Mushroom Fruiting Bodies Research Articles

Content of Biogenic Amines and Physical Properties of Lacto-Fermented Button Mushrooms

The aim of the study was to assess the content of biogenic amines and physical properties of fruiting bodies of white and brown button mushrooms subjected to the process of controlled lactic acid fermentation. Lactiplantibacillus plantarum 299v with documented probiotic properties and L. plantarum EK3, i.e., an isolate obtained from spontaneously fermented button mushrooms, were used as starter strains. Fresh, blanched, fermented, and refrigerated fruiting bodies were analysed. The mushroom samples were found to contain three amines: spermidine, putrescine, and tyramine in amounts that do not pose a threat to consumer health. The highest content of spermidine was found in the fruiting bodies of unprocessed brown and white mushrooms (367.22 ± 14.19 and 266.47 ± 13.38 mg/kg, respectively). Putrescine and tyramine were found only in the fermented mushrooms. Putrescine ranged from 0.58 ± 0.25 to 10.11 ± 0.5 mg/kg, while tyramine ranged from 1.44 ± 0.25 to 69.04 ± 1.39 mg/kg. Histamine, which is the most toxic biogenic amine, was not detected in any of the samples. The technological process caused substantial changes in all colour parameters. The blanching process resulted in a decrease in the value of parameter L* and an increase in the value of parameters a* and b*. The process of lactic acid fermentation increased the brightness parameter. It also caused a substantial increase in the yellowness parameter and reduced the redness of the fermented mushrooms. Blanching exerted a significant effect on the texture of the mushroom fruiting bodies, contributing to an increase in the puncture force value from 2.78 ± 0.22 to 4.90 ± 0.43 N and from 3.21 ± 0.23 to 5.59 ± 0.3 N in the case of the white and brown mushrooms, respectively. During the refrigerated storage, the firmness of the fermented mushrooms did not change.

Quantitative mapping of mercury and selenium in mushroom fruit bodies with laser ablation–inductively coupled plasma-mass spectrometry

This work describes the development of a novel method for quantitative mapping of Hg and Se in mushroom fruit body tissues with laser ablation coupled to inductively coupled plasma-mass spectrometry (LA-ICP-MS). Different parameters of the protocol for preparation of the standards used for quantification via external calibration were assessed, e.g., the dissolution temperature of gelatin standards and the addition of chitosan and l-cysteine as additives to the gelatin-based calibration droplets to better match the sample matrix. While chitosan was not suited for this purpose, the presence of l-cysteine considerably improved the figures of merit of the calibration, leading to limits of detection of 0.006 and 0.3 µg g−1 for Hg and Se, respectively, at a pixel size of 20 × 20 µm. Further, an in-house reference material, ideally suited for the validation of the method for application to mushroom samples, was successfully prepared from a paste of Boletus edulis. The newly developed method was used to investigate the distribution of Hg and Se in tissue sections of five porcini mushroom individuals of three different species (Boletus edulis, Boletus aereus, and Boletus pinophilus) and one sample of a parasol mushroom (Macrolepiota procera). For one sample, additional areas were ablated at higher spatial resolution, with a laser spot size down to 5 µm, which allows a detailed investigation of the spatial distribution of Hg and Se in mushrooms.Graphical abstract

Palm Press Fibre and Rice Straw for Cultivation Grey Oyster Mushroom (Pleurotus sajor-caju)

Sawdust (SD) usually sourced from rubber plant is commonly used as substrate to cultivate the grey oyster mushroom Pleurotus sajor-caju in Malaysia. However, the market price of SD is increasing because of its declining availability. Thus, this study aimed to discover an alternative substrate to cultivate grey oyster mushroom. The raw materials used in the substrate were oil palm pressed fiber (PPF), rice straw (RS) and SD, either alone or in combination with different ratios including T1: 90% SD + 10% rice bran (commercial substrate as control); T2: 30% RS + 20% SD + 50% PPF; T3: 50% RS + 50% PPF; T4: 100% RS; and T5: 100% PPF. The suitability of the substrates was determined based on growth, yield, nutrition and mineral content in the mushrooms. The growth and yield of the mushroom fruiting body on the different substrates showed significant differences. The shortest harvesting day was obtained in T4 (100% RS) after 29 days, whereas the longest harvesting day was obtained in T1 (90% SD + 10% rice bran) after 51 days. The highest yield was obtained in T4. Nutrition analysis showed significant difference among the treatments. The highest amounts of protein (26%) and ash (1.29%) were found in T5. Overall, the results showed that mushroom yield performance was good in RS but nutritional content was the highest in PPF. Therefore, RS and PPF have good impact for mushroom growers either in commercial production or in functional food industry to reduce SD dependency.

Dynamics of Changes in pH and the Contents of Free Sugars, Organic Acids and LAB in Button Mushrooms during Controlled Lactic Fermentation.

The aim of the study was to assess changes in the basic quality parameters induced by controlled lactic fermentation of fruiting bodies of the button mushroom (Agaricus bisporus). Lactiplantibacillus plantarum 299v with documented probiotic properties and L. plantarum EK3, i.e., an isolate obtained from spontaneously fermented button mushrooms, were used as starter strains. The fruiting bodies of fresh, blanched, and fermented mushrooms were analyzed at different stages of the lactic fermentation process. The content of free sugars (high-performance liquid chromatography with charged aerosol detector method; HPLC-CAD) and organic acids (high-performance liquid chromatography with diode array detector method; HPLC-DAD) was determined both in the mushroom fruiting bodies and in the brine. Five free sugars (ribose, trehalose, sucrose, glucose, and fructose), mannitol, and six organic acids (lactic, malic, succinic, citric, acetic, and fumaric acids) were detected in the samples. Lactic acid dominated in the final products. The starter cultures exhibited varying degrees of utilization of available mushroom sugars and sucrose used as an additional substrate. Sucrose was utilized at a higher rate and in greater amounts by the L. plantarum EK3 isolate. This starter culture was characterized by a significantly higher final amount of produced lactic acid, a lower pH value, and higher numbers of LAB (lactic acid bacteria). These important quality parameters largely determine the stability of fermented products. Based on the analysis results and the high scores in the organoleptic evaluation of the fermented mushrooms, the L. plantarum EK3 isolate can be recommended as an appropriate starter culture for lactic fermentation of mushroom fruiting bodies.

Detection of autophagy-related structures in fruiting bodies of edible mushroom, Pleurotus ostreatus.

Autophagy is involved in various fungal morphogenetic processes. However, there are limited reports regarding the role of autophagy in mushroom fruiting body formation. The purpose of this study was to reveal the autophagy-related structures in mushroom-forming fungi. The edible mushroom Pleurotus ostreatus was used in this study. Transmission electron microscopy revealed double-membrane bounded structures containing cytoplasmic components in the fruiting bodies of this fungus. Some of these double-membrane structures were observed to interact with the vacuoles. Additionally, curved flat cisternae of various lengths were detected in the cytoplasm. The shape, size and thickness of the limiting membrane of the double-membrane structures and the flat cisternae corresponded well with those of the autophagosomes and the isolation membranes, respectively. Regarding autophagosome formation, a membrane-bound specific zone was detected near the isolation membrane, which appeared to expand along the novel membrane. This is the first detailed report showing autophagy-related structures in P. ostreatus and provides a possible model for autophagosome formation in these filamentous fungi.

Nutritional qualities and antioxidant activity of Pleurotus floridanus grown on composted peach sawdust substrate with different composting time.

Recently, composting cultivation method is widely used in oyster mushroom production. In this study, we focused on the effects of composting processes on nutritional qualities and antioxidant activity of Pleurotus floridanus mushroom fruiting bodies. Three treatments of different composting time (2, 4, and 5 days) were performed with an atmospheric sterilization treatment as the control. The results showed that the pH value, total carbon content, and total nitrogen content of substrate were critical parameters which would significantly affect mushroom qualities and bioactivities. Fruiting bodies of the control demonstrated significantly higher crude protein content, total amino acid content, and essential amino acid content than that of composting treatments. Moreover, fruiting bodies of treatment D4 and D5 manifested significantly higher crude polysaccharide contents. Crude polysaccharide of treatment D4 represented the highest scavenging ability toward both radical 3-ethylbenzthiazoline-6-sulfonic acid (ABTS·+ ) and Hydroxyl radical (OH·). It suggests that composting processes is suitable for oyster mushroom cultivation based on nutritional and antioxidant qualities of fruiting bodies.

Pleurotus cultivation: a sustainable way to utilize agrowaste

In the present study two species of Pleurotus namely Pleurotus florida (Mont.) Singer and Pleurotus ostreatus (Jacq.) P.Kumm. were cultivated using three different agro waste substrates such as paddy straw, sugarcane bagasse, banana leaves and its mixture in equal proportion. The fastest colonization and maximum numbers of heads were produced on paddy straw substrate. Banana leaves and paddy straw substrates reported the highest yield of mushroom fruitbodies, biological efficiency and biomass loss in P. florida and P. ostreatus. It was noticed that the growth and development of fruitbodies on sugarcane bagasse was minimum and development of competitor moulds was observed on it. In the selected substrates banana leaves posses the highest percentage of nitrogen, carbon and cellulose. The results showed the possibility of utilizing different agrowaste for cultivation of oyster mushroom, which will boost the income of farmers.

Gene age shapes the transcriptional landscape of sexual morphogenesis in mushroom-forming fungi (Agaricomycetes).

Multicellularity has been one of the most important innovations in the history of life. The role of gene regulatory changes in driving transitions to multicellularity is being increasingly recognized; however, factors influencing gene expression patterns are poorly known in many clades. Here, we compared the developmental transcriptomes of complex multicellular fruiting bodies of eight Agaricomycetes and Cryptococcus neoformans, a closely related human pathogen with a simple morphology. In-depth analysis in Pleurotus ostreatus revealed that allele-specific expression, natural antisense transcripts, and developmental gene expression, but not RNA editing or a 'developmental hourglass,' act in concert to shape its transcriptome during fruiting body development. We found that transcriptional patterns of genes strongly depend on their evolutionary ages. Young genes showed more developmental and allele-specific expression variation, possibly because of weaker evolutionary constraint, suggestive of nonadaptive expression variance in fruiting bodies. These results prompted us to define a set of conserved genes specifically regulated only during complex morphogenesis by excluding young genes and accounting for deeply conserved ones shared with species showing simple sexual development. Analysis of the resulting gene set revealed evolutionary and functional associations with complex multicellularity, which allowed us to speculate they are involved in complex multicellular morphogenesis of mushroom fruiting bodies.

Proteomics Reveals the Mechanism Underlying the Autolysis of Postharvest Coprinus comatus Fruiting Bodies

Autolysis occurs widely in edible mushroom fruiting bodies after harvest, but the mechanism is still unclear. In this study, quantitative proteomics and bioinformatics analyses have been applied for revealing the autolysis mechanism of postharvest Coprinus comatus fruiting bodies. The results indicated that the autolysis mechanism of postharvest C. comatus was complicated. Before pileus opening, the carbohydrate metabolism including cell wall hydrolysis and energy biosynthesis, which were probably regulated by the ribosome, was involved in mushroom autolysis, whereas after pileus opening, the autolysis mechanism was related to the accumulated reactive oxygen species (ROS) and activated mitogen-activated protein kinase (MAPK) signaling pathway based on the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Furthermore, the changes in cell wall components and hydrolases, along with the production of ROS and the activities of oxidoreductase in C. comatus, were also verified to confirm the proteomic analysis results.

First Record of the Two Insects Megaselia halterata and Lycoriella ingenua that Attack Some Edible Mushrooms in Iraq and Assessing the Damage Caused by them

Hassan, A.A. and A.R.M. Al-Qaissi. 2022. First Record of the Two Insects Megaselia halterata and Lycoriella ingenua that Attack Some Edible Mushrooms in Iraq and Assessing the Damage Caused by them. Arab Journal of Plant Protection, 40(3): 209-214. https://doi.org/10.22268/AJPP-40.3.209214 Invading insects were found in the fruiting bodies of the edible mushrooms Agaricus bisporus and Pleurotus spp. at the mushroom farm of the Faculty of Agriculture, University of Tikrit, through the presence of holes, necrosis and tunnels in the mushroom fruits. Infested fruiting bodies containing larvae were collected and insects were reared in the laboratory for the purpose of diagnosis. The phenotypic diagnosis was confirmed by molecular diagnosis based on the nucleotide sequence of the cytochrome oxidase C subunit 1 gene (in mitochondria). The two insects were identified as Megaselia halterata and Lycoriella ingenua and were registered in the NCBI database under the accession numbers MZ021516.1 and MZ021517.1, respectively. This is the first record of these two species in Iraq. The study also included a description of the damage caused by these two insects on the fruiting bodies of the mushrooms A. bisporus and Pleurotus spp. Keywords: Molecular identification, mushroom fly, Agaricus bisporus, Pleurotus spp

Advances in Production of Medicinal Mushrooms Biomass in Solid State and Submerged Bioreactors.

Production of mushroom fruit bodies using farming technology could hardly meet the increasing demand of the world market. During the last several decades, there have been various basic and applied studies on fungal physiology, metabolism, process engineering, and (pre)clinical studies. The fundamental aspects of solid-state cultivation of various kinds of medicinal mushroom mycelia in various types of bioreactors were established. Solid-state cultivation of medicinal mushrooms for their biomass and bioactive metabolites production appear very suitable for veterinary use. Development of comprehensive submerged technologies using stirred tank and airlift bioreactors is the most promising technology for fast and large-scale production of medicinal fungi biomass and their pharmaceutically active products for human need. The potentials initiate the development of new drugs and some of the most attractive over-the-counter human and veterinary remedies. This article is to overview the engineering achievements in solid state and submerged cultivations of medicinal mushrooms in bioreactors.

Mushroom Quality Related with Various Substrates' Bioaccumulation and Translocation of Heavy Metals.

Mushrooms are popular due to the nutrition contents in the fruit bodies and are relatively easy to cultivate. Mushrooms from the white-rot fungi group can be cultivated on agricultural biomass such as sawdust, paddy straw, wheat straw, oil palm frond, oil palm empty fruit bunches, oil palm bark, corn silage, corn cobs, banana leaves, coconut husk, pineapple peel, pineapple leaves, cotton stalk, sugarcane bagasse and various other agricultural biomass. Mushrooms are exceptional decomposers that play important roles in the food web to balance the ecosystems. They can uptake various minerals, including essential and non-essential minerals provided by the substrates. However, the agricultural biomass used for mushroom cultivation is sometimes polluted by heavy metals because of the increased anthropogenic activities occurring in line with urbanisation. Due to their role in mycoremediation, the mushrooms also absorb pollutants from the substrates into their fruit bodies. This article reviews the sources of agricultural biomass for mushroom cultivation that could track how the environmental heavy metals are accumulated and translocated into mushroom fruit bodies. This review also discusses the possible health risks from prolonged uptakes of heavy metal-contaminated mushrooms to highlight the importance of early contaminants’ detection for food security.

Distribution and Origin of Major, Trace and Rare Earth Elements in Wild Edible Mushrooms: Urban vs. Forest Areas.

This paper investigates the composition of major, trace, and rare earth elements in 15 different species of wild edible mushrooms and the possible effect of urban pollution on elemental uptake. The collected mushrooms include different species from the green areas of the city, exposed to urban pollution, and from the forests, with limited anthropogenic influence. Through a comprehensive approach that included the analysis of 46 elements, an attempt was made to expand knowledge about element uptake by mushroom fruiting bodies. The results showed a wide variability in the composition of mushrooms, suggesting a number of factors influencing their element uptake capacity. The data obtained do not indicate significant exposure to anthropogenic influences, regardless of sampling location. While major elements’ levels appear to be influenced more by species-specific affinities, this is not true for trace elements, whose levels presumably reflect the geochemical characteristics of the sampling site. However, the risk assessment showed that consumption of excessive amounts of the mushrooms studied, both from urban areas and from forests, may have adverse health effects.

Nephroprotective and antioxidant effects of ethanol extract of Coprinus comatus mushroom fruit-bodies on streptozotocin-induced diabetic rat models

Diabetes mellitus (DM) is a metabolic disease with an increasing prevalence. An increase in free radicals and AGEs can cause glomerular cell damage which can lead to inflammation and DM nephropathy. WHO has recommended DM treatment by using herbal medicines that have minimal side effects and have a lot of biological activity to prevent complications. Coprinus comatus (O.F. Mull.) is known as immunomodulatory, anti-inflammatory and antidiabetic agent. The research used C. comatus cultivated in Cianjur. The basidiome of this mushroom has slightly oval and small with height of 8-12 cm and thickness of 2-3 cm. The research included six groups of male Wistar rats: Group 1 received no treatment, Groups 2–6 were administered 45 mg/kg BW streptozotocin once, Group 3 was administered 45 mg/kg BW metformin, Groups 4, 5, and 6 were administered 250, 500, and 750 mg/kg BW of C. comatus fruit body ethanol extract respectively for 14 days. The superoxide dismutase (SOD), malondialdehyde (MDA), hepcidin and β2 microglobulin (B2M) levels were evaluated. Data were analyzed using analysis of variance and Duncan’s multiple-range tests. The results showed that dose of 500 mg was effective in increasing SOD and decreasing hepcidin, B2M and MDA levels.

TMT-based quantitative proteomic analysis of postharvest Coprinus comatus fruiting body during storage

Coprinus comatus is a highly perishable edible mushroom and it is difficult to keep fresh after harvest. The high content of moisture, maintained respiration and absence of protective cuticle layer are the main factors of short shelf life. In this study, we used tandem mass tags (TMT)-based quantitative proteomic technique to investigate the proteome responses of postharvest mushrooms during 16 d of storage at 8 ± 1 ℃. The quality parameters of mushroom fruiting body including firmness, weight loss, browning index and respiration rate were determined. A total of 168 differentially abundant proteins (DAPs) were overlapped in the comparison groups (8 d-storage treatment group vs. control group and 16 d-storage treatment group vs. control group), of which 74 DAPs were up-regulated and 86 DAPs were down-regulated during the postharvest storage. Based on Gene Ontology (GO) annotation, the major biological processes were metabolic process and cellular process. Catalytic activity, binding and structural molecular activity were the predominant molecular functions. The cellular components were primarily cell and cell part. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that these DAPs were mainly involved in ribosome, glycolysis, citrate cycle and glutamate metabolism. AMPK and FOXO signaling pathways also participated in the postharvest physiology of mushrooms. Our results provide insights into the biological processes of Coprinus comatus fruiting body during storage and a theoretical basis for developing effective postharvest technology for fresh Coprinus comatus.

Uptake of perfluoroalkyl substances, pharmaceuticals, and parabens by oyster mushrooms (Pleurotus ostreatus) and exposure risk in human consumption

Organic micropollutants (MPs) pose potential threats to environmental ecosystems and human health. This study investigated uptake of perfluoroalkyl substances (PFASs), pharmaceuticals, and paraben by edible oyster mushrooms (Pleurotus ostreatus), cultivated on spiked growth substrate. Concentrations of pharmaceuticals and paraben in substrate showed a decreasing trend over a 20-day harvesting period, whereas PFAS concentrations were variable over the harvesting period. However, only propylparaben, clarithromycin, and PFASs were detected in fruiting bodies of oyster mushroom. Uptake of PFASs by oyster mushroom fruit bodies was negatively correlated with perfluorocarbon chain length. An impact of MPs on fungal colonization was observed, with decreased respiration in treatments with the highest concentration of MPs, but production of fruiting bodies was not affected by exposure level. The potential human risk from ingestion of MPs was evaluated for oyster mushrooms exposed to the highest concentration of MPs in substrate, based on acceptable daily intake (ADI).

Free Radical Scavenging Capacities and Alleviating Actions of Polysaccharides Extract of Termitomyces le-testui on Methylprednisolone-Induced Immunodepression in Rats.

Background Natural products have been said to show immunomodulatory and antioxidant activities. The research study was aimed to assess the immunomodulatory and free radical scavenging activities of crude polysaccharide from dry mushroom fruiting bodies of Termitomyces le-testui. Materials and Methods Hot water extract of polysaccharide extract of T. le-testui was prepared and tested in white albino Wister rats for its immunomodulatory activities effect on methylprednisolone-immunosuppressed animals. In addition, the radical scavenging activity of the polysaccharide was evaluated using nitrite and hydrogen peroxide. Results The result of the study showed that the polysaccharide T. le-testui increases the phagocytic index, energy metabolism of macrophages, spleen index, and nitric oxide in a concentration-dependent manner in immunosuppressed animals. Also, it was observed that the extract increased dose-dependent total oxidative stress and thymus index. Finally, the crude polysaccharide-rich extract showed nitrite and hydrogen peroxide scavenging activity in a concentration-dependent manner. Conclusion Polysaccharide-rich extract possesses immunomodulatory and antioxidant properties.

Deciphering the role of moringa leaf powder as a supplement in the cotton waste substrate for the growth and nutrition of king oyster mushroom

The addition of supplementing material in the substrate is an efficient way to increase mushroom yield and nutrition. The addition of moringa leaf powder (MLP) could boost the yield and nutrition of mushrooms. To improve the nutritional properties and yield of king oyster mushroom, moringa leaf powder as supplementing material in the cotton waste substrate was added with different ratios to standardize the supplementing material in the substrate. In the present research, the impact of moringa leaf powder at various concentrations (2%,4%,6%,8% and 10%) in cotton waste were used to check the suitability of king oyster mushroom production and quality. Results showed that addition of MLP substantially increased production and nutritional value of mushroom. The MLP at 6% on dry weight basis in cotton waste showed markedly higher total yield, biological efficiency as compared to control or cotton waste alone. The addition of 6% MLP in cotton waste also enhanced the Vitamin C, Folic acid, protein, carbohydrate, fats, fibers, ash and minerals (N, Na, K, Ca, Zn, Mg, Mn, Fe and P) of king oyster mushroom, Moreover, total phenolic contents and antioxidant activity were improved in King oyster mushroom fruit bodies. Vitamin D was quantified by HPLC-UV chromatograph in P. eryngii. Vitamin-D content was also varied with MLP addition in the substrate.. In conclusion, MLP addition in substrate could be an effective approach for the increase in production and nutritional quality of mushrooms.

ECOLOGICALLY SAFE UTILIZATION OF ORGANIC WASTES AND TECHNOLOGY OF PRODUCTION OF THE FUNGUS PLEUROTUS OSTREATUS (THE OYSTER MUSHROOM)

Background. Organic cellulose-containing wastes are represented by various substrates with diverse physicochemichal properties suitable as feedstock for obtaining new products, e.g. mycelium and fruit bodies of edible mushrooms. Industrial mushroom cultivation based on utilization of different kinds of organic wastes is one of the perspective directions in biotechnology and agriculture.
 Improving the nutrient value of substrates and increasing the reliability of mushroom culture is possible at expense of utilizing nutrient supplies originated as wastes of another industry, for example, the use of sparging grain, which is the major waste of brewery. Wet sparging grain has humidity of 70-80%, and contains in average more than 20% of dry residues with high protein level (12-15%), which is approximately three times higher its content than in barley. For that reason, as a supplement to nutrient media and substrates, for rearing mycelium and fruit bodies of the fungus Pleurotus ostreatus, we propose the use of the brewers’ industry, the wet sparging grain.
 Purpose. Evaluation of the possibility of utilising the wastes of agriculture and brewery, for growing mycelium and fruit bodies of the edible mushroom Pleurotus ostreatus.
 Materials and methods. The study was carried out in the years 2017-2021 in the laboratory of the Department of biology and technology of living systems at L.N. Tolstoy TSPU. The research objects were the fungus Pleurotus ostreatus, wet sparging grain, and various agricultural wastes (wheat straw, buckwheat chuff, sunflower seed husk).
 For mycelium extraction, we used the method of seeding superficially sterilized pieces of fruit bodies onto the hard sterile agarized nutrient medium in the Petri dishes. For obtaining the master culture of mycelium and fruit bodies of the fungus Pleurotus ostreatus we utilized substrates of various composition containing the sparging grain and agricultural wastes. The data obtained were statistically processed and analysed.
 Results. In result of the experiments, it was revealed that optimal nutrient media and substrates for obtaining and cultivation of the mycelium are those with addition of the sparging grain and cellulose-containing wastes. For obtaining fruit bodies, we used the substrate composed of the straw with addition of the brewers’ grain. Utilization of nutrient media with similar composition leads to shortening of the period of mycelium adaptation and initiation of its active growth phase when producing the master culture of the fungus. Further, for that reason, increases the speed of colonization by the fungal mycelium the substrate blocks for producing carposomes, and the fungus fruitification occurs more intensively.
 Conclusion. The use of the brewers’ draff for obtaining mycelium and fruit bodies of the edible oyster mushroom creates valuable, ecologically clean and quality nutrition product and facilitates cost effectiveness of its production. Design and application this biotechnology solve the problem of organic waste utilization, which in turn positively stimulates positive changes in the regional ecological situation and contributes partially into the task of import substitution.

Genomic DNA extraction from minimal amount of dried mushroom samples

Aims: To develop a simple and rapid genomic DNA extraction technique for dried (≈ 1 year old) mushroom fruiting bodies that yields high-quality DNA, suitable for use by post-graduate institutions. Method: Small amounts (0.04 g) of pulverized dried mushroom sample were incubated in a Tris /EDTA/SDS lysis buffer (100mM:10mM: 2%) at 65°C to lyse the chitinous fungal cell walls. Genomic DNA purification was performed using chloroform isoamyl alcohol (24:1), and DNA was precipitated using 100% ethanol. Results: Genomic DNA was successfully extracted under 70 minutes from 16 samples morphologically identified as Panaeolus, Copelandia, Gymnopilus, Pluteus and Favolus species. DNA concentrations were on average of 696.9ng/µL. PCR successfully amplified the ITS-5.8S region. The protocol has been successfully used by numerous post-graduate students in our research programme. Conclusion: The rapid and easy protocol produced high-quality genomic DNA void of any inhibitors that is suitable for downstream molecular implications across multiple mushroom genera. Noticeably, this method requires only minute quantities (0.04g) of starting material and is ideal for student training in higher academic institutions.