Genome sequence and genetic linkage analysis of Shiitake mushroom _Lentinula edodes_

_Lentinula edodes_ (Shiitake/Xianggu) is an important cultivated mushroom. Understanding the genomics and functional genomics of _L. edodes_ allows us to improve its cultivation and quality. Genome sequence is a key to develop molecular genetic markers for breeding and genetic manipulation. We sequenced the genome of _L. edodes_ monokaryon L54A using Roche 454 and ABI SOLiD genome sequencing. Sequencing reads of about 1400Mb were de novo assembled into a 40.2 Mb genome sequence. We compiled the genome sequence into a searchable database with which we have been annotating the genes and analyzing the metabolic pathways. In addition, we have been using many molecular techniques to analyze genes differentially expressed during development. Gene ortholog groups of _L. edodes_ genome sequence compared across genomes of several fungi including mushrooms identified gene families unique to mushroom-forming fungi. We used a mapping population of haploid basidiospores of dikaryon L54 for genetic linkage analysis. High-quality variations such as single nucleotide polymorphisms, insertions, and deletions of the mapping population formed a high-density genetic linkage map. We compared the linkage map to the _L. edodes_ L54A genome sequence and located selected quantitative trait loci. The Shiitake community will benefit from these resources for genetic studies and breeding.
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The biotechnology of higher fungi - current state and perspectives

This review article concisely describes methodology of biotechnological processes with the use of cultures of higher fungi, their application in bioremediation and to obtain biologically active preparations. Advantages and disadvantages of biotechnological methods used to cultivate mushrooms are analyzed. This paper contains overview of higher fungi species most commonly used in biotechnological processes, of cultivation methods applied to produce fungal biomass, of enzymes and bioactive metabolites and of the strategies for submerged cultivation of the mycelial cultures. The problems of optimization of strains and biotechnological processes are briefly discussed.Niemniej pomimo trudności, producenci substancji leczniczych pochodzenia grzybowego (Lentinan, LEM, Grifon-D, PSK, PSP), suplementów diety oraz enzymów grzybowych, wprowadzają metody biotechnologiczne do produkcji. Zgodnie ze stosowanym od dawna w biotechnologii przemysłowej (np. przez producentów antybiotyków) zwyczajem, warunki procesu rzadko są opisywane w publikacjach, a czasami nie są nawet patentowane - co ułatwia zachowanie ich w tajemnicy. W latach 90-tych XX wieku pojawiły się pierwsze informacje o możliwości stosowania metod rekombinowanego DNA dla grzybów wyższych. Współcześnie, liczne publikacje donoszą o opracowaniu metod transformacji oraz o uzyskaniu modyfikowanych genetycznie grzybów jadalnyc

微生物由来リパーゼ活性のスクリーニング : 食用キノコを中心とした担子菌類由来リパーゼの探索

キノコに代表される担子菌類は，食用や漢方など，古くから食品や医薬品として利用されてきた。しかしながら，その他の利用，特に，有用物質生産を目的とした「生体触媒」としての有用性については，ほとんど知られていなかった。そこで，本研究では，食用キノコを中心とした担子菌類の，生体触媒としての可能性を探るため，担子菌由来の細胞外分泌型リパーゼの探索を目的として研究を行った。Basidiomycetes containing edible mushrooms have been used for foods and traditional Chinese medicines since ancient times. However, the possibility of its application in other fields of the mushrooms is not well known. Therefore, in this study, in order to explore the possibility of edible mushrooms as biocatalysts, we investigated the screening for extracellular lipases derived from basidiomycetes containing edible mushroom

Sunflower seed hull: Its value as a broad mushroom substrate

Sunflower (Heliantus annus) seed hull (SSH), an abundant and cheap lignocellulosicresidue from edible oil-seed industries, was evaluated as an alternative substrate formushroom cultivation. Different edible and medicinal mushroom species were grownsuccessfully, bringing a positive use of material that could be problematic for disposal.In this review we display different studies that linked the SSH with mushroom productionand other derivations of its use as mushroom substrate.Fil: Figlas, Norma Débora. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiárida; ArgentinaFil: Gonzalez Matute, Ramiro. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiárida; ArgentinaFil: Curvetto, Nestor Raul. Universidad Nacional del Sur; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

EVALUATION OF TOTAL POLYPHENOLS AND ANTIOXIDANT CAPACITY IN MUSHROOM EXTRACTS PLEUROTUS OSTREATUS AND LENTINULA EDODES

Objective: This research was aimed at assessing the concentration of total polyphenols in ethanolic and methanolic extracts of Pleurotus ostreatus and Lentinula edodes mushrooms and antioxidant activity. Methods: Polyphenols were determined by the Folin Ciocalteu method, using lyophilized mushroom samples for the preparation of extracts and antioxidant activity by the TBARS method. Results: Extracts prepared from mushrooms showed appreciable values ​​of polyphenols, and for the ethanolic extract of Pleurotus ostreatus and Lentinula edodes values ​​of 102.78 and 81.83 mg of gallic acid/100 g of the sample respectively, comparable to those obtained in some fruits For methanolic extracts, values ​​of 100.45 and 78.92 mg of gallic acid/100 g of sample were obtained. Polyphenol concentration values ​​for the Pleurotus were higher in the two types of extracts and lower for the Lentinula edodes. Conclusion: When evaluating the antioxidant activity, high antioxidant activity was found for the two types of mushroom, Pleurotus ostreatus and Lentinula edodes, presenting peroxidase inhibition values ​​of 88.04 and 89.49% respectively

Multivariate Statistical Techniques: A new approach to Identify the commercial properties of mixtures of flours of lentinula edodes and cocoa pod husk

[ENG]Lentinula edodes is known to show high nutritional and organoleptic properties and can be mixed with different by-products in the production of new foods with important functional characteristics. Cocoa pod husk (CPH) is the main by-product in the cocoa industry and presents important bioactivities. In this research, two mixtures were applied based on Lentinula edodes mushroom flour from fifty different strains (strain 1 to strain 50) and cocoa pod husk flour (CPHF): 60% Lentinula edodes mushroom flour from each strain mixed with 40% CPHF (Mixture 1), and 80% Lentinula edodes mushroom flour from each strain mixed with 20% CPHF (Mixture 2). The parameters evaluated were moisture, proteins, fat, total dietary fiber (TDF), ash, carbohydrates, antioxidant capacity, antimicrobial activity, pH and water activity (aw). The multivariate statistical techniques evaluated those samples that exhibited the highest degree of correlation with the distinct chemical and commercial parameters, showing that sample 13 of both mixture flours (M1 and M2) obtained by the mushrooms of L. edodes (strain 13) with CPH indicated the higher significant of the parameters. In addition, the sensory test of the best flour (sample 13) was applied to 50 panelists, presenting the best sensory characteristics, such as color, aroma, texture and general acceptability

A Novel Apoptosis Correlated Molecule: Expression and Characterization of Protein Latcripin-1 from Lentinula edodes C91–3

An apoptosis correlated molecule—protein Latcripin-1 of Lentinula edodes C91–3—was expressed and characterized in Pichia pastoris GS115. The total RNA was obtained from Lentinula edodes C91–3. According to the transcriptome, the full-length gene of Latcripin-1 was isolated with 3′-Full Rapid Amplification of cDNA Ends (RACE) and 5′-Full RACE methods. The full-length gene was inserted into the secretory expression vector pPIC9K. The protein Latcripin-1 was expressed in Pichia pastoris GS115 and analyzed by Sodium Dodecylsulfonate Polyacrylate Gel Electrophoresis (SDS-PAGE) and Western blot. The Western blot showed that the protein was expressed successfully. The biological function of protein Latcripin-1 on A549 cells was studied with flow cytometry and the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyl-tetrazolium Bromide (MTT) method. The toxic effect of protein Latcripin-1 was detected with the MTT method by co-culturing the characterized protein with chick embryo fibroblasts. The MTT assay results showed that there was a great difference between protein Latcripin-1 groups and the control group (p < 0.05). There was no toxic effect of the characterized protein on chick embryo fibroblasts. The flow cytometry showed that there was a significant difference between the protein groups of interest and the control group according to apoptosis function (p < 0.05). At the same time, cell ultrastructure observed by transmission electron microscopy supported the results of flow cytometry. The work demonstrates that protein Latcripin-1 can induce apoptosis of human lung cancer cells A549 and brings new insights into and advantages to finding anti-tumor proteins

Efecto antioxidante de Chondrus crispus y Lentinula edodes en diferentes margarinas

Lipid oxidation is the reaction responsible for food degradation. To solve this problem the agri-food industry uses antioxidants, preservatives and chemical stabilizers. Currently, there is an increasingly strong demand for healthier eating. Because of this, the industry is increasingly interested in replacing chemical compounds with natural products of equal or greater effective-ness. This project studies the oxidative stability, both qualitative and quantitative, of margarines of different compositions when independently adding dehydrated material from a fungus (Len-tinula edodes) and a seaweed (Chondrus crispus) that are known for their antioxidant activity. To achieve this, Rancimat equipment was used at different temperatures: 110, 120, 130 and 140 ± 1.6 ºC. comparing treated margarine samples to their respective controls. The results varied depending on the temperatures used, the composition of margarines and the natural antioxidants added. The results show that margarines with a lower proportion of unsaturated fatty acids, and especially polyunsaturated fatty acids, have greater oxidative stability. Oxidative stability was higher when the fungus was added compared to seaweed.La oxidación lipídica es la reacción responsable de la degradación de los alimentos, para solventar este problema la industria agroalimentaria utiliza antioxidantes, conservantes y estabilizantes químicos. Actualmente hay una demanda cada vez más fuerte de una alimentación más saluda-ble; debido a esto, la industria está cada vez más interesada en sustituir los compuestos químicos por productos naturales de igual o mayor efectividad. En este proyecto se estudia la estabilidad oxidativa en margarinas de distinta composición, tanto cualitativa como cuantitativa, al adicio-nar de forma independiente material deshidratado procedente de un hongo (Lentinula edodes) y de un alga (Chondrus crispus), que se caracterizan por tener actividad antioxidante. Para ello, se utilizó un equipo Rancimat a diferentes temperaturas: 110 ºC, 120 ºC, 130 ºC y 140 ºC ± 1.6 ºC, comparando las muestras de margarinas tratadas con sus respectivos controles. Los resultados variaron según las temperaturas empleadas, la composición de las margarinas y los antioxidantes naturales añadidos. Los resultados muestran que la margarina con menor proporción en ácidos grasos insaturados, y especialmente de poliinsaturados, presenta mayor estabilidad oxidativa y que esta aumenta en mayor grado cuando se añade el hongo con respecto a la adición del alga.Ciencias Experimentale

Antioxidant properties and phenolic profile of the most widely appreciated cultivated mushrooms: a comparative study between in vivo and in vitro samples.

The present study reports a comparison of the antioxidant properties and phenolic profile of the most consumed species as fresh cultivated mushrooms and their mycelia produced in vitro: Agaricus bisporus (white and brown), Pleurotus ostreatus (oyster), Pleurotus eryngii (king oyster) and Lentinula edodes (shiitake). The antioxidant activity was evaluated through reducing power (Folin-Ciocalteu and Ferricyanide/Prussian blue assays), free radical scavenging activity (DPPH assay) and lipid peroxidation inhibition (β-carotene/linoleate and TBARS assays). The analysis of phenolic compounds was performed by HPLC/PAD. The mushroom species with the highest antioxidant potential was Agaricus bispous (brown). However, concerning to the species obtained in vitro, it was Lentinula edodes that demonstrate the highest reducing power. Generally, in vivo samples revealed higher antioxidant properties than their mycelia obtained by in vitro techniques. About the phenolic compounds researched, they were detected both in mushrooms and mycelia without any particular abundance. Results showed that there is no correlation between the studied commercial mushrooms and the corresponding mycelia obtained in vitro. Nevertheless, this study contributes to the rise of data relatively to the species consumed as fresh mushrooms and the possibility of their in vitro production as a source of bioactive compounds