Occurrence of brown root rot in Ocean Expo Park,Okinawa, Japan: Infection route and preventive control measures

Ocean Expo Park, established in 1976 on subtropical Okinawa Island, Japan, has approximately 8000 planted or naturally grown trees in an area of 77.2 ha. Windfall of these trees occurs mainly due to typhoons; wood chips made from the trunks and roots of windfall trees are commonly used as mulching material throughout the park areas, despite possible infection by pathogenic fungi such as Phellinus noxius, the pathogen causing brown root rot. P. noxius is distributed in tropical and subtropical areas worldwide and has a broad host range. The fungus can infect host plants with mycelia via root-to-root contact or with basidiospores. Here, we report the current status of tree damage caused by brown root rot in the park and infer the infection routes of P. noxius based on microsatellite markers. Among a total of 294 trees surveyed in the park, P. noxius was isolated from 13 trees of five species. P. noxius was observed more frequently on Ficus microcarpa than on other tree species. Ficus superba var. japonica was identified as a new host species, and Bombax ceiba was first recorded as a host plant of P. noxius in Japan. Among the 13 P. noxius isolates from the diseased trees, 2 isolates from adjacent diseased trees were considered to be of the same genet, while the other 11 isolates were different genets, indicating that infection by basidiospores occurred frequently among trees in the entire park, although spread via root systems also occurred between at least two adjacent trees. An identical genet was not detected at distant locations, which suggests that the disease does not spread via chips. However, mulching with chips can lead to wood decay due to decaying fungi other than P. noxius. Therefore, in accordance with the precautionary principle, the park currently prohibits the chipping and mulching of windfall or damaged trees and is considering disposal methods such as burning.     

Wood‐rotting basidiomycetes are a minor component of fungal communities associated with Acacia hybrid trees grown for sawlogs in South Vietnam

Acacia hybrid (Acacia mangium × Acacia auriculiformis) clones are widely planted in Vietnam with a total of approximately 400,000 ha to meet the demand for pulpwood, sawn timber and wood chip exports. Silvicultural techniques such as pruning and thinning have been applied to improve productivity and sawlog quality of Acacia hybrid plantations. However, those techniques may also create opportunities for wood decay fungi to enter the Acacia hybrid stems through wounds and cause stem defects that reduce sawlog quality and the value of the plantation. The presence of fungal decay agents in Acacia hybrid trees was examined in two Vietnamese plantations. In July 2011, just prior to a second thinning, discoloured wood samples were taken from a three‐year‐old Acacia hybrid plantation at Phan Truong Hai for the isolation of fungi. In July 2012, approximately 18 months after pruning and thinning treatments, discoloured wood samples were taken from a three‐year‐old Acacia hybrid plantation at Nghia Trung for the isolation of fungi. DNA sequencing of the rDNA ITS identified the isolates. In May 2015, approximately 4 years after thinning and fertilizer treatments, discoloured and decayed wood samples were taken from the above (7‐year‐old) Acacia hybrid plantation at Phan Truong Hai for fungal identification. DNA was extracted directly from discoloured and decayed wood samples and fungal rDNA ITS amplicons sequenced on a Roche 454 sequencer. The results showed that silvicultural treatments did not affect the fungal communities associated with discoloured and decayed wood of Acacia hybrid plantation at Phan Truong Hai. A total of 135 fungal species or OTUs (operational taxonomic units) were identified, including 82 members of Ascomycota and 52 Basidiomycota.     

Biodegradation of Hevea brasiliensis wood by Rigidoporus lignosus and Phellinus noxius1

In vitro wood slats degradation assays reveal that both the white root rot fungus R. lignosus and the brown root rot fungus P. noxius cause a white rot of wood. In vivo (infected tap roots) they cause the same type of decay. Nevertheless lignin determination show the rubber-tree ability to react against the parasite aggression by increased lignification of tissues.     

Interactions between Scytalidium species and brown- or white-rot basidiomycetes in birch wood

Summary Growth and subsequent decay by seven basidiomycetes in birch wood blocks was inhibited by three Scytalidium isolates. Scytalidium initially colonized the surface of the blocks and gradually overgrew the basidiomycetes. In individual wood blocks from 11 Scytalidium-basidiomycete paired treatment combinations, the basidiomycete was not inhibited throughout the entire wood block. These wood blocks demonstrated interspecific interactions and antagonism between the different fungi. The white-rot fungi responded to isolates of Scytalidium by occluding xylem cells with masses of hyphae, forming pseudosclerotial plates in the zone of initial interaction. Scytalidium appeared to gain access into portions of wood colonized by the basidiomycetes only after substantial decay had resulted by the wood decay fungus.     

The occurrence of Phellinus tremulae (Bond.) Bond. and Borisov as a primary parasite in Populus tremula L.

Phellinus tremulae was inoculated on Populus tremula. It spread in the sapwood and inner wood of a living tree without other organisms being present. Plating experiments with wood meal from different parts of a sound tree showed that there were very few, if any, propagules of micro organisms in the sound wood, neither were there any organisms found in front of the decay of P. tremulae at the top of the decay column. These results support the theory that P. tremulae is a primary parasite in aspen. Decay fungi grown on fresh and autoclaved wood of aspen had very different growth rates. Most of the fungi tested grew best on autoclaved sapvvood. The difference in growth rate on fresh and autoclaved heartwood was lcss significant, suggesting that there is a barrier in the living wood which is destroyed by autoclaving. P. tremulae was thc only fungus that grew best on fresh inner wood.     

Interactions between callus cultures of Pinus sylvestris and pine fungi with different trophic properties

Fungal virulence may be studied using tissues cultures of host plants in dual cultures in vitro, enabling analyses of interactions with undifferentiated cells of their host plants. Three genotypes of Pinus sylvestris callus, initiated by somatic embryogenesis, were used for establishing dual cultures with fungi pathogenic, endophytic or saprotrophic on pine needles or shoots. Fungal growth towards the plant callus tissue differed, depending on the life strategy of the fungus. The pathogen Gremmeniella abietina proved the slowest colonizer of callus whereas the saprotrophic Phacidium lacerum was the fastest. Gremmeniella abietina partially overgrew the callus, causing extensive necrosis and death within 10 days after inoculation. Anthostomella formosa, an endophyte of pines, did not cause evident symptoms of callus degradation: after 10 days of dual culture, the callus cells remained greenish and at least 50% of cells were alive. In dual cultures Ph. lacerum, callus remained alive until the end of the experiment, maintaining a white‐creamy colour with a loose cell structure. Electrophoresis of protein extracts from the callus showed the presence of additional bands of 25–35 kDa only in host tissues challenged with the pathogen G. abietina, possibly indicating the production of pathogenesis‐related proteins. This work has shown that pine callus does not respond equally to challenge with different fungal isolates. In general, one‐third of the isolates of each fungus examined showed greater virulence compared to other isolates.     

Anatomical characterization of decayed wood in standing light red meranti and identification of the fungi isolated from the decayed area

To further our understanding of wood decay in living light red meranti (Shorea smithiana) trees, microscopic characteristics of the cell and cell wall degradations of S. smithiana wood in the presence of the decay fungi, the identity of the causal fungi, and the decay potential and pattern by an isolated fungus were investigated. Cell wall degradations, including cell wall thinning, bore holes formation, rounded pit erosion, and eroded channel opening were clearly observed under light and scanning electron microscopy. In transverse view, many large voids resulting from a coalition of degraded wood tissue appeared in the decayed canker zone. All these observations suggest the well-known simultaneous decay pattern caused by white-rot fungi. By phylogenetic analysis based on the sequences of internal transcribed spacer region of ribosomal DNA, a basidiomycete fungus isolated from the decayed wood was identified as Schizophyllum commune. The degradation caused by this fungus on sound S. smithiana wood in an in situ laboratory decay test was classified as the early stage of simultaneous decay, and showed a similar pattern to that observed in the wood samples naturally decayed.     

In vitro interactions between bacteria isolated from Sitka spruce stumps and Heterobasidion annosum

Culture medium composition affected antagonism by bacterial isolates from Sitka spruce (Picea sitchensis) stumps against Heterobasidion annosum. Fifty percent of bacterial isolates inhibited H. annosum growth on sporulation agar or yeast–dextrose–peptone agar; only 10% of isolates caused inhibition on both media. Proportions of isolates inhibiting H. annosum varied with stump age; fewer isolates from 4‐ or 6‐year‐old stumps exhibited antagonism than isolates from older or younger stumps. Fifteen isolates showing antagonism on sporulation agar were tested against H. annosum in spruce wood cubes. None of the bacterial isolates alone caused a significant weight reduction in inoculated cubes. Relative inoculation times of bacterial isolates and H. annosum had an effect on weight loss in interactions; simultaneous inoculation with isolates and H. annosum inhibited weight loss caused by H. annosum compared with bacteria‐free controls. Inoculation with bacterial isolates 10 days before H. annosum had no effect on the decay rate. In contrast, inoculation with H. annosum 10 days before bacteria increased weight loss of cubes by 200% relative to cultures lacking bacteria. The effect of a mixed bacterial inoculum on weight change in 0.2‐mm spruce wood slips co‐inoculated with H. annosum, Resinicium bicolor, Hypholoma fasciculare, Stereum sanguinolentum or Melanotus proteus differed between different fungi.     

Growth of Phlebiopsis gigantea in wood of seven conifer species

Heterobasidion parviporum and Heterobasidion annosum are widely distributed root‐rot fungi that infect conifers throughout Europe. Infection of conifer stumps by spores of these pathogens can be controlled by treating fresh stumps with a competing non‐pathogenic fungus, Phlebiopsis gigantea. In this study, growth of three Latvian strains of P. gigantea and the biological control agent ‘Rotstop’ strain was evaluated in stem pieces of Norway spruce, Scots pine, lodgepole pine, Douglas‐fir, Weymouth pine, Siberian larch and Sitka spruce. The growth rates of one H. parviporum and one H. annosum isolate were also measured in the same stem pieces. The growth rate of P. gigantea varied greatly in wood of different conifer species. It was higher in the three pine species, lower in Norway spruce and lowest in Sitka spruce and Siberian larch, and in Douglas‐fir, this fungus did not grow. The largest area of wood occupied by P. gigantea was in lodgepole pine. Growth of Latvian isolates of P. gigantea in the wood of Pinus and Picea species was comparable to that of the Rotstop isolate. Consequently, stump treatment with local P. gigantea isolates should be recommended. However, our results suggest that Douglas‐fir stump treatment against Heterobasidion by P. gigantea may be ineffective and other stump treatment methods should be considered.     

Pinus taeda L. response to differential inoculum density of Leptographium terebrantis colonized toothpicks

Bark beetle‐vectored ophiostomatoid fungi, Leptographium terebrantis, is inoculated on the roots and lower stems of stressed Pinus species during the feeding activity of bark beetle. To determine the exact host response following inoculation, it is critical to challenge the host with a realistic amount of fungal inoculum. Thus, we designed a series of stepwise experiments using L. terebrantis colonized toothpicks which focused on the inoculum transfer from the toothpicks to excised Pinus taeda stem segments and living saplings, respectively, at different inoculum densities. The toothpicks served as a substrate for fungal growth and sporulation and the inoculation showed their utility in eliciting host's response to the pathogen. The inoculated fungus caused blue‐stain and sapwood occlusions in P. taeda stems and saplings, respectively. The volume of occluded, visually damaged sapwood increased by 1.96 cm³ per radial inoculation point on average. Fungal colonized toothpicks can be used as a suitable alternative to agar discs for studying bark beetles vectored fungi and their host interactions.     

Quantitative assessment of fungal colonization in Norway spruce after green pruning

In July and in October of 1978, 1979, 1980 five Picea abies stands in Baden-Württemberg, Germany were green-pruned up to a tree height level of c. 10 m. In 1993–94, 30 pruned trees and five control trees from each stand were harvested for investigation. The quality of pruning proved to be excellent because only few branch collar, bark injuries or too long branch stubs were detected. Based on isolations from more than 6000 wood specimens of c. 20 mm³ obtained from 175 trees, specific infection rates are given for different tree compartments. Wood decay fungi as well as important blue stain fungi were only rarely present. Tolerable discolourations were limited to the stub containing core of a few trees. Wood formed after pruning showed neither more fungal infections nor other related disadvantages compared with the respective increment from unpruned control trees. Nectria fuckeliana, proved to be the most abundant fungus in pruned trees especially in the branch stubs. Respective infection rates in and close to dry branches were compared. Bacteria could be found in the pruned branch stubs, however, they did not penetrate into the heartwood or sapwood of the stems. Both summer and autumn pruning involve only very low risk of wood deterioration. However, summer pruning shows some advantages with respect to lower infection rates by N. fuckeliana.     

Sporendocladia bactrospora associated with wounds on native broadleaved trees in Norway and Sweden

A survey to identify ophiostomatoid fungi that infect wounds on native Norwegian and Swedish broadleaved trees was undertaken during summer 2004. A fungus resembling a species of Sporendocladia was commonly isolated from the exposed cambium and inner bark of wounds. Morphological examination and comparisons of DNA sequence data for the ITS and 5.8S regions of the rRNA gene region led to its identification as Sporendocladia bactrospora. Pathogenicity trials on young Populus tremula and Betula pubescens trees showed that S. bactrospora is capable of causing lesions on these trees. There have been few previous reports of S. bactrospora, and in most cases, these have been as saprophytes on wood. In contrast, results of this study show that it is a common inhabitant of freshly made wounds on native broadleaved trees in Scandinavia, and it appears to contribute to staining of wood.     

Identification of wood‐decay fungi and assessment of damage in log depots of Western Black Sea Region (Turkey)

The aim of this study was to determine and quantify the wood‐decay fungi found on logs of forest tree species (beech, oak, hornbeam, Scots pine and fir) stored in log depots located in six different provinces in the Western Black Sea Region of Turkey. Additionally, it was aimed to determine the natural durability of some important wood species against the most commonly detected wood‐decay fungi. Eighteen families, 31 genera and 45 species belonging to the division Basidiomycota were detected; Antrodia crassa was identified for the first time in Turkey. The abundance of Panus neostrigosus, Polyporus meridionalis, Trametes hirsuta, T. versicolor and Stereum hirsutumincreased significantly with the holding time of the logs (r = 0.99, 0.87, 0.53, 0.57 and 0.78, respectively, p < 0.05). The majority of the fungal species were detected on logs stored in depots for 4–6 years (66%). The percentage of fungal species found on the logs with a holding time of three years or less was 29%, whereas the percentage for those detected on logs stored for seven or more years was 31%. Among the wood species, the greatest number of fungal species (29) and highest amount of fungi (2,539) occurred on beech wood. Natural durability tests showed that T. versicolor caused the greatest loss of wood mass, with an average of 23%. Field studies and natural durability tests performed in the laboratory showed that beech wood lost the most mass among the timber species studied.     

Ultrastructural aspects of rubber tree root rot diseases

The aggression of Hevea brasiliensis roots by two root-rot fungi, Rigidoporus lignosus and Phellinus noxius is studied at cellular level. Ultrastructural observations reveal the root penetration and tissues colonization by the fungal hyphae, showing clearly alteration in the host cell wall, leading to the root decay. This paper describes the different mechanisms used by these two fungi in degrading lignified and non lignified tissues.     

Brown root rot of trees caused by Phellinus noxius in the Ryukyu Islands,subtropical areas of Japan

The fungus Phellinus noxius has a broad host range and causes brown root rot in a variety of tree species of various ages, irrespective of their original health. The fungus is widely distributed in tropical countries of Southeast Asia, the Pacific Islands and Australia, Central America and Caribbean, and Africa. Since 1988, when brown root rot was first found on Ishigaki Island, Okinawa Prefecture, the disease has been reported on several islands in Okinawa and Kagoshima Prefectures, and it has been causing serious problems to shade, windbreak, ornamental and landscape trees in the Ryukyu Islands, located in the subtropical region of southern Japan. Here, we report on the current status of P. noxius‐caused brown root rot in the Ryukyu Islands, Japan, including symptoms, occurrence, dispersal, distribution and host tree species of the disease, pathogenicity and/or virulence of the pathogen, and disease management based on our surveys and previously published reports from Japan. Brown root rot has been confirmed in 53 tree species from 32 plant families at different sites on 10 of the 12 islands surveyed. Among the 53 tree species, 34 were first recorded in Japan as host plants of P. noxius. The disease occurs mainly in shade, ornamental and windbreak trees at sightseeing places, parks, roadsides, agricultural land such as sugar cane fields, and around residences or other places associated with human activities.     

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.     

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.     

Genotype distribution of Raffaelea quercivora in the oak galleries and its composition in the mycangia of Platypus quercivorus

Raffaelea quercivora is the pathogenic fungus that causes Japanese oak wilt. The female monogynous ambrosia beetle, Platypus quercivorus, carries this fungus in mycangia on the pronotum. These beetles bore galleries in oak trees with their partners to produce offspring, and they deposit fungus on the gallery walls from their mycangia. The offspring mature in the gallery, before loading the fungal pathogen and flying from the gallery to other healthy trees. To investigate the unloading and loading modes of the fungus within the gallery, we developed four polymorphic microsatellite markers for R. quercivora and identified the fungal genotypes in the galleries and mycangia of the beetles. Small wood chips were sampled at 5–10‐mm intervals from the walls of five galleries in a dead Quercus serrata tree. The pronota were also sampled from five female adult beetles. The genotypes of the R. quercivora isolates from the wood chips and pronota were identified using the microsatellite makers. The genotypic analysis showed that each gallery was inhabited patchily by 5–10 genotypes of R. quercivora, and the mycangia of each beetle contained 3–6 genotypes. These results indicate that diverse R. quercivora genotypes are unloaded repeatedly from the mycangia of female beetles onto the gallery wall, which results in their patchy distribution on the walls. When the offspring leave the host tree, the fungal clones that proliferate in the walls are also loaded repeatedly into the mycangia of the mature beetles.     

Fungal decomposition of sapwood and heartwood of European aspen,Populus tremula L.

In contrast to the woodrotting fungi which can only attack Populus tremula after felling, Phellinus tremulae successfully invades the central part of the living tree. Possible causes for this difference were investigated. It was found that sterile wood samples were decomposed more rapidly by saprophytic fungi than by parasitic fungi. With both types of fungi, heartwood was more resistant to decay than sapwood. It is postulated that living sapwood inhibits infection by most woodrotting fungi, but that P. tremulae can successfully overcome this barrier. This postulate cannot be confirmed by experiments using sterilized wood samples.     

Effect of pruning Acacia mangium on growth,form and heart rot

Small volumes of timber are now being produced from Acacia mangium plantations in Indonesia. These trees require pruning and thinning to increase the strength and appearance of the wood. However, cut surfaces from pruning are potential infection courts for the entry of decay-causing fungi like heart rot. This study investigated the effects of pruning on stem form and the incidence of heart rot in an 18-month-old plantation of Acacia mangium in South Sumatra. The objectives were to assess whether pruning is associated with an increase in the incidence of heart rot and whether form pruning compared to lift pruning reduced the incidence of heart rot and improved stem form. Form pruning removed 25% of leaf area by removing large branches and those subtending a narrow angle with the stem up to 3 m height, and lift pruning removed 25% of crown length from below. Trees in these treatments were singled before pruning. The third treatment, a control, was not singled and was used to assess base levels of heart rot.No significant difference in diameter increment between the two pruning treatments was found. There was strong evidence that form pruning was associated with better form 18 months after treatment. Trees in this treatment had a reduced number of branches >30 mm diameter and improved stem straightness (reduced kink). Lift pruning reduced average branch size but did not improve stem straightness. No heart rot was detected in any treatment.The results showed that form pruning is likely to have positive benefits on stem straightness and is likely to be effective to any selected pruning height. However a subsequent lift pruning is still considered a requirement. While wounds created from pruning and singling are assumed to have a large impact on the incidence of heart rot, this may not be an issue unless there is a sufficient source of fungi present in the environment to invade the wounds.