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.     

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.     

Anatomical changes in the cell-wall structure of Leucaena leucocephala (Lam.) de Wit as caused by the decay fungi Trametes versicolor and Trametes hirsuta

The structural changes in the cell wall and delignification pattern caused by Trametes versicolor and Trametes hirsuta in the sap wood of Leucaena leucocephala were examined by light and confocal laser scanning microscopy. The in vitro decay test was conducted for 12 weeks. Both species of Trametes used in this study caused anatomical characteristics specific to simultaneous white rot. Formation of boreholes, erosion troughs, erosion channels with U-shaped notches in tangential sections and thinning of cell walls were evident in the wood inoculated with each of the fungal species. Cell separation due to removal of middle lamellae occurred at the early stages of infection (30 d) with T. versicolor. In contrast, middle lamellae remained intact in wood inoculated with T. hirsuta and showed cell separation due to degradation of the outer layer of the secondary wall and degradation of the middle lamellae observed only in severely decayed wood at late stages. Confocal microscopy revealed the delignification pattern particularly from cell corners and vessel walls at an advanced stage of decay, indicating strong ligninolytic activity of both species in the sapwood of L. leucocephala.     

Micromorphological characteristics of bamboo (<Emphasis Type="Italic">Phyllostachys pubescens</Emphasis>) fibers degraded by a brown rot fungus (<Emphasis Type="Italic">Gloeophyllum trabeum</Emphasis>)

The decay pattern in bamboo fibers caused by a brown rot fungus, Gloeophyllum trabeum, was examined by microscopy. The inner part of the polylaminate secondary wall was degraded, while the outer part of the secondary wall remained essentially intact. Degradation in bamboo fiber walls without direct contact with the fungal hyphae was similar to wood decay caused by brown rot fungi. Degradation in polylaminate walls was almost confined to the broad layers whereas the narrow layers appeared resistant. The p-hydroxylphenyl unit lignin in middle lamella, particularly in the cell corner regions, was also degraded. The degradation of lignin in bamboo fibers was evidenced by Fourier transform infrared spectra. The present work suggests that the decay of bamboo fiber walls by G. trabeum was influenced by lignin distribution in the fiber walls as well as the polylaminate structures.     

Microstructural and microchemical studies of wood of Swietenia mahagoni Linn. decayed by Fomes durissimus Lloyd

Histopathological studies of wood of Swietenia mahagoni Linn. decayed by Fomes durissimus Lloyd reveal the presence of mycelia in all the wood clements and considerable thinning of cell walls and tissue disintegration due to utilization of both lignin and ccllulosc at advanccd stages of decay. Microchcmical tcsts and quantitative analysis of lignin nnd ccllulosc corroborated the results of histopathological studies indicating that the fungus mainly consumes thc lignin along with some amount of cellulose.     

Histological changes in the cell wall structure during wood decay by Trametes hirsuta and Trametes versicolor in neem (Azadirachta Indica A. Juss)

ABSTRACT

Structural alterations in the wood cell walls of neem inoculated with by Trametes hirsuta and T. versicolor were studied by microscopic methods. In vitro decayed wood showed extensive weight loss of test blocks (26.7 and 41.38% by T. versicolor and T. hirsuta, respectively) at the end of 3 months. Selective delignification in the initial phase followed by simultaneous removal of lignin was evident in test blocks inoculated with both the species. The separation of middle lamellae and patches of cellulosic polysaccharides stained blue with Astra blue in the delignified region of the fiber wall during early stages indicates selective mode of decay. In contrast, the occurrence of erosion troughs with characteristic U-notch in tangential sections is a characteristic feature of simultaneous rot that was apparent after 3 months of incubation. The decay pattern occurred concomitantly in all the xylem elements irrespective of general resistance pattern shown by vessel and axial parenchyma cells. At an advance stage, both species of Trametes showed formation of erosion channels along the microfibrils angle of cellulose which is considered as characteristics of soft rot decay type. The sharing of white rot and soft rot decay pattern by both the fungi suggest a phylogenetic link between both groups of fungi.     

Electron-dense particles in wood decayed by Ganoderma applanatum

Summary Hemlock sawdust samples degraded by Ganoderma applanatum showed no electron-dense particles either in hyphae or in wood cell walls after aldehyde/OsO₄ fixation. After KMnO₄ fixation at early stage of attack, particles were in hyphae, hyphal sheath and wood cell walls. In samples prepared by a cytochemical technique which localizes cellulase activity at the ultrastructural level, particles were in hyphae, hyphal sheath and wood cell walls. The smallest diameter range of the particles lay between 3 and 7 nm which corresponds to the size of cellulases. Larger diameter particles were peresent which are probably aggregates of the smaller units. We believe that particles present in hyphal cytoplasm and hyphal sheath are cellulolytic enzymes. Whether particles present in attacked wood cell walls are enzymes or degradation products cannot be determined by this study. Nevertheles, the particles reveal the decay pattern in wood by the white-rot fungus G. applanatum.     

Diversity and community structure of wood-inhabiting fungi found in Japanese wooden houses analyzed by the next-generation sequencing

The diversity and community structures of wood-inhabiting fungi in 16 decayed wood samples from ten wooden houses in Japan were analyzed using a next-generation sequencing (NGS) to determine the fungi responsible for wood decay. DNA of fungi in decayed wood was extracted directly, the internal transcribed spacer (ITS) region in ribosomal DNA (rDNA) was amplified by polymerase chain reaction (PCR), and then, sequences of tagged ITS fragments were analyzed by NGS. Results of sequencing indicated that 68 species of ascomycetes, 37 species of basidiomycetes, and one fungus each from Mortierellales and Mucoromycetes were detected. The fungal community structures showed diversity and included various species of ascomycetes. A microscopic examination of cell wall structure in decayed wood samples suggested that some ascomycetes were soft-rot fungi. Heat map analysis indicated that the similarity in the structures of fungal communities was influenced to a greater extent by the wood species of samples than where they were used as a component.     

Cytochemical localization of cellulases in decayed and nondecayed wood

Summary Enzyme assay showed that the commercial cellulase from Trichoderma reesei degraded several polysaccharide substrates; highest activity was on xylan. Transmission electron micrographs showed that the T. reesei cellulase degraded nondecayed wood extensively; the attack always progressed from the point of contact into the cell wall. Cytochemically prepared wood that had been decayed by Poria placenta showed uniform distribution of electron-dense particles throughout the walls; the same results were observed with added T. reesei cellulase. In wood decayed by Ganoderma applanatum without or with added T. reesei cellulase, the progress of degradation was similar to that in nondecayed wood. Negatively stained T. reesei cellulase had 3–7 nm diameter for the smallest particles; the smallest diamaters of electron-dense particles in wood ranged from 3–7 nm. This suggests that the electron-dense particles probably are cellulase molecules.     

Characterization of host–fungus interactions among wood decay fungi associated with Khaya senegalensis (Desr.) A. Juss (Meliaceae) in Singapore

Tree pruning creates wounds that are amenable for wood decay fungi colonization. To characterize the dynamic host–fungus interactions at this location in Senegal mahogany (Khaya senegalensis), in vitro and in vivo pathogenicity tests were conducted with wood decay fungi associated with this tropical tree species. Fomitiporella caryophylii, Hymenochaete murina and Phellinus noxius isolates were included in this experiment following their frequent isolation from Senegal mahogany pruning wounds. The evaluated isolates demonstrated unique host interactions in laboratory tests that suggest equally divergent prognoses for living Senegal mahoganies affected by these fungi. Although all evaluated fungal isolates successfully breached naturally induced reaction zones, P. noxius alone caused significant mass loss to incubated wood blocks. In addition, P. noxius caused extensive wood decay after inoculation in living hosts, successfully illustrating Koch's postulates for this host–fungus relationship. The wood decay ability, invasiveness and facultative parasitism demonstrated by P. noxius suggest its dominant role in wood decay columns below pruning wounds on living Senegal mahoganies. These results highlight the importance of characterizing specific host–fungus interactions and their implications for wood decay severity below pruning wounds in living trees.     

Stem-rot damage and the progress of causal fungi in old-aged Japanese larch trees at the foot of Mt. Fuji

Damage caused by stem-rot and the progress of the causal fungi in old-aged Japanese larch (Larix kaempferi (Lamb.) Carr.) was investigated at the foot of Mt. Fuji. Stem-rot was found in 75% of 108 trees investigated, and volume of rot was 6% of the total wood volume in the forest investigated. Stem-rot damage was much greater than the damage by butt-rot.Stereum sanguinolentum (Alb. and Schw. ex Fr.) Fr. infected larch trees at the greatest incidence (49.4%). However,Porodaedalea chrysoloma (Fr.) Imaz. caused the most volume loss to the trees.S. sanguinolentum infected larch stems mainly through stem wounds, and decay caused by the fungus progressed 9.75×10² cm³/year on average.P. chrysoloma infected larch stems mainly through dead branches and wounds, and the average rate of decay progress for the fungus was 2.74×10³ cm³/year.     

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     

Changes in structural and chemical components of wood delignified by fungi

Summary Cerrena unicolor, Ganoderma applanatum, Ischnoderma resinosum and Poria medulla-panis were associated with birch wood that had been selectively delignified in the forest. Preferential lignin degradation was not uniformly distributed throughout the decayed wood. A typical white rot causing a simultaneous removal of all cell wall components was also present. In the delignified wood, 95 to 98% of the lignin was removed as well as substantial amounts of hemicelluloses. Scanning and transmission electron microscopy were used to identify the micromorphological and ultrastructural changes that occurred in the cells during degradation. In delignified areas the compound middle lamella was extensively degraded causing a defibration of cells. The secondary wall, especially the S₂ layer, remained relatively unaltered. In simultaneously white-rotted wood all cell wall layers were progressively removed from the lumen toward the middle lamella causing erosion troughs or holes to form. Large voids filled with fungal mycelia resulted from a coalition of degraded areas. Birch wood decayed in laboratory soil-block tests was also intermittently delignified. Selective delignification, sparsely distributed throughout the wood, and a simultaneous rot resulting in the removal of all cell wall components were evident. Scanning electron microscopy appears to be an efficient technique for examining decayed wood for fungi with the capacity to selectively delignify wood.The authors would like to thank Kathy Zuzek for technical assistance and Dr. M. Larsen, Forest Prod. Lab., Madison, for identifying the sporophores of Poria medulla-panis. This research was founded in part by a grant from the USDA Forest Service, Forest Products Laboratory and from the Graduate School, University of Minnesota     

A comparison between decay patterns of the white‐rot fungus Pleurotus ostreatus in chestnut–leaved oak (Quercus castaneifolia) shows predominantly simultaneous attack both in vivo and in vitro

In this research, we examined decay patterns occurring in Quercus castaneifolia wood under natural conditions compared with controlled decay in vivo. Pleurotus ostreatus‐infected oak wood was obtained from the Sari forests in the north of Iran. The species causing decay was verified as P. ostreatus using rDNA‐ITS sequencing of pure cultures from infected sapwood. In addition to P. ostreatus, two wood‐inhabiting Ascomycota, Trichoderma harzianum and T. lixii, were present. Mass loss in oak sapwood samples exposed to P. ostreatus for 60 days was around 10 per cent. Samples were prepared from both naturally decayed wood and wood decayed under controlled conditions and examined using microscopy. P. ostreatus was found to produce a simultaneous white‐rot decay pattern in both conditions.     

Nursery soil cover influences germination of Thujopsis dolabrata var. hondai seeds

We examined the influence of nursery soil cover on germination of Thujopsis dolabrata var. hondai. Seeds were sown under seeding bed conditions: (1) nursery soil (andosol) without soil cover, (2) Kanuma pumice without soil cover, (3) nursery soil with nursery soil cover, and (4) nursery soil with sterilized sand cover. Germination percentages were compared between these conditions. The germination percentage (8.8%) at 10 weeks after sowing under condition 3 was significantly lower than under conditions 1, 2, and 4 (56.8%, 52.0%, and 47.2%, respectively). Of the nongerminated seeds under condition 3, 75.0% were decayed and several fungi were isolated from decayed seeds. An inoculation test of isolated fungi Cylindrocarpon tenue, Cylindrocarpon sp., Fusarium oxysporum, and Botrytis sp. to seeds showed that these fungi caused seed decay. Our results indicated that nursery soil cover may not be suitable for T. dolabrata var. hondai seeds sown in nursery soil because of seed decay by pathogens.     

The Delignification Pattern of Ailanthus excelsa Wood by Inonotus hispidus (Bull.: Fr.) P. Karst.

In vitro laboratory decay tests on Ailanthus excelsa Roxb. wood revealed that I. hispidus exhibits a combination of both white-rot and soft-rot patterns of wood decay. Early stages of wood decay showed dissolution of the middle lamella as well as defibration and localized delignification of fiber walls; vessels, axial, and ray parenchyma remained unaltered. Delignification commenced from the middle lamellae at the cell corners without any marked effect on the primary and secondary wall layers. In later stages of growth, the species produces typical soft-rot decay pattern by forming erosion channels through the S₂ layers of fiber walls, transverse bore holes in the cell walls, and erosion channels alongside/following the orientation of cellulose microfibrils. The rays showed signs of cell wall alterations only after the extensive damage to the fiber walls. After 120 days of incubation, the vessels also showed localized delignification, the erosion of pits, and separation from associated xylem elements. The extensive weight losses under natural and in vitro decayed wood as well as the very soft nature of severely degraded wood indicate that I. hispidus alters wood strength and stiffness.     

Treatment of wounds on Acer rubrum with Trichoderma viride1

Wounds on 90 Acer rubrum trees were inoculated with Trichoderma viride in July and August 1973 to determine the effect of the fungus on the development of decay. The trees were 45 years old and 15 cm diam at 1.4 m aboveground. Each tree had 12 wounds, 1.4 cm in diam and 5 cm deep. After a year 28 trees were harvested and isolations were made from the discolored, and decayed wood associated with each wound. Trichoderma viride was reisolated from all wounds that were inoculated with it. Hymenomycetes were isolated frequently from decayed wood associated with wounds not inoculated with T. viride. No Hymenomycetes were isolated from wounds inoculated with T. viride.     

Polyporus schweinitzii Fr. und Sparassis crispa (Wulf. in Jacq.) ex Fr. als Fäuleerreger in einem Douglasienbestand (Pseudotsuga menziesii (Mirb.) Franco) mit hohem Stammfäuleanteil1

Polyporus (Phaeolus) schweinitzii and Sparassis crispa as frequent decay fungi in Douglas fir. Of fifty, 55 ears old trees in a Pseudotsuga menziesii stand. 90% showed decay of the heart- wood of the butt. The possibility of transmission of the most frequent decay fungi, P. schwei- nitzii and S. crispa, which also are frequent parasites in the roots and butts of pine, from the roots of the Pinus sylvestris trees of the previous crop into the roots of the Douglas fir is discussed. A first identification of the fungi was made possible by storing the stem discs under moist conditions. The mycelia of P. schweinitzii and S. crispa grew out of the decayed wood and possessed a characteristic form and colour.     

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)     

Fomes annosus (Fr.) Cke. und andere Stammfäulepilze in einem Douglasienbestand,Pseudotsuga menziesii (Mirb.) Franco1

Fomes annosus (Fr.) Cke. and other decay fungi in a Douglas fir stand, Pseudotsuga menziesii (Mirb.) Franco. 40% of the trees in a 40 years old Pseudotsuga menziesii stand showed butt rot. 85 trees were analysed for decay fungi. Fomes annosus, the most frequent fungus, also invaded the sap wood. Factors of the soil favourable to the rot and the possibility of transmission of the most frequent decay fungus, Fomes annosus, from (a) neighbouring spruce stands, (b) from the roots of Scots pine from the previous crop arc discussed. Caniophora puteana was isolated from about 10% of the butt rots. The importance of Calocera viscosa which grew out of the central decay of twelve trees as a decay fungus is still under investigation.