The aim of this study was to investigate Rhodonia placenta expression patterns of genes involved in the depolymerisation during the non-enzymatic phase in acetylated (WAc) and furfurylated wood (WFA). During the 98-day-long exposure, WAc [22.6% weight per cent gain (WPG) on average] and WFA (69% WPG on average) lost no more than 3% mass while the untreated wood (WUn) reached 41% mass loss (ML) in 55 days. Expression of six genes putatively involved in the non-enzymatic degradation process were investigated. In conclusion, expression levels of alcohol oxidase Ppl118723 (AlOx1) and laccase Ppl111314 (Lac) were significantly higher in the modified wood materials (WMod) than in WUn, which is in accordance with previous results and may be explained by the absence of the degradation products that have been proposed to down-regulate the non-enzymatic degradation process. However, copper radical oxidase Ppl156703 (CRO1) and a putative quinate transporter Ppl44553 (PQT) were expressed at significantly lower levels in WMod than in WUn while quinone reductase Ppl124517 (QRD) and glucose oxidase Ppl108489 (GOx) were expressed at similar levels as in WUn. These results suggest that gene regulation in WMod is more complex than a general up-regulation of genes involved in the non-enzymatic degradation phase.

Brown rot fungi, such as Rhodonia placenta (previously Postia placenta), occur naturally in northern coniferous forest ecosystems and are known to be the most destructive group of decay fungi, degrading wood faster and more effectively than other wood-degrading organisms. It has been shown that brown rot fungi not only rely on enzymatic degradation of lignocellulose, but also use low molecular weight oxidative agents in a non-enzymatic degradation step prior to the enzymatic degradation. R. placenta is used in standardized decay tests in both Europe and North America. However, two different strains are employed (FPRL280 and MAD-698, respectively) for which differences in colonization-rate, mass loss, as well as in gene expression have been observed, limiting the comparability of results. To elucidate the divergence between both strains, we investigated the phenotypes in more detail and compared their genomes. Significant phenotypic differences were found between the two strains, and no fusion was possible. MAD-698 degraded scots pine more aggressively, had a more constant growth rate and produced mycelia faster than FPRL280. After sequencing the genome of FPRL280 and comparing it with the published MAD-698 genome we found 660,566 SNPs, resulting in 98.4% genome identity. Specific analysis of the carbohydrate-active enzymes, encoded by the genome (CAZome) identified differences in many families related to plant biomass degradation, including SNPs, indels, gaps or insertions within structural domains. Four genes belonging to the AA3_2 family could not be found in or amplified from FPRL280 gDNA, suggesting the absence of these genes. Differences in other CAZy encoding genes that could potentially affect the lignocellulolytic activity of the strains were also predicted by comparison of genome assemblies (e.g., GH2, GH3, GH5, GH10, GH16, GH78, GT2, GT15, and CBM13). Overall, these mutations help to explain the phenotypic differences observed between both strains as they could interfere with the enzymatic activities, substrate binding ability or protein folding. The investigation of the molecular reasons that make these two strains distinct contributes to the understanding of the development of this important brown rot reference species and will help to put the data obtained from standardized decay tests across the globe into a better biological context. © Copyright © 2020 Kölle, Horta, Nowrousian, Ohm, Benz and Pilgård.

Extractives found in the heartwood of a moderately durable conifer (Larix gmelinii var. japonica) were compared with those found in a non-durable one (Picea abies). We identified and quantified heartwood extractives by extraction with solvents of different polarities and gas chromatography with mass spectral detection (GC-MS). Among the extracted compounds, there was a much higher amount of hydrophilic phenolics in larch (flavonoids) than in spruce (lignans). Both species had similar resin acid and fatty acid contents. The hydrophobic resin components are considered fungitoxic and the more hydrophilic components are known for their antioxidant activity. To ascertain the importance of the different classes of extractives, samples were partially extracted prior to subjection to the brown-rot fungus Rhodonia placenta for 2–8 weeks. Results indicated that the most important (but rather inefficient) defense in spruce came from the fungitoxic resin, while large amounts of flavonoids played a key role in larch defense. Possible moisture exclusion effects of larch extractives were quantified via the equilibrium moisture content of partially extracted samples, but were found to be too small to play any significant role in the defense against incipient brow-rot attack.

The aim of this study was to visualize non-enzymatic oxidative degradation damages in the initial decay stage of the brown rot fungus Rhodonia placenta degradation in furfuryl alcohol (FA) modified wood cell walls and untreated wood cell walls of radiata pine (Pinus radiata) sapwood. A decay test with small wood blocks (1.5 × 1.5 × 5 mm³) of untreated and furfurylated radiata pine selected from two different furfurylation processes was performed until the first mass loss occurred. The samples were exposed to the brown rot fungus R. placenta, monitored by light microscopy and analyzed topochemically by cellular UV microspectrophotometry (UMSP). The results showed that the FA modification process directly influenced: i) the fungal colonization and hyphal growth, ii) the spectral UV behavior, and iii) degradation patterns of the entire cell wall layers. For the first time, UMSP area scans and selective line scans of individual cell wall regions provide topochemical insights into oxidative degradation at the initial decay stage of furfuryl alcohol-modified P. radiata visualizing oxidative degradation in situ. Knowledge of the initial decay stage of brown rot degradation in FA-modified wood compared to untreated wood extends our understanding of the brown rot decay processes of cell wall compounds. 

The effect of wood modification on wood-water interactions in modified wood is poorly understood, even though water is a critical factor in fungal wood degradation. A previous review suggested that decay resistance in modified wood is caused by a reduced wood moisture content (MC) that inhibits the diffusion of oxidative fungal metabolites. It has been reported that a MC below 23%-25% will protect wood from decay, which correlates with the weight percent gain (WPG) level seen to inhibit decay in modified wood for several different kinds of wood modifications. In this review, the focus is on the role of water in brown rot decay of chemically and thermally modified wood. The study synthesizes recent advances in the inhibition of decay and the effects of wood modification on the MC and moisture relationships in modified wood. We discuss three potential mechanisms for diffusion inhibition in modified wood: (i) nanopore blocking; (ii) capillary condensation in nanopores; and (iii) plasticization of hemicelluloses. The nanopore blocking theory works well with cell wall bulking and crosslinking modifications, but it seems less applicable to thermal modification, which may increase nanoporosity. Preventing the formation of capillary water in nanopores also explains cell wall bulking modification well. However, the possibility of increased nanoporosity in thermally modified wood and increased wood-water surface tension for 1.3-dimethylol-4.5-dihydroxyethyleneurea (DMDHEU) modification complicate the interpretation of this theory for these modifications. Inhibition of hemicellulose plasticization fits well with diffusion prevention in acetylated, DMDHEU and thermally modified wood, but plasticity in furfurylated wood may be increased. We also point out that the different mechanisms are not mutually exclusive, and it may be the case that they all play some role to varying degrees for each modification. Furthermore, we highlight recent work which shows that brown rot fungi will eventually degrade modified wood materials, even at high treatment levels. The herein reviewed literature suggests that the modification itself may initially be degraded, followed by an increase in wood cell wall MC to a level where chemical transport is possible.

The study of degradation and growth patterns of fungi in modified wood may increase the understanding of their mode of action and may lead to more accurate service-life predictions. The aim of this paper was to study the degradation and growth patterns of brown rot fungi in modified wood and to measure moisture content (MC), structural integrity and the acetyl content by frequent monitoring over 300 days. Mass loss (ML) in the modified wood materials increased slowly up to 3% for 50–100 days after which it flattened out and remained constant during the remainder of the test. Structural integrity and acetyl content were maintained in the modified wood materials and MC was lower compared to untreated wood throughout the decay test. ML results of untreated wood indicate that fungi in solid wood go through distinct phases; the degradation patterns in the modified wood materials were more difficult to interpret.

Furfurylation is one of the wood modification techniques via catalytic polymerization of the monomeric furfuryl alcohol (FA) in the impregnated cell wall. Little is known about the topochemistry of this process. Brown rot degradation begins with lignin modification and therefore, the reactions between FA and lignin was one focus of this research. Furfurylated radiata pine (Pinus radiata) with three different weight percent gains (WPGs of 57%, 60% and 70%) after FA uptake was observed by cellular ultraviolet microspectrophotometry (UMSP) to analyze chemical alterations of the individual cell wall layers. Moreover, light microscopy (LM) and scanning electron microscopy (SEM) analyses were performed. The ultraviolet (UV) absorbance of the modified samples increased significantly compared to the untreated controls, indicating a strong polymerization of the aromatic compounds. Highest UV absorbances were found in areas with the highest lignin concentration. The UMSP images of individual cell wall layers support the hypothesis concerning condensation reactions between lignin and FA.

The mechanism by which modified wood resists decay has long been discussed, but is still not fully understood. A better understanding of decay resistance mechanisms could improve wood protection processes and product properties. In this study, the dynamics of brown rot decay in thermally modified wood was examined through measurements of mass loss, strength loss and gene expression. Close monitoring of mass loss over 120 days in thermally modified wood exposed to Postia placenta showed a delay in the onset of degradation compared to untreated wood, and once the degradation had started, the rate was lower. Thermally modified wood did not inhibit expression of wood degradation-related genes before mass loss and was similar to that in untreated wood once mass loss could be detected. Comparing gene expression as well as strength loss at the same stage of decay rather than at the same time after exposure showed smaller differences in decay patterns between thermally modified and untreated wood than previous results indicate. It is concluded that the key to understanding degradation resistance in thermally modified wood is to compare the decay patterns in thermally modified wood and untreated wood before mass loss occurs.

Today there is an increasing demand for wood protection systems which are based on low environmental impact technologies. The aim of this study was to improve a method for investigating furfurylated Pinus radiata, both in terms of the furfuryl alcohol polymerisation process in the wood cell wall and the detection of initial signs of brown rot degradation. It is an established theory today that brown rot degradation starts with lignin modification in the outermost part of the secondary cell wall and the combined middle lamella. We adapted a method (cellular UV microspectrophotometry, UMSP) which enables direct imaging of the lignin distribution and modification within individual cell wall layers. Furfurylated P. radiata, incubated with Postia placenta, were scanned with monochromatic UV-light at 280 nm. The results showed that USMP is a promising method to study furfurylated wood and early fungal degradation damages in the cell wall of furfurylated wood.

Modified wood shows resistance against wood deteriorating fungi, but the mechanisms are still not fully understood. The aim of this paper was to summarise the molecular studies performed on modified wood with regard to brown rot decay fungi. The DNA data showed that fungi are present inside the laboratory wood test samples already after two weeks of inoculation. Generally the fungal DNA content reflects mass loss and wood moisture content. The oxidative gene expression seems to be higher in modified wood than in untreated wood and it tend to increase during incubation. Based on the gene expression data we suggest that the hypothesis of lack of substrate recognition by the fungus should be rejected. In the reviewed studies, total wood moisture content in the samples was generally not low enough to inhibit fungal colonisation. Hence, moisture distribution within the wood should be studied more closely.