Agrocybe Aegerita Research Articles

Antifungal Activity of Ageritin, a Ribotoxin-like Protein from Cyclocybe aegerita Edible Mushroom, against Phytopathogenic Fungi.

Ageritin from poplar mushrooms is a specific endonuclease that hydrolyzes a single phosphodiester bond located in the sarcin-ricin loop (SRL) of the large rRNA, thereby blocking protein synthesis. Considering the possible biotechnological use of this enzyme, here we report its antifungal activity against virulent fungi affecting crops of economic interest. Our results show that ageritin (200 µg/plug; ~13.5 nmole) inhibits the growth of Botrytis cinerea (57%), Colletotrichum truncatum (42%), and Alternaria alternata (57%), when tested on potato dextrose agar plates. At the same time, no effect was observed against Trichoderma harzianum (a fungus promoting beneficial effects in plants). To verify whether the antifungal action of ageritin against B. cinerea and T. harzianum was due to ribosome damage, we tested ageritin in vitro on partially isolated B. cinerea and T. harzianum ribosomes. Interestingly, ageritin was able to release the Endo's fragment from both tested fungal ribosomes. We therefore decided to test the antifungal effect of ageritin on B. cinerea and T. harzianum using a different growth condition (liquid medium). Differently from the result in solid medium, ageritin can inhibit both B. cinerea and T. harzianum fungal growth in liquid medium in a concentration-dependent manner up to 35.7% and 38.7%, respectively, at the highest concentration tested (~200 µg/mL; 12 µM), and the analysis of RNA isolated from ageritin-treated cells revealed the presence of Endo's fragment, highlighting its ability to cross the fungal cell wall and reach the ribosomes. Overall, these data highlight that the efficacy of antifungal treatment to prevent or treat a potential fungal disease may depend not only on the fungal species but also on the conditions of toxin application.

Preparation of polysaccharide-conjugated selenium nanoparticles from spent mushroom substrates and their growth-promoting effect on rice seedlings

Selenium nanoparticles (SeNPs) have gained significant attention in the agricultural field due to their favorable bioavailability and low toxicity, making them a highly researched subject. In this study, crude polysaccharides from spent mushroom substrate of Agrocybe aegerita (AaPs) were extracted for preparing the polysaccharide‑selenium-nanoparticles (AaPs-SeNPs) by ascorbic acid reduction method. The structure of AaPs-SeNPs was analyzed and their growth-promoting effects on rice seedlings were studied by adopting different application methods. The results revealed that AaPs-SeNPs exhibited improved free radical scavenging ability, with a lower half-maximal inhibitory concentrations compared to AaPs. Rice seedlings treated with AaPs-SeNPs showed significant enhancements in growth characteristics when compared to AaPs treatment, and foliar application exhibited a better growth-promoting effect compared to root application. Moreover, the growth performance and antioxidant enzyme activities of rice seedlings were enhanced by the addition of AaPs-SeNPs, and the absorption efficiency of essential nutrients such as N/P/K and Fe/Zn/Mn was also improved at appropriate concentrations, which could be one of the key factors contributing to the improved growth performance of plants. This study provides new aspects for the utilization of SMS, and also offers new insights from the perspective of nutrient absorption on how polysaccharide-conjugated selenium nanoparticles enhance crop growth.

Flavor, physicochemical properties, and storage stability of P. lobata-coix seed fermented beverage produced by A. aegerita

Pueraria lobata and coix seeds have complementary nutritional profiles, and their nutritional value can be enhanced through biotransformation. Agrocybe aegerita (A. aegerita) is a highly nutritious mushroom with a rich enzyme content. This study investigated the flavor, physicochemical properties, and storage stability of P. lobata-coix seed fermented beverage (PCFB) by A. aegerita. Sixty volatile compounds were detected by headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry. Compared to unfermented PCFB, fermentation with A. aegerita enhanced its physicochemical properties, with the contents of essential amino acids, γ-Aminobutyric acid, and soluble proteins increasing from 16.81%, 2.64 mg/100 mL, and 49.40% to 21.06%, 4.20 mg/100 mL, and 53.08%, respectively. Two efficient shelf-life prediction models of PCFB were established with the Arrhenius model using pH and sensory evaluation as indexes. These findings demonstrate that PCFB is a novel, high-quality beverage and provide a foundation for potential industrial production of PCFB using A. aegerita.

Peroxygenase-catalysed selective and scalable synthesis of 25-hydroxy-grundmann's ketone and its derivatives

Peroxygenases are selective catalysts for C-H bond oxyfunctionalization reactions, however, their potential for organic synthesis still needs to be explored. 25-Hydroxy-Grundmann's ketone is a key structural motifs in various biologically active vitamin D3 analogues. In this study, we report the use of unspecific peroxygenase from Agrocybe aegerita as a highly selective catalyst for the C25 hydroxylation of Grundmanns's ketone and its analogues. Under the optimised conditions, excellent conversion (> 99%) and regioselectivity (> 99%) was obtained. The selected hydroxylation processes were performed on an increased lab scale (up to 200 mL) with up to 1 mmol substrate, which led in 85.7% isolated yield.This simple biocatalytic method holds great potential in streamlining the synthetic pathways of vitamin D3 analogues.

Agrocybe cylindracea fucoglucogalactan induced lysosome-mediated apoptosis of colorectal cancer cell through H3K27ac-regulated cathepsin D

Inducing lysosomal dysfunction is emerging as a promising means for cancer therapy. Agrocybe cylindracea fucoglucogalactan (ACP) is a bioactive ingredient with anti-tumor activity, while its mechanism remains obscure. Herein, we found that ACP visibly inhibited the proliferation of colorectal cancer cells, and the IC50 value on HCT-116 cells (HT29 cells) was 490 μg/mL (786.4 μg/mL) at 24 h. RNA-seq showed that ACP regulated mitochondria, lysosome and apoptosis-related pathways. Further experiments proved that ACP indeed promoted apoptosis and lysosomal dysfunction of HCT-116 cells. Moreover, ChIP-seq revealed that ACP increased histone-H3-lysine-27 acetylation (H3K27ac) on CTSD (cathepsin D) promoter in HCT-116 cells, thus facilitating the binding of transcription factor EB (TFEB), and resulted in ascension of CTSD expression. Additionally, ACP triggered mitochondrial-mediated apoptosis by decreasing mitochondrial membrane potential and increasing pro-apoptotic protein levels. Notably, Pepstatin A (CTSD inhibitor) availably alleviated ACP-induced apoptosis. Taken together, our results indicated that ACP induced lysosome-mitochondria mediated apoptosis via H3K27ac-regulated CTSD in HCT-116 cells. This study indicates that ACP has anti-cancer potential in the treatment of colorectal cancer.

Application of the all-in-one electrode for in situ H2O2 generation in hydroxylation catalyzed by unspecific peroxygenase from Agrocybe aegerita

The versatile unspecific peroxygenase from the fungus Agrocybe aegerita (AaeUPO) is capable to perform hydroxylation of various substrates, even unactivated CH bonds. The technical application of AaeUPO is still limited due to an instability towards its co-substrate hydrogen peroxide (H2O2). Electrochemical synthesis of H2O2 is an attractive approach that provides controllable in situ generation of H2O2. The aims of this study are to promote catalyst efficiency and develop an individually tailored system for H2O2-dependent reactions. Herein, an All-in-One electrode (AiO) system with a carbon felt cathode is used to generate H2O2in situ and combined for the first time with an enzymatic hydroxylation. The AiO electrode combines the counter and working electrode in a single rod structure. This shape provides convenient integration into conventional bioreactors, thus, converting them into bioelectrochemical systems (BES). Hydroxylation of 4-ethylbenzoic acid (EBA) catalyzed by the recombinant AaeUPO (rAaeUPO) was chosen as the model reaction. Total turnover number (TTN) up to 450,000 mol mol−1 and turnover frequency (TOF) up to 7.7 s − 1 were achieved using the AiO electrode system. The H2O2 productivity was identified as the limiting factor in the hydroxylation of EBA. However, with numbering-up or surface enlargement, this process could have great potential as an optimizable platform for H2O2-dependent enzymatic reactions.

Overexpression of the Capebp2 Gene Encoding the PEBP-like Protein Promotes the Cap Redifferentiation in Cyclocybe aegerita.

Phosphatidylethanolamine-binding protein (PEBP) is widely involved in various physiological behaviors, such as the transition from vegetative growth to reproductive growth in plants, tumorigenesis in the human, etc. However, few functional studies have examined pebp genes affecting the development of fungi. In this study, Capebp2 was cloned from Cyclocybe aegerita AC0007 strains based on the genome sequence and gene prediction, and the sequence alignment of CaPEBP2 with other PEBP proteins from other biological sources including plant, animal, fungi, and bacteria indicated that PEBP had low sequence similarity in fungi, whereas all protein sequences had some conserved motifs such as DPDAP and HRY. Expression analysis showed the transcription level of Capebp2 increased approximately 20-fold in fruiting bodies compared with mycelia. To uncover the function of Capebp2 in C. aegetita development, Capebp2 was cloned into a pATH vector driven by the actin promoter for obtaining overexpression transformant lines. Fruiting experiments showed the transformed strains overexpressing Capebp2 exhibited redifferentiation of the cap on their surface, including intact fruiting bodies or partial lamella during fruiting development stage, and the longitudinal section indicated that all regenerated bodies or lamella sprouted from the flesh and shared the epidermis with the mother fruiting bodies. In summary, the sequence characterization of Capebp2, expression level during different development stages, and function on fruiting body development were documented in this study, and these findings provided a reference to study the role of pebp in the development process of basidiomycetes. Importantly, gene mining of pebp, function characterization, and the regulating pathways involved need to be uncovered in further studies.

Toward Kilogram-Scale Peroxygenase-Catalyzed Oxyfunctionalization of Cyclohexane.

Mol-scale oxyfunctionalization of cyclohexane to cyclohexanol/cyclohexanone (KA-oil) using an unspecific peroxygenase is reported. Using AaeUPO from Agrocybe aegerita and simple H2O2 as an oxidant, cyclohexanol concentrations of more than 300 mM (>60% yield) at attractive productivities (157 mM h-1, approx. 15 g L-1 h-1) were achieved. Current limitations of the proposed biooxidation system have been identified paving the way for future improvements and implementation.

Comparison of two kinds of Agrocybe cylindracea polysaccharides: Structural characteristic and antitumor activity

Agrocybe cylindracea is one of the common sources of functional polysaccharides. But the structure and function of Agrocybe cylindracea polysaccharides obtained by different preparation methods are very different. A comprehensive and detailed study of its fine structure and systematic activity is helpful for its development and utilization. In this study, a cold water-souble polysaccharide (ACP) and an alcohol-soluble polysaccharide (ACAP) from Agrocybe cylindracea were extracted. Their physicochemical property, structural feature and antitumor activity in vitro were compared and investigated. The results showed that ACP (2840.9 kDa) was mainly composed of 1,6-linked-Glcp, 1,3,6-linked-Glcp, 1,6-linked-Galp and T-Glcp and inhibited MGC-803 cells proliferation via inducing tumor cells apoptosis by the cytochrome c/caspases pathway. However, ACAP (1823.9 Da) just contained glucose. By comparison, ACAP exhibited no inhibitory effect on MGC-803 cells in vitro. This work would provide an additional explanation for the structure-activity relationship of polysaccharides from Agrocybe cylindracea and provide a theoretical support for its application in biomedicine industries.

Heterologous production of basidiomycetous peptidases to decrease sodium chloride in food

In the past, the topic of excessive salt consumption associated with various chronic diseases became a focus of nutritional science. Enzymes such as hydrolases and specifically peptidases are widely used in industry since they do not require cofactors, are often stable at high pH values, and have a broad cleavage specificity. Thus, a new approach to use peptidases would be the generation of L‐arginyl dipeptides, which do not have a taste on their own, but are able to enhance the salty taste. This thesis should focus on the isolation and characterisation of arginyl‐specific peptidase genes from basidiomycetes to generate these dipeptides.First, various peptidase genes from Trametes versicolor, Phanerochaete chrysosporium and Schizophyllum commune were isolated and expressed in Escherichia coli or Komagataella phaffii. The serine (ABB73029) peptidase from P. chrysosporium and the aspartate peptidase from T. versicolor (EIW62808) were successfully purified. Activity assays showed that the aspartate peptidase from T. versicolor is not an arginyl‐specific peptidase, while the analysis of cleavage sites indicates that the serine peptidase from P. chrysosporium is an exopeptidase. Next, 29 basidiomycetes were cultivated over 24 days in minimal medium with 1 % gluten and analysed regarding their endo‐ and exopeptidase activity. Out of 29 fungi, five interesting species with arginyl‐specific peptidase activity were chosen, namely Agrocybe aegerita, Fomitop‐ sis pinicola, Flammulina velutipes, Hypholoma sublateritium and Pleurotus eryngii. After purification of the culture supernatant using size exclusion chromatography, active fractions were used to hydrolyse the substrate casein, and cleavage products were analysed. Cleavage sites were only detected for A. aegerita, F. velutipes and P. eryngii. Beside arginyl‐specific cleavage sites also a high amount of undesired exopeptidase cleavage sites was determined assuming that these peptidases are probably exopeptidases rather than arginyl‐specific peptidases.Generation of salt taste enhancing dipeptides should finally be achieved with a dipeptidyl‐ peptidase V (DPPV) from Pleurotusfloridanus. PflDPPV was heterologously produced in E. coli, had optima at pH 8.5 and 60 °C and was not completely inhibited by the tested peptidase inhibitors.

Die Enzymologie der C8‐Oxylipinbiosynthese

Based on transcriptomic, volatilomic and metabolomic data of the model organism Cyclocybe aegerita, the endogenous and enzymatic production of the C8‐oxylipins oct‐2‐en‐1‐ol, octan‐ 1‐ol, oct‐2‐enal, octanal, octan‐3‐ol, oct‐1‐en‐3‐ol, oct‐1‐en‐3‐one and octan‐3‐ one via biotransformations with [U‐13C]linoleic acid was compared to the developmental state depending gene expression patterns of putative oxylipin‐associated enzyme classes like lipoxygenases, dioxygenases, ene‐reductases and alcohol dehydrogenases. On closer consideration of these data sets, the C8‐oxylipin production seemed to occur cluster‐like.While one cluster (ketonic‐cycle) consisted of octan‐3‐ol, oct‐1‐en‐3‐one, octan‐3‐one and oct‐1‐en‐3‐ol, which were detected through out all developmental stages, a second cluster (aldehydic‐cycle) primarily active at early developmental stages involved oct‐2‐enal, octanal, oct‐2‐en‐1‐ol and octan‐1‐ol. Studying these clusters revealed that all C8‐oxylipins of the aldehydic‐cycle and ketonic‐cycle can be derived from oct‐2‐enal and oct‐1‐en‐3‐one, respectively. A detailled consideration of the gene expression patterns revealed several enzyme classes putatively involved in the C8‐oxylipin biosynthesis. Especially the two genes AAE3 04864 and AAE3 13549 encoding the lox4‐ and enr1‐gene showed great similarities between their strongly increasing transcript counts with maturation. Subsequent recombinant production of CaeLOX4 and CaeEnR1 revealed that CaeLOX4 showed a preference for C18‐ fatty acids with the highest affinity towards linoleic acid. Analysis of the reaction product revealed that this LOX selectively oxygenated linoleic acid at position 13 to 13‐ hydroperoxyoctadecadienoic acid (13‐HPOD). CaeEnR1 showed high affinities towards the reduction of oct‐1‐en‐3‐one and oct‐2‐enal to their saturated counterparts octan‐3‐one and octanal, respectively. Due to the strong similarities between the expression patterns of CaeLOX4 and CaeEnR1 a coherence between their main reaction products 13‐HPOD and the precursor of the ketonic‐cycle oct‐1‐en‐3‐one seems very likely. This suggests a biosynthetic route from linoleic acid to 13‐HPOD, followed by a subsequent cleavage step to oct‐1‐en‐3‐ one and therefore to C8‐oxylipins of the ketonic‐cycle.

AANL6 is a new efficient tool to probe non-reducing N-acetylglucosamine of N-linked glycans

Terminal N-acetylglucosamine (GlcNAc) N-linked glycosylation is a truncated N-glycosylated modification that has been reported to be involved in various diseases, such as autoimmune diseases, cancers, and neurodegenerative diseases. New and simple tools will be always valuable for further characterization of the functions of this kind of glycosylation. Our previous paper proved that an optimized lectin created from Agrocybe aegerita GlcNAc selective lectin (AANL) named AANL6, can effectively identify O-GlcNAcylation, which is terminal GlcNAc O-linked glycosylation. We speculated that AANL6 could also be used to identify terminal GlcNAc N-linked glycosylation. Using therapeutic monoclonal antibodies as a model of terminal GlcNAc N-glycosylated proteins, we proved that AANL6 could selectively identify terminal GlcNAc N-linked glycosylation. The ratio of terminal GlcNAc N-linked glycosylation was increased by enrichment with AANL6 in human serum. Using cell membrane proteins as a complex sample, we found that AANL6 bound to the sperm surface, which expresses abundant terminal GlcNAc N-glycans, but did not bind to some tumor cell surfaces such A549 and MCF-7 cells, which is rich in high mannose glycoforms. In conclusion, AANL6 was identified as a powerful tool to probe terminal GlcNAc N-linked glycosylation and would be valuable for uncovering the function of this glycosylation.

Valorization of bioethanol by-products to produce unspecific peroxygenase with Agrocybe aegerita: Technological and proteomic perspectives

Unspecific peroxygenase (UPO) presents a wide range of biotechnological applications. This study targets the use of by-products from bioethanol synthesis to produce UPO by Agrocybe aegerita. Solid-state and submerged fermentations (SSF and SmF) were evaluated, achieving the highest titers of UPO and laccase in SmF using vinasse as nutrients source. Optimized UPO production of 331 U/L was achieved in 50% (v:v) vinasse with an inoculum grown for 14 days. These conditions were scaled-up to a 4 L reactor, achieving a UPO activity of 265 U/L. Fungal proteome expression was analyzed before and after UPO activity appeared by shotgun mass spectrometry proteomics. Laccase, dye-decolorizing peroxidases (DyP), lectins and proteins involved in reactive oxygen species (ROS) production and control were detected (in addition to UPO). Interestingly, the metabolism of complex sugars and nitrogen sources had a different activity at the beginning and end of the submerged fermentation.

Peroxygenase-Catalysed Selective Oxidation of Silanes to Silanols.

A peroxygenase-catalysed hydroxylation of organosilanes is reported. The recombinant peroxygenase from Agrocybe aegerita (AaeUPO) enabled efficient conversion of a broad range of silane starting materials in attractive productivities (up to 300 mM h-1), catalyst performance (up to 84 s-1 and more than 120,000 catalytic turnovers). Molecular modelling of the enzyme-substrate interaction puts a basis for the mechanistic understanding of AaeUPO selectivity.

Peroxygenase‐Catalysed Selective Oxidation of Silanes to Silanols

AbstractA peroxygenase‐catalysed hydroxylation of organosilanes is reported. The recombinant peroxygenase from Agrocybe aegerita (AaeUPO) enabled efficient conversion of a broad range of silane starting materials in attractive productivities (up to 300 mM h−1), catalyst performance (up to 84 s−1 and more than 120 000 catalytic turnovers). Molecular modelling of the enzyme‐substrate interaction puts a basis for the mechanistic understanding of AaeUPO selectivity.

Chemo‐Enzymatic One‐Pot Oxidation of Cyclohexane via in‐situ H2O2 Production over Supported AuPdPt Catalysts

AbstractThe introduction of dopant concentrations of Pt into supported AuPd nanoparticles, when used in conjunction with an evolved unspecific peroxygenase (UPO) from Agrocybe aegerita (PaDa‐I) is demonstrated to offer high efficacy towards the one‐pot selective oxidation of cyclohexane to KA oil (cyclohexanol and cyclohexanone), via the in‐situ synthesis of H2O2. The optimised AuPdPt/TiO2/PaDa‐I system achieves significant improvements over analogous AuPd or Pd‐only formulations or the use of commercially available H2O2, with this attributed to the increased rate of H2O2 production by the chemo‐catalyst, which results from the electronic modification of Pd species via Pt incorporation, upon the formation of trimetallic nanoalloys.

Study on nutritional components evaluation and origin differences of Agrocybe cylindracea from different regions

AbstractThe aim of this study is to investigate the nutrients of Agrocybe cylindracea from different regions and discover the law of its nutritional value. In this study, 43 A. cylindracea samples were collected from 13 provinces in China. The differences in nutritional components of this A. cylindracea were explored. The main nutritional components, including protein, dietary fiber, amino acids, fat, ash, and moisture, were evaluated. Further factor and cluster analysis were applied to analyze these nutrients. The results showed A. cylindracea had high protein (14.09–27.24 g/100 g dw) and insoluble dietary fiber (25.27–34.17 g/100 g dw), but low soluble dietary fiber (0.49–3.19 g/100 g dw) and fat (1.69–3.45 g/100 g dw). Seventeen amino acids were detected from these mushrooms, of which glutamic (18.24–46.78 g/100 g dw) was the most abundant. Based on Eta analysis, factor analysis, and cluster analysis, it was found that the comprehensive nutritional value of A. cylindracea was related to the factors of both the geographical origin and maturity stage. In addition, geographical origin had a greater influence on the nutritional value of A. cylindracea than the maturity stage. This study will serve as a useful reference for the selection of the consumption and further processing directions of A. cylindracea.

A Comparative Analysis of Biodegradation and Bioconversion of Lentinula edodes and Other Exotic Mushrooms.

Mushrooms are capable of bioconverting organic residues into food. Understanding the relationship between high-quality yields and substrate biomass from these residues is critical for mushroom farms when choosing new strains. The objective of this exploratory study was, therefore, to analyze whether exotic mushrooms, namely, Pleurotus eryngii, Flammulina velutipes, and Agrocybe aegerita, could biologically convert the substrate into edible mushrooms as effectively as Lentinula edodes (baseline). Five experiments were carried out. Biological efficiency, biodegradability coefficient, mass balance and chemical characterization of the substrate were evaluated. Strategically hydrating the sawdust enabled L. edodes to achieve the greatest biodegradability and biological efficiency of 0.5 and 94.2 kg dt-1, respectively. The values for L. edodes on wheat straw without hydration were 0.2 and 68.8 kg dt-1, respectively. From 1000 kg of fresh substrate, P. eryngii produced 150.1 kg of edible mushrooms, making it technically competitive with L. edodes on wheat straw (195.9 kg). Hence, P. eryngii was the most reliable option for scaling among the exotic mushrooms. The analytical insights from our study provide further knowledge to advance the field's prominence in high-throughput mushroom-producing systems, particularly for exotic mushrooms.

Hybrid Organometallic and Enzymatic Tandem Catalysis for Oxyfunctionalisation Reactions

AbstractUnspecific peroxygenases (UPOs) are promising biocatalysts for oxyfunctionalisation reactions, owing to their simplicity of handling, stability and robustness. A limitation of using UPOs on a large scale is their deactivation in the presence of even rather modest concentrations of H2O2, requiring a constant and controlled supply of low amount of H2O2. Herein, we report an organometallic complex [Cp*Ir(pica)NO3] {pica=picolinamidate=κ2‐pyridine‐2‐carboxamide ion (−1)} 1 capable of efficiently regenerating FMNH2 from FMN (TOF=350 h−1, 298 K), driven by NaHCOO; FMNH2, in turn, spontaneously reacts with O2 leading to H2O2. After having studied the compatibility of 1 with the UPO from Agrocybe aegerita (rAaeUPO PaDa‐I) and individuated the best experimental conditions, we applied such a hybrid catalytic tandem in some hydroxylation, epoxidation and sulfoxidation reactions. Best performances were obtained by using a 1/rAaeUPO molar ratio of 50. TONs for the biocatalyst of up to 18933 were obtained for the transformation of ethylbenzene derivatives into (R)‐1‐phenylethanols (ee>99 %). 1/rAaeUPO was found to oxidise also cis‐methyl styrene (TON=13488), leading exclusively (1R,2S)‐cis‐methyl styrene oxide (ee>99 %), cyclohexane (TON=1634) and thioanisole (TON=1369).

Lessons on fruiting body morphogenesis from genomes and transcriptomes of Agaricomycetes.

Fruiting bodies (sporocarps, sporophores or basidiomata) of mushroom-forming fungi (Agaricomycetes) are among the most complex structures produced by fungi. Unlike vegetative hyphae, fruiting bodies grow determinately and follow a genetically encoded developmental program that orchestrates their growth, tissue differentiation and sexual sporulation. In spite of more than a century of research, our understanding of the molecular details of fruiting body morphogenesis is still limited and a general synthesis on the genetics of this complex process is lacking. In this paper, we aim at a comprehensive identification of conserved genes related to fruiting body morphogenesis and distil novel functional hypotheses for functionally poorly characterised ones. As a result of this analysis, we report 921 conserved developmentally expressed gene families, only a few dozens of which have previously been reported to be involved in fruiting body development. Based on literature data, conserved expression patterns and functional annotations, we provide hypotheses on the potential role of these gene families in fruiting body development, yielding the most complete description of molecular processes in fruiting body morphogenesis to date. We discuss genes related to the initiation of fruiting, differentiation, growth, cell surface and cell wall, defence, transcriptional regulation as well as signal transduction. Based on these data we derive a general model of fruiting body development, which includes an early, proliferative phase that is mostly concerned with laying out the mushroom body plan (via cell division and differentiation), and a second phase of growth via cell expansion as well as meiotic events and sporulation. Altogether, our discussions cover 1 480 genes of Coprinopsis cinerea, and their orthologs in Agaricus bisporus, Cyclocybe aegerita, Armillaria ostoyae, Auriculariopsis ampla, Laccaria bicolor, Lentinula edodes, Lentinus tigrinus, Mycena kentingensis, Phanerochaete chrysosporium, Pleurotus ostreatus, and Schizophyllum commune, providing functional hypotheses for ~10 % of genes in the genomes of these species. Although experimental evidence for the role of these genes will need to be established in the future, our data provide a roadmap for guiding functional analyses of fruiting related genes in the Agaricomycetes. We anticipate that the gene compendium presented here, combined with developments in functional genomics approaches will contribute to uncovering the genetic bases of one of the most spectacular multicellular developmental processes in fungi. Citation: Nagy LG, Vonk PJ, Künzler M, Földi C, Virágh M, Ohm RA, Hennicke F, Bálint B, Csernetics Á, Hegedüs B, Hou Z, Liu XB, Nan S, M. Pareek M, Sahu N, Szathmári B, Varga T, Wu W, Yang X, Merényi Z (2023). Lessons on fruiting body morphogenesis from genomes and transcriptomes of Agaricomycetes. Studies in Mycology 104: 1-85. doi: 10.3114/sim.2022.104.01.