Improved molecular methods to characterise Serpula lacrymans and other Basidiomycetes involved in wood decay

Early detection of indoor wood-decay fungi is crucial to prevent building deterioration and thereby avoid considerable economic loss. Due to their increased sensitivity, two reliable DNA-based fingerprinting techniques, capillary electrophoresis single-strand conformation polymorphism (CE-SSCP) and denaturing high-performance liquid chromatography (DHPLC), were used to identify Serpula lacrymans and to profile wood-rot Basidiomycetes in the built environment. Molecular fungal diversity was assessed on 74 environmental samples, collected from 2003 to 2009 from infected buildings in France. S. lacrymans, the most widespread, indoor wood-decay fungus accounted for 64% of total wood-rot Basidiomycetes. A number of other common wood-rot fungi such as Coniophora puteana, Trametes versicolor and Donkioporia expansa were identified. Other Basidiomycetes such as Phlebiopsis gigantea and Scleroderma verrucosum were detected for the first time in the built environment. Reliable diagnostic tools were developed using two PCR-based molecular typing techniques, one for routine diagnosis and another one for community inventories. Together they provided useful data for characterising the complexity of wood-decay ecosystems and helped reveal the coexistence of different wood-decay fungi within the same microbiotope.     

Host and habitat preferences of polypore fungi in Micronesian tropical flooded forests

The distribution and ecological impacts of plant-associated fungi is determined in large part by their degree of specificity for particular host species or environmental conditions. Here we evaluate the host and habitat preferences among the Aphyllophorales, a guild of wood-decay basidiomycete fungi usually considered to be host generalists. We determined the patterns of host association in three well-defined, floristically distinct, tropical wetlands — freshwater forested wetlands, saltwater mangrove forests, and peatlands with scattered trees — on the islands of Kosrae and Pohnpei in the Federated States of Micronesia. Of 33 fungal species, 20 were locally rare. Of the 11 species sufficiently common to evaluate habitat specificity, nine showed significant habitat preferences. Of eight species common enough to evaluate within-habitat host specificity, six showed strong host preferences. All except one of the nine habitat-specialized fungi showed either statistically significant host specificity or strong numerical biases toward single host species. Our results suggest that host preferences may be important in shaping the assemblages of wood-decay fungi, and that the effect of environment on the distribution of susceptible plant species, rather on the fungi themselves, may ultimately drive the apparent habitat specificity of many fungi.     

Low temperature enhances the ability of the termite-egg-mimicking fungus Athelia termitophila to compete against wood-decaying fungi

A unique symbiosis exists between subterranean termites and the sclerotium-forming fungus Athelia termitophila, which forms termite-egg-mimicking sclerotia called ‘termite balls’. While the sclerotia gain a competitor-free habitat by being harboured by termite eggs, A. termitophila mycelia have to compete with wood-decay fungi in the life stage without termites. To understand its relationship with termites, the factors that affect the ability of A. termitophila to compete with other wood-decay fungi must be clarified. Here, we show that A. termitophila is competitive against other wood-decay fungi at low temperatures. In Petri dish experiments to evaluate the effects of the physicochemical conditions, that A. termitophila experiences in termite nests, on its competitive ability, A. termitophila overcame surrounding fungi in the winter, when termites are less active. Further studies quantifying the effects of A. termitophila on termites in winter will help us to understand this relationship.     

Induction of polygalacturonase and the formation of oxalic acid by pectin in brown-rot fungi

Extracellular polygalacturonase (PG) production was estimated in vitro, using liquid cultures of three species of brown-rot decay fungi (Postia placenta, Gloeophyllum trabeum and Serpula incrassata), by cup-plate assay, assay of reducing sugars, and decrease in viscosity. Although all three experimental assays demonstrated that PG was induced by pectin in all three fungi, decrease in viscosity gave the best correlation with decay capacity in soil block tests. PG activity, determined as an increase in reducing sugar activity, was greatest in G. trabeum and weakest in S. incrassata. The optimum pH for PG activity was between pH 2.5 and 4.5. Oxalic acid production was also enhanced by pectin and functioned synergistically with PG activity. We conclude that these fungi produce PG that is best induced by pectin and that PG activity exceeds production of xylanase and endoglucanase activity in vitro. Polygalacturonase is likely to act synergistically with oxalic acid to solubilize and hydrolyse the pectin in pit membranes and middle lamellae. Thus, production of PG and oxalic acid should facilitate early spread of hyphae and enhance the lateral flow of wood-decay enzymes and agents into adjacent tracheids and the wood cell wall, thus initiating the diffuse decay caused by brown-rot fungi.The Forest Products Laboratory is maintained in co-operation with the University of Wisconsin. This article was written and prepared by US Government employees on official time, and it is therefore in the public domain and not subject to copyright.     

Use of antagonistic bacilli for biocontrol of fungi degrading fresh wood

The species composition of micromycete complexes colonizing aspen, birch, and pine wood was studied. Calculation of the Sorens-Tchekanovsky similarity coefficients showed that these complexes shared a high degree of similarity. They were dominated by representatives of the genera Penicillium, Paecilomyces, Trichoderma, and Rhizopus. Some antagonistic bacilli inhibited growth and development of wood-decay fungi in vitro and led to the formation of spheroplasts on growing hyphae. A study of possible involvement of bacillary mycolytic enzymes in the inhibition of wood-decay fungi demonstrated selectivity of their lytic effect, which was determined by the genus and species of micromycetes and did not correlate with their relative resistance to antagonists.     

Degradation of extractive-free lignocelluloses by Coriolus versicolor and Poria placenta

Summary The wood-decay fungi Coriolus versicolor, a white-rot fungus, and Poria placenta, a brown-rot fungus, were grown on an extractive-free lignocellulose prepared from quackgrass (Agropyron repens). Their abilities to decompose this lignocellulose were compared to their abilities to decompose softwood (Picea pungens) and hardwood (Acer rubrum) lignocelluloses. The two fungi were grown on malt-extract dampened lignocelluloses at 28°C for up to 12 weeks. Replicate cultures were periodically harvested and lignocellulose decomposition was followed by monitoring substrate weight loss, lignin loss, and carbohydrate loss. Coriolus versicolor decomposed the lignin and carbohydrate components of the grass lignocellulose as efficiently as the softwood and hardwood lignocelluloses. Poria placenta, however, was not an efficient degrader of either lignin or carbohydrate in the grass lignocellulose. Poria placenta readily decomposed carbohydrate components of the softwood lignocellulose but not the hardwood lignocellulose.Paper number 81520 of the Idaho Agricultural Experiment Station     

Ectomycorrhizal and wood-decay macromycete communities along development stages of managed Pinus patula stands in Southwest Mexico

This study used fruit body surveys to evaluate the diversity and distribution of ectomycorrhizal and wood-decay macromycetes along developing stages of stands artificially reforested with Pinus patula in Southern Mexico. Three stand ages were selected: 1, 11, and 60 y old. Macromycetes were collected from June to October 2016 in 0.1 ha plots per stand age. A total of 106 taxa were recorded, 63 ectomycorrhizal and 43 wood-decay fungi. The highest diversity of ectomycorrhizal fungi was in 11-y stands, and 11- and 60-y stands had similar species composition. Wood-decay fungi showed a trend towards higher diversity in the 1- and 11-y stands, and species composition in these sites corresponded. Patterns of diversity and distribution were mainly determined by factors related to substratum availability and vegetation structure. Knowledge about the influence of artificially reforested stands on macrofungal communities is relevant to improve management plans and achieve a more sustainable use of forest timber and non-timber products.     

Regulation of low-molecular weight organic acid production in fungi

Organic acids produced by fungi have been proposed to have many roles in wood-decay processes, lignocellulose degradation or plant pathogenesis involving saprotrophic or pathogenic fungi, as well as in nutrient acquisition and metal detoxification involving mycorrhizal or rhizosphere-inhabiting fungi. In comparison with other fungi, a considerable body of work has been devoted to the comprehension of biosynthesis pathways in fungi involved in industrial production of organic acids, and also in those involved in wood-decay processes and pathogenicity. In this review we therefore focus on information available from these different types of low-molecular weight organic acid (LMWOA) producing fungi in order to better understand the environmental cues involved in regulating production of LMWOAs.     

Fungicidal properties of individual components of copper–ethanolamine-based wood preservatives

The importance of copper–ethanolamine-based wood preservatives is increasing. These preservatives usually consist of copper as a fungicide, ethanolamine as a fixative, and secondary fungicides (boron, triazoles) and other additives (water repellents, fixatives, wax emulsions, etc.). Questions arise as to how each of these ingredients interacts with wood-decay fungi, and whether there are any synergistic effects between the components. In order to elucidate these questions, Norway spruce wood specimens were impregnated with five different aqueous solutions consisting of one single component only and of complete formulation of five different concentrations. These specimens were exposed to two brown-rot fungi, Antrodia vaillantii and Gloeophyllum trabeum, as well as to the white-rot fungus Trametes versicolor for 8 weeks according to mini block procedure. In parallel, petri dishes with nutrient medium containing different quantities of ingredients and of complete wood preservative were inoculated with the same fungal species, and their growth was compared with growth on media without chemicals. The results showed that both experimental methods give similar results. In general, there was no synergistic effect determined. Ethanolamine did not decrease fungicidal properties of the system, while on the other hand octanoic acid has a positive effect on the growth of brown-rot fungi. The minimal effective concentration of tested copper–ethanolamine preservative was determined by the minimum effective concentration of the most fungi-toxic ingredient.     

Application of Bacillus-antagonists for biocontrol of fungi degrading raw wood

Species composition of micromycete complexes colonizing aspen, birch, and pine wood was studied. Calculation of the Sorens-Tchekanovsky similarity coefficients showed that these complexes shared a high degree of similarity. They were dominated by the representatives of the genera Penicillium, Paecilomyces, Trichoderma, and Rhizopus. Some antagonistic bacilli inhibited growth and development of wood-decay fungi in vitro and led to the formation of spheroplasts on growing hyphae. A study of possible involvement of bacillary mycolytic enzymes in the inhibition of wood-decay fungi demonstrated selectivity of their lytic effect, which was determined by the genus and species of micromycetes and did not correlate with their relative resistance to antagonists.     

Wood-decay fungi in fine living roots of conifer seedlings

The mycorrhizal basidiomycetes are known to have multiple, independent evolutionary origins from saprotrophic ancestors. To date, a number of studies have revealed functional resemblance of mycorrhizal fungi to free-living saprotrophs, but information on the ability of saprotrophic fungi to perform as mycorrhizal symbionts is scarce. Here, the objective was to investigate the ability of three wood-decay fungi, Phlebiopsis gigantea, Phlebia centrifuga and Hypholoma fasciculare, to colonize fine roots of conifer seedlings. For each fungus, mycorrhizal syntheses were attempted with Picea abies and Pinus sylvestris. After 24 wk, isolation of fungi and direct sequencing of fungal internal transcribed spacer (ITS) rDNA were carried out from healthy-looking surface-sterilized root tips that yielded both pure cultures and ITS sequences of each inoculated strain. Mycelial mantle of P. gigantea was frequently formed on root tips of P. abies, and microscopical examination has shown the presence of intercellular hyphae inside the roots. The results provide evidence of the ability of certain wood-decay fungi to colonise fine roots of tree seedlings.     

Indoor wood-decay basidiomycetes: damage,causal fungi,physiology, identification and characterization,prevention and control

Indoor wood-decay fungi cause considerable economical damage. Most of the structural damage to the indoors of buildings in Europe and North America is caused by brown-rot fungi that degrade conifer wood; white-rot fungi, which preferentially attack hardwoods, are less common. This review covers the approximately 80 basidiomycetes that commonly occur in buildings. Emphasis was placed on Serpula lacrymans, which is the most common indoor basidiomycete in central Europe. Meruliporia incrassata, the North American pendant to S. lacrymans, has also received considerable attention. In terms of indoor wood decay, moisture and temperature are the most important influences. Wood samples with a low moisture content can be degraded. High temperatures as an alternative control measure do not kill mycelia, with some species surviving in wood samples in the form of heat-resistant arthrospores at temperatures as high 95°C. For refurbishment and scientific purposes, the identity of the causal species should be known. More recently, several molecular techniques have been used to identify fungi; these results are often conflicting with those obtained by other, earlier applied methods. Sequencing of the internal transcribed spacers (ITS) of the rDNA is currently the best molecular tool. Among the other methods available, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS) has also been shown to be able to distinguish closely related sister taxa. For further characterization of indoor basidiomycetes, the complete sequences of the 18S, 28S rDNA and the intergenic spacers with the included 5S rDNA have been acquired for some species. If current projects involving whole funal genome sequencing are not taken into account, Antrodia vaillantii is the first basidiomycete for which the complete rDNA sequence has been deposited. The review closes with fundamentals on the prevention and control of indoor wood decay. An erratum to this article can be found at     

Differential sensitivity of fungi to lithium chloride in culture media

Forty species of fungi, representing a range of ecological and taxonomic groups, were tested for their ability to grow on agar media amended with lithium chloride (LiCl) at 1.5, 3 and 6 g l⁻¹. Species of Trichoderma varied considerably in their sensitivity to LiCl; at one week on 6 g l⁻¹ LiCl medium, the growth of seven species of Trichoderma was considerably inhibited; however, by three weeks at this level, four of the species tested were able to attain ≥30 % of control growth. Of the seven species tested, an isolate of T. viride was the most sensitive to LiCl in agar. Eleven other imperfect fungi also showed a range of ability to grow on agar amended with LiCl, from total inhibition to complete lack of inhibition. Six ascomycete fungi were greatly inhibited by LiCl at all levels; however, an isolate of Chaetomium globosum was highly tolerant of LiCl. Seven basidiomycete wood-decay fungi were quite sensitive to LiCl in agar, showing total to nearly total inhibition even at the lowest level; however, after three weeks, an isolate of Postia placenta was nearly uninhibited except at 6 g l⁻¹. Five ectomycorrhizal basidiomycete fungi were totally inhibited by all levels of LiCl; however, one ectomycorrhizal imperfect fungus (Cenococcum graniforme) was able to grow at 3 g l⁻¹ and was uninhibited at 1.5 g l⁻¹. Four zygomycete fungus isolates were nearly unaffected in their growth by all levels of LiCl.     

Fungal communities associated with acorn woodpeckers and their excavations

Wood-decay fungi soften wood, putatively providing opportunities for woodpeckers to excavate an otherwise hard substrate, yet the fungal community composition in tree cavities and the specificity of these relationships is largely unknown. We used high-throughput amplicon sequencing of the fungal ITS2 region to examine the fungal communities associated with acorn woodpeckers (Melanerpes formicivorus) and their cavities in mature valley oak (Quercus lobata) and blue oak (Q. douglasii) trees in an oak savannah of central coastal California, USA. Acorn woodpeckers and their excavations harbored over 1500 fungal taxa, including more than 100 putative wood-decay fungi. The fungal communities found on the birds were more similar to those found in excavated cavities than those found in trees without excavated holes. These results suggest that symbiotic associations between acorn woodpeckers and fungi are highly diverse, with low specificity. Symbiotic associations between cavity-excavators and fungi are likely more common and widespread than previously thought.     

Substrate specialization of wood-decay aphyllophoroid fungi in the pine forests of the right riverside of the Ob headwaters

Substrate specialization of 196 species of wood-decay aphyllophoroid fungi has been studied in the pine forests of the right riverside of the Ob headwaters. Distribution of fungi species on deciduous and coniferous trees is bearing on the types of rot caused by them and on the character of substrate. The largest number of the species of wood-deacay fungi develop on forest-forming tree species. The majority of fungi, chiefly causing white rot develop on deciduous tress.     

从白腐菌中筛选漆酶高产菌株的初步研究

从中国分离的149株木腐菌中,有30株具有漆酶活性。在添加了阿魏酸的液体培养基中,漆酶产量高(每升培养液中漆酶含量在10000单位以上)的菌株有9个:游离细胞培养的彩绒革盖菌Coriolus versicolor G30,相邻小孔菌Microporus affinis G07,血红密孔菌Pycnoporus sanguineus W006,W006-2,W3008,G05和香菇Lentinus edodes G18,以及固定细胞培养的鲑贝革盖菌Coriolus consors 98563和干酪菌Tyromyces sp. 98420。彩绒革盖菌、相邻小孔菌和血红密孔菌具有重要的生物工程研究价值。     

Spore sensitivity to sunlight and freezing can restrict dispersal in wood‐decay fungi

Assessment of the costs and benefits of dispersal is central to understanding species'' life-history strategies as well as explaining and predicting spatial population dynamics in the changing world. While mortality during active movement has received much attention, few have studied the costs of passive movement such as the airborne transport of fungal spores. Here, we examine the potential of extreme environmental conditions to cause dispersal mortality in wood-decay fungi. These fungi play a key role as decomposers and habitat creators in forest ecosystems and the populations of many species have declined due to habitat loss and fragmentation. We measured the effect of simulated solar radiation (including ultraviolet A and B) and freezing at −25°C on the spore germinability of 17 species. Both treatments but especially sunlight markedly reduced spore germinability in most species, and species with thin-walled spores were particularly light sensitive. Extrapolating the species'' laboratory responses to natural irradiance conditions, we predict that sunlight is a relevant source of dispersal mortality at least at larger spatial scales. In addition, we found a positive effect of spore size on spore germinability, suggesting a trade-off between dispersal distance and establishment. We conclude that freezing and particularly sunlight can be important sources of dispersal mortality in wood-decay fungi which can make it difficult for some species to colonize isolated habitat patches and habitat edges.     

Wood-inhabiting insects can function as targeted vectors for decomposer fungi

Most wood-inhabiting fungi are assumed to be dispersed primarily by wind, with the exception of a few species involved in mutualistic relationships with insects. In this study we tested whether several species of wood-inhabiting insects can function as dispersal vectors for non-mutualistic fungi, which would indicate that wood-inhabiting fungi can benefit from targeted animal-mediated dispersal. We sampled wood-inhabiting beetles (Coleoptera) from freshly felled wood experimentally added to forests and used DNA metabarcoding to investigate the fungal DNA carried by these insects. Staphylinid beetles rarely contained fungal DNA, while Endomychus coccineus, Glischrochilus hortensis and Glischrochilus quadripunctatus frequently carried fungal DNA with a composition specific to the insect taxon. A large proportion of the obtained fungal sequences (34%) represented decomposer fungi, including well-known wood-decay fungi such as Fomitopsis pinicola, Fomes fomentarius, Trichaptum abietinum and Trametes versicolor. Scanning electron microscopy further showed that some of the fungal material was carried as spores or yeast cells on the insect exoskeletons. Our results suggest that insect-vectored dispersal is of broader importance to wood-inhabiting fungi than previously assumed.     

嗜热毁丝霉高通量培养技术及其应用

【背景】丝状真菌是一类重要的工业发酵生产宿主菌,如何进行高通量纯菌培养和高效检测筛选性能优异的菌株是工业丝状真菌研究的重要方向。【目的】研究建立丝状真菌的高通量培养技术并测试应用效果。【方法】通过对丝状真菌培养过程中的制种、接种、培养和检测研究,建立基于孔板的高通量培养技术,并以嗜热毁丝霉为例对该技术进行验证。【结果】与传统的平板制种和摇瓶接种培养方式相比,高通量孔板的培养方式将制种通量提高24倍,单位面积产孢子能力提高350%,液体培养转接效率提高10-40倍,并建立96孔板测定乙醇含量的高通量检测技术。【结论】将丝状真菌的培养和检测通量提高1-2个数量级,为快速检测丝状真菌改造过程产生的大量性状不同菌株并获得目标菌株奠定基础,为丝状真菌高通量筛选研究提供应用指导价值。     

Soil Fungal Diversity and Community Composition in Response to Continuous Sweet Potato Cropping Practices

Soil fungi are extremely important for maintaining soil health and plant production in agricultural systems. Currently, the effect of continuous cropping of sweet potato on soil fungal communities and physiochemical parameters has not been well documented. In the present study, four sweet potato fields consecutively monocultured for 1, 2, 3, and 4 years were selected to investigate the effect of monoculture on soil fungal communities through Illumina MiSeq sequencing. Continuous cropping of sweet potatoes dramatically altered the fungal community composition, whereas fungal diversity was almost unchanged. Ascomycota and Basidiomycota were the most abundant phyla in all soil samples, accounting for 32.59% and 21.14% of the average relative abundance, respectively. The abundance of some potential pathogens, such as Ascobolus spp, specifically Ascobolus stercorarius, and some unknown fungi increased significantly as the sweet potato monoculture period increased, and their presence were highly positively correlated with disease incidence. In contrast, Basidiomycota, Bullera, Fusarium and Trichocladium most likely play roles as antagonists of sweet potato disease development, as their relative abundance decreased significantly over time and were negatively correlated with disease incidence. Redundancy and correlation analyses revealed that soil pH and organic carbon content were the most important factors driving these changes. Our findings provided a dynamic overview of the fungal community and presented a clear scope for screening beneficial fungi and pathogens of sweet potato.