Micromorphological characteristics of decayed wood and laccase produced by the brown-rot fungus <Emphasis Type="Italic">Coniophora puteana</Emphasis>

Microscopic examination showed the cell wall decay pattern produced by the brown-rot fungus Coniophora puteana to be different from the degradation pattern known to be typical for brown-rot fungi. Erosion and thinning of cell walls in patterns considered to be characteristic of white-rot decay were observed. In particular, the fungal strain COP 20242 degraded secondary cell wall layers extensively, and also degraded lignin-rich middle lamellae. Some strains of C. puteana produced soft-rot type cavities in the S2 layer. All strains of C. puteana employed in the present work showed a positive reaction to tannic acid in the Bavendamm test, indicating the production of laccase. Microscopic and enzymatic studies provided evidence to suggest that the wood decay by C. puteana is unique both in terms of micromorphological and enzymatic patterns of cell wall degradation. This is because brown-rot fungi are not generally known to form cavities in the cell walls or to produce lignin-degrading enzymes. These observations suggest that lignin degradation capacity of brown-rot fungi may be greater than previously considered.     

Bioremediation of CCA-treated wood by brown-rot fungi <Emphasis Type="Italic">Fomitopsis palustris</Emphasis>, <Emphasis Type="Italic">Coniophora puteana</Emphasis>, and <Emphasis Type="Italic">Laetiporus sulphureus</Emphasis>

This study evaluated oxalic acid accumulation and bioremediation of chromated copper arsenate (CCA)-treated wood by three brown-rot fungi Fomitopsis palustris, Coniophora puteana, and Laetiporus sulphureus. The fungi were first cultivated in a fermentation broth to accumulate oxalic acid. Bioremediation of CCA-treated wood was then carried out by leaching of heavy metals with oxalic acid over a 10-day fermentation period. Higher amounts of oxalic acid were produced by F. palustris and L. sulphureus compared with C. puteana. After 10-day fermentation, oxalic acid accumulation reached 4.2 g/l and 3.2 g/l for these fungi, respectively. Fomitopsis palustris and L. sulphureus exposed to CCA-treated sawdust for 10 days showed a decrease in arsenic of 100% and 85%, respectively; however, C. puteana remediation removed only 18% arsenic from CCA-treated sawdust. Likewise, chromium removal in F. palustris and L. sulphureus remediation processes was higher than those for C. puteana. This was attributed to low oxalic acid accumulation. These results suggest that F. palustris and L. sulphureus remediation processes can remove inorganic metal compounds via oxalic acid production by increasing the acidity of the substrate and increasing the solubility of the metals.An erratum to this article can be found at     

Relationship between degradation of wood and production of H2O2-producing or one-electron oxidases by brown-rot fungi

Summary The one-electron oxidation activity of brown-rot fungi was determined by measuring ethylene production from KTBA. Ethylene production was related to degradation of lignin, cellulose, and wood itself. Extracellular protein that catalyzed oxidation of KTBA was isolated from wood-containing cultures. This protein required H₂O₂ for KTBA oxidation. It was also found to oxidize NADH, producing H₂O₂ via in the presence of O₂. The protein showed little phenol-oxidase activity under conditions giving high activity against KTBA. The results indicate that partially reduced oxygen may play a role in the initial degradation of the cellulose and lignin in wood in cultures of brown-rot fungi.     

The action of isolated brown-rot cell-free culture filtrate,H2O2−Fe++, and the combination of both on wood

Summary Transmission electron microscopy observations of hemlock sawdust that had been incubated with concentrated filtrate from Postia placenta showed that wood was degraded despite the absence of weight loss and release of reducing groups. These results are contrary to the prevailing views that enzyme preparation from brown-rot fungi are unable to degrade wood.Maintained at Madison, WI, in cooperation with the University of Wisconsin     

In vitro decay of Aextoxicon punctatum and Fagus sylvatica woods by white and brown-rot fungi

Summary The in vitro decay of Aextoxicon punctatum and Fagus sylvatica wood by the fungi Trametes versicolor, Ganoderma australe, Phlebia chrysocrea and Lentinus cyathiformis was studied by the agar-block method, and then the decayed woods were analyzed by chemical and spectroscopic techniques. The results demonstrated the strong resistance of the A. punctatum wood to the brown-rot fungus L. cyathiformis; the resistance might be related to the low S/G lignin ratio in this Austral hardwood. Wood decay by the Austral white-rot fungi G. australe and P. chrysocrea was rather limited, and preferential degradation of lignin was not produced although all the fungi studied increased wood digestibility. The most characteristic white and brown-rot decay patterns were observed during the in vitro decay with T. versicolor and L. cyathiformis, respectively. Trametes versicolor caused high weight losses and reduced the lignin content of the wood, whereas L. cyathiformis produced a preferential removal of xylan. No important changes in the solid-state ¹³C NMR spectra were observed after wood degradation by T. versicolor, but this technique evidenced an increase in aromatic carbon by L. cyathiformis. This increase was higher than that found in the Klason lignin content, suggesting the presence of altered lignin fractions in the brown-rotted wood.The authors are indebted to Prof. H. D. Lüdemann for the facilities at the Institut für Biophysik und physikalische Biochemie (Regensburg), to A. Navarrete (INIA, Madrid) for her collaboration, and to C. F. Warren (ICE, Alcalá de Henares) for her linguistic assistance. The computer program for spectra treatment was developed by G. Almendros (Centro de Ciencias Medioambientales, CSIC, Madrid). This investigation has been funded by the Spanish Biotechnology Program (Grant BIO88-0185)     

Methods of inoculating Acer spp., Populus tremuloides, and Fagus grandifolia logs for commercial spalting applications

One of the most promising wood value-added processes currently under development is spalting, where pigment is added to wood via fungal colonization. Previous studies have shown laboratory level spalting to be achievable and highly predictable. However, large-scale spalting for potential commercial applications introduces a substantial number of additional variables which impact the spalting process. To test the potential of commercial-scale spalting, Acer saccharum, Fagus grandifolia, and Populus tremuloides logs were inoculated with multiple known spalting fungi utilizing both liquid spray cultures and live dowel pin cultures. Many of the fungi that successfully produce spalting in small, sterile cultures also produced significant amounts in large logs, with many spalting patterns identical to those found in small-scale testing. Pairings of Trametes versicolor/Scytalidium cuboideum and Xylaria polymorpha/Xylaria polymorpha (different isolates) produced significant amounts of zone lines. In addition, the method of inoculation impacted the amount of spalting: more zone lines were produced when fungi were introduced via plugs, while more stain was produced when liquid cultures were sprayed onto the logs. These results indicate that many of the standard spalting fungi are suitable for large-scale applications; however, the inoculation method appears to be a vital component for successful spalting under a restricted time schedule.     

Lignin-degrading activity of edible mushroom <Emphasis Type="Italic">Strobilurus ohshimae</Emphasis> that forms fruiting bodies on buried sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) twigs

Strobilurus ohshimae is an edible mushroom, and it specifically forms its fruiting bodies on buried sugi (Cryptomeria japonica) twigs. In this research, we studied lignindegrading activity of S. ohshimae. We isolated 18 strains of S. ohshimae from various regions of Japan, and determined their lignin degradation rates on sugi wood meal medium. All the strains of S. ohshimae degraded approximately 6%–12% of sugi lignin in 30 days, and these lignin degradation rates were 1.5–3 times higher than those of Trametes versicolor, which is a typical lignin-degrading fungus. Among the three main lignin-degrading enzymes, activity of lignin peroxidase and manganese peroxidase was not observed, while 4340U/g of laccase was produced in 30 days. To investigate the effect of wood species on lignin degradation by S. ohshimae, the lignin degradation rate and laccase productivity on sugi wood meal medium were compared with those on beech (Fagus crenata). In T. versicolor, both lignin degradation rate and laccase productivity were higher on beech than on sugi. Conversely, in S. ohshimae, lignin degradation rate and laccase productivity were higher on sugi than on beech. Therefore, it was suggested that coniferous lignin is not always difficult to degrade for the fungi that inhabit softwood. Part of this article presented at the 54th Annual Meeting of the Japan Wood Research Society, Sapporo, August 2004     

Characterization of acetylated wood decayed by brown-rot and white-rot fungi

The objective of this study was to characterize the decay of acetylated wood due to brown-rot and white-rot fungi by analysis of chemical composition, X-ray measurements, and¹³C-NMR spectroscopy. The decay by brown-rot fungus became inhibited at a weight percent gain (WPG) due to acetylation of more than 10%, and the mass loss (LOSS) due to decay became zero at a WPG of about 20%. The LOSS due to white-rot fungus decreased slowly with the increase in WPG, reaching zero at a WPG of about 12%. The losses of lignin by brown-rot decay increased initially with the decrease in LOSS owing to the progressing acetylation and then decreased at a LOSS of less than 60%. Polysaccharides were more easily decomposed than lignin during the decay of acetylated wood due to brown-rot fungus. The losses of both components due to white-rot decay decreased as the LOSS decreased with progressing acetylation. The white-rot fungus tended to preferentially decompose the lignin during the decay of acetylated wood. The brown-rot fungus decomposed the cellulose in the crystalline region to a large degree when the LOSS was more than 40%, whereas the white-rot fungus decomposed the crystalline region and the noncrystalline region in acetylated wood to the same degree. The brown-rot fungus preferentially decomposed unsubstituted xylose units in acetylated wood and partly decomposed the mono-substituted xylose units. It was suggested that the mono- and disubstituted cellulose were partly decomposed by brown-rot fungus.This paper was presented at the 46th and 47th annual meetings of the Japan Wood Research Society at Kumamoto and Kochi in April 1996 and April 1997, respectively     

Environment-friendly short-term protection of palm wood against mould and rot fungi

Felled palm trunks are susceptible to fungi as long as their moisture content is above fibre saturation. During this period, palm wood has to be protected against mould and rot fungi. The study was aimed at testing environment-friendly organic acids for their protecting efficiency. Small samples of date palm (Phoenix dactylifera) and oil palm (Elaeis guineensis) wood were treated with weak organic acids and subsequently infected by moulds and wood-decay fungi. Short dipping of the samples in solutions of 5% acetic acid and propionic acid, respectively, protected all samples for two months from colonization by Aspergillus niger, Penicillium sp., Cladosporium sp. and by a natural infection. Boric acid (4%) used in practice for protection was ineffective. Decay tests with the white-rot fungus Pleurotus ostreatus, the brown-rot species Coniophora puteana and the soft-rot fungus Chaetomium globosum showed that both acids prevented most samples from fungal colonization for three weeks and reduced the decay considerably during two months.     

The genes encoding glycoside hydrolase family 6 and 7 cellulases from the brown-rot fungus <Emphasis Type="Italic">Coniophora puteana</Emphasis>

Four genes encoding glycoside hydrolase (GH) family 6 and 7 cellulases (cel6A, cel6B, cel7A, and cel7B) were obtained from the brown-rot fungus Coniophora puteana by genomic polymerase chain reaction (PCR) using consensus degenerate hybrid oligonucleotide primers (CODEHOPs) designated from the amino acid sequence of cellobiohydrolases (CBHs) from white-rot fungi. The nucleotide sequences of four genes showed high homology with basidiomycetes CBHs, suggesting the fi rst cloning of the genes encoding Cel6 and Cel7 from brown-rot fungi. PCR using CODEHOP pairs at the catalytic domain successfully amplifi ed both cel6A and cel6B, whereas only cel6A fragment was obtained using the primers including the carbohydrate-binding modules (CBMs), suggesting lack of CBM in Cel6B. Moreover, both cel7A and cel7B were amplified by the PCR using CODEHOP pairs at the catalytic domain, but not by those including CBM, suggesting the absence of Cel7 with CBM in the fungus. From these results, three of four cellulases from C. puteana may not carry CBM, which has an important role for the degradation of crystalline cellulose.     

A histological investigation of Oriental beech wood decayed by Pleurotus ostreatus and Trametes versicolor

Chemical, light and electron microscopic studies were carried out on wood of Oriental beech (Fagus orientalis Lipsky) decayed by the white‐rot fungi Pleurotus ostreatus and Trametes versicolor for 30, 60 and 120 days according to the modified European standard EN 113. Mass loss as well as lignin, cellulose and carbohydrate content were determined before and after fungal attack. There were no significant differences of wood mass loss and chemical composition between both fungi at the end of incubation. After each incubation period, small specimens were stained for microscopic studies. The micromorphology of fungal cell wall degradation was rather similar for both fungi. Both decreased the cell wall thickness to the same extent. The accumulation of hyphae as well as the rupture of cell walls was also similar. The occurrence of hyphae, cavities in the pits and vessel walls followed nearly the same patterns. The parenchyma cells were completely destroyed. Altogether, both fungi produced a simultaneous white rot in Oriental beech wood.     

Detection of early stages of wood decay by acoustic emission technique

Acoustic emissions of pine wood samples (Pinus sylvestris L.) previously infected with the soft-rot fungi (Chaetomium globosum Kunze) and brown-rot fungi (Coniophora puteana (Schum. ex Fr.) Karst.) were measured in the perpendicular to grain compression test, in the radial direction. It was found that even a minimum mass loss (below 1%) caused by enzymatic deterioration of the wood substance resulted in an increase in acoustic emission level in the compression test measured perpendicular to grain in the radial direction. A particularly sensitive indicator of wood enzymatic destruction were cumulative counts of acoustic emission related to the value of the applied compression load. Received 4 March 1997     

木材褐腐真菌泊氏孔菌属生态学研究

木材腐朽真菌在森林生态系统中起着关键的降解还原作用,是森林生态系统物质循环中不可缺少的重要组成部分.泊氏孔菌属的种类具有降解木材中的纤维素和半纤维素,造成木材褐色腐朽的功能,而混合于针叶林土壤中的褐腐残余物是针叶林生态系统更新所必不可少的.本文对中国地区泊氏孔菌属进行生态学的研究,结果表明树木腐烂程度与泊氏孔菌属的种类分布具有高度相关性,相关系数为0.885.寄主树木选择性的研究结果表明,泊氏孔菌属的种类主要生长在针叶树倒木上,有19种,占总数的86%,其中生长在云杉属和松属树木上的种类最多,而阔叶树中最适宜该属物种生长的树种是桦属和杨属树木.     

Study on biodegradated ability of thirteen edible fungi to straw

The biodegradated abilities of 13 edible fungi to straw were studied. The results showed that all the experimental fungi except Tricholama mongolicum had definite biodegradated abilities to the lignin and cellulose of straw. The Ideal fungus for straw degradation was screened out as Pleurotus ostreatus, which showed a higher degradation ability for lignin (17.86%) and lower degradation rate for cellulose (2.24%), with a Selection Factor (SF) of 7.97. The degradation rates of lignin and cellulose for other fungi ranged from 2.30% to 16.54% and 5.60% to 17.32%, respectively, and the SF was very low in range of 0.14 to 2.24.The ratio of colony‘s diameters to the color-zone (d1/d2) and SF are negative correlation, with a correlation coefficient of -0.1476.     

Degradation of lipophilic wood extractive constituents in Pinus sylvestris by the white-rot fungi Bjerkandera sp. and Trametes versicolor

The white-rot fungi Trametes versicolor and Bjerkandera spp. are among the most frequent decomposers of angiosperm wood in forest ecosystems and in wood products in service. Wood extractives have a major impact on wood properties and wood utilization. This work evaluated the ability of two white-rot fungal strains (Bjerkandera sp. strain BOS55 and T. versicolor strain LaVec94-6) to degrade the main lipophilic extractive constituents in Scots pine (Pinus sylvestris L.). The time course of wood decay and wood extractive degradation was monitored in stationary batch assays incubated for eight weeks. The strains tested eliminated high levels of total resin, 34 to 51% in two weeks. Wood triglycerides were the most readily degraded extractive components (over 93% elimination in only two weeks). Free fatty acids and resin acids, which are potential fungal inhibitors, were also rapidly decomposed by the fungal strains. Sterols were used more slowly, nonetheless, the fungal degradation of this extractive fraction ranged from 50 to 88% after four weeks. Received 19 March 1999     

Leachability,decay, and termite resistance of wood treated with metaborates

The formation of insoluble metaborates in wood was investigated by impregnating the wood with borax and metallic salts, after which their properties (e.g., leachability in running water and biological resistance) were evaluated. The solubility of three metaborates in acidic solutions was also evaluated. Double-diffusion treatment was carried out to form the precipitates of metaborates in sapwood specimens of Japanese cedar (Cryptomeria japonica) at room temperature. Water-saturated wood specimens were first impregnated by a saturated borax solution and then diffuse-penetrated with Zn²⁺, Ca²⁺, or Pb²⁺ solution. The precipitates of the three metaborates in the wood proved to be insoluble or hardly soluble in water by the leaching test. With the decay test using a brown-rot fungus (Fomitopsis palustris) and a white-rot fungus (Trametes versicolor) and with the termite test using a virulent subterranean termite (Coptotermes formosanus), the metaborate-treated woods showed generally good decay and termite resistance with negligible mass loss of the specimens. Particularly, the lead metaborate formed in the wood provided superb biological resistance against decay and termite attacks. In addition, the precipitates of these metaborates were found to be soluble in acidic solution, suggesting a way to remove these chemicals from wood when disposing of waste materials.     

Immunogold labeling of an extracellular substance producing hydroxyl radicals in wood degraded by brown-rot fungusTyromyces palustris

A fraction containing low-molecular-weight peptides that catalyzes redox reactions between electron donors and O₂ to produce ·OH, was partially purified from wood-decaying cultures of the brown-rot fungusTyromyces palustris. Polyclonal antibodies raised to the fraction were used for immunogold labeling of transverse sections of sapwood of spruce in various stages of degradation byT. palustris to demonstrate the cellular localization of the ·OH-producing substance. Initially, the wood cell wall was attacked primarily by fungal hyphae growing in the cell lumen. During the early stages of degradation, the gold label was localized in the fungal cytoplasm and cell wall and in the extracellular slime sheath surrounding the fungal cell wall. The gold label also was found throughout the wood cell wall, although the cell wall remained almost intact so long as the fungal hyphae remained in the lumen. Thus, the ·OH-producing substance is secreted by the hyphae into the lumen, and it diffuses through the S3 layer into the S2 layer and the middle lamella. The role of this ·OH-producing system in wood degradation byT. palustris is discussed.     

东北林区64种木材腐朽菌木材分解能力的研究

采用常规重量分析法，分别测定了64种木材腐朽菌对红松、青杨、白桦木块的木材分解能力，测定了受菌侵染46d后木材样品的重量损失百分率和木块的颜色变化。结果表明这些木腐菌的木材分解能力显著不同。采自长白山林区的厚黑层孔菌Nigrofomes castaneus(Imaz.)Teng是木材分解能力最强的褐腐菌、它分别引起白桦、青杨、红松木材样品的重量损失百分率为64．9％、52．2％和16．9％。白干酪菌（Tytomyces albidus)，灵芝（Ganoderma lucidum)、冬拟多孔菌（Polyporellus brumalis)、三色革裥菌（Lenzites tricolor)、棱孔菌（Favolus alveolaris)、朱红密孔菌（Pycnoporus cinnabarinus)、彩绒革盖菌（Coriolus versicolor)等是几种木材分解能力强和较强的白腐菌。     

Interaction between Epicoccum purpurascens and xylophagous basidiomycetes on wood blocks

Epicoccum purpurascens, a component of the bark micromycota, was evaluated as a potential biocontrol agent for three decay‐causing hymenomycetes frequently found in Buenos Aires city trees, Ganoderma platense, Inonotus rickii and Rigidoporus ulmarius. Treatments consisted of dual cultures of E. purpurascens and xylophagous fungi on box elder (Acer negundo) wood blocks. Two inoculation sequences were tested: (i) the biocontrol agent as the first microorganism inoculated followed by the pathogen; and (ii) the xylophagous fungus as the first microorganism inoculated followed by the biocontrol agent. After 3 months of incubation, percentage dry weight loss was calculated. When E. purpurascens was allowed to colonize wood blocks before the introduction of the xylophagous fungus, dry weight loss was significantly reduced in all instances. However, as a secondary colonizer, E. purpurascens mildly reduced the weight loss caused by G. platense, had no effect on degradation by I. rickii and increased wood weight loss by R. ulmarius.     

Response of maple sapwood to injury and infection

In sapwood challenge experiments in Acer rubrum, columns of discolouration initiated by wounding and inoculation with pioneer fungi (Cephalosporium sp., Phialophora sp.) were similar in size to untreated wounds. Inoculation with decay fungi (Pleurotus ostreatus, Trametes versicolor) produced larger columns of wound-initiated discolouration. The removal of bark around a bore wound caused a significantly larger column to form compared to the sum of the columns inititiated by separate wounds. Stage-I discoloured wood, not associated with obviously rotted wood, had concentrations of mobile cations and soluble phenols similar to sapwood. Stage-II discoloured wood, spatially associated with rotted wood, was frequently bounded by a chemically distinct boundary layer and the discoloured wood contained significantly greater concentrations of mobile cations and soluble phenols than stage-I discoloured wood.