Sterilized pine wood strands were treated with the ascomycete Ophiostoma piliferum (Cartapip 97 (TM)) in an attempt to reduce aldehyde emissions through degradation of aldehyde-forming precursors. Wooden boards were then produced from the treated strands by means of a laboratory press. VOC emissions of these boards were characterized and the concentrations of seven major substances were followed for a period of 28 days in Markes mu-CTE micro-chambers employing Tenax-TDAS/GC/MS analytics. Boards made from treated strands showed a highly significant (P<0.001) reduction in aldehyde emissions by 70% compared to the control boards, while differences in the monoterpene emissions were not significant (P>0.05). Wood extractives from treated and untreated strands were also analyzed by GC/MS after milling and acetone/water extraction. A marked decrease of C18 unsaturated fatty acids was detected in agreement with the reduction of aldehyde emissions, and the monosaccharides as indicators of fungal growth were depleted and stilbenes and lignans were partially degraded.
Abstract Sapstain fungi, which reduce the value of pine wood, were isolated from an industrial wood yard in north-eastern Germany. The predominant wood discolouring species on the industrial wood yard and in the forests of the investigated region was Sphaeropsis sapinea (syn. Diplodia pinea), but Ophiostoma minus was also found. These fungi were challenged with antagonistic micro-organisms in laboratory trials and in field experiments. Amongst the tested microorganisms only strains of filamentous fungi (Trichoderma sp. and Phlebiopsis gigantea) could control the sapstain fungi on pine wood blocks efficiently. Although P. gigantea was unable to inhibit stain formation completely, the wood was bleached by this fungus in later incubation stages. In two field trials, sapstain on pine wood logs was successfully retarded for a period of 10 and 12 weeks, respectively, with a white sporulating mutant of Trichoderma harzianum. Thus, the concept using filamentous fungi as antagonists against sapstain developed under laboratory conditions also proved to be valid under natural conditions in forest eco-systems.
Wood is colonised and degraded by a variety of microorganisms, the most efficient ones are wood-rotting basidiomycetes. Microbial decay processes cause damage to wooden constructions, but also have great potential as biotechnological tools to change the properties of wood surfaces and of sound wood. Standard methods to evaluate changes in infected wood, e.g., EN350-1 1994, are time-consuming. Rapid FT-NIR spectroscopic methods are also suitable for this purpose. In this paper, degradation experiments on surfaces of spruce (Picea abies L. Karst) and beech (Fagus silvatica L.) were carried out with white rot basidiomycetes or the ascomycete Hypoxylon fragiforme. Experiments with brown rot or soft rot caused by Chaetomium globosum were also performed. FT-NIR spectra collected from the degraded wood were subjected to principal component analysis. The lignin content and mass loss of the specimens were estimated based on univariate or multivariate data analysis (partial least squares regression).
Summary Microbial mechanisms of lignin degradation may be utilised for solid-state fermentations other than biopulping, during which the selective conversion of lignin is required. The current paper reviews current work into selective lignin conversion, with emphasis on the contributions made by our research group, which consists of researchers from five different laboratories. Three of them cooperate within Wood K plus. The recent research of this group has focussed on fermentations utilising the unique metabolism of selective white-rot fungi to modify wood surfaces during relatively short fermentation times of less than one week and on research into the molecular mechanisms causing these modifications. Lignin degradation by selective fungi (e.g. Ceriporiopsis subvermispora and species of the genus Phlebia) on the wood surfaces was significant after three days. After seven days the overall lignin content of spruce wood shavings was reduced by more than 3.5 %. Lignin loss was accompanied by an increase of extractable substances. To evaluate small changes and to trace the fungal modification processes, Fourier transform infrared spectroscopic (FTIR) techniques and electron paramagnetic resonance (EPR) spectroscopy were applied and adapted. The spectra recorded in the near infrared region (FT-NIR) turned out to be very useful for kinetic studies of the biopulping/biomodification processes and a good method to evaluate the capabilities of fungi to modify wood surfaces within this short period.
Microbial mechanisms of lignin degradation may be utilised for solid-state fermentations other than biopulping, during which the selective conversion of lignin is required. The current paper reviews current work into selective lignin conversion, with emphasis on the contributions made by our research group, which consists of researchers from five different laboratories. Three of them cooperate within Wood K plus. The recent research of this group has focussed on fermentations utilising the unique metabolism of selective white-rot fungi to modify wood surfaces during relatively short fermentation times of less than one week and on research into the molecular mechanisms causing these modifications. Lignin degradation by selective fungi (e.g. Ceriporiopsis subvermispora and species of the genus Phlebia) on the wood surfaces was significant after three days. After seven days the overall lignin content of spruce wood shavings was reduced by more than 3.5%. Lignin loss was accompanied by an increase of extractable substances. To evaluate small changes and to trace the fungal modification processes, Fourier transform infrared spectroscopic (FTIR) techniques and electron paramagnetic resonance (EPR) spectroscopy were applied and adapted. The spectra recorded in the near infrared region (FT-NIR) turned out to be very useful for kinetic studies of the biopulping/biomodification processes and a good method to evaluate the capabilities of fungi to modify wood surfaces within this short period.
The most efficient wood-rotting fungi are basidiomy cetes and ascomycetes. In general, microbial decay processes go along with a loss of wood quality. For example, brown rot decay leads to a rapid decrease in wood strength a lready in early stages of growth. On the other hand, a projected bi odegradation may enhance the quality when sound woo d or surface properties of wood are modified by means of biotech nology. The well-known standard methods to assess c hanges in infected wood are time-consuming, and therefore una cceptable for efficient process-control and quality -assurance. We could show, that fungal infestation of hardwood (Fagus sylvatica L.) and softwood (Picea abies L. Karst) can be assessed by means of FT-NIR spectroscopy in combina tion with uni- and multivariate data analysis. The rapid method is shown to be suitable for these purposes, presumed t he regression models are well chosen. The results o f degradation experiments of wood blocks, veneers, and wood shavi ngs demonstrated that different types and stages of decay could be distinguished and lignin content after decay could be estimated from FT-NIR spectra collected from the degraded wood surfaces and milled wood. Spectral data were subjec ted to principal component analysis (PCA) and uni- and multivariate regression models were calculated. Mass loss after degradation and lignin content could be estimated f rom degraded wood surfaces and it was possible to estimate the select ivity of white rot fungi for the preferential degradation of lignin.
Seventeen isolates from white rotted beech wood and six strains from a local culture collection were evaluated for their capability to delignify beech and spruce wood selectively. Six peroxidase-positive isolates were found using a colorimetric agar plate test (Poly R-478), and genetically identified by their internal transcribed spacer (ITS1) or 28S rDNA sequences. Colonised on beech and spruce wood veneers, some of the peroxidase-positive isolates caused selective white rot on both wood species. Weight loss and lignin content of the degraded veneers were estimated from FT-NIR spectra with established linear regression models and multivariate models based on partial least squares regression (PLSR). Weight loss of the samples was also determined gravimetrically. A measure for the relative selectivity of the strains for lignin degradation was formulated and the values were calculated. Two strains that were identified as Oxyporus latemarginatus and Trametes cervina exhibited high selectivity on spruce wood, but the lignin content of the decayed wood was higher than that degraded by the reference strain Ceriporiopsis subvermispora. One strain – identified as Phlebia tremellosa – led to a lower lignin content of beech wood but caused also comparably high weight loss and thus exhibited an overall lower selectivity. The NIR spectroscopic method proved to be convenient for the quick screening of selective white rot fungi. Furthermore, the results revealed that high selectivity for lignin degradation is much more pronounced in early degradation stages.
Beech wood (Fagus sylvatica L.) veneers were cultivated with white and brown rot fungi for up to 10 weeks. Fungal wood modification was traced with Fourier transform near infrared (FT-NIR) and Fourier transform mid infrared (FT-MIR) methods. Partial least square regression (PLSR) models to predict the total lignin content before and after fungal decay in the range between 17.0% and 26.6% were developed for FT-MIR transmission spectra as well as for FT-NIR reflectance spectra. Weight loss of the decayed samples between 0% and 38.2% could be estimated from the wood surface using individual PLSR models for white rot and brown rot fungi, and from a model including samples subjected to both degradation types.
Due to their outstanding capability of degrading the recalcitrant biomacromolecule lignin, white rot fungi have been attracting interest for several technological applications in mechanical pulping and wood surface modification. However, little is known about the time course of delignification in early stages of colonisation of wood by these fungi. Using a Fourier transform near infrared (FT-NIR) spectroscopic technique, lignin loss of sterilised spruce wood shavings (0.4–2.0mm particle size) that had been degraded by various species of white rot fungi could be monitored already during the first 2 weeks. The delignification kinetics of Dichomitus squalens, three Phlebia species (Phlebia brevispora, Phlebia radiata and Phlebia tremellosa), three strains of Ceriporiopsis subvermispora as well as the white rot ascomycete Hypoxylon fragiforme and the basidiomycete Oxyporus latemarginatus were determined. Each of the fungi tested was able to reduce the lignin content of spruce wood significantly during the first week. The amount of delignification achieved by the selected white rot fungi after 2 weeks ranged from 7.2% for C. subvermispora (FPL 105.752) to 2.5% for P. radiata. Delignification was significant (P=95%) already after 3 days treatment with C. subvermispora and P. tremellosa. Activities of extracellular ligninolytic enzymes (laccase, manganese peroxidase and/or lignin peroxidase), expressed by each of the tested fungi, were determined. Lignin was degraded when peroxidase activity was detected in the fungal cultures, but only a low level of correlation between enzyme activities and the extent of delignification was found.
We describe or redescribe species of Hypocrea/Trichoderma (Ascomycetes, Hypocreales) having hyaline ascospores and pachybasium-like conidiophores. Teleomorphs are reported for Trichoderma minutisporum (Hypocrea minutispora sp. nov.) and T. polysporum (H. pachybasioides). Hypocrea pilulifera/T. piluliferum is redescribed. Trichoderma croceum is synonymized with T. polysporum. The new species H. parapilulifera, H. stellata and H. lacuwombatensis are described. All of these species fall within the morphological concept of Trichoderma sect. Pachybasium and within the phylogenetic group pachybasium B5 of Kullnig-Gradinger et al (2002). Parsimony analysis of nucleotide sequences from three unlinked loci-ITS1 and 2, endochitinase (ech42) and translation elongation factor 1-alpha (tef1)-detects two distinct phylogenetic lineages within the group pachybasium B5. One comprises H. pachybasioides/T. polysporum, H. pilulifera/T. piluliferum, H. parapilulifera and H. stellata; this group, the "polysporum" lineage, is characterized by having conidia that are white in mass and is the only lineage within Hypocrea characterized by such conidia. The second group includes the green conidial T. minutisporum and H. lacuwombatensis. The partition homogeneity test reveals significant recombination within the "polysporum" lineage but not within the "minutisporum" lineage.
Seven new species of Trichoderma, isolated from soil or tree bark from Siberia, Nepal, northern India, Taiwan, Thailand, Cambodia, and Malaysia, are described based on morphological and physiological characters, and from their phylogenetic position inferred from parsimony analyses of nucleotide sequences of the internal transcribed spacer (ITS) regions of the rDNA cluster (ITS1 and 2) and partial sequences of translation elongation factor 1-alpha (tef1α). Trichoderma sinensis sp. nov. and Trichoderma effusum sp. nov. are additions to Trichoderma section Longibrachiatum. Trichoderma helicum sp. nov., Trichoderma rossicum sp. nov., Trichoderma velutinum sp. nov., and Trichoderma cerinum sp. nov. are additions to section Pachybasium. Trichoderma erinaceum sp. nov. is described in section Trichoderma. The contribution of sequence data in resolving species in Trichoderma is demonstrated by Trichoderma helicum, which is morphologically indistinguishable from Trichoderma tomentosum Bissett, even though the two species are not phylogenetically closely related.Key words: Hypocrea, systematics, taxonomy, phylogeny.