Fruiting Bodies Research Articles

Structural clarification of mannoglucan GSBP-2 from Ganoderma sinense and its effects on triple-negative breast cancer migration and invasion

Ganoderma sinense, known as Lingzhi in China, is a medicinal fungus with anti-tumor properties. Herein, crude polysaccharides (GSB) extracted from G. sinense fruiting bodies were used to selectively inhibit triple-negative breast cancer (TNBC) cells. GSBP-2 was purified from GSB, with a molecular weight of 11.5 kDa and a composition of α-l-Fucp-(1→, β-d-Glcp-(1→, β-d-GlcpA-(1→, →3)-β-d-Glcp-(1→, →3)-β-d-GlcpA-(1→, →4)-α-d-Galp-(1→,→6)-β-d-Manp-(1→, and →3,6)-β-d-Glcp-(1→ at a ratio of 1.0:6.3:1.7:5.5:1.5:4.3:8.0:7.9. The anti-MDA-MB-231 cell activity of GSBP-2 was determined by methyl thiazolyl tetrazolium, colony formation, scratch wound healing, and transwell migration assays. The results showed that GSBP-2 could selectively inhibit the proliferation, migration, and invasion of MDA-MB-231 cells through the regulation of genes targeting epithelial–mesenchymal transition (i.e., Snail1, ZEB1, VIM, CDH1, CDH2, and MMP9) in the MDA-MB-231 cells. Furthermore, Western blotting results indicated that GSBP-2 could restrict epithelial–mesenchymal transition by increasing E-cadherin and decreasing N-cadherin expression through the PI3K/Akt pathway. GSBP-2 also suppressed the angiogenesis of human umbilical vein endothelial cells. In conclusion, GSBP-2 could inhibit the proliferation, migration, and invasion of MDA-MB-231 cells and showed significant anti-angiogenic ability. These findings indicate that GSBP-2 is a promising therapeutic adjuvant for TNBC.

Study on isolating, breeding, and cultivation of Xylaria nigripes (Klotzsch) Cooke

The Xylaria nigripes fungus is one of the medicinal materials that was used in traditional medicine. Extracts from the mycelium and fruiting bodies of the X. nigripes showed protecting cells, preventing DNA damage, and reducing the programmed death cell rate. In this study, the fruiting bodies of the X. nigripes were collected to isolate, propagate in the liquid medium, and then cultured to induce the fruiting body formation in the rice medium. The results showed that the X. nigripes was successfully isolated on PDAY (Potato Dextrose Agar Yeast Extract) medium, and the mycelium and spore of the X. nigripes had unique characteristics of the species. The biomass of X. nigripes grew well in a liquid nutrient medium containing 20g glucose, 1g MgSO4.7H2O, 2g KH2PO4, and 5g peptone, reaching the best mass after 21 to 28 days. Using an initial cultured liquid of X. nigripes to culture on rice medium, fruiting bodies were formed and developed after 14 days of culture. This result could potentially be applied to the production of X. nigripes biomass via mycelium and fruit bodies for application in medicine.

Dynamics of α-glucan from Agrocybe cylindracea water extract at different developmental stages and its structure characteristics

Polysaccharides are the important bioactive macromolecules in Agrocybe cylindracea, but their changes are as yet elusive during developmental process. This study investigated the dynamic changes of polysaccharides from A. cylindracea fruiting body water extract at four developmental stages and its structure characteristics. Results revealed that the polysaccharides from A. cylindracea water extract significantly increased at the pileus expansion stage and the increased fraction could be α-glucan. The further purification and identification indicated that this α-glucan was a glycogen. It had typical morphology of β particles with a molecular weight of 1375 kDa. Its backbone comprised α-D-(1 → 4)-Glcp and α-D-(1 → 4,6)-Glcp residues at a ratio of 5:1, terminated by α-D-Glcp residue. Rheological behavior suggested that it was a Newtonian fluid at the concentration of 1 %. In addition, despite both the glycogen and natural starch were composed of D-glucose, they exhibited the entirely distinct Maltese cross characteristic and unique crystalline structure. This study is the first to demonstrate the presence of abundant glycogen in the pileus expansion stage of A. cylindracea, which provides new insights on the change patterns of fungal polysaccharides.

Pulvinic Acid Derivative Pigments in Lanmaoa asiatica and L. macrocarpa.

Lanmaoa asiatica G. Wu & Zhu L. Yang and L. macrocarpa N. K. Zeng, H. Chai & S. Jiang are two important gourmet bolete in China, and locally named "Jian Shou Qing" meaning their fruiting bodies turn blue after bruising. The genus represents a distinct lineage in Boletaceae. The pigment(s) associated with the discoloration in Lanmaoa has not been identified. The aim of this study was to determine the pigment(s) underpinning the bluing reaction of L. asiatica and L. macrocarpa when bruised. Potential compounds were isolated by HPLC and identified by LC-HRMS and NMR. In total five to six pigments of hydroxylated pulvinic acid derivatives were detected with similar distribution patterns in both L. asiatica and L. macrocarpa, which by abundance were variegatic acid, variegatorubin, xerocomic acid (and/or isoxerocomic acid), xerocomorubin, and atromentic acid. Variegatic acid, the most abundant pigment, was isolated by HPLC, and the structure was further characterized by NMR. The amount of variegatic acid increased after regular cooking, which may suggest its enhanced health benefit as human diet. The types of pigments that cause bluing reactions often differ among families of Boletales. Our results showed that the pigments in Lanmaoa belong to the category of hydroxylated pulvinic acid derivatives, the major bluing compounds in Boletaceae.

Comprehensive untargeted lipidomics study of black morel (Morchella sextelata) at different growth stages

The black morel (Morchella sextelata) is a valuable edible and medicinal mushroom appreciated worldwide. Here, lipidomic profiles and lipid dynamic changes during the growth of M. sexletata were analyzed using ultra-performance liquid chromatography coupled with mass spectrometry. 203 lipid molecules, including four categories and fourteen subclasses, were identified in mature fruiting bodies, with triacylglycerol being the most abundant (37.00 %). Fatty acid composition analysis revealed that linoleic acid was the major fatty acid among the free fatty acids, glycerolipids and glycerophospholipids. The relative concentration of lipids in M. sextelata changed significantly during its growth, from which 12 and 29 differential lipid molecules were screened out, respectively. Pathway analysis based on these differential lipids showed that glycerophospholipid metabolism was the major pathway involved in the growth of M. sextelata. Our study provides a comprehensive understanding of the lipids in M. sextelata and will facilitate the development and utilization of M. sextelata.

Biological characteristics of Cordyceps militaris single mating-type strains.

Cordyceps militaris has been extensively cultivated as a model cordyceps species for commercial purposes. Nevertheless, the problems related to strain degeneration and breeding technologies remain unresolved. This study assessed the physiology and fertility traits of six C. militaris strains with distinct origins and characteristics, focusing on single mating-type strains. The results demonstrated that the three identified strains (CMDB01, CMSY01, and CMJB02) were single mating-type possessing only one mating-type gene (MAT1-1). In contrast, the other three strains (CMXF07, CMXF09, and CMMS05) were the dual mating type. The MAT1-1 strains sourced from CMDB01, CMSY01, and CMJB02 consistently produced sporocarps but failed to generate ascospores. However, when paired with MAT1-2 strains, the MAT1-1 strains with slender fruiting bodies and normal morphology were fertile. The hyphal growth rate of single mating-type strains (CMDB01, CMSY01, and CMJB02) typically surpassed that of dual mating-type strains (CMXF07, CMXF09, and CMMS05). The growth rates of MAT1-2 and MAT1-1 strains were proportional to their ratios, such that a single mating-type strain with a higher ratio exhibited an increased growth rate. As C. militaris matured, the adenosine content decreased. In summary, the C. militaris strains that consistently produce sporocarps and have a single mating type are highly promising for production and breeding.

Comprehensive Analysis of Bioactive Compounds in Wild Ganoderma applanatum Mushroom from Kerala, South India: Insights into Dietary Nutritional, Mineral, Antimicrobial, and Antioxidant Activities.

The present study focused on the mushroom Ganoderma, which has been used in Eastern countries for centuries as a food and medicinal source. Specifically, the fruiting bodies of Ganoderma applanatum from the Kerala Forest Research Institute in Thirussur, Kerala, India, were analyzed for their nutritional and medicinal properties. The methanolic extracts of G. applanatum were used to examine secondary metabolites and proximate profiles, revealing the presence of various phytochemicals such as terpenoids, phenolics, glycosides, alkaloids, flavonoids, and saponins. Further analysis revealed the presence of significant amounts of calcium, sodium, phosphorus, and manganese. The compounds were characterized using chromatographic analysis, FTIR, and GC-MS, which revealed potential therapeutic compounds with C-H and C-O bonds in the amide group, β-glycosides, and C-C/C-O vibrations of phenolic substances. Mushroom extract at a concentration of 100 µg mL-1 exhibited potent antimicrobial activity against various pathogens. This study suggests that G. applanatum has a rich biochemical composition and pharmacological potential, making it a promising candidate for drug development and traditional medicine, and contributes valuable insights into its diverse therapeutic applications.

No Evidence Was Found for the Presence of Terreolides, Terreumols or Saponaceolides H-S in the Fruiting Bodies of Tricholoma terreum (Basidiomycota, Agaricales).

Two different collections of the gilled wild fungus Tricholoma terreum, collected in Italy, were subjected to phytochemical analysis. The fungal material was confidently identified by analysis of the ITS genomic sequences. Using both HR-LC-MS and NMR techniques, no evidence was found for the presence in the fruiting bodies of terreolides, terreumols or saponaceolides H-S, in striking contrast with the isolation of these terpenoids by Chinese authors from a mushroom collected in France and identified as T. terreum. The main cytotoxic terpenoid identified and isolated from the extracts of the specimens investigated in this work was the C30 derivative saponaceolide B, which had been previously isolated from T. saponaceum and other T. terreum collections. Although saponaceolide B is a rather labile molecule, easily degradable by heat or in acidic conditions, our study indicated that none of the extraction protocols used produced saponaceolide H-S or terreolide/terreumol derivatives, thus excluding the possibility that the latter compounds could be extraction artifacts. Considered together, these findings point to the need for the unambiguous identification of mushroom species belonging to the complex genus Tricholoma, characterized by high variability in the composition of metabolites. Moreover, based on our data, T. terreum must be considered an edible mushroom.

Pelecocera (Pelecocera) tricincta and Pelecocera (Chamaesyrphus) caledonica (Diptera, Syrphidae) reared from Rhizopogon fungal host in Finland.

MtDNA COI barcodes have frequently been used in identification to associate an unknown life stage in insects with a known species. This study reports the discovery of hoverfly larvae in the fungal fruit bodies of Rhizopogonluteolus Fr. & Nordholm, 1817 in Finland. The identity of the larvae was firstly resolved using mtDNA COI barcodes generated from the larvae and tree-based identification confirming the species Pelecocera (Pelecocera) tricincta Meigen, 1822 and Pelecocera (Chamaesyrphus) caledonica (Collin, 1940) (Diptera, Syrphidae). Obtained pupae were reared into adult flies and produced the same two species. The morphological features of these mycophagous larvae are compared with those of other fungus-feeding hoverfly species. This study confirms Rhizopogonluteolus as fungal host for these Pelecocera species in the Western Palaearctic Region.

Analysis of bioactive compounds in Cordyceps militaris fruiting bodies and dietary supplements: in vitro bioaccessibility determination in artificial digestive juices

SummaryIn recent years, Cordyceps militaris has garnered increased attention in both scientific research and popular culture, exemplified by references in the entertainment industry, such as ‘The Last Of Us’. This study aims to investigate the qualitative and quantitative composition of bioactive compounds (such as cordycepin, ergothioneine, L‐phenylalanine, lovastatin, sterols, indole and phenolic compounds) in various samples. These samples encompass commercially available dietary supplements (in the form of capsules or powder), featuring standardised extracts or non‐standardised C. militaris material. Additionally, fruiting bodies cultivated in Poland and China were examined. The bioactive compounds were subsequently analysed after extraction into artificial digestive juices. Each sample underwent extraction, evaporation and dissolution in methanol, followed by qualitative and quantitative analyses using RP‐HPLC (with UV and DAD detectors) for the determination of bioactive compounds. The results of this analysis revealed that extracts with cordycepin contents of 7% and 1% exhibited the highest cordycepin concentrations, as well as significant ergothioneine levels (4405 and 928 mg/100 g d.w. and 234 and 77.6 mg/100 g d.w. respectively). However, on analysis of simulated digestive juices, it became evident that the fruiting bodies of C. militaris are superior sources of bioactive compounds. Consequently, this study underscores the efficacy of unprocessed material, such as fruiting bodies, as the preferred form of supplementation for potential consumers of C. militaris.

Molecular phylogenetic analyses support the validity of Ceratiomyxa porioides (Amoebozoa, Eumycetozoa) at species level

The frequently encountered macroscopic slime molds of the genus Ceratiomyxa have long been recognized by mycologists and protistologists for hundreds of years. These organisms are amoebozoan amoebae that live and grow inside and on the surface of decaying wood. When conditions are favorable, they form subaerial sporulating structures called fruiting bodies which take on a variety of forms. These forms are typically some arrangement of column and/or branches, but one is uniquely poroid, forming folds instead. Originally, this poroid morphology was designated as its own species. However, it was not always clear what significance fruiting body morphology held in determining species. Currently, Ceratiomyxa fruticulosa var. porioides, the poroid form, is considered a taxonomic variety of Ceratiomyxa fruticulosa based on morphological designation alone. Despite its long history of observation and study, the genus Ceratiomyxa has been paid little molecular attention to alleviate these morphological issues. We have obtained the first transcriptomes of the taxon C. fruticulosa var. porioides and found single gene phylogenetic and multigene phylogenomic support to separate it from C. fruticulosa. This provides molecular evidence that fruiting body morphology does correspond to species level diversity. Therefore, we formally raise Ceratiomyxa porioides to species level.

Ganonorsterone A, a norsteroid from the medicinal fungus Ganoderma lingzhi

Phytochemical investigation on the fruiting bodies of the medicinal fungus Ganoderma lingzhi led to the isolation of a new norsteroid, namely ganonorsterone A (1), together with one known steroid, cyathisterol (2). The structure and absolute configuration of compound 1 were assigned by extensive analysis of MS, NMR data, and quantum-chemical calculations including electronic circular dichroism (ECD) and calculated 13C NMR-DP4+ analysis. Bioassay results showed that compound 1 displayed moderate inhibition on NO production in RAW 264.7 macrophages.

Differences in Soil Microflora between the Two Chinese Geographical Indication Products of “Tricholoma matsutake Shangri-la” and “T. matsutake Nanhua”

Tricholoma matsutake is a kind of ectomycorrhizal fungus. In addition to its vital influence on symbiotic plants, the impact of the soil microbial community on the growth and development of T. matsutake has been garnering attention. To clarify the differences in soil microflora between “T. matsutake Shangri-la” and “T. matsutake Nanhua”, and the effects of soil bacteria and fungi on the growth and development of T. matsutake, this study took the soil of “T. matsutake Shangri-la” and “T. matsutake Nanhua” at different developmental stages. A total of 7694 bacterial OTUs and 2170 fungal OTUs were obtained through microbial omics based on amplicon sequencing. The results indicate that the α diversity and composition of the soil microorganisms in the T. matsutake Shangri-la shiro were higher than those of the T. matsutake Nanhua. It is affected by species, geographical locations, and the growth period of matsutake. Matsutake mycelia also recruit certain types of bacteria and fungi in the stage of fruiting body development. Both bacteria and fungi positively and negatively regulate the fruiting body development of matsutake mushrooms. This study will provide a basis for the semi-artificial cultivation of matsutake.

Decomposition of Fomes fomentatius fruiting bodies - transition of healthy living fungus into a decayed bacteria-rich habitat is primarily driven by Arthropoda.

Fomes fomentarius is a widespread, wood-rotting fungus of temperate, broadleaved forests. Although the fruiting bodies of F. fomentarius persist for multiple years, little is known about its associated microbiome or how these recalcitrant structures are ultimately decomposed. Here we used metagenomics and metatranscriptomics to analyse the microbial community associated with healthy living and decomposing F. fomentarius fruiting bodies to assess the functional potential of the fruiting body-associated microbiome and to determine the main players involved in fruiting body decomposition. F. fomentarius sequences in the metagenomes were replaced by bacterial sequences as the fruiting body decomposed. Most CAZymes expressed in decomposing fruiting bodies targeted components of the fungal cell wall with almost all chitin-targeting sequences, plus a high proportion of beta-glucan-targeting sequences, belonging to Arthropoda. We suggest that decomposing fruiting bodies of F. fomentarius represent a habitat rich in bacteria, while its decomposition is primarily driven by Arthropoda. Decomposing fruiting bodies thus represent a specific habitat supporting both microorganisms and microfauna.

Wickerhamomyces corioli f.a., sp. nov., a novel yeast species discovered in two mushroom species.

Two yeast strains, designated as 19-39-3 and 19-40-2, obtained from the fruiting bodies of Trametes versicolor and Marasmius siccus collected in Yunwu Mountain Forest Park, PR China, have been identified as representing a novel asexual ascomycetous yeast species. From the results of phylogenetic analyses of the sequences of the D1/D2 domains of the large subunit (LSU) rRNA, small subunit (SSU) rRNA and translation elongation factor 1-α (TEF1) genes, it was determined that these strains represent a member of the genus Wickerhamomyces, with Wickerhamomyces alni and Candida ulmi as the closest relatives. The novel species exhibited 6.6 and 6.7% differences in the D1/D2 domains compared with W. alni and C. ulmi, respectively. Additionally, distinct biochemical and physiological differences were observed between the novel species and its related counterparts. No sexual reproduction was observed in these strains, leading to the proposal of the name Wickerhamomyces corioli f.a., sp. nov. for this newly discovered species.

Identification, Nutrient Composition, and Evaluation of a Wild Pleurotus citrinopileatus Strain (X21156) from Tibet for Antioxidant and Cytotoxic Activities

A fungal strain (X21156) collected in Tibet was used as the material, identified based on its morphological characteristics and internal transcribed spacer (ITS) sequence; its optimal culture conditions were analyzed by single-factor experiments; artificial domestication and cultivation were carried out; its nutrient composition was determined; and the bioactivities of its polysaccharides were detected using chemical antioxidant assays and MTT assays. The results showed that the strain was Pleurotus citrinopileatus Sing. Its optimal culture conditions were a pH of 7, a temperature of 25 °C, glucose (20 g·L−1) as the carbon source, and yeast powder (20 g·L−1) as the nitrogen source. The fresh weight of a single domesticated fruiting body was 41.16 g. The strain had high protein (28.5%), high fiber (34%), and low fat (1.4%) contents, with high proportions of fresh and sweet amino acids. Polysaccharides had good scavenging ability on ABTS+, DPPH, and OH free radicals (EC50 0.06 mg/mL, 1.21 mg/mL, and 3.62 mg/mL, respectively), and the cytotoxicity of polysaccharides to hepatocellular carcinoma cells (HepG2) (IC50: 1.69 mg/mL) was higher than that of triple-negative breast cancer cells (MDA-MB-468) (IC50: 1.76 mg/mL). In conclusion, the study provides a reference on the optimal culture conditions, domestication and cultivation, and dietary and medicinal values of wild P. citrinopileatus Sing.

Ectomycorrhizal fungi are influenced by ecoregion boundaries across Europe

AbstractAimEcoregions and the distance decay in community similarity are fundamental concepts in biogeography and conservation biology that are well supported across plants and animals, but not fungi. Here we test the relevance of these concepts for ectomycorrhizal (ECM) fungi in temperate and boreal regions.LocationEurope.Time Period2008–2015.Major Taxa StudiedEctomycorrhizal fungi.MethodsWe used a large dataset of ~24,000 ectomycorrhizas, assigned to 1350 operational taxonomic units, collected from 129 forest plots via a standardized protocol. We investigated the relevance of ecoregion delimitations for ECM fungi through complementary methodological approaches based on distance decay models, multivariate analyses and indicator species analyses. We then evaluated the effects of host tree and climate on the observed biogeographical distributions.ResultsEcoregions predict large‐scale ECM fungal biodiversity patterns. This is partly explained by climate differences between ecoregions but independent from host tree distribution. Basidiomycetes in the orders Russulales and Atheliales and producing epigeous fruiting bodies, with potentially short‐distance dispersal, show the best agreement with ecoregion boundaries. Host tree distribution and fungal abundance (as opposed to presence/absence only) are important to uncover biogeographical patterns in mycorrhizas.Main ConclusionsEcoregions are useful units to investigate eco‐evolutionary processes in mycorrhizal fungal communities and for conservation decision‐making that includes fungi.

THE EFFECT OF USING SAGU DRUGS (Metroxylon sagu) AND DRIED BANANA LEAVES (Musa paradisiaca) ON THE GROWTH OF WHITE OYSTER MUSHROOM (Pleurotus ostreatus)

Oyster mushrooms (Pleurotus ostreatus) are wood fungi that grow sideways on rotting wood stems. Mushrooms take food that has been made by other dead organisms (saprophyte), because they do not have chlorophyll, all types of saprophyte, especially those that grow on wood, can be easily cultivated. White oyster mushroom cultivation is easy to do because it does not require expensive maintenance costs and does not require spacious land. The aim of this research was to test the effect of using sago dregs and dried banana leaves on the growth of white oyster mushrooms. The results of this research indicate that there are differences in the growth of white oyster mushrooms based on different concentrations of growing media. The average mycelium growth, stem height, pileus diameter, number of fruit bodies, wet weight and dry weight of the best white oyster mushrooms were found in media with a concentration of 100% sawdust, while pind heads appeared in a concentration of 100% sago dregs

Pinus sibirica Du Tour в условиях интродукции на примере Полярно-альпийского ботанического сада-института

The study has been conducted at the Polar-Alpine Botanical Garden-Institute, the northernmost botanical garden in Russia (67°38′ N). The aim of this work has been a comprehensive analysis of the condition of Siberian stone pine trees (Pinus sibirica Du Tour) during their introduction in the Arctic. The condition of 8 specimens of the species cultivated at the experimental plot in the town of Apatity has been examined. Their phenological development has been analyzed: the swelling and breaking of the vegetative buds, the end of the elongation and maturation of the needles, the beginning and end of growth of the annual shoots, the lignification of the shoots, the beginning and end of pollen dispersion. The degree of lignification of the annual shoot, the winter hardiness, the habit, the shootforming capacity, the height increment, the generative development and the reproduction in the plantation have also been assessed. The risk of a tree falling has been determined by considering its surroundings and exposure to wind. The root zones, the butts, the trunks, the crown bases, the boughs and the crowns have been examined for the presence of hollows, cracks, decomposition, fungal fruit bodies, damage, weak forks, dead branches, etc. The application of an approach combining the analysis of seasonal development, viability and risk of tree fall is promising for a comprehensive and objective assessment of the condition of Siberian stone pine trees by morphological and phenological characteristics during the introduction to the Arctic zone of the Russian Federation. The phenological rhythm of the studied trees corresponds to the environmental conditions during the introduction to the Polar-Alpine Botanical Garden-Institute named after N.A. Avrorin. Vegetation begins in late May. By the end of August the annual shoots lignify, which indicates the readiness of the plants for the winter period. The close timing of the onset of phenophases and their low variation may indicate a low dependence of the seasonal development of Siberian stone pine on weather conditions during the introduction to the central part of the Kola Peninsula. The trees are winter-hardy, retain the life form inherent to the species, have a high shoot-forming capacity and annual height increment. They reach the generative stage of development. For their propagation in the plantation, it is necessary to attract seed material from other regions. The trees are monocormic, unimucronate and bimucronate. They are characterized by dense crowns. Their apical and lateral shoots are developed. In rare cases, the increment of lateral shoots is reduced. The V-shaped forks have been noted on the stems and a hollow in the butt area has been found on one of the trees. An additional instrumental examination is needed to determine the effect of diseases or pathogens on the stability of the trees.

MORPHOLOGICAL IDENTIFICATION OF WILD EDIBLE MUSHROOM (Termitomyces striatus) AT IPB UNIVERSITY CAMPUS FOREST

Indonesia is a tropical country which is famous for its 2 seasons, namely the rainy season and the dry season. In the rainy season, the humidity level will increase and induce the development of mushroom’s fruiting body, including the edible kind. The research on wild edible mushroom diversity should be carried out massively in various regions in Indonesia. One of the potential areas is the forest area of the IPB University Campus (HKIU). This study aims to explore and identify wild edible mushroom in HKIU. The mushroom exploration was done using an opportunistic sampling method. Selected wild mushroom was obtained and identified for macroscopic and microscopic morphological characters. The identification results based on the morphological characters of the fruiting bodies indicated that the mushrooms obtained from the current study is Termitomyces striatus which is acknowledged as an edible mushroom. Our finding can provide the additional information on distribution of T. striatus in Indonesia and the potential of this mushroom cultivation in foreseeable future.