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     

Efficacy of radio frequency treatment and its potential for control of sapstain and wood decay fungi on red oak,poplar, and southern yellow pine wood species

The effectiveness of radio frequency (RF) treatment in the control of wood decay fungi (Gloeophyllum trabeum, Ganoderma lucidum, and Irpex lacteus) and sapstain fungus (Ceratocystis fimbriata) in red oak (Quercus spp.), poplar (Populus alba), and southern yellow pine (Pinus spp.) was evaluated in the laboratory as an alternative to methyl bromide (MB) treatment. Wood samples (15.5 x 10 x 10 cm) were inoculated with fungi from a 7-day culture by dipping them to a depth of one face deep (2 cm) into inoculum and incubating them at 25°C for 14 days. Identical wood samples were left uninoculated as controls. Subsequent to incubation, the wood blocks were exposed to RF radiation in an industrial 40-kW dielectric oven at temperatures between 60° and 70°C for 2 min. The test fungi were recovered and reisolated from all of the control wood blocks but not from RF-treated wood blocks. RF treatment resulted in complete inhibition of the fungus in 98%-100% of the wood samples. Moisture content loss (≥1%) was noted after wood had been exposed to RF treatment. Moisture content may be an important factor to consider with RF treatments. RF treatment can, therefore, potentially provide an effective and rapid quarantine treatment as an alternative to MB fumigation for certain pathogen-wood combinations. This article reports the results of research only. Mention of a product does not constitute an endorsement or recommendation by USDA for its use.     

木材天然耐腐性室内试验标准方法的研究

采用三种培养基、两种白腐菌、两种褐腐菌对马尾松、毛白杨的天然耐腐性进行了试验。结果表明:砂子锯屑培养基法对采绒革盖菌(Corjolus versicolor)和绵腐卧孔菌(Poiraplacenta)的生长较快,对木材腐朽力,即重量损失较明显。     

木腐菌及其腐朽机理研究进展

木腐菌可以有效地改善针叶材心材的渗透性,可用于辅助阔叶材散孔材年轮的鉴定,木腐菌及其所分泌的酶还可作为生物技术应用于木材工业,如生物制浆预处理、生物漂白以及木材酶解为糖的预处理等领域。本文从褐腐、白腐、软腐、竹材及人造板等几个方面出发,对国内外腐朽机理的研究现状进行了较为系统、全面的介绍,旨在使学者了解真菌选择性地降解木竹材的化学成分和细胞各部分的知识,为木竹材的保护和合理利用提供参考。     

Defense mechanisms in the sapwood of living trees against microbial infection

When pathogenic microorganisms invade living sapwood of woody plants, a series of defense responses occurs at the lesion margin. Putative active defense mechanisms include constitutive and induced inhibitory compounds, cell wall alterations, and occlusion of xylem elements. Active defenses play an important role in the sapwood, while constitutive and induced microenvironmental conditions in the wood might also constrain pathogen development. It is necessary to develop a unified understanding, in which these factors could act synergistically and provide effective defense barriers.     

Distribution of stem decay in the beech trees of the Azerbaijan and its impact on the output of commercial wood

To determine the distribution of stem decay in the beech stands of Azerbaijan, we sought to identify the species, number, and height of attachment of fruiting bodies;the extent of decay in tree trunks growing under different conditions;and its influence on the trees’ commercial wood. The research was conducted on three farms representing the most common forest types of the Greater Caucasus within Azerbaijan. Examination of the presence of fruiting bodies in the tree trunk revealed that stem decay is the most common infection in beech (Fagus orientalis) stands, especially in fresh, moist areas, with less infection under dry growing conditions. In this work, the length, diameter, and volume of decay were studied by infecting the trunks of 93 model trees affected with mushrooms in 1–4 m cuttings. Our results showed that as the age of the stand increased, extent, diameter, and volume of decay increased significantly: the extent of decay from 1.47 to 6.43 m;the diameter of the decay from 8.15 to 32.7 cm;and the volume of decay from 2.5 to 13.2%. The relationship between age and the specified indicators is presented graphically. Using data obtained from the same sample of trees, we determined the expected and actual yield of commercial wood by age class. We determined that stem decay in beech stands leads to a decrease in the yield of commercial wood on average from 25.1% in the middle-aged to 14.8% in overmature plantations, respectively, with an estimated yield of 40.8–62.7%. Here we present mathematical models of the output of commercial wood from the infected part of the stands by age classes with the use of average data on the extent of decay in model trees.     

Decay resistance of sapwood and heartwood of untreated and thermally modified Scots pine and Norway spruce compared with some other wood species

Abstract

Thermal modification has been developed for an industrial method to increase the biological durability and dimensional stability of wood. In this study the effects of thermal modification on resistance against soft- and brown-rot fungi of sapwood and heartwood of Scots pine and Norway spruce were investigated using laboratory test methods. Natural durability against soft-rot microfungi was determined according to CEN/TS 15083-2 (2005) by measuring the mass loss and modulus of elasticity (MOE) loss after an incubation period of 32 weeks. An agar block test was used to determine the resistance to two brown-rot fungi using two exposure periods. In particular, the effect of the temperature of the thermal modification was studied, and the results were compared with results from untreated pine and spruce samples. The decay resistance of reference untreated wood species (Siberian larch, bangkirai, merbau and western red cedar) was also studied in the soft-rot test. On average, the soft-rot and brown-rot tests gave quite similar results. In general, the untreated heartwood of pine was more resistant to decay than the sapwood of pine and the sapwood and heartwood of spruce. Thermal modification increased the biological durability of all samples. The effect of thermal modification seemed to be most effective within pine heartwood. However, very high thermal modification temperature over 230°C was needed to reach resistance against decay comparable with the durability classes of “durable” or “very durable” in the soft-rot test. The brown-rot test gave slightly better durability classes than the soft-rot test. The most durable untreated wood species was merbau, the durability of which could be evaluated as equal to the durability class “moderately durable”.     

Evaluation of a stress-wave timer for the minimally destructive detection of decay in living trees in Northern-Japanese forests

The use of a stress-wave timer as a minimally destructive device for detecting decay or defect in living trees was evaluated. Measurements were conducted on five tree species (Picea jezoensis, P.glehnii, Betula platyphylla var.japonica, Abies sachalinensis, andLarix kaempferi) with or without decay. Except in sap-rottedA. sachalinensis, the apparent stress-wave velocity in most decayed trees was considerably lower than the value obtained from healthy trees. Our results showed that defect or decay in the trees was detectable more effectively by the method used in the field survey, although the device occasionally failed to detect decay that was incipient, of small extent or confined to sapwood. Other disadvantages of this method are briefly discussed in this paper.     

Efficacy of wood preservatives formulated from okara with copper and/or boron salts

As a substitute for high-cost copper azole (CuAz) and alkaline copper quaternary (ACQ) wood preservatives, alternative wood preservatives were formulated with okara, which is an organic waste from the production of tofu, and copper chloride and/or borax. Each preservative was used in treatment of wood blocks in a reduced-pressure method to measure its treatability. The treated wood blocks were placed in hot water for 3 days to examine the stability of the preservatives against hot-water leaching. The preservatives successfully penetrated into wood blocks, probably due to the use of ammonium hydroxide as a dissociating agent. However, the stability of okara-based preservatives dropped as the concentration of acid in the solutions used for hydrolysis of okara increased. The treatability and leachability of the preservatives were not affected by hydrolysis temperature but were negatively affected by the addition of borax. Leached wood blocks treated with okara-based preservatives and exposed to decay fungi Gloeophyllum trabeum and Postia placenta over 12 weeks showed good decay resistance. Okara-based wood preservatives can protect wood against fungal attack as effectively as CuAz, and have potential for use as environmentally friendly wood preservatives.     

Immunogold labeling of an extracellular substance producing hydroxyl radicals in wood degraded by brown-rot fungusTyromyces palustris

A fraction containing low-molecular-weight peptides that catalyzes redox reactions between electron donors and O₂ to produce ·OH, was partially purified from wood-decaying cultures of the brown-rot fungusTyromyces palustris. Polyclonal antibodies raised to the fraction were used for immunogold labeling of transverse sections of sapwood of spruce in various stages of degradation byT. palustris to demonstrate the cellular localization of the ·OH-producing substance. Initially, the wood cell wall was attacked primarily by fungal hyphae growing in the cell lumen. During the early stages of degradation, the gold label was localized in the fungal cytoplasm and cell wall and in the extracellular slime sheath surrounding the fungal cell wall. The gold label also was found throughout the wood cell wall, although the cell wall remained almost intact so long as the fungal hyphae remained in the lumen. Thus, the ·OH-producing substance is secreted by the hyphae into the lumen, and it diffuses through the S3 layer into the S2 layer and the middle lamella. The role of this ·OH-producing system in wood degradation byT. palustris is discussed.     

Leachability,decay, and termite resistance of wood treated with metaborates

The formation of insoluble metaborates in wood was investigated by impregnating the wood with borax and metallic salts, after which their properties (e.g., leachability in running water and biological resistance) were evaluated. The solubility of three metaborates in acidic solutions was also evaluated. Double-diffusion treatment was carried out to form the precipitates of metaborates in sapwood specimens of Japanese cedar (Cryptomeria japonica) at room temperature. Water-saturated wood specimens were first impregnated by a saturated borax solution and then diffuse-penetrated with Zn²⁺, Ca²⁺, or Pb²⁺ solution. The precipitates of the three metaborates in the wood proved to be insoluble or hardly soluble in water by the leaching test. With the decay test using a brown-rot fungus (Fomitopsis palustris) and a white-rot fungus (Trametes versicolor) and with the termite test using a virulent subterranean termite (Coptotermes formosanus), the metaborate-treated woods showed generally good decay and termite resistance with negligible mass loss of the specimens. Particularly, the lead metaborate formed in the wood provided superb biological resistance against decay and termite attacks. In addition, the precipitates of these metaborates were found to be soluble in acidic solution, suggesting a way to remove these chemicals from wood when disposing of waste materials.     

Testing and evaluation of natural durability of wood in above ground conditions in Europe – an overview

Natural durability of wood is determined by the European standard EN 252 for specimens in ground contact and EN 113 for basidiomycetes in the laboratory, but no test exists for above ground conditions. For above ground conditions, the European prestandard ENV 12037 and EN 330 are used to determine the durability of treated wood. The most important factors for fungal establishment on the surface and within wood are the moisture content, the surrounding temperature, and the relative humidity. Strength tests are the most sensitive for decay detection, but neither strength tests nor identification of fungi responsible for the decay are included in the standards of above ground durability in field tests. To detect decay, visual examination, pick or splinter tests, and mass loss determination are used. Identifying fungi with traditional methods, e.g., growth on solid medium, is time consuming and complicated. Molecular methods like polymerase chain reaction and sequencing do not require mycological skill for identification to species level, and furthermore the methods do not depend on the subjective judgement like most traditional methods, but are based on the objective information of the target organism (e.g., nucleotide sequences). The next generation of standard field tests will probably consider the drawbacks of standard tests today and be rapid and include both quality tests like molecular identification and nondestructive quantitative tests, e.g., acoustic tests. Laboratory tests can be improved by using fungi identified from field trials and by combining different fungi in the same test and thus simulate degradation in practice.     

Seasonal and height-dependent fluctuation of starch and free glucose contents in moso bamboo (Phyllostachys pubescens) and its relation to attack by termites and decay fungi

The potential of termite (Coptotermes formosanus Shiraki) and fungal attacks [Fomitopsis palustris (Berkeley et Curtis) Murrill (FFPRI 0507) and Trametes versicolor (L. ex Fr.) Quel (FFPRI 1030)] against moso bamboo (Phyllostachys pubescens Mazel) were evaluated with regard to the seasonal and height-dependent changes of the free glucose and starch contents, which were measured by a newly developed method. The free glucose contents were generally lower in autumn and winter than in spring and summer, whereas the lowest starch contents were obtained in August, and the contents increased almost linearly up to February and March. In terms of the height-dependent fluctuation, the free glucose contents tended to decrease as the sampling heights increased. There was no special correlation between the free glucose or starch contents and the consumption by termites, even though higher mortalities were obtained in the bamboo-fed termites than in the wood-fed termites. However, a positive correlation between the free glucose contents and mass losses of the samples at 4-m and 8-m heights from the bottom by the decay fungi was observed. For starch, no influence on fungal attack was found.     

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.     

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.     

A simple technique for measuring the permeability of coniferous wood and its application to the study of water conduction in living trees

A method is described for measuring the permeability to water of fresh coniferous wood samples in accordance with the provisions of Darcy's Law describing fluid flow through porous media. Departures from this law are also described and briefly discussed.     

Relationship between production of hydroxyl radicals and degradation of wood, crystalline cellulose, and a lignin-related compound or accumulation of oxalic acid in cultures of brown-rot fungi

The degradation of wood, filter paper cellulose, and a lignin-substructure model, was measured in cultures of seven fungi usually regarded as brown-rot fungi. Hydroxyl radical production and the accumulation of oxalic acid in the cultures were also measured. Four of the fungi, Gloeophyllum trabeum, Tyromyces palustris, Laetiporus sulphureus, and Postia placenta, were typical brown-rot fungi, in that they preferentially degraded and eliminated the polysaccharides in wood and produced large amounts of hydroxyl radical. The rates of hydroxyl radical generation in cultures of the four fungi were directly proportional to the degradation rates of wood, cellulose, and the lignin-related compound, and inversely proportional to the amount of oxalic acid in the cultures. Two of the fungi, Daedalea dickinsii and Lentinus lepideus, did not degrade any of the substrates significantly and produced very little hydroxyl radical. Coniophora puteana had the highest rate of cellulose degradation, but did not degrade wood or the lignin model significantly and produced only negligible amounts of hydroxyl radical. These results indicate that brown-rot fungi produce large amounts of hydroxyl radical for the degradation of wood and crystalline cellulose.     

Stocks and decay dynamics of above- and belowground coarse woody debris in managed Sitka spruce forests in Ireland

Coarse woody debris (CWD) has become recognised as an important component of the carbon (C) pool in forest ecosystems. In Ireland, managed Sitka spruce (Picea sitchensis (Bong) Carr.) forests account for 52.3% of the total forest estate. To determine the stock and decay dynamics of above and belowground CWD, field surveys using fixed area sample plots, were conducted in six even-aged Sitka spruce stands, representing the young, intermediate and mature stages of a typical commercial rotation. The volume, mass, density loss and C:N ratio of all CWD types (logs, stumps, and coarse roots) were determined using a five-decay class (DC) system. The decay rates and half life of CWD was also determined. To estimate CWD coarse root mass; roots associated with stumps classified in different decay classes were excavated. The coarse roots were categorised into small (2-10 mm), medium (10-50 mm) and large (>50 mm) diameter classes.CWD C-mass ranged from 6.98 to 18.62 Mg ha⁻¹ and was highest in an intermediate forest (D35), while the aboveground volume varied from 6.31 to 42.27 m³ ha⁻¹. Coarse roots accounted for 21% to 85% of the total CWD C-pool in the surveyed stands. The total CWD C-mass was poorly correlated with the number of thinning events (R² = 0.29), when data from D35 was excluded. The density loss was significant in logs (45%), stumps (58%), and small- (38%), medium- (50%) and large roots (38%) as decay progress from DC 0 to 4. There was a 46%, 41%, 51%, 72% and 57% decline in C:N ratio of logs, stumps, small-, medium- and large roots, respectively, as decay progressed from DC 0 to 4. The density decay rates were 0.059, 0.048 and 0.036 kg m⁻³ year⁻¹ for logs, stumps and coarse roots, respectively. The size classification of roots did not significantly affect their decay rate. The half life (50% decomposition) of CWD was estimated has 12-, 14- and 19 years for logs, stumps and roots of Sitka spruce. Regression curves showed a strong correlation between the density and C:N ratio (R² = 0.69, 0.74 and 0.93 for logs, stumps and coarse roots, respectively). The long term storage of C and its slow rate of decomposition make CWD a vital structural and functional component of the CWD C-pool and a major controller of forest ecosystem C-retention.     

6种江西常见速生阔叶材纤维形态及材性比较

为发展和科学利用速生阔叶工业用材种,对6种江西常见速生阔叶树木荷、苦楝、鹅掌楸、深山含笑、东京野茉莉及檫树的纤维形态及材性进行测定与分析。结果表明：木荷、苦楝、深山含笑、东京野茉莉的纤维都属长细型纤维,具有良好的制浆造纸性能,尤其苦楝属制浆造纸优等材,纤维长度最长,平均纤维长度达1.43 mm。檫树和鹅掌楸制浆造纸性能略差。11～12年生木荷和檫树密度已达中等密度木材范围,其中木荷平均密度最大,平均密度为0.69 g/cm3,但干缩性相对略大,深山含笑尺寸稳定性相对较好;木荷的综合强度最高,抗压强度50.52 MPa,属中级,抗弯强度和抗弯弹性模量分别为141.7 MPa、17.88 GPa,均属高等水平;鹅掌楸综合强度中等,顺纹抗压强度为50.52 MPa、抗弯强度84.99 MPa,抗弯弹性模量14.55 GPa;深山含笑抗弯强度和抗弯弹性模量均可达中等水平,但抗压强度略差;苦楝、东京野茉莉在材性综合强度方面稍差,抗压强度、抗弯强度和抗弯弹性模量方面均属低等水平。     

Wood colour variation in sapwood and heartwood of young trees of Tectona grandis and its relationship with plantation characteristics, site, and decay resistance

? Wood colour of Tectona grandis produced from fast-growth plantations is highly variable and the causes of this variation are relatively unknown.

? With the purpose of understanding the colour variation, different fast-growth plantations were sampled with different growth rates, tree ages, and sites.

? Wood colour was measured with a CIELab system, where three variables are estimated: coordinate L* for lightness, coordinate a* defines redness and coordinate b* defines yellowness.

? Results showed only a negative correlation between L* and a*. L* and a* were negatively and positively respectively correlated with pith distance in heartwood, but not for b*. No correlations were found between L* and b* in sapwood and plantation characteristics, while a* was positively correlated with age and height of tree and growth rate. In heartwood, tree age and diameter at breast height were correlated with all colour parameters, but tree height and plantation density were correlated with a* and b*. Cluster site had correlation with L*. Multiple correlation analysis showed that the heartwood is increasing darker (L*) and redder (a*) when the trees are older and bigger. Correlation coefficient shown that sapwood and heartwood with lighter colour (L*) is less resistance to fungal attack, but redness colour (a*) increasing decay resistance.