Matsutake Research Articles

Impact of polyphenols extracted from Tricholoma matsutake on UVB-induced photoaging in mouse skin.

Despite Tricholoma matsutake has been used as natural health products with multiple medicinal properties, detailed information about its polyphenolic composition as sources of anti-photoaging agents remains to be determined. To investigate the impact of polyphenols extracted from Tricholoma matsutake (TME) on Ultraviolet B (UVB)-induced skin photoaging. Various factors of oxidative stress and inflammation as well as histological and immunohistochemical analysis in the mouse dorsal skin were determined after UVB radiation. Topical administration with TME suppressed the UVB-induced skin thickness, wrinkles and erythema, and increased skin collagen content. Furthermore, TME decreased reactive oxygen species (ROS) level, upregulated glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT), and glucose-6-phosphate dehydrogenase (G6PDH) activities and inhibited the expression of IL-1, IL-6, IL-8, and TNF-α in mice irradiated with UVB. TME could reduce UVB-induced p38 mitogen-activated protein kinase (p38 MAPK) phosphorylation and effectively inhibited the activity of the transcriptional factor nuclear factor-kappa B (NF-κB), thereby reducing the cyclooxygenase-2 (COX-2) expression, which is an important mediator of inflammatory cascade leading to the inflammatory response. Our data demonstrated that TME had various beneficial effects on UVB-induced skin photoaging due to its antioxidant and anti-inflammatory activities, and it might be exploited as a promising natural product in skin care, anti-photoaging and the therapeutic intervention of skin disorders related to both oxidative stress and inflammation.

A Practical Method for Determination of Nine Nucleosides in Tricholoma matsutake by UPLC/MS and Quantitative Analysis of Multicomponents Using Single Marker Method.

Nucleosides can be used as quality evaluation indicators of Tricholoma matsutake. In this work, a new ultra-performance liquid chromatography-tandem mass spectrometry (UPLC/MS) strategy for quantitative analysis of multiple components using a single marker (QAMS) was proposed to determine nine nucleosides (adenosine, cytidine, guanosine, inosine, uridine, 2′-deoxyadenosine, 2′-deoxycytidine, 2′-deoxyguanosine, and 2′-deoxyuridine) in T. matsutake. Guanosine was set as the internal reference substance, whose content in T. matsutake was determined using the conventional external standard method. Relative correction factors (RCFs) between guanosine and eight other nucleosides were measured. The concentrations of the eight components were calculated with the obtained RCFs by QAMS. An ultrasonic extraction method is used for sample preparation. This method was validated to be sensitive, precise, and accurate with the LODs of 0.31–1.9 ng, the overall intraday and interday variations less than 4.08%, and the overall recovery over 89.0%. The correlation coefficients (r2) of the calibration curves were higher than 0.9918. The values of vector angle analysis were above 0.99845, which indicates no significant differences between the conventional external standard method and the present QAMS method. As far as we know, this is also the first report of UPLC/MS analysis of nucleoside compounds by QAMS, providing an efficient and feasible quality assessment method for other natural products containing nucleosides.

Identification and Differentiation of Wide Edible Mushrooms Based on Lipidomics Profiling Combined with Principal Component Analysis.

Mushroom, as a kind of higher fungus, is a precious homology resource of medicine and foods. In this study, total lipids were extracted from eight wild edible mushrooms and subsequently characterized by ultra-high-performance liquid chromatography-Quadrupole-Exactive Orbitrap mass spectrometry. 20 lipid classes and 173 molecular species were identified and quantified. Lipid molecules and their concentrations in Boletus speciosus, Boletus bainiugan, and Tricholoma matsutake exhibited significantly different behaviors compared with the remaining mushrooms. Hierarchical cluster analysis revealed that lipid profiles of B. bainiugan were most similar to B. speciosus followed by T. matsutake, Canthar-ellus cibarius, Sarcodon aspratu, Termitomyces eurrhizus, Laccaria laccata, and Thelephora ganbajun. In addition, several differential lipids can be considered as potential biomarkers to distinguish different mushroom species, for instance, lysophosphatidylethanolamine (16:1) and ceramide non-hydroxy fatty acid-dihydrosphingosine (d23:0-10:0). This study provided a new perspective to discriminate the mushroom species from the perspective of lipidomics.

Distinct Compartmentalization of Microbial Community and Potential Metabolic Function in the Fruiting Body of Tricholoma matsutake.

The uniquely compartmentalized fruiting body structure of the ectomycorrhizal fungus (EMF) Tricholoma matsutake, is a hotspot of microbial habitation and interaction. However, microbial diversity within this microniche structure of the EMF is rarely investigated. Furthermore, there is limited information concerning microbiomes associated with sporomes belonging to the ubiquitous fungal phylum Basidiomycota, particularly with respect to fungus-EMF interactions. In this study, we conducted high throughput sequencing, using ITS (fungal) and 16S rRNA (bacterial) marker genes to characterize and compare fruiting body microbiomes in the outer (pileipellis and stipitipellis) and inner layers (pileum context, stipe context, and lamellae) of the fruiting body of T. matsutake. Our results show the number of unique bacterial operational taxonomic units (OTUs) among the different compartments ranged from 410 to 499 and was more than double that of the shared/common OTUs (235). Micrococcales, Bacillales, Caulobacter, and Sphingomonas were the primary significant bacterial taxa within the different compartments of the dissected T. matsutake fruiting body. Non-parametric multivariate analysis of variance showed significant compartmental differences for both the bacterial and the fungal community structure within the T. matsutake fruiting body. The metabolic profiling revealed putative metabolisms (of amino acids, carbohydrates, and nucleotides) and the biosynthesis of secondary metabolites to be highly enriched in outer layers; in the inner parts, the metabolisms of energy, cofactors, vitamins, and lipids were significantly higher. This study demonstrates for the first time the distinct compartmentalization of microbial communities and potential metabolic function profiles in the fruiting body of an economically important EMF T. matsutake.

Pollution level and risk assessment of lead, cadmium, mercury, and arsenic in edible mushrooms from Jilin Province, China.

To evaluate the pollution level of toxic elements in edible mushrooms from Jilin Province, China, the lead, cadmium, mercury, and arsenic content present in them were monitored and evaluated. A total of 610 edible mushroom samples, including fresh and dried, were collected from nine cities in the Jilin Province. The concentrations of lead, cadmium, mercury, and arsenic ranged from 0.007 to 3.31, 0.006 to 48.52, 0.003 to 0.56, and 0.008 to 57.34mg/kg, respectively, in dried samples, and 0.007 to 0.06, 0.006 to 0.17, 0.003 to 0.06, and 0.008 to 0.12mg/kg, respectively, in fresh samples. Theconcentration of cadmium and arsenic exceeded the standard (Cd ≤ 0.2mg/kg, Cd of Lentinus edodes ≤ 0.5mg/kg, and As ≤ 0.5mg/kg) in some mushroom samples. The bioconcentration levels of lead, cadmium, mercury, and arsenic depended on the edible mushroom species, environment, and the physicochemical properties of lead, cadmium, mercury, and arsenic. The results of the single factor pollution index, a method used to calculate the level of single elements in substances, showed that all edible mushrooms were at the level of unpolluted except Tricholoma matsutake and Pleurotus eryngii. The comprehensive factor (P) for T. matsutake was 1.093. The comprehensive factor pollution index, a method used to calculate the level of combined mixture in substances, was in the unpolluted level, except for T. matsutake. The pollution index results showed that cadmium and arsenic concentrations were lightly polluting in some mushroom samples. The health risk index for arsenic was the highest. Therefore, more attention should be paid to arsenic contamination in T. matsutake and P. eryngii in the Jilin Province, China. PRACTICAL APPLICATION: In this paper, we evaluated the contamination levels and associated safety issues of four toxic elements, lead, cadmium, mercury, and arsenic, in edible mushrooms. The results showed that cadmium and arsenic concentrations were lightly polluting in some mushroom samples.

Metagenomic Analysis of Bacterial and Fungal Communities Inhabiting Shiro Dominant Soils of Two Production Regions of Tricholoma Matsutake S. Ito & S. Imai in Korea

Tricholoma matsutake is an ectomycorrhizal fungus that has obligate symbiotic relationships with Pinus densiflora. Its fruiting body has a distinctive flavor and is traded at a high price. Thus, it has been a significant source of income for rural communities in Korea. We hypothesized that biotic factors considerably influence the formation of the T. matsutake mushroom, and the soils producing T. matsutake share similar microbial characteristics. Therefore, the present study aimed to detect the specific fungal and bacterial groups in T. matsutake production soils (shiro+) and nonproduction soils (shiro−) of the Bonghwa and Yanyang regions via next-generation sequencing. In a total of 15 phyla, 36 classes, 234 genera of bacteria, six phyla, 29 classes, and 164 genera of fungi were detected from four samples at both sites. The species diversity of shiro+ soils was lower than the shiro− samples in both the fungal and bacterial groups. In addition, we did not find high similarities in the microbial communities between the shiro+ soils of the two regions. However, in the resulting differences between the fungal communities categorized by their trophic assembly, we found a distinguishable compositional pattern in the fungal communities from the shiro+ soils and the shiro− soils of the two sites. Thus, the similarity among the microbial communities in the forest soils may be due to the fact that the microbial communities in the T. matsutake dominant soils are closely associated with biotic factors and abiotic factors such as soil properties.

Potential Mechanisms Mediating the Protective Effects of Tricholoma matsutake-Derived Peptides in Mitigating DSS-Induced Colitis.

Intestinal barrier dysfunction and inflammatory cytokine secretion play crucial roles in inflammatory bowel disease (IBD). Herein, we investigated the protective effects of Tricholoma matsutake-derived peptides SDIKHFPF and SDLKHFPF against dextran sulfate sodium-induced colitis. Both peptides alleviated colitis signs, including diarrhea, weight loss, bloody stools, colon shortening, and histopathological changes, while reducing mucus destruction, goblet cell exhaustion, and intestinal permeability. SDIKHFPF and SDLKHFPF protected the barrier function by promoting the expression of tight junction (TJ) zonula occludens-1 and occludin within the colon, as well as attenuating colonic inflammation through myeloperoxidase and pro-inflammatory cytokine suppression. Western blotting indicated that the peptides suppressed myosin light chain kinase (MLCK) and nuclear factor kappa B (NF-κB) levels, inhibiting MLC phosphorylation. SDLKHFPF was more potent than SDIKHFPF. These findings suggest that peptide SDLKHFPF mitigates colitis by regulating TJ protein expression and pro-inflammatory cytokine production via NF-κB/MLCK/p-MLC signaling, improving the barrier function.

Expansion Process of ‘Shiro’ of <i>Tricholoma matsutake</i> at the Kyotamba-cho Test Site

Expansion Process of ‘Shiro’ of <i>Tricholoma matsutake</i> at the Kyotamba-cho Test Site

Pyrosequencing and Taxonomic Composition of the Fungal Community from Soil of Tricholoma matsutake in Gyeongju.

Tricholoma matsutake is an ectomycorrhizal fungus that has a symbiotic relationship with the root of Pinus densiflora. Soil microbial communities greatly affect the growth of T. matsutake, however, few studies have examined the characteristics of these communities. In the present study, we analyzed soil fungal communities from Gyeongju and Yeongdeok using metagenomic pyrosequencing to investigate differences in fungal species diversity, richness, and taxonomic composition between the soil under T. matsutake fruiting bodies (Sample 2) and soil where the fairy ring of T. matsutake was no longer present (Sample 1). The same spot was investigated three times at intervals of four months to observe changes in the community. In the samples from Yeongdeok, the number of valid reads was lower than that at Gyeongju. The operational taxonomic units of most Sample 2 groups were less than those of Sample 1 groups, indicating that fungal diversity was low in the T. matsutakedominant soil. The soil under the T. matsutake fruiting bodies was dominated by more than 51% T. matsutake. From fall to the following spring, the ratio of T. matsutake decreased. Basidiomycota was the dominant phylum in most samples. G-F1-2, G-F2-2, and Y-F1-2 had the genera Tricholoma, Umbelopsis, Oidiodendron, Sagenomella, Cladophialophora, and Phialocephala in common. G-F1-1, G-F2-1, and Y-F1-1 had 10 genera including Umbelopsis and Sagenomella in common. From fall to the following spring, the amount of phyla Basidiomycota and Mucoromycota gradually decreased but that of phylum Ascomycota increased. We suggest that the genus Umbelopsis is positively related to T. matsutake.

Flavor Changes of Tricholoma matsutake Singer under Different Processing Conditions by Using HS-GC-IMS.

Headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) was used to analyze the changes to volatile compounds in fried Tricholoma matsutake Singer under different heating temperatures and times. A total of 40 signals that corresponded to 24 compounds were identified through this approach. Differences in volatile compounds of T. matsutake samples were shown in topographic plots and fingerprints. The heating temperatures were more important than the heating times in affecting the volatile compounds. Moreover, changes to the main volatile compounds in T. matsutake under different processing conditions were based on the thermal decomposition and a series of chemical reactions of C8 compounds. Principal component analysis (PCA) results showed that samples under different processing conditions could be effectively distinguished. Hence, the combination of HS-GC-IMS and PCA can identify and classify the volatile compounds of T. matsutake quickly and sensitively. This study provides a new supplementary analytical method for the rapid identification of T. matsutake. The above results can provide a theoretical basis for the quality control and change mechanism of flavor in the processing of traditional edible fungi products.

Potential mechanisms underlying the protective effects of Tricholoma matsutake singer peptides against LPS-induced inflammation in RAW264.7 macrophages

This study aimed to investigate the protective effects of a < 3 kDa Tricholoma matsutake Singer peptide (TMWP) on lipopolysaccharide (LPS)-induced inflammation in RAW264.7 macrophages. The results showed that TMWP significantly upregulated superoxide dismutase (SOD) activity and reduced reactive oxygen species (ROS) generation in RAW264.7 macrophages. Western blotting and immunofluorescence analysis indicated that TMWP inhibited the activation of the mitogen-activated protein kinase (MAPK) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathways, thereby reducing the secretion of IL-1β and IL-6 and the expression of TNF-α, COX-2, and iNOS. Additionally, TMWP improved mitochondrial respiration in LPS-stressed macrophages, counteracting the harmful effects of LPS treatment on mitochondrial function. Three peptides (SDIKHFPF, SDLKHFPF, and WFNNAGP) with the highest predicted scores for potential anti-inflammatory activity were identified using nano-HPLC-MS/MS. These data indicated that T. matsutake peptides could be an attractive natural ingredient for developing novel functional foods.

How the Matsutake Mushroom Become a Luxury Food during the Postwar Period:

How the Matsutake Mushroom Become a Luxury Food during the Postwar Period:

Effects of Tricholoma matsutake (Agaricomycetes) Extracts on Promoting Proliferation of HaCaT Cells and Accelerating Mice Wound Healing.

Tricholoma matsutake is popular in Asian countries because of its edibility and medicinal use. T. matsutake is a precious natural medicinal fungus, and it is widely used in food and biological products. This study aimed to explore the mechanism of T. matsutake on promoting proliferation of human immortalized keratinocyte (HaCaT) cells and accelerating wound healing in mice. The MTT assay was used to test the effects of three different T. matsutake extracts (0, 62.5, 125, 250, 500, and 1000 μg/mL) on HaCaT cell viability. HaCaT cells were treated with the three T. matsutake extracts (100, 500 μg/mL) and morphological and biophysical properties were detected by atomic force microscopy with JPK data processing. Western blot analysis detected Notch signaling pathways of HaCaT cells treated with 50% ethanol extract of T. matsutake (50%T) for 24 h (100, 500 and 1000 μg/mL). Mouse wounds were treated with 50%T for 15 days. Wound healing effects were observed on the back skin of mice at different times. The quality of wound healing was estimated by histological staining (hematoxylin and eosin and Masson's trichrome). All data were counted by GraphPad Prism 5 software. The increased concentration of T. matsutake remarkably promoted HaCaT cell proliferation. The Young's modulus of HaCaT cells showed the biggest increase from 1.73 ± 0.13 kPa (0 μg/mL) to 4.57 ± 0.16 kPa (500 μg/mL) in the 50%T group. The Notch1/Jagged1 pathways were upregulated with an increase in concentration (0, 100, 500, and 1000 μg/mL). Moreover, compared with the negative and positive control groups, T. matsutake promoted wound healing in mice by epidermal regeneration, subepidermal tissue formation, and collagen deposition. The results showed that T. matsutake promotes not only proliferation of HaCaT cells but also wound healing in mice.

Industrial applications of selected JFS articles

Industrial applications of selected <i>JFS</i> articles

Genetic structure of Tricholoma matsutake in Japan: conservation of genetic resources of domestic “matsutake” mushrooms

ABSTRACT Tricholoma matsutake is an ectomycorrhizal fungus found in Pinaceae forests in Japan. It produces the “matsutake” mushroom, which has been consumed as an important part of Japanese cuisine since ancient times. Domestic matsutake are among the most valuable special forest products; however, their production in Japan has decreased over the past several decades. In the present study, to facilitate the conservation of genetic resources of domestic matsutake, we analyzed the fruiting bodies of T. matsutake collected from 13 prefectures, including one Picea glehnii forest and 12 Pinus densiflora forests in the Japanese archipelago, using SSR markers and elucidated their genetic structures . In total, 123 genotypes were detected in these populations. The genetic differentiation between populations was low (F ST = 0.048 and G’ST = 0.052). Moreover, no significant IBD was detected on the spatial scale, where the distance between the populations was up to approximately 1300 km. The genetic differences among T. matsutake populations in different regions of Japan are small. Three genetic clusters were identified, although no clear structure was confirmed based on geographical and genetic distributions However, F IS>0 was confirmed in several populations (p < 0.01), and AMOVA showed a significant genetic variation of approximately 5% among the groups, indicating a weak genetic structure of Japanese T. matsutake. The genetic resource conservation area could not be uniformly determined, because the correlation between genetic structure and geographical distribution is unclear.

Antifatigue effect of functional cookies fortified with mushroom powder (Tricholoma Matsutake) in mice.

Bakery products made by cereal and edible fungi powder have a unique flavor and health benefits, dramatically enhancing the nutritional value of the products. In this study, we investigated the antifatigue effect of a novel Tricholoma matsutake cookie (TMC) by the exhaustive swimming test. Male Kunming ICR mice were randomly divided into seven groups (each group, n = 10), fed with saline, ordinary cookies (4, 8, 16g/kg B.W./day), and TMC (4, 8, 16g/kg B.W./day) by gavage. After 30-day administration, the weight-loaded swimming test was carried out on the mice to evaluate the antifatigue effect of TMC. In comparison with the effect of ordinary cookies, the intake of TMC significantly prolonged the exhaustive swimming time of mice and increased the level of muscle glycogen and liver glycogen, accompanied by the reduction of lactic acid and urea nitrogen level in serum. Additionally, TMC dramatically improved the activity of superoxide dismutase and glutathione peroxidase in serum and largely decreased the level of malondialdehyde. All in all, TMC could enforce exhaustive swimming tolerance, accelerate the decomposition of sports-related metabolites such as lactic acid and urea nitrogen, and increase the activity of the antioxidant enzyme, thereby improving sports-related energy storage and relieving fatigue. Our findings broadened the application of T. matsutake in the processing of bakery products and provided the theoretical basis and technical support for the development of antifatigue products. PRACTICAL APPLICATION: In this study, we investigated the antifatigue effect of a novel Tricholoma matsutake cookie by the exhaustive swimming test. Collectively, the results of the present study suggested that the cookies fortified with T. matsutake could be considered as an antifatigue bakery product. Furthermore, our findings broadened the application of T. matsutake in the processing of bakery products and provided the theoretical basis and technical support for the development of antifatigue products.

Argon-ion beam induced mutants of the ectomycorrhizal agaricomycete Tricholoma matsutake defective in β-1,4-endoglucanase activity promote the seedling growth of Pinus densiflora in vitro

Tricholoma matsutake is an ectomycorrhizal agaricomycete that produces the prized matsutake mushroom in conifer forests. Here, we isolated mutants generated by the argon-ion beam irradiation of T. matsutake NBRC 33136, designated as Ar 5002 and Ar 5012, which are defective in β-1,4-endoglucanase activity, that promoted the above- and below-ground growth of the symbiotic partner Pinus densiflora in vitro, compared with the control. Pine seedlings associated with Ar 5002, compared with NBRC 33136, had significantly higher above-ground dry mass and significantly greater plant growth, while those associated with Ar 5012 had significantly higher above- and below-ground dry mass. There were no significant differences between the numbers of mycorrhizal root-tips synthesized by the wild-type and mutants, indicating that β-1,4-endoglucanase did not influence the mycorrhization of T. matsutake. In seedlings inoculated with Ar 5002- and Ar 5012, the Hartig nets resembled those of seedlings inoculated with NBRC 33136. The data suggest that T. matsutake lacking β-1,4-endoglucanase activity confer beneficial effects on the growth of P. densiflora. Further studies are necessary to determine whether the symbiotic relationship could be strengthened by such a mutated fungal trait.

Successional Change of the Fungal Microbiome Pine Seedling Roots Inoculated With Tricholoma matsutake.

The pine mushroom (Tricholoma matsutake; Agaricales, Tricholomataceae) is an ectomycorrhizal fungus that produces a commercially valuable, edible mushrooms. Attempts to artificially cultivate T. matsutake has so far been unsuccessful. One method used to induce T. matsutake to produce fruiting bodies of in the wild is shiro (mycelial aggregations of T. matsutake) transplantation. In vitro ectomycorrhization of T. matsutake with seedlings of Pinus densiflora has been successful, but field trials showed limited production of fruiting bodies. Few studies have been done to test what happens after transplantation in the wild, whether T. matsutake persists on the pine seedling roots or gets replaced by other fungi. Here, we investigated the composition and the interaction of the root fungal microbiome of P. densiflora seedlings inoculated with T. matsutake over a 3 year period after field transplantation, using high-throughput sequencing. We found a decline of T. matsutake colonization on pine roots and succession of mycorrhizal fungi as P. densiflora seedlings grew. Early on, roots were colonized by fast-growing, saprotrophic Ascomycota, then later replaced by early stage ectomycorrhiza such as Wilcoxina. At the end, more competitive Suillus species dominated the host roots. Most of the major OTUs had negative or neutral correlation with T. matsutake, but several saprotrophic/plant pathogenic/mycoparasitic species in genera Fusarium, Oidiodendron, and Trichoderma had positive correlation with T. matsutake. Four keystone species were identified during succession; two species (Fusarium oxysporum, and F. trincintum) had a positive correlation with T. matsutake, while the other two had a negative correlation (Suillus granulatus, Cylindrocarpon pauciseptatum). These findings have important implications for further studies on the artificial cultivation of T. matsutake.

Analysis of flavor-related compounds in four edible wild mushroom soups

Four edible wild mushrooms, ie., Tuber aestivum, Cantharellus cibarius, Tricholoma matsutake and Boletus edulis were used as experiment materials to cook soup, and the difference of flavor profiles among the four soups were observed. Sensory evaluation results showed that C. cibarius soup had the highest score. The content of total fatty acids was the most in T. matsutake soup. Palmitic, stearic, oleic and linoleic acids were the main in four soups. The water-soluble low molecular weight (WSLMW) components from four soups were detected by liquid chromatography-mass spectrometry (LC-MS) based metabolomics approach, and then betaine, isopentenyladenine, choline and other 11 differential WSLMW components were screened out. 41 volatile compounds were identified by gas chromatography-ion mobility spectrometry (GC-IMS) coupled with multivariate statistical analysis, and the profiles of volatile compounds were obviously different among the four soups. Every studied soup had a unique taste, which could be completely distinguished by the electronic-tongue (ET).

Changes in the key aroma compounds of matsutake mushroom (Tricholoma matsutake Sing.) from Canada during pan-frying elucidated by application of the sensomics approach

A distillate obtained by solvent extraction and solvent-assisted flavor evaporation (SAFE) from raw, unprocessed matsutake mushrooms (Tricholoma matsutake Sing.) from Canada were analyzed by application of the aroma extraction dilution analysis (AEDA). Twenty-eight aroma-active compounds were detected in the flavor dilution (FD) factor range of 4–4096 among which (E)-methyl cinnamate (sweet, fruity), 4-methylquinazoline (minty), 1-octen-3-one (mushroom-like), trans-4,5-epoxy-(E)-2-decenal (metallic), and 3-methylbutanoic (sweaty) appeared with the highest FD factors. Quantitation of 18 odour-active compounds by stable isotope dilution assays, and a calculation of odour activity values (OAV ratio of concentration to odour threshold) confirmed 1-octen-3-one and (E)-methyl cinnamate as well as (E,E)-2,4-nonadienal, and 3-methylbutanal with OAVs ≥ 390 as key aroma compounds of the raw matsutake. A quantitative study on changes in the concentrations of (E)-methyl cinnamate over time suggested its rapid enzymatic formation within a few minutes. After pan-frying, among the 36 odour-active compounds located by AEDA, the highest FD factors were determined for 4-hydroxy-2,5-dimethyl-3(2H)-furanone (caramel-like), (E)-methyl cinnamate, 4-methylquinazoline, 2H-chromen-2-one (woodruff, almond-like), 1-octen-3-one, (Z)-methyl cinnamate (fruity), trans-4,5-epoxy-(E)-2-decenal, and 2-aminoacetophenone (foxy). Based on the quantitative data, the highest OAVs ≥ 278 were calculated for (E)-methyl cinnamate, (E,E)-2,4-nonadienal, (Z)-1,5-octadien-3-one, and 3-methylbutanal. The overall aroma profile of the pan-fried matsutake could be simulated by an aroma recombinate of 17 odorants in their natural concentrations occurring in the mushrooms. A total of 14 compounds were newly identified in matsutake.