Sequencing, assembly, and genomic annotation of Leucoagaricus gongylophorus LEU18496, a dikarya mutualistic species.

The basidiomycete fungus Leucoagaricus gongylophorus is able to grow in the fungus garden of leaf-cutter ants. This mutualistic interaction has driven the evolutionary adaptation of L. gongylophorus, shaping its metabolism to produce enzymes adept at lignocellulosic biomass degradation. In this study, we undertook the comprehensive sequencing, assembly, and functional annotation of the genome of L. gongylophorus strain LEU18496, mutualistic fungus of the Atta mexicana. Our genomic analyses revealed a distinctive bimodal nature to the genome: a predominant region characterized by AT enrichment and low genetic density, alongside a smaller region exhibiting higher GC content and higher genetic density. The presence of transposable elements (TEs) within the AT-enriched region suggests genomic compartmentalization, facilitating differential evolutionary rates. With a gene count of 6748, the assembled genome of L. gongylophorus LEU18496 surpasses previous reports for this fungal species. Inspection of genes associated with central metabolism unveiled a remarkable abundance of carbohydrate-active enzymes (CAZymes) and fungal oxidative lignin enzymes (FOLymes), underscoring their pivotal roles in the life cycle of this fungus.

Unroughing the cat's tongue mushrooms: Four new species of Pseudohydnum from Brazil based on morphological and molecular phylogenetic evidence.

Pseudohydnum, commonly known as cat's tongue mushrooms, is a monophyletic assemblage within Auriculariales, which encompasses species with gelatinous basidiomata, spathulate, flabellate, or shell-shaped pileus, hydnoid hymenophore, globose to ellipsoidal basidiospores, and longitudinally cruciate-septate basidia. According to the available literature, 16 species have been described in Pseudohydnum, mostly represented in temperate-boreal forests of the Northern Hemisphere. However, the limited morphological, molecular, and ecological information, especially from the Southern Hemisphere ecosystems, does not presently allow a reliable assessment of its taxonomic boundaries nor provide a complete picture of the species diversity in the genus. In an ongoing effort to examine specimens collected in dense and mixed ombrophilous forest fragments (Atlantic Rainforest domain) from Southeastern and Southern Brazil, additional taxa assigned to Pseudohydnum were identified. Four new species are recognized based mostly on characters of the pileus surface, stipe, hymenium, and basidiospores. Molecular phylogenetic analyses based on nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS barcode), partial nuc rDNA 28S, and partial RNA polymerase II largest subunit (RPB1) sequences supported the description of these new taxa. Here, we propose Pseudohydnum brasiliense, P. brunneovelutinum, P. cupulisnymphae, and P. viridimontanum as new species. Morphological descriptions, line drawings, habitat photos, and comparisons with closely related taxa are provided. A dichotomous key for identification of currently known Southern Hemisphere Pseudohydnum species is presented.

Evaluation of nutritional, technological, oxidative, and sensory properties of low-sodium and phosphate-free mortadellas produced with bamboo fiber, pea protein, and mushroom powder.

This study examined the effects of replacing alkaline phosphate (AP) with bamboo fiber (BF), isolated pea protein (PP), and mushroom powder (MP) on the nutritional, technological, oxidative, and sensory characteristics of low-sodium mortadellas. Results indicated that this reformulation maintained the nutritional quality of the products. Natural substitutes were more effective than AP in reducing water and fat exudation. This led to decreased texture profile analysis (TPA) values such as hardness, cohesiveness, gumminess, and chewiness. The reformulation reduced the L* values and increased the b* values, leading to color modifications rated from noticeable to appreciable according to the National Bureau of Standards (NBS) index. Despite minor changes in oxidative stability indicated by increased values in TBARS (from 0.19 to 0.33 mg MDA/kg), carbonyls (from 2.1 to 4.4 nmol carbonyl/mg protein), and the volatile compound profile, the sensory profile revealed a beneficial increase in salty taste, especially due to the inclusion of MP, which was enhanced by the synergy with BF and PP. In summary, the results confirmed the potential of natural alternatives to replace chemical additives in meat products. Incorporating natural antioxidants into future formulations could address the minor oxidation issues observed and enhance the applicability of this reformulation strategy.

Psilocybin fungi microdose treatment in major depressive disorder: a case report / Tratamiento con microdosis de hongos con psilocibina en trastorno depresivo mayor: reporte de un caso.

Major depression disorder is an entity with high prevalence and worldwide impact. Current treatments present a non-response rate of 15-30%, and certain adverse effects are seen like apathy syndrome and lack of emotional response. It is stated that the treatment with psilocybin fungi allows the possibility of dose reduction and suspension of classic psychotropic drugs and entails changes on an emotional and behavioral level that result benefic in patients with major depressive syndrome. We present a case of a 19 years old patient with major depressive syndrome diagnosis. Accompaniment and patient advice was made appealing to the right of autonomy, on the psilocybin microdose self-administration process, aiming to reducing health risks and potentiate probable beneficial effects, with weekly evaluations, for a period of 7 months; using clinical anamnesis, laboratory tests and the Hamilton depression scale. As a result of this intervention, a symptomatic complete remission was proven, alongside with the suspension of conventional pharmacological treatment without discontinuation symptoms and improvements at the communicational level, social interaction and general well-being. These findings support the idea that psilocybin microdose treatments are promising tools in depression treatments. Scientific studies are needed in order to certify these findings.

La depresión mayor es una enfermedad de gran prevalencia e impacto mundial. Los tratamientos actuales presentan una tasa de no respuesta del 15 al 30 %, mientras que en casos de eficacia se suelen observar efectos adversos como el síndrome de apatía y la falta de respuesta emocional. Se postula que el tratamiento con hongos psilocibios genera la posibilidad de reducción de dosis y suspensión de psicofármacos clásicos y ocasiona cambios a nivel emocional y comportamental benéficos en pacientes con trastorno depresivo mayor. Este es un caso de un paciente no binario de 19 años de edad con diagnóstico de trastorno depresivo mayor. Se realizó unacompañamiento y asesoramiento del paciente apelando al derecho de autonomía, en el proceso de autoadministración de microdosis de psilocibina, para disminución de riesgos en salud y potenciar efectos benéficos probables, con evaluación semanal, durante un periodo de 7 meses; utilizando la anamnesis clínica, análisis de laboratorio y la escala validada de depresión de Hamilton. Como resultado de esta intervención se evidenció una remisión completa sintomática, la suspensión del tratamiento farmacológico convencional, sin síntomas de discontinuación y mejorías a nivel comunicacional, de interacción social y bienestar general. Estos hallazgos apoyan la idea de que los tratamientos con microdosis de psilocibina son una herramienta prometedora en los tratamientos de depresión. Se necesitan más estudios que aporten evidencia científica para comprobar dichos hallazgos.

New Cortinariaceae species associated with Dicymbe, Aldina, and Pakaraimaea in Guyana.

Species of the ectomycorrhizal (ECM) family Cortinariaceae (Agaricales, Agaricomycetes, Basidiomycota) have long been considered impoverished or absent from lowland tropical rainforests. Several decades of collecting in forests dominated by ECM trees in South America's Guiana Shield is countering this view, with discovery of numerous Cortinariaceae species. To date, ~12 morphospecies of this family have been found in the central Pakaraima Mountains of Guyana. Here, we describe three of these as new species of Cortinarius and two as new species of Phlegmacium from forests dominated by the ECM tree genera Dicymbe (Fabaceae subfam. Detarioideae), Aldina (Fabaceae subfam. Papilionoideae), and Pakaraimaea (Cistaceae). Macromorphological, micromorphological, habitat, and DNA sequence data are provided for each new species.

A cascade approach to sustainable agriculture: From mushroom mycelium to lettuce harvest.

Sustainable agriculture involves adopting best practices in food production to promote environmental and economic sustainability. Its implementation primarily aims to utilise organic residues to increase yield, diversify production, and reduce costs. In this context, the objective of this study was to investigate different substrates for Hypsizygus ulmarius production and, from its residual substrate, to develop formulations for lettuce seedling growth and subsequent greenhouse cultivation. For mushroom production, substrates were prepared from sawdust with the addition of wheat bran, rice bran, soybean meal, and calcite, resulting in four distinct substrate formulations. The spent mushroom substrate (SMS), obtained at the end of cultivation, was used for lettuce seedling production along with the commercial substrate Carolina Soil® and the soil conditioner BacSol®. The top five formulations were selected for transplanting in the greenhouse. Regarding mushroom production, substrates with higher carbon/nitrogen ratios, around 66: 1, resulted in higher yields. For seedling production, SMS showed lower efficiency compared to the commercial substrate Carolina Soil®, which also benefited from the addition of the soil conditioner BacSol®. However, after transplanting lettuce seedlings, the formulation containing SMS showed superior results in almost all evaluated parameters. Therefore, we concluded that despite the inefficiency of using H.ulmarius SMS for lettuce seedling production, it favours the establishment of seedlings in greenhouse cultivation environments.

Anodic stripping voltammetry of arsenic determination with edible mushroom-nafion-modified glassy carbon electrode.

An edible Mushroom-Nafion modified glassy carbon electrode (M2N5-GCE) was prepared using a homogeneous mixture varying the concentrations of these, in addition to the origin of the mushroom (Shiitake, Lentinula edodes, M1 and Abrantes, Agariscus bisporus, M2) and applied to the As(III) determination by anodic stripping voltammetry. After choosing the optimal conditions in the preparation of the electrode, the second stage was to study the effects of various parameters such as supporting electrolyte, pH, accumulation potential, and time (Eacc, tacc). The optimum experimental conditions chosen were Britton Robinson buffer 0.01 mol L-1 pH:4.6; Eacc: -1.0 and tacc: 60 s obtaining a signal of oxidation of As(0) to As(III) about 0.08 V. Peak current was proportional to arsenic concentration over the 19.6-117.6 µg L-1 range, with a 3σ detection limit of 13.4 µg L-1. The method was validated using As(III) spiked tap water from the laboratory with satisfactory results (RE:3.0 %). Finally, the method was applied to the determination of As(III) in water samples from the Loa River (Northern Chile) in the presence of As(V) in a concentration >20 times higher (RE: 2.3 %).

Enhancing biodegradation of vegetable oil-contaminated soil with soybean texturized waste, spent mushroom substrate, and stabilized poultry litter in microcosm systems.

Industrial activities contribute to environmental pollution, particularly through unregulated effluent discharges, causing adverse effects on ecosystems. Vegetable oils, as insoluble substances, exacerbate this pollution, forming impermeable films and affecting the oxygen transfer, leading to serious habitat disruption. Organic wastes, such as soybean texturized waste, spent mushroom substrate, and stabilized poultry litter, were assessed for their efficacy in enhancing the degradation of vegetable oil in contaminated soil. For this purpose, contaminated soil was amended with each of the wastes (10% w/w) using microcosm systems, which were monitored physico-chemically, microbiologically and toxicologically. Results indicate that the wastes promoted significant oil degradation, achieving 83.1, 90.7, and 86.2% removal for soybean texturized waste, spent mushroom substrate, and stabilized poultry litter, respectively, within a 90-day period. Additionally, they positively influenced soil microbial activity, as evidenced by increased levels of culturable microorganisms and hydrolytic microbial activity. While bioassays indicated no phytotoxicity in most cases, soybean texturized waste exhibited inhibitory effects on seed germination and root elongation of Lactuca sativa. This study significantly enhances our comprehension of remediation techniques for sites tainted with vegetable oils, highlighting the critical role of organic waste as eco-friendly agents in soil restoration. Emphasizing the practical implications of these findings is imperative to underscore the relevance and urgency of addressing vegetable oil contamination in soil. Moving forward, tailored strategies considering both contaminant characteristics and soil ecosystem traits are vital for ensuring effective and sustainable soil remediation.

Microencapsulation of apricot kernel oil: Utilization of mushroom by-product as an emulsifier in oil-in-water emulsion.

The present study investigated mushroom by-products as a substitute for emulsifiers in the microencapsulation of apricot kernel oil. Mushroom by-product emulsions were more viscous and had higher centrifugal (85.88±1.19 %) and kinetic (90.52±0.98 %) stability than control emulsions (Tween 20 was used as emulsifier). Additionally, spray-drying mushroom by-product emulsions yielded a high product yield (62.56±1.11 %). Furthermore, the oxidative stability of powder products containing mushroom by-products was observed to be higher than that of the control samples. For an accelerated oxidation test, the samples were kept at various temperatures (20, 37, and 60 °C). TOTOX values were assessed as indicators of oxidation, with values exceeding 30 indicating oxidation of the samples. Of the samples stored at 60 °C, the non-microencapsulated apricot kernel oil oxidized by the fifth day (41.12±0.13 TOTOX value), whereas the powder samples containing the mushroom by-products remained unoxidized until the end of the tenth day (37.05±0.08 TOTOX value). This study revealed that mushroom by-products could be a viable alternative for synthetic emulsifiers in the microencapsulation of apricot kernel oil. It has been observed that using mushroom by-products instead of synthetic emulsifiers in oil microencapsulation can also delay oxidative degradation in microencapsulated powders.

Mushrooms and Fungi and Their Biological Compounds with Antidiabetic Activity: A Review.

Mushrooms have been used by humans for centuries as food and medicine because they have been shown to affect certain diseases. Mushrooms for medicinal purposes have been consumed in the form of extracts and/or biomass of the mycelium or fruiting body. The beneficial health effects of mushrooms are due to their content of bioactive compounds (polysaccharides, proteins, ergosterol, lectins, etc.). On the other hand, diabetes is one of the metabolic diseases that affects the population worldwide, characterized by hyperglycemia that involves a defective metabolism of insulin, a hormone secreted by ß cells and that mainly stimulates glucose absorption by the cells. However, it also affects the metabolism of carbohydrates, fats and proteins; poor control of this disease leads to serious damage to eyesight, kidneys, bones, heart, skin, blood vessels, nerves, etc. It has been reported that the consumption of some mushrooms helps control and treat diabetes, since among other actions, they promote the secretion of insulin by the pancreas, help reduce blood glucose and have α-glucosidase inhibitory activity which improves glucose uptake by cells, which are effects that prescription medications have for patients with diabetes. In that sense, this manuscript shows a review of scientific studies that support the abilities of some mushrooms to be used in the control and/or treatment of diabetes.

Revealing the 1H-NMR Profiling of Six Edible Mushrooms Consumed in the Northeastern Highlands of Puebla, Mexico.

The 1H-NMR metabolomics profiling of six edible mushrooms consumed in the northeastern highlands of Puebla, Mexico is presented. These fungi were morpho- and molecularly identified as Infundibulicybe squamulosa, Amanita jacksonii, Lepista nuda, Russula delica, Russula brevipes, and Lactarius indigo. The chemical profiling confirmed the presence of eight essential amino acids and their derivatives, six organic acids, six nucleosides, low amounts of reducing sugars, and valuable nutraceuticals such as betaine, carnitine, glycero-3-phosphocholine and O-acetylcarnitine which were differentially determined and quantified in the six mushrooms by qNMR. Principal component analysis (PCA) and orthogonal projections to latent structures discriminant analysis (OPLS-DA) generated four different groups. Two of these groups were constituted by fungal species with phylogenic relationships whereas non-phylogenetic related species were separated from each other. The potential use of 1H-NMR metabolomics and chemometrics to group macromycetes and determine the nutritional and nutraceutical potential of these local foods is demonstrated.

Linking the protease activity to the nematicidal action of edible mushroom.

Biological control using edible mushrooms as natural enemies is a sustainable alternative for pest management. Despite the well-established literature on toxins and secondary metabolites produced by these fungi in the biochemical control of nematodes, the nematicidal activity of proteases from different Pleurotus species is yet to be investigated. Therefore, this study aimed to correlate protease to the nematicidal activity of different mushrooms, Pleurotus sp., P. ostreatus (SB), P. ostreatus (Pearl), and P. djamor. For such a purpose, we performed motility assays of Panagrellus sp. at different time intervals, 6, 12, and 24 h for each of the mushrooms. In addition, the protease activity was measured using different pH (5, 7, and 9) and fermentation time intervals (45 and 75 days). Furthermore, we also evaluated the effect of this cell-free extract on Panagrellus sp. In response to these experiments, all edible mushrooms showed a reduction over 82% for the nematode-feeding activity (p < 0.01). The cell-free crude extract of each of the fungi studied showed nematocidal activity (p < 0.01). For the 45-day fermentation, P. djamor exhibited statistical significance (p < 0.01) compared with the others, reaching a reduction percentage of 73%. For the 75-day fermentation, Pleurotus sp. and P. ostreatus (Pearl) showed significant differences compared with the other fungi (p < 0.01), with reduction percentages of 64 and 62%, respectively. Herein, protease activity was associated with the nematicidal action of different Pleurotus species in controlling Panagrellus sp.

Variation in traditional knowledge of culturally important macromycete species among three indigenous communities of Oaxaca, Mexico.

BACKGROUND: For centuries, wild mushrooms have been a forest resource of significant cultural value in several ethnic groups worldwide. In Mexico, extensive traditional knowledge on the use of fungal resources has been developed and deeply rooted. Mexico is the second country in the world in which the most species of wild mushroom are consumed, and it is considered a pioneer in ethnomycology. Nonetheless, there are still many indigenous groups in this country that have not been studied from an ethnomycological approach. The present study aimed to record the traditional knowledge on wild mushrooms in three indigenous groups of the state of Oaxaca, Mexico, and assess the variation in this knowledge within and across the studied groups. METHODS: The data were recorded from April to October 2022 within three communities belonging to the indigenous groups Chatino, Chontal, and Chinanteco. Through 84 interviews, information related to their knowledge of wild mushrooms was obtained. The cultural significance index of wild edible mushrooms was calculated for each community. Regression analyses, analysis of variance and covariance, t test, and non-metric multidimensional scaling analysis were performed to assess the distribution of traditional knowledge in the communities. RESULTS: A total of 32 culturally important mushroom species were recorded for the three indigenous groups (30 edible, 2 medicinal); 23 used by Chatinos, 16 by Chontales, and 6 by Chinantecos. Only Chatinos and Chinantecos use wild mushrooms in medicine. The cultural significance of wild edible mushrooms differed among groups. Traditional knowledge about wild mushrooms declines when the level of schooling increases and age decreases, especially in the Chatino group. This knowledge distributes more homogeneously in the Chontal and Chinanteco groups. Their age determines the difference in knowledge between men and women. CONCLUSION: Documenting how traditional knowledge differs among ethnic groups is relevant for preserving cultural and biological diversity. Factors such as level of schooling and age can affect traditional knowledge of wild mushrooms, but the effects of these factors vary within and across communities. Conducting studies encompassing a broader range of variables is of interest for a better understanding of the human-mushroom relationship.

Crude Extracts of Pleurotus Spp. and the Presence of Their Proteins in the Ovicidal and Larvicidal Activity of Haemonchus contortus.

Mushrooms of the genus Pleurotus have shown nematophagous activity as it produces many chemical compounds and enzymes affecting parasitic nematodes. This study aimed to extract the inhibitory activity of the five strains of the fungus Pleurotus spp. It was evaluated against eggs and larvae of Haemonchus contortus. The extract of P. ostreatus obtained the highest level of inhibition of eggs at 97.6% (1341 µg/mL) followed by P. pulmonarius (EPP) at 81.2% (774 µg/mL). The extract selected for evaluation against larvae was P. pulmonarius, showing no effect for L3 larvae, but for L4 larvae an immobility effect of 56.93% was observed at 900 µg/mL. The protein profile showed the presence of 23 protein bands in the extract. The crude extract of P. pulmonarius showed degradation of tissues both inside the eggs and larvae L1. Metabolites produced by Pleurotus mushrooms can consider using in agriculture sustainable by utilizing in producing of ovicidal and larvicidal against H. contortus instead of chemical compounds.

Ectomycorrhizal fungal communities in natural and urban ecosystems: Quercus humboldtii as a study case in the tropical Andes.

Worldwide urban landscapes are expanding because of the growing human population. Urban ecosystems serve as habitats to highly diverse communities. However, studies focusing on the diversity and structure of ectomycorrhizal communities are uncommon in this habitat. In Colombia, Quercus humboldtii Bonpl. is an ectomycorrhizal tree thriving in tropical montane forests hosting a high diversity of ectomycorrhizal fungi. Q. humboldtii is planted as an urban tree in Bogotá (Colombia). We studied how root-associated fungal communities of this tree change between natural and urban areas. Using Illumina sequencing, we amplified the ITS1 region and analyzed the resulting data using both OTUs and Amplicon Sequence Variants (ASVs) bioinformatics pipelines. The results obtained using both pipelines showed no substantial differences between OTUs and ASVs for the community patterns of root-associated fungi, and only differences in species richness were observed. We found no significant differences in the species richness between urban and rural sites based on Fisher's alpha or species-accumulation curves. However, we found significant differences in the community composition of fungi present in the roots of rural and urban trees with rural communities being dominated by Russula and Lactarius and urban communities by Scleroderma, Hydnangium, and Trechispora, suggesting a high impact of urban disturbances on ectomycorrhizal fungal communities. Our results highlight the importance of urban trees as reservoirs of fungal diversity and the potential impact of urban conditions on favoring fungal species adapted to more disturbed ecosystems.

The perception of fungi among Karajá indigenous children and adolescents from Brazil.

BACKGROUND: Environmental perception involves the interpretation and interaction of individuals with their surroundings, influenced by cultural, social, and individual factors. Analyzing the environmental perception of children and adolescents contributes to fostering awareness and ethical behavior toward the environment. Indigenous communities, such as the Karajá from Brazil, possess significant environmental knowledge due to their connection with nature, providing distinctive insights into biodiversity and natural interconnections. In this study, the perception of fungi among Karajá indigenous children and adolescents was investigated. METHODS: The study involved 229 elementary school students from the Macaúba, Fontoura and Santa Isabel do Morro communities, located on Bananal Island, Brazil. Students were encouraged to draw what they knew about fungi and answer where they learned about fungi and what name they give to these organisms. The drawings were analyzed considering seven categories. RESULTS: The term most used to refer to fungi was hedoro(u) (56%), followed by fungus (21%) and mushroom (11%). Most students said they had learned about fungi in nature (38%) and at school (36%). The most represented organisms were in fact fungi (93%), mainly being portrayed in nature (94%). Most participants did not attribute any ecological function to fungi (83%), although 16% of them recognized fungi as decomposers and 1% as phytopathogenic agents. Negative aspects, particularly food contamination, were more frequently represented (13%) than positive aspects (4%). The drawings identified two morphological types: mushrooms (87%) and mold (13%). Among these mushrooms, 68% possibly represent the Amanita muscaria species. CONCLUSIONS: Although the children and adolescents showed that they noticed the fungi around them, the group's concept and understanding were limited to the figure of the mushroom and the negative aspects related to food contamination. Strong association of the fungi with the A. muscaria is noteworthy, since it does not occur in the environment in which the participants live, suggesting that external stimuli, such as TV or the internet, can influence their perception more than the nature they are exposed to.

Production of Escovopsis conidia and the potential use of this parasitic fungus as a biological control agent of leaf-cutting ant fungus gardens.

This study was carried out to investigate the use of different substrates for the production of Escovopsis conidia and verify the virulence of four different isolates cultured on four types of substrates using a novel bioassay. Escovopsis isolates were molecularly identified, based on Internal Transcribed Spacer (ITS) nucleotide sequences. To evaluate conidial production, suspensions (1 × 106 conidia mL-1) of each Escovopsis isolate were inoculated onto four substrates (parboiled rice, white rice, rolled oats, and corn grits). After 14 days, conidial yields were assessed. The virulence of each isolate cultured on the four substrates was tested against Leucoagaricus fungus garden fragments, by directly applying 500 µL of each conidial suspension (1 × 107 conidia mL-1), and the development of the parasite was monitored daily until it completely colonized the fungus garden. It was observed that rolled oats were the best substrate for conidial production, with a yield of 1.7 × 107 to 2.0 × 108 conidia mL-1. Furthermore, isolate AT-01 produced the highest number of conidia when compared with the other isolates. Regardless of the substrate used to produce AT-01 conidia, this isolate completely colonized the fungus garden 6 days post inoculation (dpi), followed by AT-02, AC-01, and AC-2. High levels of both conidial production and virulence against the leaf-cutting ant fungus garden were observed here.

The effect of casing and gypsum on the yield and psychoactive tryptamine content of Psilocybe cubensis (Earle) Singer.

Psychedelic fungi have experienced a surge in interest in recent years. Most notably, the fungal secondary metabolite psilocybin has shown tremendous promise in the treatment of various psychiatric disorders. The mushroom species that produce this molecule are poorly understood. Here we sought to examine for the first time, the response of a psilocybin-producing species Psilocybe cubensis to casing (peat moss and vermiculite) and supplementation with gypsum (calcium sulfate dihydrate), two common practices in commercial mushroom cultivation. Mycelial samples of genetically authenticated P. cubensis were used to inoculate popcorn grain bags. The fully colonized bags of popcorn grain (0.15 kg) were transferred to bins of 0.85 kg pasteurized horse manure, with or without 1 cm thick layer of casing and/or 5 % gypsum. Our results indicate that the use of a casing layer significantly increases the biological efficiency (161.5 %), by approximately four fold, in comparison to control (40.5 %), albeit with a slight delay (∼2 days) for obtaining fruiting bodies and a somewhat reduced total tryptamine content (0.85 %) as gauged by High Performance Liquid Chromatography measurements. Supplementation with both casing and gypsum, however, appears to promote maximal yields (896.6 g/kg of dried substrate), with a biological efficiency of 89.6 %, while also maintaining high total tryptamine expressions (0.95 %). These findings, revealing methods for maximizing yield of harvest and expressions of psychoactive tryptamines, may prove useful for both home growers and commercial cultivators of this species, and ultimately support the growth of a robust industry with high quality natural products.

The selenium-independent phospholipid hydroperoxide glutathione peroxidase from Theobroma cacao (TcPHGPX) protects plant cells against damages and cell death.

Proteins from the glutathione peroxidase (GPX) family, such as GPX4 or PHGPX in animals, are extensively studied for their antioxidant functions and apoptosis inhibition. GPXs can be selenium-independent or selenium-dependent, with selenium acting as a potential cofactor for GPX activity. However, the relationship of plant GPXs to these functions remains unclear. Recent research indicated an upregulation of Theobroma cacao phospholipid hydroperoxide glutathione peroxidase gene (TcPHGPX) expression during early witches' broom disease stages, suggesting the use of antioxidant mechanisms as a plant defense strategy to reduce disease progression. Witches' broom disease, caused by the hemibiotrophic fungus Moniliophthora perniciosa, induces cell death through elicitors like MpNEP2 in advanced infection stages. In this context, in silico and in vitro analyses of TcPHGPX's physicochemical and functional characteristics may elucidate its antioxidant potential and effects against cell death, enhancing understanding of plant GPXs and informing strategies to control witches' broom disease. Results indicated TcPHGPX interaction with selenium compounds, mainly sodium selenite, but without improving the protein function. Protein-protein interaction network suggested cacao GPXs association with glutathione and thioredoxin metabolism, engaging in pathways like signaling, peroxide detection for ABA pathway components, and anthocyanin transport. Tests on tobacco cells revealed that TcPHGPX reduced cell death, associated with decreased membrane damage and H2O2 production induced by MpNEP2. This study is the first functional analysis of TcPHGPX, contributing to knowledge about plant GPXs and supporting studies for witches' broom disease control.

Evaluation of spontaneous exploratory and anxiety-related behaviors of mice treated with gymnopilins obtained from the mushroom Gymnopilus imperialis (Agaricomycetes, Basidiomycota).

Gymnopilins are long chain oligoisoprenoids produced through the condensation of isoprene units from MEV and MEP biosynthetic pathways. In Gymnopilus, these carotenoid-like molecules are recognized as major compounds in some species. In the present study, oligoisoprenoids derived from gymnopilins were dereplicated from Gymnopilus imperialis, a mushroom-forming basidiomycete, using liquid chromatographic coupled with high-resolution mass spectrometry (tandem LC-HRMS/MS) and GNPS. From the dichloromethane extract (Gym-DCM) of G. imperialis we annotated 3 oligoisoprenoids from the GNPS molecular library spectra and 15 analogs from the curation of the molecular networking. Data from NMR spectroscopic of the extract confirmed the annotation of the metabolites. Based on the literature data suggesting the neurotoxic effect of gymnopilins, we investigated the effects of the administering different doses of gymnopilin extracts (1, 4 or 10 mg/kg) and diazepam (4 mg/kg) on the acquisition of object recognition memory (ORM) in mice. By studying novel object recognition memory (ORM), a type of non-aversive memory. ORM was assessed based on the total time of spontaneous exploration of both objects, the discrimination index (DI), and the frequency of contact with both objects. Our present findings reveal, for the first time, that gymnopilins treatment before training modulates ORM in a dose-dependent manner. It is also suggested that differential effects on memory might be related to differential effects on GABAA receptors but do not exclude its effects in other neurotransmitter systems. Another class of secondary metabolites, alkaloids, might modulate AChR, which is essential for maintaining object recognition memory over time.