4-羟基-2氢-吡喃-2-酮｝、phelligridin D和phelliribsin A.本研究结果为粗毛纤孔菌子实体药效物质基础研究和开发利用提供参考.
竹荪(Phallusspp.)是我国常见食用菌,部分物种已实现规模化栽培.市场上销售的多数竹荪产品因外观与长裙竹荪(P.indusiatus)非常接近,被标注为"长裙竹荪"或"竹荪".笔者采用形态鉴定方法,并基于ITS-LSU联合序列构建系统发育树,发现多数市售竹荪产品为红托竹荪(P.rubrovolvatus)和棘托竹荪(P.echinovolvatus),有些为白赭竹荪(P.cremeo-ochraceus),而长裙竹荪很可能在我国无自然分布.
分析4℃处理24、48 h的草菇(Volvariella volvacea)渗出液代谢组;用200 μmol·L-1黄芪多糖溶液浸泡草菇后在4℃放置24、48 h,测定草菇β-葡萄糖苷酶活性和转录组,检测糖基转移酶基因VVO_02307表达量.结果表明:4℃处理24、48 h草菇渗出液代谢组分明显不同;与4℃处理24 h草菇比较,处理48 h的草菇中蔗糖含量较低,亚油酸含量较高.与对照比较,黄芪多糖溶液浸泡的草菇在4℃处理24 h时自溶不明显,β-葡萄糖苷酶活性显著降低.转录组分析结果显示,黄芪多糖增强草菇对低温胁迫的耐受性,可能是通过提高N-聚糖生物合成通路活力实现的.研究结果可为草菇低温保鲜技术研发提供参考.
通过单因素实验和响应面实验优化粗毛纤孔菌(Inonotus hispidus)多糖提取工艺,纯化获得均一多糖(INP);采用ELISA法测定NO和细胞因子IL-6、TNF-α含量,采用RT-PCR方法测定炎症相关基因表达量,采用蛋白免疫印迹方法检测p38MAPK/NF-κB、AKT信号通路相关蛋白表达量,研究INP(低、中、高剂量分别为125、250、500μg·mL-1)对脂多糖(LPS)诱导小鼠巨噬细胞RAW 264.7炎症的影响.结果表明:多糖提取工艺为料液比1∶18(g∶mL)、提取温度80.5℃、提取时间1.9 h,在此条件下多糖得率为(0.612±0.057)％.与模型组相比,INP中剂量组的NO、TNF-α含量显著降低,高剂量组非常显著降低;高剂量组的IL-6含量显著降低.INP中、高剂量组的IL-1β表达量均非常显著降低,高剂量组的TNF-α表达量非常显著降低,高剂量组的IL-6表达量显著降低.INP中、高剂量组的p-p38表达量分别非常、极显著降低,高剂量组的p-Jnk1/2表达量极显著降低,低、高剂量组的p-Erk1/2表达量均极显著降低,高剂量组的p-p65表达量非常显著降低,中、高剂量组的IκBα表达量均非常显著降低,低、中、高剂量组的p-AKT表达量均极显著降低.研究结果表明INP可减轻LPS诱导的细胞炎症损伤,将为粗毛纤孔菌多糖进一步开发利用提供参考.
为探讨六妹羊肚菌(Morchella sextelata)栽培外源营养袋代谢物变化特征,采用液相色谱-质谱(liquid chromatography mass spectrometry,LC-MS)联用分别检测未放置于畦面的外源营养袋(初始营养袋,CK)、播种覆土后80 d未污染(NC)和污染外源营养袋(C)的代谢物;采用非靶向代谢组学分析外源营养袋代谢物变化特征,通过正交偏最小二乘判别分析(osrthogonal partial least squares discriminant analysis,OPLS-DA)结合模型变量重要性投影及差异倍数,筛选NC vs.CK和C vs.NC的差异代谢物;基于京都基因与基因组百科全书注释进行富集分析及拓扑学分析,获得差异代谢物显著富集通路.结果表明:外源营养袋中鉴定到的代谢物主要包括脂类、类脂分子和有机酸及其衍生物;NC vs.CK和C vs.NC的主要差异代谢物分别有202种和163种,主要为脂肪酰基、异戊烯醇脂、甘油磷脂和类固醇及类固醇衍生物;C中脂肪酰基和异戊烯醇脂相对含量显著上调,甘油磷脂和类固醇及类固醇衍生物相对含量显著下调;NC vs.CK和C vs.NC的差异代谢物分别富集到197条和148条通路,基于校正后P值获得NC vs.CK和C vs.NC中显著富集(P＜0.05)的代谢通路分别有29条和12条,差异代谢物参与较多的通路有代谢途径、次级代谢产物的生物合成、微生物代谢和甘油磷脂代谢.
以64个毛木耳(Auricularia cornea)菌株为材料,对20个性状(6个菌丝性状、14个子实体性状)进行筛选,经表型观测、菌丝生长速度的差异显著性分析,筛选出3个菌丝性状(菌丝体胶质物、菌落背面色素和菌棒颜色)作为DUS测试性状.在14个子实体性状中,抽取2个数量性状(鲜耳片长度、宽度)进行6个取样量(20、40、60、100、150、200朵)的方差分析,发现测量结果无显著性差异,选取40朵作为子实体性状测试的取样数量.对14个子实体性状中4个数量性状(鲜耳片长度、宽度、厚度和耳片干湿比)进行变异分析,变异系数范围为16.00％～38.17％,表明各性状的遗传变异较丰富;进而进行正态性检验和频次分布分析,4个数量性状基本符合正态性分布;再比较概率分级法、传统分级法和最小显著差法3种分级方法,确定概率分级法作为4个数量性状的分级方法,性状分为3～5级.经聚类分析,在遗传相似系数为1.82时,供试菌株分为4个类群,鲜耳片腹面主要颜色是类群Ⅲ和类群Ⅳ划分的代表性状,2个类群供试菌株的占比近97％,适合做分组性状.通过以上评价分析,筛选的17个测试性状(3个菌丝性状和14个子实体性状)适合作为毛木耳品种DUS测试评价.
采用生物信息学方法对金针菇(Flammulina filiformis)3个锌簇转录因子的基本性质、锌簇域和分子进化进行分析,并利用荧光定量PCR研究其在菌丝、原基、菌柄和菌盖中的表达模式.结果表明,Fvzcp1、Fvzcp2和Fvzcp3分布在不同染色体片段重叠群,分别含有9、14、10个外显子,编码序列(coding sequence,CDS)长度分别为2 553、2 634、1 743 bp,对应蛋白质FVZCP1、FVZCP2和FVZCP3的相对分子质量分别为93 763.19、98 333.89、64 251.26,理论等电点分别为7.10、6.54、6.64,均为不稳定的疏水性蛋白质,亚细胞定位于细胞核;FVZCP1、FVZCP2和FVZCP3均具有典型的锌簇域,且其锌簇域类型不同.就表达量而言,与菌丝相比,Fvzcp1在原基和菌盖中较高,Fvzcp2在原基和菌柄中较高,Fvzcp3仅在菌柄中较高;3个基因在菌柄伸长区的表达量均显著高于非伸长区.
在中国发现的广义虫草据统计至少有299个,可形成子实体或棒束孢的种类至少有40个;虽然广义虫草物种在中国丰富多样,但只有4种可实现商业化栽培;系统总结可形成子实体或孢梗束的广义虫草的研究进展,并指出广义虫草在菌种、菌群复合群、驯化等方面存在的问题,提出未来研究重点的建议,以期为中国广义虫草的研究提供参考.
