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Lentinus edodes-Stropharia rugosoannulata composite biscuits were produced with low-gluten flour and L. edodes-S. rugosoannulata powder as the main raw materials. The single factor test and Box-Behnken response surface test were carried out to determine the optimal process formula with sensory score as the index, and the quality characteristics of the composite biscuits were investigated. The results showed that the optimum formula of L. edodes-S. rugosoannulata composite biscuits was 9.4 g/100 g mushroom powder, 90.6 g/100 glow gluten flour, on this basis, 32.5 g butter, 30.6 g white sugar, 22.5 g water, 0.8 g salt and 0.6 g baking soda. The sensory score of composite biscuits optimized according to this formula reached 89.8 points, with rich mushroom flavor and crisp taste. The moisture and alkalinity were 2.82 % and 0.22 g/100 g (calculated as Na2CO3 ), respectively. The incorporation of L. edodes-S. rugosoannulata powder did not have a significant adverse impact on the textural properties of the composite biscuits. However, it resulted in a darker color of the composite biscuits. The contents of protein, dietary fiber and total phenol were 5.64 g/100 g, 1.57 g/100 g and 1.76 mg/ g, respectively, which were higher than those of the control group, and the antioxidant capacity was significantly improved (P<0.05). [ABSTRACT FROM AUTHOR]

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Stropharia rugosoannulata is a valuable medicinal and food fungus with high nutritive value. Freeze-drying addresses the storage and transportation challenges of fresh Stropharia rugosoannulata, expanding its market while preserving its flavor and quality more effectively than other drying methods. This study optimizes the vacuum freeze-drying process for Stropharia rugosoannulata using an orthogonal experiment method. The process parameters were optimized to determine their effects on the quality of the vacuum freeze-dried product, including pre-freezing temperature, pre-freezing time, and freeze-drying time. The optimal conditions were identified as a pre-freezing time of 60 h, a pre-freezing temperature of −80 °C, and a freeze-drying time of 72 h. The optimal product exhibited a bright color close to its natural state, with minimal browning and its natural white color maintained post-drying. During the drying process, the internal structure of the raw materials remained intact. After drying, the finished product retained its natural form, making it suitable for sale on the market. The soluble protein content of the vacuum freeze-dried Stropharia rugosoannulata reached 68 mg/g. Optimizing the freeze-drying process can better preserve the tissue structure and bioactive substances of Stropharia rugosoannulata, providing a reference for high-quality food processing and showing potential for sustainable development. [ABSTRACT FROM AUTHOR]

The volatile components of Stropharia rugosoannulata and Tricholoma lobayense were analyzed by mass spectrometry, and the relative contents of each component were determined by the internal standard method. In Stropharia rugosoannulata, 58 volatile components were identified, including 10 alcohols, 14 esters, 9 aldehydes, 5 ketones, 1 phenol, 3 alkanes, 4 amines, 5 acids, 1 furan, 1 pyrazine, 1 olefin, 1 oxime, 1 pyrazole, and 2 other components. Among them, the most abundant volatile component was hexanal (1655.40 ng/g), followed by pentanoic acid (1503.10 ng/g) and phenylethyl alcohol (1215.94 ng/g). In Tricholoma lobayense, 68 volatile components were identified, including 17 aldehydes, 11 ketones, 8 acids, 10 esters, 5 alcohols, 1 phenol, 2 furans, 4 alkanes, 1 alkene, 1 oxime, 1 pyrazole, 1 amine, and 6 other components. Among them, the most abundant volatile component was benzaldehyde (2190.67 ng/g), followed by hexanal (1708.81 ng/g) and furan, 2-pentyl- (838.45 ng/g). There were 21 common volatile components between the two different edible fungi, including 7 aldehydes, 5 esters, 3 ketones, 2 alcohols, 2 acids, 1 oxime, and 1 furan. [ABSTRACT FROM AUTHOR]

This study examined the volatile organic compounds (VOCs) in Stropharia rugosoannulata Farlow that were cultivated on four substrates formulated with agricultural and forestry wastes. The VOCs were analyzed by an electronic nose (E-nose), gas chromatography-ion mobility spectroscopy (GC-IMS), principal component analysis (PCA), and an orthogonal partial least squares discriminant analysis (OPLS-DA). A4(40% sawdust, 30% camellia shells, 20% rice husk, 8% bran, and 2% lime) was the most effective overall at determining the quality of flavor. The E-nose showed that there were similar profiles of aromas for A2(100% Eleusine coracana (L.) Gaertn straw) and A3(70% bamboo chips, 20% rice husk, 8% bran, and 2% lime). A total of 91 VOCs, including 82 known compounds, such as formaldehyde, alcohols, esters, and ketones, and 9 unknown compounds, were detected in each sample by GC-IMS. The relative contents of formaldehyde, ketones, alcohols, and esters in the samples was more than 80%. Among the 29 VOCs with variable importance in projection (VIP) values > 1 and P < 0.05, formaldehyde, heptagonal(dimer), 2-methyl-E-2-butenal-M", 3-methyl-2-butenal-M(dimer), 1-octen-3-ol, butyl acetate(dimer), ethyl 3-methylbutanoate, and 2-pentylfuran were the markers that distinguished the volatiles in S. rugosoannulata cultivated with different groups of raw substrate materials.

Introduction: Currently, straw biodegradation and soil improvement in rice)mushroom rotation systems have attracted much attention. However, there is still a lack of studies on the effects of rice-mushroom rotation on yield, soil properties and microbial succession. Methods: In this study, no treatment (CK), green manure return (GM) and rice straw return (RS) were used as controls to fully evaluate the effect of Stropharia rugosoannulata cultivation substrate return (SRS) on soil properties and microorganisms. Results: The results indicated that rice yield, soil nutrient (organic matter, organic carbon, total nitrogen, available nitrogen and available potassium) and soil enzyme (urease, saccharase, lignin peroxidase and laccase) activities had positive responses to the rice-mushroom rotation. At the interannual level, microbial diversity varied significantly among treatments, with the rice-mushroom rotation significantly increasing the relative alpha diversity index of soil bacteria and enriching beneficial microbial communities such as Rhizobium, Bacillus and Trichoderma for rice growth. Soil nutrients and enzymatic activities were significantly correlated with microbial communities during rice-mushroom rotation. The fungal-bacterial co-occurrence networks were modular, and Latescibacterota, Chloroflexi, Gemmatimonadota and Patescibacteria were closely related to the accumulation of nutrients in the soil. The structural equation model (SEM) showed that fungal diversity responded more to changes in soil nutrients than did bacterial diversity. Discussion: Overall, the rice-mushroom rotation model improved soil nutrients and rice yields, enriched beneficial microorganisms and maintained microbial diversity. This study provides new insights into the use of S. rugosoannulata cultivation substrates in the sustainable development of agroecosystems. [ABSTRACT FROM AUTHOR]

Objective: To avoid browning and spoilage caused by microbial growth, there is an urgent need to carry out researches related to the spoilage species of Stropharia rugosoannulata, and to provide a theoretical basis for exploring storage and preservation techniques. Methods: The spoilage fungi of S. rugosoannulata were isolated and purified, and the dominant spoilage fungi were selected by corrosivity analysis and identified by morphology and molecular biology methods. Results: The spoilage fungi from S. rugosoannulata were carried out to 16 strains of Fusarium, 14 strains of Aspergillus and 4 strains of Rhizopus among them. Four decay-causing fungi were selected to have a rot-causing effect, and identified by morphology and molecular biology as Fusarium pseudoanthophilum, Aspergillus niger, Rhizopus azygosporus and Talaromyces purpureogenus, respectively. The growth characteristics showed that the optimal temperature was 25~28 ℃. The spoilage fungi grew slowly with the temperature of 4~10 ℃ and 35~60 ℃, and the lethal temperature was 60 ℃. The optimal pH was about 7.0 and the spoilage fungi could not grow when the pH was 13.0. Conclusion: Post-harvest storage at low temperature and suitable alkaline environment could inhibit the growth and reproduction of spoilage fungi, which was conducive to the storage and preservation of S. rugosoannulata.

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Stropharia rugosoannulata is abundant in natural polysaccharides, offering not only high nutritional value but also significant medicinal properties, potentially useful in managing hyperlipidemia. This study aimed to assess the hypolipidemic effects of S. rugosoannulata polysaccharide in mice with hyperlipidemia caused by a high-fat diet (HFD). We examined the influence of S. rugosoannulata polysaccharide on protein expression in the LKB1/AMPK pathway using Western blot analysis. The results showed that S. rugosoannulata polysaccharide increased the expressions of LKB1 and AMPK, decreased the expressions of CD36, SREBP1, ACC1, FAS, and HMGCR, and effectively alleviated the accumulation of liver lipids. Furthermore, in HepG2 cells exposed to oleic acid, mirroring lipid accumulation, we elaborated on these animal study findings. Our results demonstrated that weight gain was inhibited, body fat content was decreased, and dyslipidemia and liver steatosis were improved in mice fed with an HFD after treatment with S. rugosoannulata polysaccharide. The findings of this study indicate that S. rugosoannulata polysaccharide can regulate the liver LKB1/AMPK signaling pathway and inhibit lipid accumulation, thus alleviating hyperlipidemia. [ABSTRACT FROM AUTHOR]

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Objective: This study aims to investigate the effect of different drying methods on the drying characteristics and volatile flavor of Stropharia rugosoannulata as raw material. Methods： The fresh S.rugosoannulata was dried by freeze-drying, heat pump drying, hot air drying, and microwave drying, and then the physical and chemical indexes were investigated. Results: The contents of crude protein, crude fat, ash, potassium, calcium, total flavonoids, essential amino acids, flavor amino acids and total amino acids are the highest in freeze drying. The six teenamino acids achieved 16.88 g/100 g in the freeze-drying. The rehydration rate and color all play the best performance in freeze drying. The scavenging rate of DPPH radical of S. rugosoannulata polysaccharide reached 61.14% in hot air drying; The rate was hot air drying>heat pump drying>microwave drying>freeze drying. Similarly, the scavenging rate of ABTS radical reached 67.97% in heat pump drying; The rates were heat pump drying>hot air drying>microwave drying>freeze drying. The monosaccharide of S. rugosoannulata is mainly composed of glucose Glc and Galactose gal, among which the content of freeze-dried glucose Glc is the most, accounting for 36.79% of the total sugar; The maximum molecular weight of S.rugosoannulata was freeze-drying; The volatile flavor of S.rugosoannulata different under different drying methods, and the volatile flavor of S. rugosoannulata after drying is quite different from that of fresh S. rugosoannulata. The volatile components in freeze-dried S. rugosoannulata are the most abundant, and some components are high. Conclusion: Freeze-drying has advantages over other drying methods in terms of nutritional composition, rehydration rate, color, and flavor. It has the least impact on the quality of S. rugosoannulata and is the most suitable drying method.

Stropharia rugosoannulata is a valuable medicinal and food fungus with high nutritive value. Freeze-drying addresses the storage and transportation challenges of fresh Stropharia rugosoannulata, expanding its market while preserving its flavor and quality more effectively than other drying methods. This study optimizes the vacuum freeze-drying process for Stropharia rugosoannulata using an orthogonal experiment method. The process parameters were optimized to determine their effects on the quality of the vacuum freeze-dried product, including pre-freezing temperature, pre-freezing time, and freeze-drying time. The optimal conditions were identified as a pre-freezing time of 60 h, a pre-freezing temperature of −80 °C, and a freeze-drying time of 72 h. The optimal product exhibited a bright color close to its natural state, with minimal browning and its natural white color maintained post-drying. During the drying process, the internal structure of the raw materials remained intact. After drying, the finished product retained its natural form, making it suitable for sale on the market. The soluble protein content of the vacuum freeze-dried Stropharia rugosoannulata reached 68 mg/g. Optimizing the freeze-drying process can better preserve the tissue structure and bioactive substances of Stropharia rugosoannulata, providing a reference for high-quality food processing and showing potential for sustainable development.

Flavonoids are a diverse family of natural compounds that are widely distributed in plants and play a critical role in plant growth, development, and stress adaptation. In recent years, the biosynthesis of flavonoids in plants has been well-researched, with the successive discovery of key genes driving this process. However, the regulation of flavonoid biosynthesis in fungi remains unclear. Stropharia rugosoannulata is an edible mushroom known for its high nutritional and pharmacological value, with flavonoids being one of its main active components. To investigate the flavonoid content of S. rugosoannulata, a study was conducted to extract and determine the total flavonoids at four stages: young mushroom (Ym), gill (Gi), maturation (Ma), and parachute-opening (Po). The findings revealed a gradual increase in total flavonoid concentration as the fruiting body developed, with significant variations observed between the Ym, Gi, and Ma stages. Subsequently, we used UPLC-MS/MS and transcriptome sequencing (RNA-seq) to quantify the flavonoids and identify regulatory genes of Ym, Gi, and Ma. In total, 53 flavonoid-related metabolites and 6726 differentially expressed genes (DEGs) were identified. Through KEGG pathway enrichment analysis, we identified 59 structural genes encoding flavonoid biosynthesis-related enzymes, most of which were up-regulated during the development of the fruiting body, consistent with the accumulation of flavonoids. This research led to the establishment of a comprehensive transcriptional metabolic regulatory network encompassing flavonoids, flavonoid synthases, and transcription factors (TFs). This represents the first systematic exploration of the molecular mechanism of flavonoids in the fruiting of fungi, offering a foundation for further research on flavonoid mechanisms and the breeding of high-quality S. rugosoannulata. [ABSTRACT FROM AUTHOR]

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China possesses abundant wild germplasm resources and a wide range of cultivated strains of Stropharia rugosoannulata. However, issues such as genetic diversity and unclear genetic relationships have had an impact on the classification and preservation of germplasm resources, the breeding of new varieties, and the promotion of superior strains. There is an urgent need for genetic diversity analysis and assessment of germplasm resources. In this study, we conducted whole-genome resequencing of 50 cultivated and wild strains collected from various regions across the country. After applying a series of filtering parameters, we obtained 888,536 high-quality Single Nucleotide Polymorphism (SNP) markers. Using these SNP markers, we performed principal component analysis, population structure analysis, and phylogenetic clusters analysis on the 50 strains. Most cultivated strains exhibited high genetic similarity, while significant genetic diversity was observed among wild strains. Based on factors such as marker distribution throughout the entire genome and marker quality, we selected 358 core SNP markers to construct SNP fingerprints. Two-dimensional barcodes were generated for each strain to enable specific identification. Additionally, the mycelial growth rate of strains was evaluated on Potato Dextrose Agar (PDA) and substrate culture media. We also assessed their lignin degradation capability using guaiacol agar plates assay. It was observed that the mycelial growth rate on PDA and substrate culture medium exhibited a significant correlation with the diameter of the mycelial colony on guaiacol agar medium. Additionally, the correlation between the mycelial growth rate on the substrate medium and that on the guaiacol agar plate was stronger than that on PDA medium. This study provided molecular-level identification and assessment of germplasm resources. It clarified the genetic relationships among strains and the characteristics of mycelium growth-related agronomic traits of each strain. This research contributed to the enrichment and development of utilizable germplasm resources and breeding materials for S. rugosoannulata, offering a scientific basis for further research. [ABSTRACT FROM AUTHOR]

The potential of Stropharia rugosoannulata as a microbial remediation material for cadmium (Cd)-contaminated soil lies in its capacity to absorb and accumulate Cd in its mycelia. This study utilized the TMT and LC−MS techniques to conduct integrated proteomic and metabolomic analyses with the aim of investigating the mycelial response mechanisms of S. rugosoannulata under low- and high-Cd stresses. The results revealed that mycelia employed a proactive defense mechanism to maintain their physiological functions, leading to reduced sensitivity to low-Cd stress. The ability of mycelia to withstand high levels of Cd stress was influenced primarily by the comprehensive regulation of six metabolic pathways, which led to a harmonious balance between nitrogen and carbohydrate metabolism and to reductions in oxidative stress and growth inhibition caused by Cd. The results provide valuable insights into the molecular mechanisms involved in the response of S. rugosoannulata mycelia to Cd stress. [ABSTRACT FROM AUTHOR]

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Abstract A novel polysaccharide PSRa-2 was purified from Stropharia rugosoannulata fruiting bodies using high pressure homogenization-assisted dual enzyme method, ion exchange, and gel chromatography. The PSRa-2 was characterized via FT-IR, HPAEC, SEM, Congo red test, SEC–MALLS-RI, methylation analysis, and NMR analysis. Structural characterization revealed that PSRa-2 was an α-glucan with a Mw 455.6 kDa. The backbone of PSRa-2 was composed of →4)-α-d-Glcp-(1→ and →3)-α-d-Glcp-(1→ and branches of α-d-Glcp-(1→ at position O-6 of →4,6)-α-d-Glcp-(1→. PSRa-2 induced splenocyte proliferation and protected splenocytes against 5-Fu-induced immunosuppression by restoring the proliferation and secretion of cytokines (TNF-α and IL-2) secretion levels. Thus, PSRa-2 exhibits obviously immunomodulatory activity and represents a potential natural immunomodulator. Graphical Abstract

The ultrasonic-assisted extraction process of polysaccharides from Stropharia rugosoannulata in Bo'ai County and the antibacterial and antioxidant activities were analyzed. On the basis of single factor experiments, with the polysaccharide yield as response value, the ultrasonic-assisted extraction process of polysaccharides from Stropharia rugosoannulata in Bo'ai County was optimized using Box-Behnken experimental design with three factors and three levels. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the polysaccharides extracted from Stropharia rugosoannulata in Bo'ai County were determined using the double dilution method and the plate coating method, respectively. The antioxidant activity was determined using ABTS method and FRAP methods. The results showed that the optimal ultrasonic-assisted extraction conditions were 62 ℃ (ultrasonic temperature), 1:30 g/mL (solid-liquid ratio) and 62 min (ultrasonic time). Under these conditions, the average yield of polysaccharide was 13.25%, which agreed well with the predicted value of 13.35%, the relative error was 0.75%. Among the five common pathogenic bacteria tested, the polysaccharide extract showed the strongest inhibitory effect on Salmonella typhimurium with the minimum inhibitory concentration of 6.25 mg/mL and the minimum bactericidal concentration of 12.5 mg/mL. Meanwhile, the polysaccharide extract showed strong ability to scavenge ABTS+ free radicals and reduce Fe3+, and showed dose-effect relationship within a certain concentration range, although weaker than the positive control (VC) at the same concentration. The results suggested that the regression models were stable and feasible for polysaccharide extraction from Stropharia rugosoannulata in Bo'ai County and the extract had strong antioxidant and antibacterial activities, but its antibacterial effect was selective.

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Objective: This study aimed to optimize the processing technology of edible fungi and evaluate its application potential as flavoring agents in the food industry. Methods: The content of non-volatile flavor components of the samples was determined by ion chromatography and high-performance liquid chromatography, and the umami of the samples was evaluated by equal umami concentration. The volatile aroma components in the samples were detected by GC-MS, electronic nose and electronic tongue. The microstructure before and after Nano-pulverization was observed, and the experiment of increasing freshness and reducing salt was carried out. Results: After Nano-pulverization, the contents of umami amino acids, flavor nucleotides and organic acids in Stropharia rugosoannulata and A. bisporus increased significantly, and the types and quantities of volatile compounds increased significantly, while Nano-pulverization had little effect on the flavor components of Volvariella volvacea. The results of the electronic tongue showed that the pulverized samples were significantly improved in both umami and saltiness, the addition of 0.5% Nano-pulverization A. bisporus could increase the salt concentration by 0.28%, and the umami value was increased by 145%. Conclusion: Nano-pulverization technology can improve the taste of food. Nano-pulverized edible fungus powder can be used as an effective alternative to salt and flavor enhancers in the food manufacturing industry.

A novel polysaccharide PSRa-2 was purified from Stropharia rugosoannulata fruiting bodies using high pressure homogenization-assisted dual enzyme method, ion exchange, and gel chromatography. The PSRa-2 was characterized via FT-IR, HPAEC, SEM, Congo red test, SEC–MALLS-RI, methylation analysis, and NMR analysis. Structural characterization revealed that PSRa-2 was an α-glucan with a Mw 455.6 kDa. The backbone of PSRa-2 was composed of →4)-α-d-Glcp-(1→ and →3)-α-d-Glcp-(1→ and branches of α-d-Glcp-(1→ at position O-6 of →4,6)-α-d-Glcp-(1→. PSRa-2 induced splenocyte proliferation and protected splenocytes against 5-Fu-induced immunosuppression by restoring the proliferation and secretion of cytokines (TNF-α and IL-2) secretion levels. Thus, PSRa-2 exhibits obviously immunomodulatory activity and represents a potential natural immunomodulator. [ABSTRACT FROM AUTHOR]

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ABSTRACT: An acidic extracellular polysaccharide (EPS) was gained from Stropharia rugosoannulata fermentation broth by the alcohol precipitate method. Sulfated derivative of EPS (S-EPS) was gained by chlorosulfonic acid-pyridine method and the conditions were as follows: pyridine as the reaction solvent, temperature 80 °C, and reaction time 90 min. The chemical structure and chain conformation of EPS and S-EPS were characterized by FT-IR, NMR and size exclusion chromatography (SEC) coupled with multi-angle laser light scattering (MALLS) detection. FT-IR spectra indicated the sulfated group was linked to the polymer. The EPS mainly consists of mannose, glucose and glucuronic acid, and the glycosidic bond type is mainly (1→4)-linked and (1→6)-linked. C6 and C2 were substituted by sulfate groups. The EPS was degraded into smaller molecular weight (Mw) polysaccharides with the larger molecular size (Rg) after sulfation and S-EPS has better hydrophilicity. Moreover, the sulfated derivative S-EPS was found to have a better scavenging ability on hydroxyl radicals than EPS. It proved that the chemical modification of S. rugosoannulata polysaccharides by sulfation effectively enhanced their antioxidant activity.

In order to explore the flavor composition profiles of fresh and dried Stropharia rugosoannulata and the key components determining its good flavor, the aroma and flavor components of fresh and dried samples prepared by four different drying methods, namely hot air drying (HAD), vacuum freeze drying (VFD), microwave drying (MWD), natural drying (ND) were analyzed. A total of 118 aroma compounds were detected. After drying, the aroma richness was significantly improved (P HAD > ND > fresh sample > MWD. A partial least squares regression model was established based on the sensory scores and the component data. The R2X, R2Y and Q2 of the first three principal components were 79.4%, 98.9%, and 0.974, respectively. The loading information showed that the interaction between aroma and taste characteristics was obvious. Variable importance in projection scores showed that sweet amino acids, 1-octene-3-ol, butyl-2-octenal and EUC were crucial for the good characteristic flavor of S. rugosoannulata. This study will provide a theoretical basis for subsequent screening of sample pretreatment methods and the development of flavor related products.

In order to study the effect of both greenhouse and forest cultivating environments on Stropharia rugosoannulata, its volatile aroma compounds were measured by a headspace solid phase micro extractions—gas chromatograph—mass spectrometer (SPME–GC–MS). The optimal adsorption temperature was 75 °C and the optimal adsorption time was 40 min. A total of 36 volatile aroma compounds were identified by GC–MS, including 8 aldehydes, 2 ketones, 4 alcohols, 15 alkenes, and 4 alkanes. Hexanal, 3-Octanone, 2-Undecanone, (E)-Nerolidol, and (Z)-β-Farnesene made great aromatic contributions. Among them, Hexanal, 3-Octanone, 2-Undecanone were the key aroma compounds for which odor activity values (OAVs) were more than 1. (E)-Nerolidol showed odor modification in the forest samples and showed a key aroma effect in greenhouse samples. (Z)-β-Farnesene showed odor modification in greenhouse samples. 3-Octanone was the largest contributing compound for which the OAV was more than 60. The total content of volatile aroma compounds first increased and then decreased with growth time; it reached the highest level at 48 h: 2203.7 ± 115.2 μg/kg for the forest environment and 4516.6 ± 228.5 μg/kg for the greenhouse environment. The aroma was the most abundant at this time. All samples opened their umbrella at 84 h and become inedible. Principal component analysis (PCA), hierarchical cluster analysis (HCA), and orthogonal partial least squares discriminant analysis (OPLS–DA) were combined to analyze the aroma difference of S. rugosoannulata under two cultivation modes. PCA and HCA could effectively distinguish the aroma difference in different growth stages. Under different culturing methods, the aroma substances and their changes were different. The samples were divided into two groups for forest cultivation, while the samples were divided into three groups for greenhouse cultivation. At the end of growth, the aroma of S. rugosoannulata with the two cultivation modes was very similar. OPLS–DA clearly distinguished the differences between the two cultivation methods; 17 key aroma difference factors with variable importance projection (VIP) > 1 were obtained from SPLS–DA analysis. [ABSTRACT FROM AUTHOR]

Stropharia rugosoannulata has been used in environmental engineering to degrade straw in China. The nitrogen and carbon metabolisms are the most important factors affecting mushroom growth, and the aim of this study was to understand the effects of different nitrogen levels on carbon metabolism in S. rugosoannulata using transcriptome analysis. The mycelia were highly branched and elongated rapidly in A3 (1.37% nitrogen). GO and KEGG enrichment analyses revealed that the differentially expressed genes (DEGs) were mainly involved in starch and sucrose metabolism; nitrogen metabolism; glycine, serine and threonine metabolism; the MAPK signaling pathway; hydrolase activity on glycosyl bonds; and hemicellulose metabolic processes. The activities of nitrogen metabolic enzymes were highest in A1 (0.39% nitrogen) during the three nitrogen levels (A1, A2 and A3). However, the activities of cellulose enzymes were highest in A3, while the hemicellulase xylanase activity was highest in A1. The DEGs associated with CAZymes, starch and sucrose metabolism and the MAPK signaling pathway were also most highly expressed in A3. These results suggested that increased nitrogen levels can upregulate carbon metabolism in S. rugosoannulata. This study could increase knowledge of the lignocellulose bioconversion pathways and improve biodegradation efficiency in Basidiomycetes. [ABSTRACT FROM AUTHOR]

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China possesses abundant wild germplasm resources and a wide range of cultivated strains of Stropharia rugosoannulata. However, issues such as genetic diversity and unclear genetic relationships have had an impact on the classification and preservation of germplasm resources, the breeding of new varieties, and the promotion of superior strains. There is an urgent need for genetic diversity analysis and assessment of germplasm resources. In this study, we conducted whole-genome resequencing of 50 cultivated and wild strains collected from various regions across the country. After applying a series of filtering parameters, we obtained 888,536 high-quality Single Nucleotide Polymorphism (SNP) markers. Using these SNP markers, we performed principal component analysis, population structure analysis, and phylogenetic clusters analysis on the 50 strains. Most cultivated strains exhibited high genetic similarity, while significant genetic diversity was observed among wild strains. Based on factors such as marker distribution throughout the entire genome and marker quality, we selected 358 core SNP markers to construct SNP fingerprints. Two-dimensional barcodes were generated for each strain to enable specific identification. Additionally, the mycelial growth rate of strains was evaluated on Potato Dextrose Agar (PDA) and substrate culture media. We also assessed their lignin degradation capability using guaiacol agar plates assay. It was observed that the mycelial growth rate on PDA and substrate culture medium exhibited a significant correlation with the diameter of the mycelial colony on guaiacol agar medium. Additionally, the correlation between the mycelial growth rate on the substrate medium and that on the guaiacol agar plate was stronger than that on PDA medium. This study provided molecular-level identification and assessment of germplasm resources. It clarified the genetic relationships among strains and the characteristics of mycelium growth-related agronomic traits of each strain. This research contributed to the enrichment and development of utilizable germplasm resources and breeding materials for S. rugosoannulata, offering a scientific basis for further research.

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Abstract Low temperature is an important environmental factor that restricts the growth of Stropharia rugosoannulata; however, the molecular mechanisms underlying S. rugosoannulata responses to low-temperature stress are largely unknown. In this study, we performed a transcriptome analysis of a high-sensitivity strain (DQ-1) and low-sensitivity strain (DQ-3) under low-temperature stress. The liquid hyphae of S. rugosoannulata treated at 25 °C and 10 °C were analyzed by RNA-Seq, and a total of 9499 differentially expressed genes (DEGs) were identified. GO and KEGG enrichment analyses showed that these genes were enriched in “xenobiotic biodegradation and metabolism”, “carbohydrate metabolism”, “lipid metabolism” and “oxidoreductase activity”. Further research found that carbohydrate enzyme (AA, GH, CE, and GT) genes were downregulated more significantly in DQ-1 than DQ-3 and several cellulase activities were also reduced to a greater extent. Moreover, the CAT1, CAT2, GR, and POD genes and more heat shock protein genes (HSP20, HSP78 and sHSP) were upregulated in the two strains after low-temperature stress, and the GPX gene and more heat shock protein genes were upregulated in DQ-3. In addition, the enzyme activity and qRT–PCR results showed trends similar to those of the RNA-Seq results. This result indicates that low-temperature stress reduces the expression of different AA, GH, CE, and GT enzyme genes and reduces the secretion of cellulase, thereby reducing the carbohydrate metabolism process and mycelial growth of S. rugosoannulata. Moreover, the expression levels of different types of antioxidant enzymes and heat shock proteins are also crucial for S. rugosoannulata to resist low-temperature stress. In short, this study will provide a basis for further research on important signaling pathways, gene functions and variety breeding of S. rugosoannulata related to low-temperature stress.

This study systematically determined the nutrient components, bioactive substances and heavy metal content of Stropharia rugosoannulata in Bo’ai County, and its nutritional value was analyzed and evaluated. The nutrient components, mineral elements, vitamins, amino acid composition, bioactive substances and heavy metal content of Stropharia rugosoannulata were determined using national standards and conventional inspection methods. The results showed that the contents of moisture, ash, protein, fat, crude fiber, carbohydrates, total sugars, reducing sugars, polysaccharides and total triterpenes in the tested Stropharia rugosoannulata were 11.43%±0.15%, 8.47%±0.15%, 34.17%±3.17%, 1.33%±0.25%, 7.30%±0.30%, 45.17%±3.75%, 36.67%±2.40%, 3.53%±0.31%, 13.18%±0.05% and 1.420%±0.005%, respectively. It contained 18 kinds of amino acids, of which isoleucine was as high as 6.32±0.42 mg/100 g, accounting for 33.54%±2.92% of the total amino acids, and the content of 8 essential amino acids for the human body was 11.70±0.91 mg/100 g. The E/T was 61.94% and the E/N was 162.73%, which were better than the ideal protein model proposed by FAO/WHO. It contained many kinds of mineral elements, and was rich in content including some constant elements necessary for the human body such as Na, K, Mg, P, Ca, etc. and some trace elements such as Fe, Zn, Mn, Se, Cu, etc. The content of the constant elements was more than 1000 mg/kg, and selenium as high as 2.57±0.47 mg/kg. It was rich in six group-B vitamins including B2, B6, B12, folic acid, niacin, especially the content of niacin was as high as 392.053±17.200 mg/kg. The content of total lead, total mercury and cadmium were all within the limits range of national food safety standards except total arsenic. Bo'ai Stropharia rugosoannulata was a good raw material for eating and developing into health food, which was high in protein, low in fat and fiber, and rich in mineral elements and B-group vitamins, had a reasonable amino acid composition, and contained water-soluble polysaccharides, triterpenoids and other biologically active substances.

The basidiomycetous edible mushroom Stropharia rugosoannulata has excellent nutrition, medicine, bioremediation, and biocontrol properties. S. rugosoannulata has been widely and easily cultivated using agricultural by-products showing strong lignocellulose degradation capacity. However, the unavailable high-quality genome information has hindered the research on gene function and molecular breeding of S. rugosoannulata. This study provided a high-quality genome assembly and annotation from S. rugosoannulata monokaryotic strain QGU27 based on combined Illumina-Nanopore data. The genome size was about 47.97 Mb and consisted of 20 scaffolds, with an N50 of 3.73 Mb and a GC content of 47.9%. The repetitive sequences accounted for 17.41% of the genome, mostly long terminal repeats (LTRs). A total of 15,726 coding gene sequences were putatively identified with the BUSCO score of 98.7%. There are 142 genes encoding plant cell wall degrading enzymes (PCWDEs) in the genome, and 52, 39, 30, 11, 8, and 2 genes related to lignin, cellulose, hemicellulose, pectin, chitin, and cutin degradation, respectively. Comparative genomic analysis revealed that S. rugosoannulata is superior in utilizing aldehyde-containing lignins and is possible to utilize algae during the cultivation. [ABSTRACT FROM AUTHOR]

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The isolation of Stropharia rugosoannulata polysaccharide (SRP) by three-phase extraction was optimized, and its structure and biological activities were identified. The optimal extraction conditions were: mass fraction of ammonium sulfate, 20%; volume ratio of sample solution to t-butanol, 1:1.5; extraction temperature, 35°C. Under these conditions, the yield of SRP was 6.85% ± 0.13%. SRP was found to be composed of glucose (35.79%), galactose (26.80%), glucuronic acid (9.92%), fructose (8.65%), xylose (7.92%), fucose (4.19%), arabinose (3.46%) and rhamnose (3.26%), with the molecular weight of 27.52 kDa. The results of DPPH, hydroxyl, ABTS+ radical scavenging and reducing power tests showed that SRP had good antioxidant capacities. SRP had no cytotoxic effect on RAW264.7 macrophages at the concentrations of 25-200 μg/mL, and could significantly promote phagocytosis activity and cell migration according to CCK-8 assay, phagocytosis assay and cell scratch experiment. SRP can significantly stimulate the transcript expression levels of TNF-α, IL-1β and IL-6, as determined by RT-PCR and Western blot assays. SRP activated the TLR4/NF-κB signaling pathway, and autophagy also occurred. These results suggest that SRP is a safe antioxidant and immunomodulator, and that it can be used in the development of functional foods and/or pharmaceuticals.

Stropharia rugosoannulata is a widely grown edible mushroom with a high nutritional value. S. rugosoannulata polysaccharides is one of the most important bioactive components of S. rugosoannulata and has a wide range of activities. A S. rugosoannulata polysaccharides, named SRF-3, was derived from the S. rugosoannulata extraction by freeze-thaw combine with hot water extraction method, then prepareed with DEAE-cellulose column and Sephacryl S-200 HR gel column, and its hypolipidemic activity was determined. The structural characteristics of SRF-3 were analyzed by infrared spectral scanning (FT-IR), ultra-high performance liquid chromatography (UHPLC), acid hydrolysis, methylation analysis, nuclear magnetic resonance (NMR), and Gas Chromatography-Mass Spectrometer (GC-MS). SRF-3 is composed of mannose, galactose, methyl galactose and fructose with ratios of 16, 12, 58 and 12, respectively. In addition, the average relative molecular mass of SRF-3 is approximately 24 kDa. The main chain of SRF-3 is mainly composed of repeating α-D-1,6-Galp and α-D-1,6-Me-Galp units, with branches in the O-2 position of Gal. The structure is presumed to be a mannogalactan, with a small amount of t-β-D-Manp present as a side chain. Hypolipidemic activity assay showed that SRF-3 had good antioxidant and hypolipidemic effects in vitro, suggesting that SRF-3 have potential application in reducing liver fat accumulation.

Through virtual screening, electronic tongue verification, and molecular docking technology, the structure-taste activity relationship of 47 kinds of umami peptides (octapeptide - undecapeptide) from Stropharia rugosoannulata prepared by simultaneous ultrasonic-assisted directional enzymatic hydrolysis was analyzed. The umami peptides of S.rugosoannulata can form hydrogen bond interaction and electrostatic interaction with umami receptors T1R1/T1R3. The amino acid residues at the peptides' N-terminal and C-terminal play a vital role in binding with the receptors to form a stable complex. D, E, and R are the primary amino acids in the peptides that easily bind to T1R1/T1R3. The basic amino acid in the peptides is more easily bound to T1R1, and the acidic amino acid is more easily bound to T1R3. The active amino acid sites of the receptors to which the peptides bind account for 42%−65% of the total active amino acid residues in the receptors. ASP147 and ASP219 are the critical amino acid residues for T1R1 to recognize the umami peptides, and ARG64, GLU45, and GLU48 are the critical amino acid residues for T1R3 to recognize the umami peptides. The increase in the variety and quantity of umami peptides is the main reason for improving the umami taste of the substrate prepared by synchronous ultrasound-assisted directional enzymatic hydrolysis. This study provides a theoretical basis for understanding simultaneous ultrasound-assisted directional enzymatic hydrolysis for preparing umami peptides from S.rugosoannulata, enhancing the flavor of umami, and the relationship between peptide structure and taste activity.

In recent years, a rare edible mushroom Stropharia rugosoannulata has become popular. S. rugosoannulata has the characteristics of easy cultivation, low cost, high output value, and low labor requirement, making its economic benefits significantly superior to those of other planting industries. Accumulating research demonstrates that cultivating edible fungus is advantageous for farming soil. The present experiment used idle croplands in winter for S. rugosoannulata cultivation. We explored the effects of S. rugosoannulata cultivation on soil properties and soil microbial community structure in paddy and dry fields, respectively. We cultivated S. rugosoannulata in the fields after planting chili and rice, respectively. The results showed that Chili-S. rugosoannulata and Rice-S. rugosoannulata planting patterns increased the yield, quality and amino acid content of S. rugosoannulata. By analyzing the soil properties, we found that the Chili-S. rugosoannulata and Rice-S. rugosoannulata cropping patterns increased the total nitrogen, available phosphorus, soil organic carbon, and available potassium content of the soil. We used 16s amplicons for bacteria and internal transcribed spacer (ITS) region for fungi to analyze the microbial communities in rhizosphere soils. Notably, S. rugosoannulata cultivation significantly increased the abundance of beneficial microorganisms such as Chloroflexi, Cladosporium and Mortierella and reduce the abundance of Botryotrichumin and Archaeorhizomyces. We consider S. rugosoannulata cultivation in cropland can improve soil properties, regulate the community structure of soil microorganisms, increase the expression abundance of beneficial organisms and ultimately improve the S. rugosoannulata yield and lay a good foundation for a new round of crops after this edible mushroom cultivation. [ABSTRACT FROM AUTHOR]

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In order to study the effect of both greenhouse and forest cultivating environments on Stropharia rugosoannulata, its volatile aroma compounds were measured by a headspace solid phase micro extractions—gas chromatograph—mass spectrometer (SPME–GC–MS). The optimal adsorption temperature was 75 °C and the optimal adsorption time was 40 min. A total of 36 volatile aroma compounds were identified by GC–MS, including 8 aldehydes, 2 ketones, 4 alcohols, 15 alkenes, and 4 alkanes. Hexanal, 3-Octanone, 2-Undecanone, (E)-Nerolidol, and (Z)-β-Farnesene made great aromatic contributions. Among them, Hexanal, 3-Octanone, 2-Undecanone were the key aroma compounds for which odor activity values (OAVs) were more than 1. (E)-Nerolidol showed odor modification in the forest samples and showed a key aroma effect in greenhouse samples. (Z)-β-Farnesene showed odor modification in greenhouse samples. 3-Octanone was the largest contributing compound for which the OAV was more than 60. The total content of volatile aroma compounds first increased and then decreased with growth time; it reached the highest level at 48 h: 2203.7 ± 115.2 μg/kg for the forest environment and 4516.6 ± 228.5 μg/kg for the greenhouse environment. The aroma was the most abundant at this time. All samples opened their umbrella at 84 h and become inedible. Principal component analysis (PCA), hierarchical cluster analysis (HCA), and orthogonal partial least squares discriminant analysis (OPLS–DA) were combined to analyze the aroma difference of S. rugosoannulata under two cultivation modes. PCA and HCA could effectively distinguish the aroma difference in different growth stages. Under different culturing methods, the aroma substances and their changes were different. The samples were divided into two groups for forest cultivation, while the samples were divided into three groups for greenhouse cultivation. At the end of growth, the aroma of S. rugosoannulata with the two cultivation modes was very similar. OPLS–DA clearly distinguished the differences between the two cultivation methods; 17 key aroma difference factors with variable importance projection (VIP) > 1 were obtained from SPLS–DA analysis.