大型担子菌分布广泛，种类繁多，它们是重要的食药用资源的宝库。萜类化合物是其主要活性成分之一，包括倍半萜、二萜和三萜等，这些化合物具有预防、缓解或治疗癌症、抑郁症、糖尿病和高血脂症等多种疾病的功效。目前，从担子菌中分离出的二萜类化合物基本骨架结构特征主要为鸟巢烷 （cyathanes） 型、截短侧耳素 （pleuromutilins） 型、guanacastanes型、海松烷 （pimaranes） 型、松香烷 （abietanes） 型和毛皮伞烷 （crinipellins） 型6大类型。本文综述了担子菌中二萜类化合物的结构特点、生物活性和生物合成的研究进展，对参与担子菌中二萜化合物生物合成的二萜合成酶进行了分类，对两种重要的二萜化合物生物合成途径进行了系统总结和论述。本文将为未知二萜化合物生物合成途径及关键基因功能解析提供参考。
本研究对盐地碱蓬内生菌Neocamarosporium sp. ZLM-26的活性次级代谢产物进行了挖掘。采用薄层色谱、硅胶柱色谱、高效液相色谱等分离方法，从该菌株大米培养基发酵产物的乙酸乙酯萃取物中分离得到6个单体化合物，经波谱学解析和文献数据对比，鉴定6个化合物分别为：5-butyl-6-（hydroxymethyl）-2H-pyran-2-one（1）、（7S）-xylariolide E（2）、（6S）-xylariolide D（3）、diaporpyrone A（4）、4-hydroxybenzaldehyde（5）和2-（2-hydroxyethyl）phenol（6）。分别采用CCK-8法和96孔板法评估了6个化合物的抗肿瘤和抗菌活性。结果显示，6个化合物（50 μmol/L）对胰腺癌细胞株SW1990、PANC-1的增殖无明显抑制作用；而化合物4对大肠杆菌有较好的增殖抑制活性，化合物5、6对铜绿假单胞菌有较好的增殖抑制活性。化合物1为1个新的天然产物，本研究首次报道了其核磁数据和抗菌活性。此外，化合物1–4均为首次从新凸轮孢菌属真菌中分离得到。
研究广叶绣球菌多糖（polysaccharides， SLPs）对小鼠CD8 + T细胞活化、增殖分化以及效应阶段的影响，探索广叶绣球菌多糖对CD8 + T细胞的免疫应答调控机制。通过腹腔注射环磷酰胺溶液（80 mg/kg）构建免疫低下小鼠模型，灌胃广叶绣球菌多糖（低、中、高剂量分别为100、200、400 mg/kg），连续饲养20 d后处死，取脾脏和胸腺。血细胞分析仪测定血常规；MTT检测脾T淋巴细胞的扩增能力；流式细胞术分析脾脏CD8 + T细胞表面CD28、CTLA-4和PD1以及树突状细胞表面MHC Ⅰ、CD80和PD-L1的表达量；ELISA测定外周血中TNF-α、IFN-γ、IL-2、IL-10、穿孔素、颗粒酶B的含量；RT-PCR测定脾脏Psmb9、Psmd9、TAP1、TAP2、IFN-γ、IL-2、JAK1、JAK2、STAT1、STAT3、SOCS-3和Blimp-1的mRNA表达量；Western blot测定pJAK2/JAK2、pSTAT3/STAT3和Blimp-1的蛋白表达量。试验表明广叶绣球菌多糖高剂量组脾脏指数显著升高，外周血中红细胞、白细胞、淋巴细胞、单核细胞、血小板的含量显著升高（P＜0.05或P＜0.01）；多糖中、高剂量组脾T淋巴细胞的扩增能力显著增强（P＜0.05）；中、高剂量组CD8 + T细胞含量显著降低，CD8 + T细胞表面PD-1分子含量显著降低，树突状细胞表面PD-L1分子含量显著降低、MHC Ⅰ细胞含量显著升高（P＜0.05或P＜0.01）；多糖高剂量组TNF-α、IFN-γ、IL-2、穿孔素和颗粒酶B的含量显著极显著升高（P＜0.05或P＜0.01），IL-10的含量显著降低（P＜0.05）；中、高剂量组Psmb9、Pmsd9、TAP1、TAP2和Blimp-1的mRNA含量显著升高（P＜0.05或P＜0.01），JAK1、JAK2、STAT1和STAT3的mRNA和蛋白表达量显著降低（P＜0.05或P＜0.01）。多糖中高剂量组Blimp-1蛋白表达量显著升高（P＜0.05），多糖组与阳性对照组趋势一致。因此，认为广叶绣球菌多糖能够调节MHC Ⅰ类抗原加工呈递分子，并通过JAK2/STAT3信号通路促进CD8 + T细胞的活化、增殖分化和效应杀伤过程，从而调控CD8 + T细胞的免疫应答，起到改善小鼠免疫低下的作用。
金针菇是重要的食药用菌，具有很高的经济价值。随着金针菇全基因组序列的公布、多组学分析、转基因技术、基因编辑技术的发展，越来越多的研究者开始关注金针菇重要性状（如生长速度、菌柄长度、生物活性物质含量等）相关的分子调控机制以及关键基因的发掘。本文综述了分子生物学技术在金针菇研究中的应用并分析了不同技术的利弊；同时，系统介绍了金针菇菌丝和子实体生长发育、温度应答、生物活性物质代谢、蓝光响应等重要生物学过程中调控基因的最新研究进展，并对未来金针菇分子生物学研究的方向进行了展望，旨在为我国金针菇产业的发展提供有价值的参考。
The polysaccharide content, total phenol content, weight average molecular weight distribution, Fourier transform infrared spectroscopy, monosaccharide composition, antioxidant activities and immune activities in vitro of graded ethanol precipitation derived crude polysaccharides of Sanghuangporus vaninii fruiting bodies were compared. Pearson correlation analysis was used to analyze the compositional characteristics and activities of graded ethanol precipitation derived crude polysaccharides. The results showed that the physicochemical properties and activities of different ethanol components derived crude polysaccharides were quite different. The polysaccharide content of graded ethanol precipitation derived crude polysaccharides increased with the increase of ethanol precipitation concentration. The content of total phenol was higher in 20% ethanol gradients, and lower in 50% ethanol gradients. The weight average molecular weight decreased with the increase of ethanol precipitation concentration. The infrared spectrum showed that the hydroxyl vibrational peak of the polysaccharide in the 70%alcoholic fraction was not significant and correlated with low molecular weight distribution of the polysaccharide. In terms of monosaccharide composition, fructose and mannose contents were higher in 50% ethanol gradients, while glucose content was higher in 20% and 70% ethanol gradients. Correlation analysis results show that total phenolic content in crude polysaccharides of the basidiomata showed a significant positive correlation with DPPH and ABTS free radical scavenging activity. All the three crude polysaccharide fractions could activate NO release from RAW 264.7 cells and promote their secretion of immunomodulatory factors such as tumor necrosis factor-α（TNF-α）, interleukin-6（IL-6） and interleukin-1β（IL-1β）. There was a significant positive correlation between NO release and polysaccharide content, and S50 with lower total phenolic content and higher galactose and fructose content had better immune activity in vitro.This study provides a reference for the study of polysaccharide effective components of Sanghuangporus vaninii.
Sanghuangporus vaninii contains many phenolic compounds, and the polyphenols show high antioxidant activities. In this study, deep eutectic solvent（DES） was used for extraction of polyphenolic compounds of S. vaninii, and the optimal process parameters for the purification using macroporous resin were determined by a single-factor test. The experimental results showed that HPD-100 macroporous resin gave the best results for purification of S. vaninii polyphenols,and the optimal parameters for static adsorption-desorption were adsorption time of 4 h, loading solution concentration of 10 mg/mL, pH value of 4.0 and desorption ethanol concentration of 70%; the optimal parameters for dynamic adsorption-desorption were loading sample volume of 100 mL and elution volume of 110 mL. The purity of polyphenols increased from 14.56% to 33.81%, and the purified polyphenols had good antioxidant activities. DPPH and ABTS free radical scavenging capacities were 92.75% and 93.03% respectively, effectively preventing oxidative damage to bovine serum proteins and inhibiting the senescence of L929 cells.
Comparative studies of the effects of water extracts of wild Sanghuangporus sanghuang, cut-log cultivation Sanghuangporus vaninii, sack cultivation S. vaninii and sack cultivation Inonotus hispidus on the immune function of immunocompromised mice were carried out, and relationship between the polysaccharide content and the immunomodulatory efficacy of sanghuang fungi was clarified on the basis of analysis of polysaccharide content of the fungi. The experimental samples of the fungi were extracted by hot water, and then concentrated and freeze-dried to obtain the water extract. Cyclophosphamide was used to construct the immunosuppressive model in mice. After continuous administration for 21 days, the index of peripheral immune organs in mice, proliferative and transformed ability of spleen lymphocytes induced by ConA（concanavalinA） and LPS（lipopolysaccharide）, and the content of TNF-α and IL-2 in serum were determined, and the influence of water extract of different varieties and cultivation methods of sanghuang fungi on the immune function of mice was comparatively examined to explore the relationship between the content of polysaccharide and immunomodulatory efficacy of sanghuang fungi. The results showed that the water extracts of experimental samples of sanghuang fungi could regulate the immune function of immunocompromised mice to varying degrees, and improve the immune organ index, spleen lymphocyte proliferation and serum cytokine content of mice, of which the water extract of sack cultivation Inonotus hispidus had the strongest immunomodulatory effect. The results showed that the higher content of polysaccharide led to the better immunomodulatory effect on mice,and the immunomodulatory efficacy is positively correlated with the polysaccharide content.Polysaccharide of sanghuang fungi is the main material basis of the immunomodulatory function.
Sanguangporus spp. are a group of fungi with a wide range of pharmacological activities, and their excellent antioxidant activity has attracted people’s extensive attention.Antioxidant activities and chemical components of ethanol extracts of fermentation products from Sanghuangporus sanghuang and S. vaninii were compared. The scavenging rate of 1,1-diphenyl-2-trinitrophenylhydrazine（DPPH）, the protective effect of DNA damage caused by hydroxyl radical and L929 cell anti-ageing effect were used to compare the differences of antioxidant activity. The results showed that the extracts of fermentation products from S.sanghuang and S. vaninii both showed effective antioxidant activities, and the antioxidant activity of S. sanghuang fermented product was significantly stronger than that of S. vaninii. A total of 11polyphenols was identified from S. sanghuang by ultra performance liquid chromatography tandem triple quadrupole time-of-flight mass spectrometry（UPLC-triple-TOF-MS）. The components of the fermentation products of S. vaninii were relatively simple.
Currently, sanghuang mushroom products tend towards diversification in the market,and the industry is booming. More and more provinces have issued quality standards or processing standards for sanghuang mushroom, which play an important role in regulating and guiding the quality control of sanghuang products. However, due to the lack of national-level quality control standards, there are still differences in the quality control evaluation indicators and detection methods among provincial sanghuang grading standards, resulting in disadvantage to the horizontal comparison of sanghuang quality and market supervision. In addition, the limits of harmful heavy metal elements are not prescribed in provincial sanghuang grading standards. In this study, the “one-off extraction and multiple detection” method was established and optimized for the extraction process and efficient detection of the content of polysaccharides, total flavonoids, total phenols, and triterpenes and sterols in three species of sanghuang mushrooms.Two different methods determining total flavonoids in current provincial standards, the NaNO 2 -Al（NO 3 ） 3 -NaOH method and the AlCl 3 method, were compared. Referring to the methods published in the 2020 edition of the Chinese Pharmacopoeia, the content of Pb, Cd, As, Hg, and Cu was determined by microwave digestion-inductively coupled plasma mass spectrometry（ICP-MS）. The results showed that the determination method used was accurate and met the requirements of methodological validation. The AlCl 3 method was selected for the determination of total flavonoid content due to its better stability. Basidiomata of artificially cultivated Sanghuangporus vaninii had high levels of active ingredients and low levels of harmful heavy metal elements, providing a certain guarantee for the quality of widely applicative sanghuang in the current market. Sanghuangporus baumii had a relatively low level of active ingredients, while Porodaedalea mongolica had a relatively higher level of active ingredients and significantly higher Pb and Cu content than other sanghuang samples. It is suggested that, more attention should be paid to monitoring the content of active ingredients and harmful heavy metals in the future application and research of sanghuang mushrooms. The evaluation method established in this study might provide a reference for scientific and efficient quality evaluation of sanghuang mushrooms.
A Phellinus orientoasiaticus strain was isolated from basidiomata collected in Shunping County of Hebei Province. The biological characteristics and domestication of this strain were studied. Temperature, initial pH, nitrogen and carbon source were optimized in single factor test. Three optimal levels of four optimal factors were selected for orthogonal experiment to achieve appropriate mycelial culture conditions. Single factor test showed that the optimum growth temperature, initial pH, carbon source, and nitrogen source were 30℃, pH 5.0, soluble starch, and yeast extract respectively. The orthogonal test indicated that the best combination was temperature of 30℃, initial pH of 5.0, nitrogen source of yeast extract, and carbon source of sucrose. The appropriate cultivation spawn formula for P. orientoasiaticus was 80% chestnut sawdust, 17% bran, 1% sucrose, 1% quicklime and 1% gypsum. Mycelia were sackful after 45–50 d of cultivation at 25℃ in dark. Primordia differentiated after 7–10 d under the temperature of 25–28℃ and relative air humidity of 90%–95%. The conditions for fruiting body growth stage were kept at temperature of 25–30℃ and relative air humidity of 90%–95%. Fruiting bodies can be harvested after 25–35 d of growth stage.
The inhibitory effects of EtOAc fraction and ethanolic extracts of Sanghuangporus baumii basidiomata on the proliferation of tumour cells were detected by CCK-8 method. The results showed that the EtOAc fraction exhibits good inhibitory activities against NCI-H1299,SGC-7901, HepG2, MDA-MB-231 and HeLa cells after 48 h treatment, while the ethanolic extract shows good inhibitory activity against NCI-H1299 cell. The major chemical constituents of the extracts were analyzed by using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry（UPLC-Q-TOF-MS） in the negative ion mode. In total,26 phenolic compounds and 12 fatty acids were identified, and the main phenolic compounds were hispidin and its derivatives. The phenolic components with higher content in EtOAc fraction were mainly composed of hispidin, hispolon, davallialactone, osmundacetone, phelligridins C and D, and pinillidine.
Based on morphological and molecular data, a new species Sanghuangporus mongolicus T. Bau sp. nov. is described from Horqin sandy land, Inner Mongolia. Morphologically, it is characterized by perennial, semicircular basidiomata with thin edges, a dimitic hyphal system, and elliptic to broadly elliptic, thick-walled basidiospores measuring 4.2–5×2.7–3.5 μm. The ITS and nrLSU phylogeny shows that Sanghuangporus mongolicus belongs to Sanghuangporus and forms an independent clade. Illustrated description and taxonomic discussion of this new species is given.
Mulberry sawdust was used as the cultivation substrate to cultivate Sanghuangporus vaninii. The mycelium growth, yield and the content of polysaccharides, flavonoids and polyphenols in fruiting bodies were measured. The antioxidant activity of active ingredients was evaluated by 1,1-diphenyl-2-picrylhydrazyl（DPPH） free radical scavenging ability, hydroxyl free radical scavenging ability and superoxide anion free radical scavenging ability. The results showed that there were significant differences in mycelium growth, yield, the content of active components and antioxidant activity between S. vaninii cultivates with mulberry sawdust and that cultivated with traditional sawdust substrate. With the increase of the content of mulberry sawdust in cultivation substrate, the mycelium growth, the content of active components and the antioxidant activity of S. vaninii increased. Except for the mycelium growth rate, the active components and antioxidant activity of S. vaninii cultivated with mulberry sawdust were higher than those of S. vaninii cultivated with traditional ordinary sawdust. The yield of S. vaninii is the highest when cultivation substrate contains 40%–60% mulberry sawdust. Pearson analysis showed that flavonoids and polyphenols were positively correlated with DPPH free radical,superoxide anion free radical and hydroxyl radical scavenging ability, while polysaccharides were positively correlated with superoxide anion scavenging ability and hydroxyl radical scavenging ability. Comprehensive analysis of yield, the content of active ingredients and antioxidant activity indicated that the addition quantities of mulberry sawdust should be controlled in the range of 40%–60%. The research results provide a scientific reference for optimization of S. vaninii cultivation substrate.
To study the uric acid-lowering effects of Sanghuangporus vaninii extracts, the active parts of S. vaninii basidiomata were selected and the functional mechanism was explored. Rat hyperuric acid animal model was established, and SPF male SD rats were randomly divided into 8 experimental groups: normal control group（NC group）, model group（Model group）, positive control group（PC group）, alcohol extract treatment group（CT group）, water extract treatment group（ST group）, treatment group of water extraction precipitated by 20% ethanol（ST20 group）,treatment group of water extraction precipitated by 50% ethanol（ST50 group） and treatment group of water extraction precipitated by 70% ethanol（ST70 group）. After continuous intragastric administration for 8 weeks, all rats were euthanized. The levels of uric acid, creatinine, blood urea nitrogen, AST（alanine aminotransferase）, and ALT（aspartate aminotransferase） in the serum were determined, and the uric acid-lowering effect and protective effect on kidneys of S. vaninii extract were analyzed on the basis of liver-kidney pathological section and immunohistochemical method. Compared with the model group, the levels of serum uric acid, blood urea nitrogen, and ALT in each treatment group of S. vaninii extract were significantly decreased（P＜0.05）. HE and Masson staining showed that the kidney structure was clear, and the pathological phenomena such as cell necrosis and fibrosis were significantly improved. The uric acid-lowering activity of S.vaninii basidiomata were mainly displayed by the water extract, and the ST20 component of crude polysaccharide had the best uric acid-lowering activity. Immunohistochemical results showed that S. vaninii polysaccharide significantly enhanced the expression level of uric acid transporter ABCG2 in rat kidney tissue. The uric acid-lowering active site of S. vaninii basidiomata is mainly water extraction, and the macromolecular water-extracted polysaccharide ST20 has the best uric acid lowering effect. It is preliminarily proved that S. vaninii polysaccharide can promote uric acid excretion and alleviate the pathological damage of the kidney by activating the ABCG2transporter in the kidney. S. vaninii polysaccharide as a pharmacological inducer of uric acid transporter ABCG2 can be used as a potential drug for the treatment of gouty arthritis.
昆虫激素不仅能调控昆虫的生长发育,而且也是昆虫免疫系统的重要调节因子,因此当虫生真菌入侵昆虫寄主体腔并定殖时可能会受其影响.但定殖过程中虫生真菌与昆虫激素之间的直接作用关系尚不清楚.本研究利用不同浓度的蜕皮激素(20E)和保幼激素(JH)模拟鳞翅目昆虫家蚕体内的主要激素水平,对虫生真菌鹿儿岛被毛孢Hirsutella satumaensis进行培养,观察其表观形态特征的变化.结果表明,两种昆虫激素对鹿儿岛被毛孢的表观形态均有不同程度的影响.添加20E和JH培养后,鹿儿岛被毛孢菌落直径和菌丝生物量的变化差异不大;孢子的萌发率稍有下降;而菌落产生的色素和产孢量与对照组相比则呈现显著性变化.菌落色素圈和分生孢子的粘液厚度会随两种激素浓度的增加而增大,而产孢量随激素浓度增加呈先增加后降低的趋势,这说明鹿儿岛被毛孢定殖昆虫血体腔的过程中两种昆虫激素会抑制真菌的生长,而虫生真菌会适应性地做出应答反应,这对于进一步理解虫生真菌在昆虫血体腔中的定殖机理有重要意义.
涠洲岛火山口生态环境特殊,蕴藏着丰富且独具特色的微生物资源.关于涠洲岛火山口海洋真菌来源的次级代谢产物鲜有报道.本研究采用两种培养基从涠洲岛火山口海洋植物中分离真菌,对菌株的代谢产物进行分离纯化,并通过波谱等方法鉴定化合物结构.从4种涠洲岛火山口附近的海洋植物样本中分离真菌共31株,从菌株青霉菌Penicillium sp.TX-M1-3和Penicillium sp.LW-2-1的发酵物中纯化获得2个主要次级代谢产物,并鉴定为核丛青霉素和弯孢霉菌素.活性评价表明核丛青霉素对NPC1L1蛋白具有一定的抑制作用,暗示其在治疗心血管疾病方面的潜力.本结果拓展了涠洲岛火山口微生物资源及其次生代谢产物方面的研究,为该地区微生物与其次生代谢产物的研究提供了相关基础.
本研究以收集的105份金针菇种质资源经遗传性分析后,选出'上研1号'与'0747'菌株作为亲本,采用单孢杂交技术共得到168个杂交菌株.经实验室初筛和工厂化小中试复筛,最终获得产量高、出芽整齐和商品性状优良的金针菇新品种'上研1820'.该品种为浅黄色,菌盖呈球形,菌柄基部颜色加深不显著,子实体干品氨基酸总含量为1.82×104 mg/kg.工厂化栽培条件下,菌丝最适培养温度为20℃,子实体生长发育温度为5-15℃,瓶栽菌丝培养周期为22 d,子实体生育时间为25 d,平均单瓶产量为314.3 g/瓶.研究结果表明,利用分子标记技术和传统选育手段,可有效提高育种效率,对筛选出具有遗传性状稳定、商品性状优良和适用于工厂化栽培等特性的目标菌株具有重要的理论指导意义.
采用平板培养和基因表达等方法,研究了精氨酸对斑玉蕈菌丝生长的影响及其机理.结果表明添加1-2 mmol/L精氨酸能明显提高菌丝的生长速度.添加精氨酸促进精氨酸代谢,提高GCN2介导的翻译通路活力以及空泡氨基酸转运载体基因AVT3的表达.营养补充实验证实在营养匮乏的情况下,单一添加精氨酸能促进斑玉蕈菌丝良好生长.研究结果证实外源添加精氨酸能作为主要营养物质促进斑玉蕈菌丝生长.
为了优化草菇子实体多肽的提取工艺和探究其抗氧化活性,以草菇子实体为原料,采用酶解法提取草菇子实体多肽,通过单因素试验得出最佳的酶解工艺,并使用Box-Behnken设计试验组合.结果表明:草菇子实体提取多肽的最佳工艺为料液比1∶52(g/mL)、加酶量7 200 U/g、酶解温度43℃,此工艺条件下的多肽得率为67.76％.从1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除能力、铁离子还原能力、超氧阴离子自由基清除能力和羟自由基清除能力4个方面研究其体外抗氧化能力,结果表明,草菇子实体多肽对DPPH自由基清除率为74.11％,超氧阴离子自由基和羟自由基清除率分别在69.64％和91.83％达到稳定,草菇子实体多肽还具有一定的还原力,说明草菇子实体多肽可以作为优质抗氧化肽的良好来源.该研究为草菇多肽的高效制备和抗氧化肽等高附加值产品的研发提供理论依据.
已有研究报道灵芝栽培生长的最适pH在中性偏酸环境,在碱性范围的生长及代谢情况鲜见报道.本研究主要探究广泛pH对灵芝液态发酵代谢物及其抗氧化活性的影响.采用摇瓶液态培养后分析代谢物中灵芝三萜、胞内外多糖、菌丝体蛋白及抗氧化活性等指标,系统比较灵芝菌丝体在pH值2-11的生长和代谢情况.研究结果表明,灵芝菌丝体生长、合成灵芝三萜、胞内多糖、30E胞外多糖、菌丝体蛋白和菌丝体水解氨基酸的最适pH值分别为10、3、2、7、2和2.对应结果分别为 17.13 g/L、33.86 mg/g、72.73 mg/g、7.86 g/L、71.42 mg/g 和 107.10 mg/g.比对照分别提高28.5％、77.3％、22.4％、96.5％、97.1％和70.8％.胞内多糖组分1和组分2最高分子量均在初始pH 4,分别为1.016×108g/mol和9.280×104 g/mol,胞外多糖组分1最高分子量在初始pH 10,为4.946×106 g/mol;对菌丝体的总抗氧化能力、1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除能力、羟自由基清除能力分析结果表明最佳的初始pH分别为3、7、9.本研究为液态发酵方式下灵芝生长及其代谢物定向调控发酵的工艺优化提供参考依据,同时发现灵芝菌丝体中优质蛋白及抗氧化活性可在功能性食品和化妆品领域推广应用.
