Combined Proteomic and Metabolomic Analyses Reveal the Comprehensive Regulation of Stropharia rugosoannulata Mycelia Exposed to Cadmium Stress.

The potential of Stropharia rugosoannulata as a microbial remediation material for cadmium (Cd)-contaminated soil lies in its capacity to absorb and accumulate Cd in its mycelia. This study utilized the TMT and LC-MS techniques to conduct integrated proteomic and metabolomic analyses with the aim of investigating the mycelial response mechanisms of S. rugosoannulata under low- and high-Cd stresses. The results revealed that mycelia employed a proactive defense mechanism to maintain their physiological functions, leading to reduced sensitivity to low-Cd stress. The ability of mycelia to withstand high levels of Cd stress was influenced primarily by the comprehensive regulation of six metabolic pathways, which led to a harmonious balance between nitrogen and carbohydrate metabolism and to reductions in oxidative stress and growth inhibition caused by Cd. The results provide valuable insights into the molecular mechanisms involved in the response of S. rugosoannulata mycelia to Cd stress.

Potential use of propolis-loaded quaternized chitosan/pectin hydrogel films as wound dressings: Preparation, characterization, antibacterial evaluation, and in vitro healing assay.

Quaternized chitosan (QCS) was blended with pectin (Pec) to improve water solubility and antibacterial activity of the hydrogel films. Propolis was also loaded into hydrogel films to improve wound healing ability. Therefore, the aim of this study was to fabricate and characterize the propolis-loaded QCS/Pec hydrogel films for use as wound dressing materials. The morphology, mechanical properties, adhesiveness, water swelling, weight loss, release profiles, and biological activities of the hydrogel films were investigated. Scanning Electron Microscope (SEM) investigation indicated a homogenous smooth surface of the hydrogel films. The blending of QCS and Pec increased tensile strength of the hydrogel films. Moreover, the blending of QCS and Pec improved the stability of the hydrogel films in the medium and controlled the release characteristics of propolis from the hydrogel films. The antioxidant activity of the released propolis from the propolis-loaded hydrogel films was ∼21-36 %. The propolis-loaded QCS/Pec hydrogel films showed the bacterial growth inhibition, especially against S. aureus and S. pyogenes. The propolis-loaded hydrogel films were non-toxicity to mouse fibroblast cell line (NCTC clone 929) and supported the wound closure. Therefore, the propolis-loaded QCS/Pec hydrogel films might be good candidates for use as wound dressing materials.

Coprinopsis cinerea dioxygenase is an oxygenase forming 10(S)-hydroperoxide of linoleic acid, essential for mushroom alcohol, 1-octen-3-ol, synthesis.

1-Octen-3-ol is a volatile oxylipin found ubiquitously in Basidiomycota and Ascomycota. The biosynthetic pathway forming 1-octen-3-ol from linoleic acid via the linoleic acid 10(S)-hydroperoxide was characterized 40 years ago in mushrooms, yet the enzymes involved are not identified. The dioxygenase 1 and 2 genes (Ccdox1 and Ccdox2) in the mushroom Coprinopsis cinerea contain an N-terminal cyclooxygenase-like heme peroxidase domain and a C-terminal cytochrome P450-related domain. Herein, we show that recombinant CcDOX1 is responsible for dioxygenation of linoleic acid to form the 10(S)-hydroperoxide, the first step in 1-octen-3-ol synthesis, whereas CcDOX2 conceivably forms linoleic acid 8-hydroperoxide. We demonstrate that KO of the Ccdox1 gene suppressed 1-octen-3-ol synthesis, although added linoleic acid 10(S)-hydroperoxide was still efficiently converted. The P450-related domain of CcDOX1 lacks the characteristic Cys heme ligand and the evidence indicates that a second uncharacterized enzyme converts the 10(S)-hydroperoxide to 1-octen-3-ol. Additionally, we determined the gene KO strain (ΔCcdox1) was less attractive to fruit fly larvae, while the feeding behavior of fungus gnats on ΔCcdox1 mycelia showed little difference from that on the mycelia of the WT strain. The proliferation of fungivorous nematodes on ΔCcdox1 mycelia was similar to or slightly worse than that on WT mycelia. Thus, 1-octen-3-ol seems to be an attractive compound involved in emitter-receiver ecological communication in mushrooms.

Impact of Cultivation Substrate and Microbial Community on Improving Mushroom Productivity: A Review.

Lignocellulosic materials commonly serve as base substrates for mushroom production. Cellulose, hemicellulose, and lignin are the major components of lignocellulose materials. The composition of these components depends upon the plant species. Currently, composted and non-composted lignocellulosic materials are used as substrates in mushroom cultivation depending on the mushroom species. Different substrate compositions can directly affect the quality and quantity of mushroom production yields. Consequently, the microbial dynamics and communities of the composting substrates can significantly affect mushroom production. Therefore, changes in both substrate composition and microbial diversity during the cultivation process can impact the production of high-quality substrates and result in a high degree of biological efficiency. A brief review of the current findings on substrate composition and microbial diversity for mushroom cultivation is provided in this paper. We also summarize the advantages and disadvantages of various methods of mushroom cultivation by analyzing the microbial diversity of the composting substrates during mushroom cultivation. The resulting information will serve as a useful guide for future researchers in their attempts to increase mushroom productivity through the selection of suitable substrate compositions and their relation to the microbial community.

A Taxonomic Appraisal of Bambusicolous Fungi in Occultibambusaceae (Pleosporales, Dothideomycetes) with New Collections from Yunnan Province, China.

During our ongoing studies of bambusicolous fungi in southwest China and Thailand, three saprobic pleosporalean taxa were discovered on bamboos in Yunnan Province of China. Occultibambusa hongheensis and Seriascoma bambusae spp. nov. are introduced based on morphological characteristics coupled with multi-locus phylogenetic analyses of combined LSU, SSU, TEF1-α, RPB2 and ITS sequence data. Occultibambusa kunmingensis is also reported from a terrestrial habitat for the first time. Comprehensive descriptions, color photo plates of micromorphology, and a phylogenetic tree showing the placements of these three taxa are provided. In addition, synopsis tables of Occultibambusa and Seriascoma with morphological features are also provided.

Morphological and phylogenetic appraisal of Ophioceras (Ophioceraceae, Magnaporthales).

Ophioceras is accommodated in the monotypic family Ophioceraceae (Magnaporthales, Sordariomycetes), and the genus is delimited based on molecular data. During an ongoing survey of bambusicolous fungi in southwest China, we collected a submerged decaying branch of bamboo from Sichuan Province, China and an Ophioceras species occurring on this substrate was observed and isolated. An Ophioceras taxon was delimited based on morphological characteristics and combined LSU, RPB1 and ITS sequence analyses and is described as Ophioceras sichuanense sp. nov. The species formed a well-supported clade basal to Ophioceras (100% ML, 1.00 PP). Based on the updated phylogenetic tree of Magnaporthales, Ceratosphaerella castillensis (generic type) and C. rhizomorpha formed a clade within Ophioceras and morphologically resemble Ophioceras. Therefore, Ceratosphaerella is synonymized under Ophioceras. The phylogenetic relationships of Ophioceras are discussed in relation to morphological similarities of genera in Magnaporthales. The generic circumscription of Ophioceras is emended.

Reappraisal of Immotthia in Dictyosporiaceae, Pleosporales: Introducing Immotthia bambusae sp. nov. and Pseudocyclothyriella clematidis comb. et gen. nov. Based on Morphology and Phylogeny.

Immotthia is a poorly known genus, and currently, no DNA sequence data are available to ascertain its proper phylogenetic placement and evolutionary relationships with other bitunicate fungi. To date, there are only two species accepted in the genus. During our ongoing research study of bambusicolous fungi in southwest China and Thailand, a fungus associated with stromata of Hypoxylon sp. was found on dead bamboo culms in Loei Province, Thailand. Preliminary morphological identification revealed that the fungal collection belongs to Immotthia. A novel species, Immotthia bambusae, is introduced herein based on a comparison of morphological characteristics with the type specimen of I. hypoxylon (≡ Amphisphaeria hypoxylon Ellis and Everh.), a synonym of I. atrograna (Cooke and Ellis) M. E. Barr. Phylogenetic analyses of a concatenated ITS, LSU, SSU, and TEF1-α DNA sequence matrix showed that Immotthia belongs to Dictyosporiaceae, Pleosporales. Despite I. bambusae strains constituting a supported subclade, they are nested with the genus Pseudocoleophoma. Pseudocoleophoma clematidis is morphologically different from all other Pseudocoleophoma species, while its conidial characteristics are similar to Cyclothyriella. Multigene phylogenetic analyses showed that P. clematidis formed a clade basal to Immotthia, separated from Pseudocoleophoma with strong statistical support. Therefore, we introduce a monotypic genus, Pseudocyclothyriella Phukhams. and Phookamsak, gen. nov. to accommodate the single species, Pseudocyclothyriella clematidis (Phukhams. and K. D. Hyde) Phukhams. and Phookamsak, comb. nov. Detailed descriptions, color micrographs, and phylogenetic trees to show the placement of the new taxa are provided. In addition, an updated taxonomic treatment of the genera Immotthia and Pseudocyclothyriella is also provided based on the study of the type materials and phylogeny generated from DNA sequence data.

Cultivation of Mushrooms and Their Lignocellulolytic Enzyme Production Through the Utilization of Agro-Industrial Waste.

A large amount of agro-industrial waste is produced worldwide in various agricultural sectors and by different food industries. The disposal and burning of this waste have created major global environmental problems. Agro-industrial waste mainly consists of cellulose, hemicellulose and lignin, all of which are collectively defined as lignocellulosic materials. This waste can serve as a suitable substrate in the solid-state fermentation process involving mushrooms. Mushrooms degrade lignocellulosic substrates through lignocellulosic enzyme production and utilize the degraded products to produce their fruiting bodies. Therefore, mushroom cultivation can be considered a prominent biotechnological process for the reduction and valorization of agro-industrial waste. Such waste is generated as a result of the eco-friendly conversion of low-value by-products into new resources that can be used to produce value-added products. Here, we have produced a brief review of the current findings through an overview of recently published literature. This overview has focused on the use of agro-industrial waste as a growth substrate for mushroom cultivation and lignocellulolytic enzyme production.

Bioprocess for Production, Characteristics, and Biotechnological Applications of Fungal Phytases.

Phytases are a group of enzymes that hydrolyze the phospho-monoester bonds of phytates. Phytates are one of the major forms of phosphorus found in plant tissues. Fungi are mainly used for phytase production. The production of fungal phytases has been achieved under three different fermentation methods including solid-state, semi-solid-state, and submerged fermentation. Agricultural residues and other waste materials have been used as substrates for the evaluation of enzyme production in the fermentation process. Nutrients, physical conditions such as pH and temperature, and protease resistance are important factors for increasing phytase production. Fungal phytases are considered monomeric proteins and generally possess a molecular weight of between 14 and 353 kDa. Fungal phytases display a broad substrate specificity with optimal pH and temperature ranges between 1.3 and 8.0 and 37-67°C, respectively. The crystal structure of phytase has been studied in Aspergillus. Notably, thermostability engineering has been used to improve relevant enzyme properties. Furthermore, fungal phytases are widely used in food and animal feed additives to improve the efficiency of phosphorus intake and reduce the amount of phosphorus in the environment.

Formation of 1-octen-3-ol from Aspergillus flavus conidia is accelerated after disruption of cells independently of Ppo oxygenases, and is not a main cause of inhibition of germination.

Eight-carbon (C8) volatiles, such as 1-octen-3-ol, are ubiquitous among fungi. They are the volatiles critical for aroma and flavor of fungi, and assumed to be signals controlling germination of several fungi. In this study, we found that intact Aspergillus flavus conidia scarcely synthesized C8 volatiles but repeated freeze-thaw treatment that made the cell membrane permeable promoted (R)-1-octen-3-ol formation. Loss or down regulation of any one of five fatty acid oxygenases (PpoA, PpoB, PpoC, PpoD or lipoxygenase) hypothesized contribute to 1-octen-3-ol formation had little impact on production of this volatile. This suggested that none of the oxygenases were directly involved in the formation of 1-octen-3-ol or that compensatory pathways exist in the fungus. Germination of the conidia was markedly inhibited at high density (1.0 × 10(9)spores mL(-1)). It has been postulated that 1-octen-3-ol is an autoinhibitor suppressing conidia germination at high density. 1-Octen-3-ol at concentration of no less than 10 mM was needed to suppress the germination while the concentration of 1-octen-3-ol in the suspension at 1.0 × 10(9) mL(-1) was under the detection limit (<1 µM). Thus, 1-octen-3-ol was not the principal component responsible for inhibition of germination. Instead, it was evident that the other heat-labile factor(s) suppressed conidial germination.

Characterization of volatile compounds in Astraeus spp.

Astraeus spp. is consumed in several regions of Southeast Asia. C(8) compounds, including 1-octen-3-ol, are the main volatile compounds in fresh Astraeus spp. Other compounds typical of edible mushrooms, such as terpenoids and sulfur-containing compounds, were not detected in fresh Astraeus spp. The amounts of fatty acids, including linoleic acid, substantially decreased after homogenization of fresh Astraeus spp. This high metabolic activity possibly correlates with formation of the C(8) volatiles. Heating the mushrooms produced cyclohexene compounds, 2-n-pentyl-furan, furanyl compounds, and benzaldehyde.

Volatile oxylipins and related compounds formed under stress in plants.

Plants form volatile oxylipins and related compounds under stress. Some of them are important flavor chemicals and give big impact on the flavor quality of food made from plant materials. They are also involved in defense responses of plants against pathogens and herbivores. Furthermore, in some instances, they cause harmful effects on plants themselves. Because of these significances of volatile oxylipins and related compounds, demands to perform comprehensive analyses of these compounds are increasing. In this chapter, we describe the simple but efficient procedures to reveal profiles of volatile oxylipins and related compounds by using HPLC and GC-MS. They are simple and can be performed in biochemical laboratories equipped with common facilities.

Development of a screening system for the evaluation of soybean volatiles.

Flavor properties are important factors of soybean seeds in their utilization as food materials. In order to isolate novel varieties and mutants of soybean having preferable flavor properties, a simple and efficient screening system was established using an automated headspace sampler coupled to gas chromatography. With this system, five major volatile compounds were analyzed within 12 min. By applying this screening system to 626 soybean varieties collected worldwide, we isolated four soybean varieties that showed unique compositions of volatile compounds. Through biochemical analysis, it was found that the uniqueness of three of them was possibly independent of lipoxygenase enzyme, and thus perhaps this screening system can expand the subject of flavor properties beyond lipoxygenase and thus be useful in discovering new types of soybeans.