Assessment of the decay risk of airborne wood-decay fungi III: decay risks at different sampling sites

The decay risk of airborne wood-decay fungi in the same volume of air was investigated by using an air sampler over the course of a year at three different sampling sites. Japanese cedar disks measuring 7.8 cm in diameter and about 3 mm in thickness, and with a moisture content of about 100 % were placed in a “BIOSAMP” air sampler and then exposed to 1000 l of air in the northern, central, and southwest parts of Japan. The exposed disks were incubated for 20 weeks in a damp container maintained at 26 ± 2 °C and degraded by fungi trapped on the disks. The decay risk was calculated from the mass loss during incubation, and the factors affecting the said risk were explored. The results showed that sampling sites apparently do not affect decay risk, even though the Scheffer’s climate indexes of the sites were quite different. The relation between the sampling month and decay risk reveals that decay risk remains virtually the same year-round. Relative humidity on a sampling day is one of the key factors affecting decay risk in sampling conducted at the central or southwest site. In contrast, no weather factors influenced decay risk at the northern sampling site.     

Molecular identification of decay fungi in the wood of urban trees

Trees are valuable for urban areas, however, are also susceptible to wood rot fungi. For accurate and fast assessment of the severity and evolution of decay in standing trees, a molecular technique was used to identify the causal agents of wood rot. Fruit bodies of wood decay fungi were collected from infected trees in various stands in Germany. Thirty-six species were identified by traditional methods. The DNA of fruit bodies was extracted, ITS-rDNA amplified by PCR, and ITS regions sequenced. Wood samples from infected urban trees were collected, the entire DNA extracted from affected wood parts, and fungal ITS amplified and sequenced. Fungal species were identified by comparing sequence data with the fruit body data. The technique enables an accurate and rapid identification of causal rot fungi in urban trees.     

Assessment of decay risk of airborne wood-decay fungi

The decay risk of airborne wood-decay fungi was investigated by using an air sampler. Japanese cedar disks 7.8 cm in diameter and about 3 mm in thickness with moisture content of about 100% were placed in a “BIOSAMP” air sampler and exposed to 1000 l air. Air sampling was carried out from June to September at the same sampling site in Tsukuba, Japan. The exposed disks were then incubated for 16 weeks in a damp container kept at 26° ± 2°C. During the incubation period, wood mass loss ranged from −15 to 807 mg with a mean mass loss of 244 mg. Factors affecting mass loss were explored. Wood moisture content and ratio of heartwood area proved to be significant factors. In addition, six weather factors were found to influence mass loss. Disks that were sampled on a cloudy day showed significantly higher mean mass loss compared to those sampled on a sunny day. Subculturing of filamentous fungi from 16-week incubated disks suggested one-third of the isolated fungi produced ligninolytic enzymes.     

A molecular diagnostic assay for the detection and identification of wood decay fungi of conifers

Ten taxon‐specific primers were designed to amplify the Internal Transcribed Spacer of the rRNA operon of several important decay fungi of coniferous wood, including Armillaria spp., Echinodontium spp., Fomitopsis pinicola, Fuscoporia torulosa, Heterobasidion annosum sensu lato (s.l.), Onnia spp., Phaeolus schweinitzii, Phellinus weirii s.l., Pholiota spp. and Porodaedalea spp. Primers designed in this study and in a previous one for the identification of Laetiporus sulphureus and Stereum spp. were combined in two multiplex PCRs, which were tested for efficiency and specificity, and detected at least 1 pg of fungal target DNA. Target DNA at concentrations of 10^?1 pg or lower can be detected with this assay using SYBR^® Green Real‐Time PCR. Validation assays performed on 129 naturally infected wood samples or fruiting bodies confirmed the reliability of the multiplex PCR‐based diagnostic method. This method represents a simple and rapid diagnostic tool for the detection of a number of destructive wood decay fungi of conifer wood.     

Root-associated fungi of healthy-looking Pinus sylvestris and Picea abies seedlings in Swedish forest nurseries

The aim of this study was to assess fungal communities in roots of healthy-looking Pinus sylvestris L. and Picea abies (L.) Karst. seedlings in nine forest nurseries in Sweden using a combination of traditional culturing and direct sequencing of internal transcribed spacer of fungal ribosomal RNA (ITS rRNA) from the roots. Culturing from 1800 surface-sterilised root segments resulted in 2387 fungal cultures representing 42 different taxa. Direct sequencing from 180 root segments resulted in 119 ITS rRNA sequences representing 25 different taxa. In total, 55 different fungal taxa were detected using both methods. Although direct sequencing was more efficient than culturing in detecting different fungal taxa, both methods provided complementary information about fungal communities in roots since each detected rather different groups of fungi. The most dominant taxa detected by culturing were Trichoderma viride Pers. (19.5%), Phoma mucivora Davey & Currah (19.1%), Phialocephala fortinii Wang & Wilcox (17.4%) and Meliniomyces variabilis Hambl. & Sigler (10.2%), while Thelephora terrestris Ehrh. (26.1%), Unidentified sp. NS126 (25.2%) and Heliotales sp. C20 (10.1%) were most commonly detected by direct sequencing. In conclusion, results showed that forest nurseries in Sweden harbour diverse communities of fungi associated with the roots of healthy-looking P. sylvestris and P. abies seedlings. Although fungal communities were often dominated by saprotrophs and endophytes, several facultative pathogens were also detected indicating that under suitable conditions they may be a potential threat to the plants.     

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.     

Biocontrol of decay in seasoning utility poles. I. Growth rate and colonizing ability of bluestain and decay fungi in vivo and in vitro

Seasoning (air drying) of utility poles for 6–12 months is essential before preservative treatment can be achieved. However, during seasoning, pine sapwood is often colonized by decay fungi, thereby compromising the performance and service life of the poles. This study investigated the potential of bluestain fungi to act as short‐term biocontrol agents against decay during seasoning. An important attribute for biocontrol is rapid growth, so growth rates of common bluestain (Ceratocystis coerulescens, Ophiostoma minus, Ophiostoma piceae, Ophiostoma piliferum, Sphaeropsis sapinea) and decay fungi (Heterobasidion annosum, Phlebiopsis gigantea, Stereum sanguinolentum) were compared on agar medium and pine in logs at various temperatures. On agar, the growth temperature optimum of most bluestain fungi and all the decay fungi was ～25°C, with little growth at ≤5°C or above 32.5°C. Overall, the fastest growing were S. sapinea and O. minus. In logs, the most effective colonizers were S. sapinea and O. minus with pathogenic abilities that made them well fitted to colonize the sapwood of freshly felled pine. Within these species, certain isolates produced much larger lesions in phloem and the sapwood tangential plane than all the decay fungi. Notably, there was significant variation in colonizing ability between different isolates within a species, emphasizing the need for testing a range of isolates when selecting a potential biocontrol agent.     

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.     

Occurrence and pathogenicity of fungi in necrotic and non-symptomatic shoots of declining common ash (<Emphasis Type="Italic">Fraxinus excelsior</Emphasis>) in Sweden

Currently, massive dieback of Fraxinus excelsior is observed in countries of eastern, northern and central Europe, and the reasons for it are unclear. The aims of the present work were (a) to study fungal communities in declining F. excelsior crowns; (b) to clarify role of fungi in the decline. Shoots from symptomatic crowns were collected in four localities in central Sweden, and distributed into the following categories: (a) visually healthy; (b) initial necroses; (c) advanced necroses; (c) dead tops. The most frequently isolated fungi were Gibberella avenacea, Alternaria alternata, Epicoccum nigrum, Botryosphaeria stevensii, Valsa sp., Lewia sp., Aureobasidium pullulans and Phomopsis sp., and these taxa were consistently found in shoots of all four symptomatic categories. Forty-eight taxa of other fungi were isolated, and fungal diversity was not exhausted by the sampling effort. The same taxa of fungi were dominant in F. excelsior shoots of different symptomatic categories, and moderate to high similarity of fungal communities was observed in shoots despite the symptoms. Forty-four isolates from 24 fungal taxa were used for artificial inoculations of 277 1-year-old F. excelsior seedlings in bare root nursery. After 2 years, only four fungi caused symptomatic necroses of bark and cambium: A. alternata, E. nigrum, Chalara fraxinea and Phomopsis sp. The most pathogenic was C. fraxinea, inducing symptoms on 50% of inoculated trees, while three other fungi caused necroses on 3–17% of inoculated trees. Infection biology of C. fraxinea and environmental factors determining susceptibility of F. excelsior to decline deserve future investigations.     

Occurrence and decay patterns of common wood‐decay fungi in hazardous trees felled in the Helsinki City

To improve the management of ageing urban trees, the role of wood‐decay fungi as potential causes of stem breakage was investigated among hazardous trees removed in the Helsinki City area during 2001–2004. The study material comprised 194 trees, and included 76 Tilia spp. trees, 58 Betula spp. and 60 Acer spp. Thirteen species or genera of commonly occurring decay fungi were identified on the basis of fruiting bodies and pure cultures. The occurrence of the fungi was investigated in terms of frequency, visibility and as potential causes for stem breakage. Most hazardous fungi caused extensive horizontal decay in the stem; such fungi were Ganoderma lipsiense on Tilia and Acer, Phellinus igniarius on Acer, Inonotus obliquus and Cerrena unicolor on Betula and Kretzschmaria deusta on Acer, Tilia and Betula. Typically, Rigidoporus populinus was frequently present in weak fork formations on Acer trees. Agaric fungi (Pholiota, Armillaria, Pleurotus and Hypholoma) were frequently recorded but were of minor importance from the point of view of tree breakage hazard.     

Use of fungal biosystematics and molecular genetics in detection and identification of wood‐decay fungi for improved forest management

Advances in fungal biosystematics and molecular genetics have clarified relationships among the wood‐decay fungi and are providing new tools for their detection and identification. Species complexes of forest pathogens, including those within Heterobasidion, Armillaria, Laetiporus, and Phellinus, are being resolved. The ability to isolate fungal DNA directly from wood without in intermediate culturing step will greatly facilitate sampling and disease detection and has applications in forest disease management, hazard tree assessment, invasive species detection, and carbon cycling, sequestration and climate change research. Recent changes in fungal nomenclature and their application to forest pathology are discussed.     

Ophiostomatoid fungi isolated from three different pine species in Argentinian Patagonia

Pine plantations in Argentinian Patagonia cover ca. 95,000 ha in Chubut, Río Negro and Neuquén provinces. Exotic bark beetles (Orthotomicus laricis, Hylastes ater and Hylurgus ligniperda) commonly occur in freshly cut logs, stumps and slash. These beetles are vectors of “ophiostomatoid” fungi which include primary tree pathogens as well as important agents of blue stain. The aim of this study was to identify these beetle‐associated fungi. Sawing mills and pine plantations were surveyed three consecutive years. Fungal isolates from stained logs, processed wood and insect galleries were identified based on morphological and DNA sequence comparisons of ITS and β‐tubulin gene regions. Two Grosmannia, one Graphilbum and three Ophiostoma species were identified. Ophiostoma piliferum and O. peregrinum sp. nov. were the most frequently isolated taxa. O. peregrinum occurred in all provinces, colonizing different conifer species and, interestingly, also the native broadleaved species Nothofagus dombeyi. Pine plantation forestry in southern South America includes Argentina, Brazil, Chile, Paraguay and Uruguay. Emerging data from Argentina, Chile and Uruguay revealed some coincidences between these countries, but also several differences, probably, as a result of multiple introduction events.     

The effect of bagging branches on levels of endophytic fungal infection in Japanese beech leaves

The species composition of the endophytic mycobiota in leaves of Japanese beech trees (Fagus crenata) and the sources for leaf infections were studied in a forest reserve situated in central eastern Honshu, Japan. To clarify the mechanism of infection of leaves, half of the branches were covered with polyethylene bags and species composition and levels of endophytic fungal infection were then compared with those of unbagged controls. Isolations were carried out from the leaves, petioles, and current‐year twigs of both, bagged and unbagged branches. Additionally, species composition was detected in overwintered terminal buds of beech trees and in the leaves of potted seedlings that had been placed in the field in different seasons. The species assemblage of the unbagged leaves, petioles, and current‐year twigs was dominated by Mycosphaerella buna, Ascochyta fagi, Periconiella sp., and Tritirachium sp. Other frequently recovered species were Xylaria sp., Phomopsis sp., and Tubakia dryina. Mycosphaerella buna and A. fagi were never isolated from leaves on bagged branches. A. fagi was, however, detected on both bagged and unbagged petioles and current‐year twigs at comparatively low isolation frequencies. The detection of Periconiella sp. on all occasions in both bagged and unbagged leaves was a characteristic feature that differs from those of the other three dominant endophytic fungi. The fungus was also detected without significant differences in bagged and unbagged petioles and current‐year twigs on most sampling dates. Furthermore, Periconiella sp. was isolated from immature twigs inside the bud scales. Tritirachium sp. was frequently detected in unbagged leaves and petioles and in both bagged and unbagged current‐year twigs, and rarely in bagged leaves and petioles, but was never recovered from terminal buds. The results of the potted seedling experiments revealed that all four dominant species had airborne inocula. The infection of leaves by M. buna occurs exclusively by airborne propagules, i.e. ascospores in spring and conidia in autumn. In Periconiella sp. hyphal growth of the fungus from immature twigs inside the buds into the leaf tissues was suggested in addition to infection by airborne inocula. Tritirachium sp. hyphae were suggested to grow from previous‐ to current‐year twigs. Ascochyta fagi was present in the outermost scales of overwintered terminal buds, but no systemic growth of the fungus into the petioles and current‐year twigs was observed. Our technique of covering the branches before new leaves unfolded was effective in preventing infection by airborne inocula of endophytic fungi.     

Mycorrhizal status of some fungi fruiting beneath indigenous trees in Burkina Faso

During the rainy season, putative ectomycorrhizal fungi were observed under three species of Caesalpinioideae Afzelia africana Sm., Isoberlinia doka Craib. and Stapf. and Isoberlinia dalziellii Craib. and Stapf., one species of Dipterocarpaceae Monotes kerstingii Gilg. and two species of Euphorbiaceae Uapaca guineensis Müll. Arg. and Uapaca somon Aub. and Lean. The fungi belong to the orders of Agaricales, Aphyllophorales, Boletales, Cantharellales, Gautieriales, Hymenogastrales, Russulales and Sclerodermatales. Some of these fungal species e.g. Lactarius gymnocarpus Heim., Cantharellus pseudofriesii Heinem., Scleroderma dictyosporum Pat., Scleroderma verrucosum Pers., Scleroderma sp2 and Russula sp1, are common to the tropical tree families known to form ectomycorrhizas. A few species of these fungi e.g. Amanita hemibapha (Berk. and Br.) Sacc., Inocybe sp1, Boletellus sp3, Lactarius sp2 and Xerocomus subspinulosus Heinem., are observed only under mycorrhizal trees belonging to the group of Caesalpinioideae legumes. However, seven fungal species Coltricia cinnamomea (Pers.) Murr., Boletellus sp5, Austrogautiera sp. (hypogeous), Lactarius sp1, Russula annulata Heim., Russula sp2 and Scleroderma sp1 seem more specific to the Uapaca species. In all, twenty-seven putative ectomycorrhizal fungi were recovered and only five fungal species were isolated from sporocarps and maintained in pure culture. Four Scleroderma species were confirmed as mycorrhizal fungi on A. africana, Isoberlinia spp. and two East African trees, Brachystegia speciformis Benth. and Afzelia quanzensis Welw. The two Uapaca species, B. speciformis and M. kerstingii possess both arbuscular mycorrhizas and ectomycorrhizas.     

Utility pole decay

Summary The Basidiomycetes associated with decay in pine, Douglas-fir, and cedar utility poles within various geographic regions of North America were investigated. On the basis of 313 isolations from these poles, 9 fungi appeared to be of major importance in internal pole decay in the United States. These fungi were the following: Lentinus lepideus, Lenzites saepiaria, L. trabea, Peniophora A., P. gigantea, and Poria radiculosa in pine, and L. lepideus, Poria carbonica, P. monticola, and P. xantha in Douglas-fir poles. Lentinus lepideus was overwhelmingly predominant in pine poles, whereas Poria carbonica was similarly predominant in Douglas-fir poles. Lenzites trabea was the fungus most often isolated from cedar poles; however, it is believed to be associated primarily with shell rot of cedar poles and to be of little significance, therefore, in causing internal decay of cedar. The fungi associated with western red-cedar and red and jack pine poles in Canada are listed, although the frequency of their occurrence is not included.This work was in cooperation with the Navy Department, Naval Facilities Engineering Command.The author is indebted to the following for furnishing cultures and pole sections or both for culturing or for information on the identities of the species of decay fungi found in Canadian poles: Joe Clark and John Kulp, U. S. Forest Products Laboratory, Madison, Wisconsin; Robert Graham and John Mothershead, Forest Research Laboratory, Oregon State University, Corvallis, Oregon; O. Floyd Hand, Bonneville Power Administration, Vancouver, Washington; John Shields, Canadian Forest Products Laboratory, Ottawa, Ontario; and John Roff, Canadian Forest Products Laboratory, Vancouver, British Columbia. The author is especially indebted to members of the Forest Disease Laboratory, Laurel, Maryland, for their invaluable aid in identifying representative cultures of many of the fungi discussed in this investigation.The Laboratory is maintained at Madison in cooperation with the University of Wisconsin.     

Qualitative and quantitative PCR methods using species-specific primer for detection and identification of wood rot fungi

Species-specific oligonucleotide primers for detecting wood rot fungi, Gloeophyllum trabeum, Trametes versicolor, Coniophora puteana, and Serpula lacrymans, and the primer detecting a group of related fungi to G. sepiarium were developed. These primer sequences were picked up from the internal transcribed spacer region between small-subunit rDNA and large-subunit rDNA. The species selectivities of the developed primers were checked. Real-time polymerase chain reaction (PCR) was carried out using these highly specific primers to quantitatively detect at least of 0.01 ng genome DNA of the target species. This quantitative PCR was also used to differentiate the target species DNA from mixed species DNA. A PCR-based technique using the species-specific primers would be applicable to multiple-sample assay in diagnosis of wood decay and to investigation of environmental fungal populations. Part of this article was presented at the International Symposium on Wood Science and Technology (IAWPC 2005), Yokohama, November 2005     

Occurrence of common phyllosphere fungi of horse-chestnut (Aesculus hippocastanum) is unrelated to degree of damage by leafminer (Cameraria ohridella)

The horse chestnut leaf-miner (Cameraria ohridella Deschka & Dimi?, Lepidoptera, Gracillariidae) is an invasive pest causing extensive damage to leaves of the horse-chestnut (Aesculus hippocastanum L.) in Europe. In Lithuania, C. ohridella invaded in 2002 causing wilting, browning and premature fall of A. hippocastanum leaves. The aim was to get a better understanding of possible linkages between foliar fungal communities and leaf-miner damage in A. hippocastanum. Leaves of A. hippocastanum, differentially damaged by C. ohridella, were collected in 10 sites in Lithuania. The fungal communities were described through DNA isolation and amplification using an ITS rRNA marker and Ion Torrent-sequencing. Clustering of 214,897 high-quality sequences resulted in 1017 non-singleton fungal taxa, among which Aureobasidium pullulans (28.2% of all fungal sequences), Endoconidioma populi (27.7%), Phoma fungicola (11.3%), Cladosporium ramotenellum (7.6%) and Cryptococcus sp. 2185_4 (5.0%) were most common. Correspondence analysis showed that fungal communities from heavily and slightly damaged leaves were largely intermingled, showing that in both types of samples fungal communities were similar. In conclusion, the study demonstrated that the phyllosphere of A. hippocastanum is inhabited by a high diversity of fungal species, the majority of which constitute generalist endophytes, epiphytes and saprotrophic fungi. The occurrence of common phyllosphere fungi was unrelated to the degree of damage by C. ohridella.     

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.     

Scale insects,decay and canker fungi in American beech

Tripartite interactions among phytophagous insects, pathogens and their host plants provide insight into the role of host physiology in determining susceptibility to attack. American beech (Fagus grandifolia) often is simultaneously attacked by beech scale (Cryptococcus fagisuga), one or more Neonectria pathogens and Xylococculus betulae that can result in beech bark disease (BBD). Additionally, beech is frequently infected by heartrot‐decay fungi. Cursory observations in 2011 suggested that beech scale and Neonectria lesion densities were lower and greater, respectively, on trees with decay. In 2012, digital image analysis was used to quantify densities of these organisms on 123 beech from the Adirondack region of New York. Three groups of study trees (n = 41) were used: Inonotus glomeratus‐infected, Phellinus igniarius‐infected and non‐decay trees. Trees infected by decay pathogens supported lower densities of beech scale and higher densities of Neonectria. Densities of X. betulae did not significantly vary among decay groups. These results may be explained by decay‐induced changes in host physiology. Additional work is needed to elucidate the potential role of host bark chemistry in the BBD complex.     

Fungi associated with the fir bark beetle Cryphalus piceae in Poland

The aim of this study was to identify fungi associated with Cryphalus piceae on European silver fir (Abies alba) in Poland and to test the pathogenicity of selected isolates. Fungi were isolated from five populations of overwintered adults and their galleries. A great diversity of taxa was associated with C. piceae. In total, 2487 isolates, including 58 species distributed in 25 genera, were obtained. The two most frequently isolated fungi, an undescribed species of the genus Geosmithia and Ophiostoma piceae, appeared to be specifically associated with C. piceae, whereas Pesotum fragrans, Pesotum sp. and Sporothrix sp. were sporadically associated. Two‐year‐old seedlings of silver fir were wound‐inoculated with three species of fungi (Geosmithia sp., O. piceae and Pesotum sp.) recovered from C. piceae. Only Pesotum sp. showed pathogenic ability, but we do not consider it to be an important pathogen of A. alba.