Occurrence of fungi on submerged pine and teak blocks in Hong Kong coastal waters

An investigation of the lignicolous marine fungi in Hong Kong coastal waters was carried out for 18 months in order to determine successional patterns of fungal occurrence. Submerged blocks of pine (Pinus massoniana Lambert) and teak (Tectona grandis Linnaeus) were used as baits and were scraped at intervals to remove surface fouling organisms. Occurrence of fungi is described in terms of the composition of the fungal flora and the number of fungal colonies developed on the test blocks before and after incubation. The scraped teak and pine blocks supported slightly different fungal floras, and the selective effect of these substrates on fungal colonization is discussed.     

Reciprocal interactions between Scots pine and soil food web structure in the presence and absence of ectomycorrhiza

Mycorrhizal plants are commonly believed to direct much more of their photosynthates into the soil than non-mycorrhizal plants. As the growth of most organisms of the detrital food web is limited by energy, the flow of C through mycorrhizal plants into the below-ground milieu is widely assumed to nourish a variety of decomposer organisms in soils. In the current experiment, I explored whether some representatives of soil mesofauna, either fungivores or microbi-detritivores, derive benefit from the presence of ectomycorrhizal (EM) fungi growing on the roots of Scots pine (Pinus sylvestris). I also investigated whether the role of soil mesofauna in affecting pine growth depends on the presence of EM fungi in the pine rhizosphere. The study was established in microcosms with a mixture of raw humus and sand. The soil was defaunated, reinoculated with 10 species of soil bacteria and 11 species of saprophytic soil fungi, and pine seedlings, either infected or non-infected with four taxa of EM fungi, were planted in the microcosms. Five treatments with different food web configurations were established: (1) saprophytic microbes alone, (2) as (1) but with the omnivorous enchytraeid species Cognettia sphagnetorum present, (3) as (1) but with Collembola (Hypogastrura assimilis), (4) as (1) but with four species of oribatid mites (Acari) involved, and (5) as 1) but with C. sphagnetorum, H. assimilis and the Acari. The microcosms were incubated in a climate chamber with varying temperature and illumination regimes for two growing periods for the pine. After 60 weeks, pine biomass production was significantly greater in the mycorrhizal systems, the total biomass being 1.43 times higher in the presence than absence of EM fungi. Similarly, almost ten times more fungal biomass was detected on pine roots growing in the mycorrhizal than in the non-mycorrhizal systems. The presence of EM fungi was also associated with significantly lowered pH and percent organic matter of the soil. Despite the clearly larger biomass of both the pines and the fungi on the pine roots, neither the numbers nor biomasses of the mesofauna differed significantly between the EM and non-EM systems. The presence of Collembola and C. sphagnetorum had a positive influence on pine growth, particularly in the absence of EM fungi, whereas oribatid mites had no effects on pine growth. The complexity of the mesofaunal community was not related to the biomass production of the pines in a straightforward manner; for example, the complex systems with each faunal group present did not produce more pine biomass than the simple systems where C. sphagnetorum existed alone. The results of this experiment suggest that the short-term role of EM fungi in fuelling the detrital food web is less significant than generally considered, but that their role as active decomposers and/or stimulators of the activity of saprophytic microbes can be more important than is often believed. Received: 22 December 1999 / Accepted: 14 April 2000     

Defoliation effects on enzyme activities of the ectomycorrhizal fungus <Emphasis Type="Italic">Suillus granulatus</Emphasis> in a <Emphasis Type="Italic">Pinus contorta</Emphasis> (lodgepole pine) stand in Yellowstone National Park

Ectomycorrhizal (EM) basidiomycete fungi are obligate mutualists of pines and hardwoods that receive fixed C from the host tree. Though they often share most recent common ancestors with wood-rotting fungi, it is unclear to what extent EM fungi retain the ability to express enzymes that break down woody substrates. In this study, we tested the hypothesis that the dominant EM fungus in a pure pine system retains the ability to produce enzymes that break down woody substrates in a natural setting, and that this ability is inducible by reduction of host photosynthetic potential via partial defoliation. To achieve this, pines in replicate blocks were defoliated 50% by needle removal, and enzyme activities were measured in individual EM root tips that had been treated with antibiotics to prevent possible bacterial activity. Results indicate that the dominant EM fungal species (Suillus granulatus) expressed all enzymes tested (endocellulase D-glucosidase, laccase, manganese peroxidase, lignin peroxidase, phosphatase and protease), and that activities of these enzymes increased significantly (P < 0.001) in response to defoliation. Thus, this EM fungus (one of the more specialized mutualists of pine) has the potential to play a significant role in C, N and P cycling in this forested ecosystem. Therefore, many above-ground factors that reduce photosynthetic potential or divert fixed C from roots may have wide-reaching ecosystem effects.     

Selective Degradation of Wood Components by White-Rot Fungi

In order to find naturally occurring white-rot fungi which preferentially degrade lignin. 25 different species of such fungi were cultivated on pine wood blocks and on kraft lignin agar plates with and without cellulose. Due to differences in phenol oxidase reactions on the kraft lignin agar plates, the 25 fungi could be divided into two groups, 1 and 2, which also differed in other properties. The three Group I fungi Sporotrichum pulverulentum, Phanerochaete sp. L1 and Polyporus dichrous produced high levels of endo-l,4-β-glucanase and cellobiose:quinone oxidoreductase in shaking cellulose flasks and a low level of phenol oxidase in standing wood meal flasks, The four fungi Merulius tremellosus, Phlebia radiata, Pycuoporus cinnabarinus and Pleurotus ostreatus from Group 2, on the other hand, produced low levels of endo-1,4-β-glucanase and cellobiose:.quinone oxidoreductase in the cellulose. flasks and a high level of phenol oxidase in the wood meal flasks. Analyses of pine wood blocks degraded by the above-mentioned fungi in the presence of either malt extract, asparagine or NH₄H₂PO₄ revealed that malt extract gave good lignin degradation. In the presence of this nutrient source. P. cinnabarinus, at 3.4% weight loss, even degraded 12.5% lignin without loss of cellulose or mannan. No common degradation pattern was, however, obtained using mall extract, asparagine or NH₄H₂PO₄, It is suggested that while-rot fungi, which preferentially degrade lignin, may be found among Group 2 fungi producing large amounts of phenol oxidases.     

Seasonal patterns of primary colonization by lignicolous marine fungi in Hong Kong

During a 14-month investigation of the seasonal pattern of primary colonization by lignicolous marine fungi in the coastal waters of Hong Kong, a pine block (Pinus massoniana Lamb.) was submerged each month at each of the five stations and recovered in the following month. Thirty-eight species of fungi were isolated, three of which were not identified. In this first recorded attempt to describe fungal colonization on submerged blocks quantitatively, five species (Periconia prolifica, Cirrenalia macrocephala, Ceriosporopsis halima, Trichocladium achrasporum and Halosphaeria quadricornuta) were the most frequent and most abundant fungi on the monthly blocks. Statistical analyses of seasonal abundance of these fungi are presented and their occurrence in relation to varying temperature and salinity throughout the year, including the Phoma pattern, are discussed.     

Factors affecting the distribution of lignicolous marine fungi in Hong Kong

An investigation into the effect of environmental factors on the general distribution and occurrence of lignicolous marine fungi using submerged blocks of pine (Pinus massoniana Lamb.) and teak (Tectona grandis L.) was carried out for 18 months in the coastal waters of Hong Kong. Five test sites, with environmental conditions varying from estuarine to oceanic, and from polluted to non-polluted, were selected. During each collection, salinity, temperature, pH, dissolved oxygen, biochemical oxygen demand, nitrate-nitrogen, inorganic phosphate-phosphorus and light transmission were measured. A total of 51 species of fungi were recorded among which only 28 were either obligate or facultative marine forms. Neither the general distribution pattern nor the distribution of the more frequent fungi could be solely accounted for by differences in salinity at the test sites and it is suggested that other ecological factors such as heavy sediments in the waters, low pH, and the presence of an abundant source of inocula may be important.     

Cellulolytic activity of white, brown and gray wood rot fungi

Culture fluids obtained from submerged cultures of white, brown and gray wood rot fungi were assayed for the presence of cellulolytic activity complexes against the model substrated carboxymethylcellulose-Na and Standard Whatman cellulose and natural substrates, i.e. celluloses isolated from pine bark and sawdust. The cellulolytic activity of the examined fungal species was highly differentiated. The use of model and natural substrates allowed determination of the high substrate specificity of the cellulase complexes produced by the fungi. Not all the fungi were found to produce EC 3.2.1.4. endo-1, 4-beta-glucanase under the culture conditions employed. All the fungi were, however, able to produce a complex of EC 3.2.1.4. exo-1, 4-beta-glucanases. All the examined fungi were also able to degrade, although to a varied extent, such higher forms of cellulose as Standard Whatman cellulose or natural celluloses isolated from pine bark and sawdust. Determination of the cellulolytic activity of fungi against the above-mentioned specific natural substrates affords the possibility of their practical use.     

Correlation between infection by ophiostomatoid fungi and the presence of subterranean termites in loblolly pine (Pinus taeda L.) roots

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The effects of toxic metals,content of nutrients and inoculation with mycorrhizal fungi on the level of phenolics in roots and growth of Scots pine seedlings

The effects of two substrates and several strains of mycorrhizal fungi on the content of soluble phenolics in roots and growth of Scots pine seedlings was investigated. The first substrate was fertile and contaminated with copper, zinc and lead, whereas the second one displayed nutrient deficiency. The dry weights of needles, trunks, roots and the total biomass were higher in groups of seedlings inoculated with mycorrhizal fungi on fertile and polluted substrate. Inoculation of pine seedlings on this substrate resulted in a decrease in concentration of phenolics in roots and except for seedlings inoculated with Laccaria laccata negatively influenced the above-ground part: root ratio.     

Biodegradation of wood in crude oil-polluted soil

A field investigation (April–November) in Nigeria showed that biodegradation of obeche (Triplochiton scleroxylon) wood blocks was initially retarded in crude oil-contaminated soil but later became enhanced as indicated by loss of compression resistance. Further indication of this pattern was the detection of soft-rot cavities and basidiomycete fungi after 2–3 months exposure when compared to control blocks in uncontaminated soil. Laboratory tests with Pleurotus sp., Trametes sp., Gloeophyllum sp. (basidiomycetes) and Chaetomium sp. (soft-rot fungus) confirmed that degradation of crude oil-coated obeche blocks was markedly retarded without the presence of hydrocarbon-degrading bacteria. The filtrate of hydrocarbon-degrading Pseudomonas sp. grown in mineral salt/crude oil medium for 3–4 weeks supported growth of the test fungi better than in carboxymethyl cellulose medium but less than in potato dextrose broth. Similarly, wood blocks immersed in the filtrate became significantly more susceptible to fungal degradation. Pseudomonas sp. from stationary phase growth in crude oil medium depleted residual sugar in basidiomycete-degraded sawdust with a concomitant marked increase in its population. It may be concluded that readily metabolizable products of crude oil degradation by soil organisms and the removal of residual sugar which may have prevented catabolite repression of cellulases, culminated in increased attack on the wood by soil-borne wood-decomposing organisms.     

Oxalic acid overproduction by copper-tolerant brown-rot basidiomycetes on southern yellow pine treated with copper-based preservatives

Accumulation of oxalic acid (OA) by brown-rot fungi and precipitation of copper oxalate crystals in wood decayed by copper-tolerant decay fungi has implicated OA in the mechanism of copper tolerance. Understanding the role of OA in copper tolerance is important due to an increasing reliance on copper-based wood preservatives. In this study, four copper-tolerant brown-rot fungi were evaluated for decay capacity and OA production in early stages of exposure to four waterborne copper-based wood preservatives (ammonical copper quat type B and D, ammonical copper citrate, and chromated copper arsenate, type C) and one oilborne copper-based wood preservative (oxine copper) in southern yellow pine blocks. Weight losses were less than 14% during the 4-week incubation. The presence of copper in waterborne preservatives uniformly stimulated OA production by the test fungi within 2 weeks of exposure of the treated blocks to test fungi; 66% to 93% more OA was produced in treated blocks than untreated controls. Oxine copper, a nickel-containing oilborne preservative, prevented both weight loss and OA production in all fungi tested.     

Impact of white-rot fungi on numbers and community composition of bacteria colonizing beech wood from forest soil

White-rot fungi are important wood-decomposing organisms in forest ecosystems. Their ability to colonize and decompose woody resources may be strongly influenced by wood-inhabiting bacteria that grow on easily utilizable compounds e.g. oligomers of wood-polymers released by fungal enzymes. However, so far, it is not known how white-rot fungi deal with the presence of potential competing bacteria. Here, the effects of two white-rot fungi, Hypholoma fasciculare and Resinicium bicolor, on the numbers and composition of bacteria colonizing sterile beech wood blocks from forest soil are reported. Both total numbers (microscopic counts) and the numbers of cultivable wood-inhabiting bacteria were considerably lower in wood blocks that became colonized by the white-rot fungi than in control blocks. This points to the fungi out-competing the opportunistic bacteria. The presence of white-rot fungi resulted in a change in the relative abundance of families of cultivable bacteria in wood and also in a change of denaturing gradient gel electrophoresis patterns of directly amplified 16S rRNA gene fragments. Analysis of the bacterial community structure in soil adhering to exploratory mycelium (cords) indicated that fungal species-specific effects on bacterial community composition were also apparent in this fungal growth phase.     

Occurrence of Bursaphelenchus mucronatus (Nematoda: Aphelenchoididae) and the microhabitat distribution of fungi in declining pine trees in a locale in Korea

A total of 33 pine trees with symptoms of decline were collected in Jeonnam Province, South Korea, and were examined for the presence of nematodes. About 20% of the trees sampled were positive with Bursaphelenchus species. All Bursaphelenchus species were found in recently dead or dying trees. Based on morphological observations, the nematode extracted from the declining pine trees was identified as B. mucronatus. The highly pathogenic pine wood nematode B. xylophilus was not found in any pine trees sampled. B. mucronatus was easily reared on fungus Botrytis cinerea. Twenty one fungal isolates were isolated from dead trees, fallen twigs, and healthy pine trees. The fungal isolates belonged to Trichoderma genus and were dominant in the wood of partially declining pines. The blue‐stain fungi transmitted by the Monochamus beetle were not detected. The B. mucronatus population decreased markedly on Auxarthron reticulatum DY‐2 isolated from soils. The number of nematodes also reduced on Verticillium saksenae A‐1, a nematophagous fungus, and Beauveria bassiana, an entomopathogenic fungus. This observation suggested the fungal production of nematicidal activity against B. mucronatus. When the fungal culture filtrates were also used for nematicidal activity on B. mucronatus, the culture filtrates of A‐1, DY‐2 and B. bassiana showed over 50% mortality within 48 h exposure. The fungi BC4, BC5 and BC6 isolated from declining pine trees inhibited the reproduction of B. mucronatus, and their culture filtrates also expressed nematicidal activity, indicating a possible interaction between the fungi in pine trees and nematodes at microhabitat level.     

Simulated nitrogen deposition affects wood decomposition by cord-forming fungi

Anthropogenic nitrogen (N) deposition affects many natural processes, including forest litter decomposition. Saprotrophic fungi are the only organisms capable of completely decomposing lignocellulosic (woody) litter in temperate ecosystems, and therefore the responses of fungi to N deposition are critical in understanding the effects of global change on the forest carbon cycle. Plant litter decomposition under elevated N has been intensively studied, with varying results. The complexity of forest floor biota and variability in litter quality have obscured N-elevation effects on decomposers. Field experiments often utilize standardized substrates and N-levels, but few studies have controlled the decay organisms. Decomposition of beech (Fagus sylvatica) blocks inoculated with two cord-forming basidiomycete fungi, Hypholoma fasciculare and Phanerochaete velutina, was compared experimentally under realistic levels of simulated N deposition at Wytham Wood, Oxfordshire, UK. Mass loss was greater with P. velutina than with H. fasciculare, and with N treatment than in the control. Decomposition was accompanied by growth of the fungal mycelium and increasing N concentration in the remaining wood. We attribute the N effect on wood decay to the response of cord-forming wood decay fungi to N availability. Previous studies demonstrated the capacity of these fungi to scavenge and import N to decaying wood via a translocating network of mycelium. This study shows that small increases in N availability can increase wood decomposition by these organisms. Dead wood is an important carbon store and habitat. The responses of wood decomposers to anthropogenic N deposition should be considered in models of forest carbon dynamics.     

The Effects of Seaweed Secondary Metabolites on Biofouling

Antifouling activity is one poorly investigated property of seaweed natural products. To determine, in the field, whether seaweeds contain chemicals able to influence the settlement of fouling organisms, crude organic extracts from Stypopodium zonale, Dictyota menstrualis (Phaeophyceae) and Laurencia obtusa (Rhodophyceae) were incorporated at natural volumetric concentrations, into hard stable gels that served as substrata for fouling in the experiments. Fouling organisms settled at a significantly higher rate on plates treated with S. zonale extracts than on control gels, while settlement was strongly inhibited on gels containing L. obtusa extracts. Fouling on gels treated with the D. menstrualis extract was not significantly different from the fouling found on control gels. The findings suggest that the broad antifouling properties of the crude extract of L. obtusa inhibit the settlement of fouling as well as hinder the development of settled fouling species, thereby reducing the richness of species. The results imply that L. obtusa possibly harbours powerful agents that can be explored for the development of antifouling technology.     

Influence of hydrodynamic stress on the frictional drag of biofouling communities

The role of hydrodynamic wall shear stresses on the development of the fouling community structure and resulting frictional drag were examined using a commercially available fouling release coating. Immersed test panels were exposed to three different hydrodynamic treatments, one static and two dynamic (corresponding to an estimated wall shear stress of 7.0 and 25.5 Pa). The drag of the panels was measured in a hydrodynamic test chamber at discrete time intervals over 35 days. The fouling community composition on the static panels was significantly different from the organisms observed on the dynamic panels. Despite different fouling community composition, the drag forces measured on the panels were very similar. This suggests that the frictional drag of low form and soft fouling communities are similar and that there may be a stepwise increase in frictional drag associated with the presence of mature calcareous organisms.     

The effect of vibration frequency and amplitude on biofouling deterrence

The use of vibration is proposed as a means of controlling the settlement of marine fouling organisms. In this study, panels with embedded lead zirconate titanate, known as PZT, were placed in the field over 3?months. The panels were vibrated at different velocity levels at frequencies between 70 and 445?Hz. It was found that barnacles (Amphibalanus variegatus Darwin and Elminius sp.) were the only fouling organisms affected by the applied vibration, and these organisms settled in significantly lower numbers when the plates were excited at specific frequencies and amplitudes. Panels vibrating at relatively higher frequencies, greater than 260?Hz, exhibited reduced barnacle settlement, whilst lower frequencies in the 70–100?Hz range had little or no effect. The settlement of other fouling organisms such as tubeworms, bryozoans, ascidians and algae did not appear to be affected by the applied excitation. The experimental results showed that increasing the velocity amplitude of vibration was a contributing factor in inhibiting barnacle settlement.     

Characterization of main sulfur source of wood-degrading basidiomycetes by S K-edge X-ray absorption near edge spectroscopy (XANES)

The main wood degraders in aerobic terrestrial ecosystems belong to the white- and brown-rot fungi, where their biomass can be created on wood decay only. However, total sulfur (S) concentration in wood is very low and only little is known about the different sulfur compounds in wood today. Sulfur-starved brown-rot fungi Gloeophyllum trabeum and Oligoporus placenta were incubated on sterilized pine wood blocks whereas Lentinus cyathiformis and the white-rot fungi Trametes versicolor were incubated on sterilized beech wood blocks. After 19 weeks of incubation, the S oxidation status was analyzed in wood, in degraded wood, and in biomass of wood-degrading fungi by synchrotron based S K-edge XANES, and total S and sulfate were quantified. Total sulfur and sulfate content in pine wood blocks were approximately 50 and 1 ??g g⁻¹, respectively, while in beech wood approximately 100 and 20 ??g g⁻¹ were found, respectively. Sulfur in beech was dominated by sulfate-esters. In contrast, pine wood also contained larger amounts of reduced S. Three out of four selected fungi caused a reduction of the S oxidation state in wood from oxidized S (sulfate-ester, sulfate) to intermediate S (sulfonate, sulfoxide) or reduced S (thiols, e.g., proteins, peptides, enzyme cofactors). Only O. placenta shifted thiol to sulfonate. Growth experiments of these fungi on selective minimal media showed that in particular cysteine (thiol), sulfonates, and sulfate enhanced total mycelium growth. Consequently, wood-degrading fungi were able to utilize a large variety of different wood S sources for growth but preferentially transformed in vivo sulfate-esters and thiol into biomass structures.     

Activity of dehydroabietic acid derivatives against wood contaminant fungi

The antifungal activity of 10 dehydroabietic acid derivatives with different configuration in A and B rings (cis/trans A/B junction) and different substituents and/or functionalities was evaluated in bioassays in vitro and in situ (pine wood blocks).

The test compounds dissolved in acetone were assayed at several concentrations w/w (test compound/culture medium) against the fungi. The Relative Inhibition (RI) was determined by measuring the radial growth of colonies of the fungi treated with the test compounds by comparison with those of control cultures; the results are expressed as EC₅₀.

The results of bioassays in vitro have shown that hydroxyl and aldehyde functions are required for antifungal activity in this group of compounds and deisopropylation can increase the activity. Our assay of antifungal activity in situ (in pine wood blocks) provides a means to investigate the preservative activities of these antifungal compounds under actual conditions of use.

The dehydroabietic acid derivative cis-deisopropyldehydroabietanol (10) inhibited the growth of several of the fungi tested, in vitro and in situ.

The results obtained in situ with the test compound (10) at 6% and 8% were not significantly different from the reference products and a good level of protection of the wood against the organisms tested was achieved.

The results in wood bioassays present new possibilities in the search for natural new compounds in the wood protection, as an alternative to conventional fungicides.