Brown Rot Fungus Fomitopsis Pinicola Research Articles

Mycelium-based biomimetic composite structures as a sustainable leather alternative

Fungi, the main destructors of bio-waste, are opening up a new field for bio-based materials that contribute to a more eco-friendly future. In the fashion industry, using mycelium as a substitute for leather seems particularly promising. However, research has not yet been able to imitate the properties of leather. To this end, the mycelium of various white-rot fungi (Fomesfomentarius, Pleurotuseryngii, Trametesversicolor, and, as a control, the brown-rot fungus Fomitopsispinicola) was characterised. The biomimetic techno-pull approach was used to find suitable reinforcements like needle felts or woven fabrics made from regenerated cellulose fibres for the mycelium that increases tensile strength. Two methods to produce composite materials were tested: mycelial pulp grown on a liquid medium and mycelium grown directly on a reinforcement soaked with the same liquid. Overall, mechanical characteristics from purely cultured mycelium will not necessarily reflect the same characteristics of the fungus in an embedded fibre reinforcement. Nevertheless, both methods have resulted in promising materials, with F.fomentarius pulp and T.versicolor on needle felt, outperforming the tensile strength of artificial leather and Reishi™ from MycoWorks.

Complete mitochondrial genome sequence of the Agaricomycetes brown rot fungus Fomitopsis pinicola isolate FBCC1181.

The mitochondrial genome of the brown rot fungus Fomitopsis pinicola isolate FBCC 1181 is a 66.5 kbp circular chromosome. It contains 64 predicted genes, including a set typical for Basidiomycota Agaricomycetes mitogenomes. Introns of cox and cob genes contain several homing endonucleases of both LAGLIDADG and GIY-YIG types.

Decolorization and Transformation of Synthetic Dye Methylene Blue by Brown-Rot Fungus <i>Fomitopsis pinicola</i>

Methylene blue (MB) is a synthetic dye widely used in industries that is difficult to degrade in the environment due to its stability. Brown-rot fungus Fomitopsis pinicola is an organism that is known to be able to degrade some organic pollutants such as DDT, aldrin, dieldrin, and methyl orange dye. This study aimed to explore the ability of F. pinicola on MB biodecolorization, to identify metabolites and propose a biodecolorization pathway. F. pinicola was evaluated for MB biodecolorization on PDA agar and PDB liquid media. The metabolites were determined by using LCTOF/MS. The PDA agar medium's largest index decolorization (ID) was 0.915%. The MB decolorization in liquid PDB medium showed the highest percentage of decolorization of 92.56% at MB concentration of 100 mg/L after 14-days incubation. The analysis using LC-TOF/MS showed metabolites from MB biodecolorization, namely 3-amino-7-(methylamino) phenothiazin-5-ium (Azure C), 3-(dimethylamino)-7-(methylamino) phenothiazin-5-ium (Azure B), and 3,7-bis(dimethylamino)-4aH-phenothiazin-5-one. The MB degradation pathway was proposed through demethylation and oxidation reactions based on the detected product metabolites. These results indicated that F. pinicola is a suitable agent for biodecolorization of MB, and can potentially be used for bioremediation of MB waste in the environment.

Role of Fe2+-dependent Reaction in Biodecolorization of Methyl Orange by Brown-rot Fungus Fomitopsis pinicola

The involvement of Fenton reaction on biodegradation of methyl orange (MO) by brown-rot fungus Fomitopsis pinicola was investigated based on Fe2+-dependent reaction. The degradation of MO (final concentration 75 mg/L) was performed in mineral salt media with and without Fe2+ with incubation period at 0, 7, 14, 21, and 28 days. Degradation analysis was performed using UV-Vis Spectrophotometer and LC-TOF/MS. F. pinicola decolorized MO in a medium containing Fe2+ and a medium that lacked the mineral, at percentages of 89.47% and 80.08%, respectively. The optimum decolorization occurred after 28 days of incubation with the fungus on the presence of Fe2+, indicated that the presence of Fe2+ enhanced MO degradation with assumed to correlate with Fenton reaction. Two metabolites were detected through the LC-TOF/MS analysis, namely 4-(2-(4-(dimethyliminio)-2-hydroxycyclohexa-2,5-dien-1-ylidene) hydrazinyl) phenolate (m/z 258, RT: 1.28 min, compound 1) and 4-(2-(4-(dimethyliminio) cyclohexa-2,5-dien-1-ylidene) hydrazinyl) benzenesulfonate (m/z 391, RT: 2.70 min, compound 2). Compound 1 was a transformation product of hydroxylation and methylation, compound 2 was a product of dehydroxylation and desulfonation. This study indicated that the transformation of the metabolite structures was involved hydroxyl radical (OH.) and enzymatic mechanisms, which involved Fe2+-dependent reaction.

Retracted and Republished from: "Substrate-Specific Differential Gene Expression and RNA Editing in the Brown Rot Fungus Fomitopsis pinicola"

Wood-decaying fungi tend to have characteristic substrate ranges that partly define their ecological niche. Fomitopsis pinicola is a brown rot species of Polyporales that is reported on 82 species of softwoods and 42 species of hardwoods. We analyzed gene expression levels of F. pinicola from submerged cultures with ground wood powder (sampled at 5 days) or solid wood wafers (sampled at 10 and 30 days), using aspen, pine, and spruce substrates (aspen was used only in submerged cultures). Fomitopsis pinicola expressed similar sets of wood-degrading enzymes typical of brown rot fungi across all culture conditions and time points. Nevertheless, differential gene expression was observed across all pairwise comparisons of substrates and time points. Genes exhibiting differential expression encode diverse enzymes with known or potential function in brown rot decay, including laccase, benzoquinone reductase, aryl alcohol oxidase, cytochrome P450s, and various glycoside hydrolases. Comparing transcriptomes from submerged cultures and wood wafers, we found that culture conditions had a greater impact on global expression profiles than substrate wood species. These findings highlight the need for standardization of culture conditions in studies of gene expression in wood-decaying fungi. IMPORTANCE All species of wood-decaying fungi occur on a characteristic range of substrates (host plants), which may be broad or narrow. Understanding the mechanisms that allow fungi to grow on particular substrates is important for both fungal ecology and applied uses of different feedstocks in industrial processes. We grew the wood-decaying polypore Fomitopsis pinicola on three different wood species—aspen, pine, and spruce—under various culture conditions. We found that F. pinicola is able to modify gene expression (transcription levels) across different substrate species and culture conditions. Many of the genes involved encode enzymes with known or predicted functions in wood decay. This study provides clues to how wood-decaying fungi may adjust their arsenal of decay enzymes to accommodate different host substrates.

The addition effect of Pseudomonas aeruginosa on biodegradation of methyl orange dye by brown-rot fungus Fomitopsis pinicola

The addition effect of Pseudomonas aeruginosa on methyl orange (MO) biodecolorization by Fomitopsis pinicola had been investigated. P. aeruginosa was added into F. pinicola culture at 2, 4, 6, 8, 10 mL (1 mL × 5.05 χ 1012 CFU). The addition of 4 ml of P. aeruginosa showed the highest MO biodecolorization approximately 99.53%, while by F. pinicola only was 77.22% in potato dextrose broth (PDB) medium for 7 days incubation. Four metabolites were detected from MO degradation by mixed cultures such as 4-(4-(dimethylimino) hydroxy-cyclohexa-2,5-dien-1-ylidene) hydrazinyl) phenolate; 4-(hydroxy-4-iminio-cyclohexa-2,5-dien-1-ylidene) hydrazinyl) benzene sulfonate; 4-(4-(dimethylimino) methoxy-cyclohexa-2,5-dien-1-ylidene) hydrazinyl) hydroxy-benzene sulfonate; and 4-(hydroxy-4-iminio-cyclohexa-2,5-dien-1-ylidene) hydrazinil) hidroxy-benzene sulfonate. The mixed cultures transformed MO via three pathways: (1) desulfonylation, (2) demethylation, and (3) hydroxylation. These results indicated that P. aeruginosa can enhance MO biodecolorization by F. pinicola.

Synergistic interaction of a consortium of the brown-rot fungus Fomitopsis pinicola and the bacterium Ralstonia pickettii for DDT biodegradation

1,1,1-Trichloro-2,2-bis (4-chlorophenyl) ethane (DDT) is a toxic and recalcitrant pesticide that has been greatly used to eradicate malaria mosquitos since the 1940s. However, the US Environmental Protection Agency banned and classified DDT as priority pollutants due to its negative impact on wildlife and human health. Considering its negative effects, it is necessary to develop effective methods of DDT degradation. A synergistic interaction of a consortium consisting of the brown-rot fungus Fomitopsis pinicola and the bacterium Ralstonia pickettii was adopted to degrade DDT. For the microbial consortia, F. pinicola was mixed with R. pickettii at 1, 3, 5, 7 and 10 ml (1 ml ≈ 1.44 × 1013 CFU) in a potato dextrose broth (PDB) medium to degrade DDT throughout the seven days incubation period. The degradation of DDT by only the fungus F. pinicola was roughly 42%, while by only R. pickettii was 31%. The addition of 3 ml of R. pickettii into F. pinicola culture presented appropriate optimization for efficient DDT degradation at roughly 61%. The DDT transformation pathway by co-inoculation of F. pinicola and R. pickettii showed that DDT was converted to 1,1-dichloro-2,2-bis(4-chlorophenyl) ethane (DDD), further transformed to 1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene (DDE), and then ultimately transformed to 1-chloro-2,2-bis(4-chlorophenyl) ethylene (DDMU). These metabolites are less toxic than DDT. This research showed that R. picketti synergistically interacts with F. pinicola by enhancing DDT degradation.

Enzyme Activity Profiles Produced on Wood and Straw by Four Fungi of Different Decay Strategies.

Four well-studied saprotrophic Basidiomycota Agaricomycetes species with different decay strategies were cultivated on solid lignocellulose substrates to compare their extracellular decomposing carbohydrate-active and lignin-attacking enzyme production profiles. Two Polyporales species, the white rot fungus Phlebia radiata and brown rot fungus Fomitopsis pinicola, as well as one Agaricales species, the intermediate “grey” rot fungus Schizophyllum commune, were cultivated on birch wood pieces for 12 weeks, whereas the second Agaricales species, the litter-decomposing fungus Coprinopsis cinerea was cultivated on barley straw for 6 weeks under laboratory conditions. During 3 months of growth on birch wood, only the white rot fungus P. radiata produced high laccase and MnP activities. The brown rot fungus F. pinicola demonstrated notable production of xylanase activity up to 43 nkat/mL on birch wood, together with moderate β-glucosidase and endoglucanase cellulolytic activities. The intermediate rot fungus S. commune was the strongest producer of β-glucosidase with activities up to 54 nkat/mL, and a notable producer of xylanase activity, even up to 620 nkat/mL, on birch wood. Low lignin-attacking but moderate activities against cellulose and hemicellulose were observed with the litter-decomposer C. cinerea on barley straw. Overall, our results imply that plant cell wall decomposition ability of taxonomically and ecologically divergent fungi is in line with their enzymatic decay strategy, which is fundamental in understanding their physiology and potential for biotechnological applications.

Review of Chemical Constituents and Pharmacology of Brown-Rot Fungus Fomitopsis Pinicola

Fomitopsis pinicola is a well-known medicinal mushroom commonly used in Chinese traditional medicine and Korean folk medicine for many years, and it is believed as a non-toxic medicinal mushroom that has many clinical effects on the human body with no sub-acute toxic impacts. In recent years, the use of medicinal mushrooms is enhanced greatly. Several different principal chemical constituents are present in the fruiting bodies of F.pinicola . These principal compounds were extracted from fruiting bodies of F.pinicola , have medicinal qualities and give different pharmacological impacts in the human body. The motive of the current review paper was to intricate the principal chemical constituents that are extracted from F.pinicola fruiting bodies and demonstrate the pharmacological effects of these extracts. F.pinicola extracts have no toxic side effects on vital organs such as liver, kidney, heart, spleen, pancreas, lungs and brain. The present study describes that both low molecular weight and high molecular weight compounds that are present in the fruiting bodies of F.pinicola . This included many useful enzymes, steroids, triterpenes and triterpenes derivatives, anti-tumor active constituents, and health beneficial nutritious compounds. These principal compounds showed important medicinal effects on the human body by providing a shield effect to the internal organs against diseases and also heal the damaged tissues and organs. The pharmacological effects of F.pinicola active constituents include anti-tumor, anti-diabetic, anti-hyperlipidemic by controlling obesity, anti-oxidant effect, anti-microbial, anti-inflammatory and also showed an industrial waste treatment effect. Keywords: Fomitopsis pinicola, Chinese traditional medicine, chemical constituents, pharmacological effects, waste treatment. DOI : 10.7176/JNSR/10-2-07 Publication date: January 31 st 2020

Substrate-Specific Differential Gene Expression and RNA Editing in the Brown Rot Fungus Fomitopsis pinicola.

Wood-decaying fungi tend to have characteristic substrate ranges that partly define their ecological niche. Fomitopsis pinicola is a brown rot species of Polyporales that is reported on 82 species of softwoods and 42 species of hardwoods. We analyzed the gene expression levels and RNA editing profiles of F. pinicola from submerged cultures with ground wood powder (sampled at 5 days) or solid wood wafers (sampled at 10 and 30 days), using aspen, pine, and spruce substrates (aspen was used only in submerged cultures). Fomitopsis pinicola expressed similar sets of wood-degrading enzymes typical of brown rot fungi across all culture conditions and time points. Nevertheless, differential gene expression and RNA editing were observed across all pairwise comparisons of substrates and time points. Genes exhibiting differential expression and RNA editing encode diverse enzymes with known or potential function in brown rot decay, including laccase, benzoquinone reductase, aryl alcohol oxidase, cytochrome P450s, and various glycoside hydrolases. There was no overlap between differentially expressed and differentially edited genes, suggesting that these may provide F. pinicola with independent mechanisms for responding to different conditions. Comparing transcriptomes from submerged cultures and wood wafers, we found that culture conditions had a greater impact on global expression profiles than substrate wood species. In contrast, the suites of genes subject to RNA editing were much less affected by culture conditions. These findings highlight the need for standardization of culture conditions in studies of gene expression in wood-decaying fungi.IMPORTANCE All species of wood-decaying fungi occur on a characteristic range of substrates (host plants), which may be broad or narrow. Understanding the mechanisms that enable fungi to grow on particular substrates is important for both fungal ecology and applied uses of different feedstocks in industrial processes. We grew the wood-decaying polypore Fomitopsis pinicola on three different wood species, aspen, pine, and spruce, under various culture conditions. We examined both gene expression (transcription levels) and RNA editing (posttranscriptional modification of RNA, which can potentially yield different proteins from the same gene). We found that F. pinicola is able to modify both gene expression and RNA editing profiles across different substrate species and culture conditions. Many of the genes involved encode enzymes with known or predicted functions in wood decay. This work provides clues to how wood-decaying fungi may adjust their arsenal of decay enzymes to accommodate different host substrates.

Abilities of Co-cultures of Brown-Rot Fungus Fomitopsis pinicola and Bacillus subtilis on Biodegradation of DDT.

DDT (1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane) is one of the pesticides that are hazardous for the environment and human health. Effective environmental-friendly treatment using co-cultures of fungi and bacteria is needed. In this study, the bacteria Bacillus subtilis at various volumes of 1, 3, 5, 7, and 10mL (1mL≈6.7×108CFU) were mixed into 10mL of the brown-rot fungus Fomitopsis pinicola culture for degrading DDT during a 7-days incubation period. DDT was degraded by approximately 42% by F. pinicola during the 7-days incubation period. The addition of 10mL of B. subtilis into F. pinicola culture showed the highest DDT degradation of approximately 86% during the 7-days incubation period. DDD (1,1-dichloro-2,2-bis(4-chlorophenyl)ethane), DDE (1,1-dichloro-2,2-bis(4-chlorophenyl)ethylene), and DDMU (1-chloro-2,2-bis(4-chlorophenyl)ethylene) were detected as metabolic products from DDT degradation by co-cultures of F. pinicola and B. subtilis. Transformation pathway was proposed in which DDT was transformed into three pathways as follows: (1) dechlorination to DDD, (2) dehydrochlorination to DDE, and (3) formation of DDMU.

Enhancement of β-Glucosidase Activity from a Brown Rot Fungus Fomitopsis pinicola KCTC 6208 by Medium Optimization

β-Glucosidase, which hydrolyzes cellobiose into two glucoses, plays an important role in the process of saccharification of the lignocellulosic biomass. In this study, we optimized the activity of β-glucosidase of brown-rot fungus Fomitopsis pinicola KCTC 6208 using the response surface methodology (RSM) with various concentrations of glucose, yeast extract and ascorbic acid, which are the most significant nutrients for activity of β-glucosidase. The highest activity of β-glucosidase was achieved 3.02% of glucose, 4.35% of yeast extract, and 7.41% ascorbic acid where ascorbic acid was most effective. The maximum activity of β-glucosidase predicted by the RSM was 15.34 U/mg, which was similar to the experimental value 14.90 U/mg at the 16th day of incubation. This optimized activity of β-glucosidase was 23.6 times higher than the preliminary activity value, 0.63 U/mg, and was also much higher than previous values reported in other fungi strains. Therefore, a simplified medium supplemented with a cheap vitamin source, such as ascorbic acid, could be a cost effective mean of increasing β-glucosidase activity.

Optimization of Endoglucanase Production from Fomitopsis pinicola Mycelia

The culture conditions to maximize the production of endoglucanase (EC 3.2.1.4) from the brown rot fungus Fomitopsis pinicola MKACC 54347 mycelia were investigated. Among the tested media for endoglucanase production, Mandel's mineral salts medium (MSM; 1% cellulose, 0.1% peptone, 0.14% , 0.03% urea, 0.2% , 0.03% , 0.03% , and 0.1% trace metal solution (19.8 mM , 13.0 mM , 12.2 mM , and 15.4 mM )) produced the highest activity of the enzyme. To optimize the medium composition for enzyme activity, the effects of various carbon, nitrogen, phosphorus, and inorganic sources were investigated in MSM. Maximal enzyme production was accomplished using a medium containing 2% carboxymethyl cellulose (CMC), 2% yeast extract, 0.2% , 0.03% , and 0.3% trace metal solution. Different physiological conditions, like incubation period and temperature, were also examined to assess their influence on enzyme production. Enzyme production from F. pinicola reached its highest level after cultivation for 8 days at . Nondenaturing polyacrylamide gel electrophoresis (PAGE), followed by the endoglucanase activity staining using CMC as the substrate, was performed to identify the endoglucanase under the culture conditions studied. Zymogram analysis of the culture supernatant revealed an endoglucanase band with a molecular mass of 52 kDa. The optimum pH and temperature for enzyme activity were and pH 5.0, respectively.

Degradation of cellulose by the major endoglucanase produced from the brown-rot fungus Fomitopsis pinicola

An endoglucanase that is able to degrade both crystalline and amorphous cellulose was purified from the culture filtrates of the brown-rot fungus Fomitopsis pinicola grown on cellulose. An apparent molecular weight of the purified enzyme was approximately 32 kDa by SDS-PAGE analysis. The enzyme was purified 11-fold with a specific activity of 944 U/mg protein against CMC. The partial amino acid sequences of the purified endoglucanase had high homology with endo-beta-1,4-glucanase of glycosyl hydrolase family 5 from other fungi. The K(m) and K(cat)values for CMC were 12 mg CMC/ml and 670/s, respectively. The purified EG hydrolyzed both cellotetraose (G4) and cellopentaose (G5), but did not degrade either cellobiose (G2) or cellotriose (G3).

Oxalate production by wood-rotting fungi growing in toxic metal-amended medium

In this report, we have identified oxalic acid as an important metabolite elaborated in the response of wood-rotting fungi to toxic metal stress. The formation of oxalate crystals by white rot fungi ( Bjerkandera fumosa, Phlebia radiata and Trametes versicolor) and the brown rot fungus Fomitopsis pinicola, grown on media containing high levels of toxic metal ions has been visualized using scanning electron microscopy (SEM) with energy-dispersive X-ray micro-analysis (EDXA) and HPLC. There were no significant differences between the growth of controls (metal-free) and on the 0.5% CaCO 3, Co 3(PO 4) 2 or Zn 3(PO 4) 2-amended plates. ZnO inhibited the growth of all strains. Crystals were not detected in Zn 3(PO 4) 2-amended plates. The four examined strains displayed the formation of crystals on ZnO, Co 3(PO 4) 2 and CaCO 3-amended plates.

Mineralization of alachlor by lignin-degrading fungi

White rot fungi were able to mineralize the aromatic ring carbon of alachlor to CO2. After 122 days, 14 and 12% of the alachlor that was initially present in malt extract cultures supplemented with a wood substrate was mineralized at room temperature by Ceriporiopsis subvermispora and Phlebia tremellosa, respectively. Although Phanerochaete chrysosporium mineralized alachlor at 25 degrees C, it did so more slowly than the other two white rot fungi. The brown rot fungus Fomitopsis pinicola did not mineralize alachlor.

Hyphal Deterioration in Ganoderma Applanatum

Older conks of Ganoderma applanatum were observed to have a whitened appearance in the context and older tramal tissues and were similar in this respect to white-rotted wood. Hyphae taken from whitened areas of the context became greatly expanded when mounted in 2% KOH and showed signs of deterioration when viewed microscopically. Holes and spore wall erosion were present in basidiospores trapped in the current year's pore layer. Nitrogen levels and relative densities were lower in the whitened areas than in the brown current year's growth. Comparisons of relative densities between older and newer tissues of the brown rot fungus Fomitopsis pinicola indicated little difference. However, as in G. applanatum, N levels were lower in older tissues of F. pinicola. Recycling of nutrients from older nonfunctional tissues in G. applanatum and F. pinicola conks may be a mechanism whereby nutrients are transferred to the current year's growth. However, the whitening phenomenon and reduction of hyphal number was peculiar only to some older G. applanatum conks. Hyphal decay (autolysis) may be a mechanism of N utilization in conks of this fungus.

Transmission electron microscope observations of brown rot caused by Fomitopsis pinicola with respect to osmiophilic particles

Changes in pinewood tracheids during decay by the brown rot fungus Fomitopsis pinicola were observed in the transmission electron microscope for a cultivation period of 80 days. It was found that F. pinicola normally caused degradation of carbohydrates over the entire cell wall with the cell wall contours maintained, but more rarely was capable of a complete degradation of the cell walls including the middle lamella and the cell corners. This indicates that some hyphae may have a ligninolytic enzyme system. The occurrence of osmiophilic particles in the protoplasm of hyphae and in wood cell walls was observed from early decay stages. They were randomly distributed in the wood and their distribution, being an indicator of carbohydrate depolymerization, permits this type of decay to be observed in the transmission electron microscope. The origin and function of the osmiophilic particles is discussed with respect to what is known about the enzymes of wood decay. Furthermore, the curve of weight loss is discussed in connection with earlier findings concerning wood decay and with the appearance of the osmiophilic particles.