Blue-stain Fungi Research Articles

Protection of Wood from Microorganisms by Laccase-Catalyzed Iodination

In the present work, Norway spruce wood (Picea abies L.) was reacted with a commercial Trametes versicolor laccase in the presence of potassium iodide salt or the phenolic compounds thymol and isoeugenol to impart an antimicrobial property to the wood surface. In order to assess the efficacy of the wood treatment, a leaching of the iodinated and polymerized wood and two biotests including bacteria, a yeast, blue stain fungi, and wood decay fungi were performed. After laccase-catalyzed oxidation of the phenols, the antimicrobial effect was significantly reduced. In contrast, the enzymatic oxidation of iodide (I(-)) to iodine (I(2)) in the presence of wood led to an enhanced resistance of the wood surface against all microorganisms, even after exposure to leaching. The efficiency of the enzymatic wood iodination was comparable to that of a chemical wood preservative, VP 7/260a. The modification of the lignocellulose by the laccase-catalyzed iodination was assessed by the Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) technique. The intensities of the selected lignin-associated bands and carbohydrate reference bands were analyzed, and the results indicated a structural change in the lignin matrix. The results suggest that the laccase-catalyzed iodination of the wood surface presents an efficient and ecofriendly method for wood protection.

Nutritional and pathogenic fungi associated with the pine engraver beetle trigger comparable defenses in Scots pine

Conifer bark beetles are often associated with fungal complexes whose components have different ecological roles. Some associated species are nutritionally obligate fungi, serving as nourishment to the larvae, whereas others are pathogenic blue-stain fungi known to be involved in the interaction with host defenses. In this study we characterized the local and systemic defense responses of Scots pine (Pinus sylvestris L.) against Ophiostoma brunneo-ciliatum Math. (a blue-stain pathogen) and Hyalorhinocladiella macrospora (Franke-Grosm.) Harr. (a nutritional fungus). These fungi are the principal associates of the pine engraver beetle, Ips acuminatus (Gyll.). Host responses were studied following inoculation with the fungi, singly and as a fungal complex, and by identifying and quantifying terpenoids, phenolic compounds and lignin. Although the length of the necrotic lesions differed between control (wound) and fungal treatments, only two compounds (pinosylvin monomethyl ether and (+)-α-pinene) were significantly affected by the presence of the fungi, indicating that Scots pine has a generic, rather than specific, induced response. The fact that both nutritional and blue-stain fungi triggered comparable induced defense responses suggests that even a non-pathogenic fungus may participate in exhausting host plant defenses, indirectly assisting in the beetle establishment process. Our findings contribute to the further development of current theory on the role of associated fungal complexes in bark beetle ecology.

Effect of water availability and fertilization on water status, growth, vigour and the resistance of Scots pine to fungal mass inoculation with Ophiostoma ips

The effect of water and nutrient availability on the performance of Scots pine (Pinus sylvestris L.) against Ophiostoma ips (Rumb.), a bark beetle-associated phytopathogenic blue-stain fungus, was investigated. Field-grown trees were subjected for 18 months to water-stress and/or fertilization, and the effects of such treatments on the needle nutrient status, tree vegetative growth and vigour were examined. At the end of the experimental period, the trees were mass-inoculated (800 inocula m−2) with the fungus, and the relationship between resource availability and tree performance against pathogen attack was also tested. Predawn shoot water potential (ΨPD) of irrigated trees was significantly higher than that of water-stressed trees, and fertilized trees had a significantly lower C/N ratio. The ΨPD values and needle nitrogen content suggest that resource-limited trees were under moderate stress. Improved nutrient availability significantly increased tree growth and tree vigour. However, no evidence for an effect of improved nutrient availability on tree fungal resistance was found in our study.

What is Next in Bark Beetle Phylogeography?

Bark beetle species within the scolytid genera Dendroctonus, Ips, Pityogenes and Tomicus are known to cause extensive ecological and economical damage in spruce and pine forests during epidemic outbreaks all around the world. Dendroctonus ponderosae poses the most recent example having destroyed almost 100,000 km2 of conifer forests in North America. The success and effectiveness of scolytid species lies mostly in strategies developed over the course of time. Among these, a complex system of semiochemicals promotes the communication and aggregation on the spot of infestation facilitating an en masse attack against a host tree’s defenses; or an association with fungi that evolved either in the form of nutrition (ambrosia fungi) or even by reducing the resistance of host trees (blue-stain fungi). Although often specific to a tree genus or species, some bark beetles are polyphagous and have the ability to switch on to new hosts and extend their host range (i.e., between conifer genera such as Pityogenes chalcographus or even from conifer to deciduous trees as Polygraphus grandiclava). A combination of these capabilities in concert with life history or ecological traits explains why bark beetles are considered interesting subjects in evolutionary studies. Several bark beetle species appear in phylogeographic investigations, in an effort to improve our understanding of their ecology, epidemiology and evolution. In this paper investigations that unveil the phylogeographic history of bark beetles are reviewed. A close association between refugial areas and postglacial migration routes that insects and host trees have followed in the last 15,000 BP has been suggested in many studies. Finally, a future perspective of how next generation sequencing will influence the resolution of phylogeographic patterns in the coming years is presented. Utilization of such novel techniques will provide a more detailed insight into the genome of scolytids facilitating at the same time the application of neutral and non-neutral markers. The latter markers in particular promise to enhance the study of eco-physiological reaction types like the so-called pioneer beetles or obligate diapausing individuals.

Hydraulic limits preceding mortality in a piñon–juniper woodland under experimental drought

Drought-related tree mortality occurs globally and may increase in the future, but we lack sufficient mechanistic understanding to accurately predict it. Here we present the first field assessment of the physiological mechanisms leading to mortality in an ecosystem-scale rainfall manipulation of a piñon-juniper (Pinus edulis-Juniperus monosperma) woodland. We measured transpiration (E) and modelled the transpiration rate initiating hydraulic failure (E(crit) ). We predicted that isohydric piñon would experience mortality after prolonged periods of severely limited gas exchange as required to avoid hydraulic failure; anisohydric juniper would also avoid hydraulic failure, but sustain gas exchange due to its greater cavitation resistance. After 1 year of treatment, 67% of droughted mature piñon died with concomitant infestation by bark beetles (Ips confusus) and bluestain fungus (Ophiostoma spp.); no mortality occurred in juniper or in control piñon. As predicted, both species avoided hydraulic failure, but safety margins from E(crit) were much smaller in piñon, especially droughted piñon, which also experienced chronically low hydraulic conductance. The defining characteristic of trees that died was a 7 month period of near-zero gas exchange, versus 2 months for surviving piñon. Hydraulic limits to gas exchange, not hydraulic failure per se, promoted drought-related mortality in piñon pine.

The Ratio and Concentration of Two Monoterpenes Mediate Fecundity of the Pinewood Nematode and Growth of Its Associated Fungi

The pinewood nematode (PWN) Bursaphelenchus xylophilus, vectored primarily by the sawyer beetle, Monochamus alternatus, is an important invasive pest and causal agent of pine wilt disease of Chinese Masson pine, Pinus massoniana. Previous work demonstrated that the ratios and concentrations of α-pinene∶β-pinene differed between healthy trees and those trees containing blue-stain fungus (and M. alternatus pupae). However, the potential influence of the altered monoterpene ratios and concentrations on PWN and associated fungi remained unknown. Our current results show that low concentrations of the monoterpenes within petri dishes reduced PWN propagation, whereas the highest concentration of the monoterpenes increased PWN propagation. The propagation rate of PWN treated with the monoterpene ratio representative of blue-stain infected pine (α-pinene∶β-pinene = 1∶0.8, 137.6 mg/ml) was significantly higher than that (α-pinene∶β-pinene = 1∶0.1, 137.6 mg/ml) representative of healthy pines or those damaged by M. alternatus feeding, but without blue stain. Furthermore, inhibition of mycelial growth of associated fungi increased with the concentration of the monoterpenes α-pinene and β-pinene. Additionally, higher levels of β-pinene (α-pinene∶β-pinene = 1∶0.8) resulted in greater inhibition of the growth of the associated fungi Sporothrix sp.2 and Ophiostoma ips strains, but had no significant effects on the growth of Sporothrix sp.1, which is the best food resource for PWN. These results suggest that host monoterpenes generally reduce the reproduction of PWN. However, PWN utilizes high monoterpene concentrations and native blue-stain fungus Sporothrix sp.1 to improve its own propagation and overcome host resistance, which may provide clues to understanding the ecological mechanisms of PWN's successful invasion.

Bioincising of Norway spruce wood using wood inhabiting fungi

Norway spruce (Picea abies) is one of the most important wood species in Europe. Its wood is frequently used for commercial applications. More extensive use is limited due to insufficient durability and low permeability. Particularly heartwood is difficult to impregnate. The aim of this preliminary study was to use wood inhabiting fungi for pre-treatment of spruce wood prior to impregnation. Therefore Norway spruce wood specimens (2.0 × 2.0 × 5.0 cm) with end sealed axial surfaces were exposed to one brown rot (Antrodia vaillantii) one white rot (Hypoxylon fragiforme) and one blue stain fungus (Sclerophoma pithyophila) for periods between 14 and 60 days. Afterward mass loss, compression strength, uptake of preservative solutions and retention were determined. The results showed that fungal pre-treatment increased the uptakes of preservative solutions. Among tested fungi, H. fragiforme was found particularly effective. The highest loadings were determined after 30 days of exposure. However, prolonged pre-treatment periods did not improve the impregnability of the specimens but resulted in more prominent losses of mechanical properties.

Influencia del azulado (mancha azul) en la densidad y estabilidad dimensional de la madera de Pinus sylvestris

Pinus sylvestris timber is commercially important, but its value is considerably decreased by the effects of blue stain fungi. Seven pine trees from Spanish plantations were felled for the study, and 14 discs were sampled from different heights in their stems. When the discs showed partial discolouration due to fungal infection, they were cut to produce small defect-free specimens for determining density and dimensional stability (in volume and in the three main dimensions of wood). The physical properties of fully stained and stain free specimens were compared, in each disc, by one-factor analysis of variance. This method enabled analysis of the specific influence of the blue stain fungi while avoiding other sources of variation: tree factor, height in the stem, ring width, presence of sapwood and cambial age. However, the only significant differences between discoloured and unstained wood were in the density and axial shrinkage, in some discs (the density of the blue-stained wood was 1% higher and the axial shrinkage 13% higher than in the unstained wood).

Impact of Systemic Fungicides on Lesions Formed by Inoculation with the Bluestain Fungus (Ophiostoma minus) in Loblolly Pine (Pinus taeda L.)

Ophiostomatoid fungi are important components within the highly damaging insect-fungal complexes attacking trees in North America. This group includes the disease agents responsible for Dutch elm disease and laurel wilt, as well as a number of associates of tree-killing bark beetles. While systemic fungicides have been applied against ophiostomatoids in certain fungus-host systems, the breadth of their utility and their management implications for a wide array of fungus and host species remain uncertain. To evaluate the impact of commercially available fungicides against the bluestaining fungus Ophiostoma minus, an associate of the southern pine beetle, researchers conducted two experiments with loblolly pines (Pinus taeda; mean dbh 14.2 and 18.5 cm, respectively) in central Louisiana, U.S. The first experiment screened three fungicides (Alamo®, Arbotect® 20-S, and PHOSPHO-jet) for their systemic impacts on O. minus, and the second further evaluated the best performing product (Alamo) alone and in combination with Arbotect 20-S. In all cases, loblolly pines were stem-injected basally and challenged with inoculations of cultured O. minus at time periods ranging from 28 days to 738 days post-treatment. In both experiments, treatment of loblolly pines with Alamo produced the smallest lesion areas, indicating that this treatment was the most effective for limiting the within-tree growth of O. minus. This effect was still present more than two years post-treatment. The study authors conclude that Alamo is the most effective product of those evaluated for prophylactic treatment against O. minus, and suggest that evidence is building for the effectiveness of this product generally against ophiostomatoid fungi.

The effects of α‐cellulose extraction and blue‐stain fungus on retrospective studies of carbon and oxygen isotope variation in live and dead trees

Tree-ring carbon and oxygen isotope ratios from live and recently dead trees may reveal important mechanisms of tree mortality. However, wood decay in dead trees may alter the δ(13)C and δ(18)O values of whole wood obscuring the isotopic signal associated with factors leading up to and including physiological death. We examined whole sapwood and α-cellulose from live and dead specimens of ponderosa pine (Pinus ponderosa), one-seed juniper (Juniperous monosperma), piñon pine (Pinus edulis) and white fir (Abies concolor), including those with fungal growth and beetle frass in the wood, to determine if α-cellulose extraction is necessary for the accurate interpretation of isotopic compositions in the dead trees. We found that the offset between the δ(13)C or δ(18)O values of α-cellulose and whole wood was the same for both live and dead trees across a large range of inter-annual and regional climate differences. The method of α-cellulose extraction, whether Leavitt-Danzer or Standard Brendel modified for small samples, imparts significant differences in the δ(13)C (up to 0.4‰) and δ(18) O (up to 1.2‰) of α-cellulose, as reported by other studies. There was no effect of beetle frass or blue-stain fungus (Ophiostoma) on the δ(13)C and δ(18)O of whole wood or α-cellulose. The relationships between whole wood and α-cellulose δ(13)C for ponderosa, piñon and juniper yielded slopes of ~1, while the relationship between δ(18)O of whole wood and α-cellulose was less clear. We conclude that there are few analytical or sampling obstacles to retrospective studies of isotopic patterns of tree mortality in forests of the western United States.

Effects of chemical modification with glutaraldehyde on the weathering performance of Scots pine sapwood

Scots pine sapwood was treated with glutaraldehyde (GA) in aqueous solution using magnesium chloride as a catalyst in order to evaluate the durability towards weathering. Infrared spectroscopy suggested that GA treatment increased the photo-stability of lignin during artificial weathering of micro-veneers in a QUV over 168 h; photo-protection increased with increasing GA concentration. In comparison with the unmodified controls, GA-modified pine micro-veneer strips exhibited a lower tensile strength loss measured in a zero-span mode in the course of weathering. During 18 months of outdoor exposure, GA-modified pine wood boards exhibited a lower moisture content and water uptake than the unmodified ones. GA treatment also clearly restricted the penetration of blue stain fungi into deeper layers of wood. On the macroscopic scale, the surface of the GA-modified boards was significantly smoother due to less erosion, cracking and minor peeling of tracheids. Scanning electron microscopy further revealed that individual tracheids were detached from the cell compound and then washed away from the unmodified wood surface, whereas tracheids on surfaces of GA-modified wood remained in the tissue compound but displayed many axial and transversal cracks.

Blue-stain infections in roots, stems and branches of declining Pinus sylvestris trees in a dry inner alpine valley in Switzerland

Summary Increased mortality rates in Scots pine (Pinus sylvestris) forests have recently been observed in the inner alpine Swiss Rhone valley. Drought, in combination with stand competition, mistletoe infections as well as nematode and insect infestations, appears to be the main factor for the decline. In focus of this study was the occurrence and role of fungal pathogens in the decline dynamics. Branches, stems and roots of 208 trees in five different crown transparency classes were collected and inspected for blue stain and fungal infections. Neither Armillaria species nor Heterobasidon annosum s. str. were detected, but blue stain was commonly observed. Visible blue stain increased with increasing crown transparency. Among the recently dead trees, 80% showed visible blue stain in the branches, 90% in the roots and 100% in the stems. In the crown transparency classes 2 and 3 (25–60% crown transparency), five of the 103 trees showed visible blue stain in the roots, one of 130 trees in the stem but none in the branches. Blue-stain fungi were isolated from all parts of the trees and from all crown transparency classes. Overall incidence of blue stain was highest in the roots and lowest in the branches. In class 2, roots of 60% of the trees were visibly blue-stained or developed blue stain in culture, but stems of only 24% and branches of 14% of the trees. In the roots Leptographium species, mostly L. serpens, dominated. From stems and branches, mainly Ophiostoma species were isolated. The positive relationship between the incidence of blue stain and crown transparency, in combination with the high infection levels of roots of fairly vigorous Scots pines, indicates the pathogenic potential of the blue-stain fungi. Hence, these fungi together with their insect vectors may well act as an important contributing factor involved in pine decline.

Production and Characterization of Palm Fruit Shell Bio-Oil for Wood Preservation

Pyrolytic liquid (bio-oil), produced by pyrolyzing the shell of the palm fruit, was characterized, and its preservative properties were examined using drywood termites (Cryptotermes spp.) and blue stain fungi (Ceratocystis spp.). The yield from shell bio-oil production ranged from 35 to 37 percent relative to the mass of the raw material. The shell bio-oil had the following properties: specific gravity (ranging from 1.0365 to 1.0431), refractive index (ranging from 1.3594 to 1.3613), wood absorption level (ranging from 0.0088 to 0.0625 g/cm3), and retention (ranging from 0.0022 to 0.0141g/cm3). The shell bio-oil also had termicidal activity as shown by drywood termite mortalities: 25 percent between days 3 and 6, 50 percent between days 3 and 20, and 100 percent between days 5 and 49 in termite resilience tests. Using a fluorescence microscope, we demonstrated that the shell bio-oil–treated wood completely inhibited the growth of blue stain fungi on both pine (Pinus merkusii) and Sengon (Paraserianthes falcataria) wood.

Abundance-dependent transmission of the pinewood nematode, Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae), to the Japanese pine sawyer, Monochamus alternatus (Coleoptera: Cerambycidae), adult in its pupal chamber

The transmission ratio of the pinewood nematode (PWN), Bursaphelenchus xylophilus, to the emerging adult Japanese pine sawyer (JPS), Monochamus alternatus, in its pupal chamber is a determinant of the number of the nematodes carried by JPS beetles. To investigate the factors affecting the transmission ratio, we counted the number of the nematodes carried by 36 newly emerged JPS beetles and the number remaining in and around their pupal chambers, and then estimated the transmission ratio (the number of nematodes carried by a JPS adult as a percentage of the total number of nematodes aggregating in and around its pupal chamber). The total number of nematodes aggregating in and around a pupal chamber ranged from 0 to 19,041, and the number of nematodes carried by a beetle ranged from 0 to 18,920. The transmission ratio correlated with neither the water content of the wood around the pupal chamber nor the degree of wood discoloration caused by blue-stain fungus. The transmission ratio varied with the abundance of the nematodes aggregating in and around the pupal chamber. In pupal chambers with more than 1,000 nematodes, almost all the nematodes were transmitted to the beetle. However, in pupal chambers with fewer than 1,000 nematodes, the transmission ratio varied greatly, from 0 to 100%. These results suggest that aggregation of many PWNs in the pupal chamber might stimulate transmission of the PWNs to the JPS adult and that this abundance-related transmission might contribute to the large variation in the number of PWNs carried by the JPS beetle.

Migration of blue stain fungi within wax impregnated wood

The colonization of wax impregnated pine sapwood (Pinus sylvestris L.) and beech (Fagus sylvatica L.) by blue stain fungi and their hyphal migration within the treated wood was investigated. Neither the hydrophobation effect nor the physical presence of the congealed wax deposits could impede fungal growth on the surface. However, in contrast to the untreated controls, there was a lower occurrence of hyphae in wax-treated wood. Blue stain fungi grew mainly near the cross cut as well as in empty spaces such as micro-capillaries, parenchyma tissue or other wax free zones of treated wood.

Combining land surface temperature and shortwave infrared reflectance for early detection of mountain pine beetle infestations in western Canada

The current mountain pine beetle (Dendroctonus ponderosae Hopkins) outbreak, whichbeganin1999,continuestobetheleadingcauseofpinetreemortalityinBritishColumbia. Information regarding the location and spatial extent of the current attack is required for mit- igating practices and forest inventory updates. This information is available from spaceborne observations. Unfortunately, the monitoring of the mountain pine beetle outbreak using remote sensing is usually limited to the visible stage at which the expansion of the attack beyond its initial hosts is unpreventable. The disruption of the sap flow caused by a blue-staining fungi carried by the beetles leads to: 1. a decrease in the amount of liquid water stored in the canopy, 2. anincrease incanopy temperature, and 3. anincrease inshortwave infraredreflectance shortly after the infestation. As such, the potential for early beetle detection utilizing thermal remote sensing is possible. Here we present a first attempt to detect a mountain pine beetle attack at its earliest stage (green attack stage when the foliage remains visibly green after the attack) using the temperature condition index (TCI) derived from Landsat ETM+ imagery over an affected area in British Columbia. The lack of detailed ground survey data of actual green attack areas limits the accuracy of this research. Regardless, our results show that TCI has the ability to differentiate between affected and unaffected areas in the green attack stage, and thus it provides information on the possible epicenters of the attack and on the spatial extent of the outbreak at later stages (red attack and gray attack). Furthermore, we also developed a moisture condition index (MCI) using both shortwave infrared and thermal infrared measurements. The MCI index is shown to be more effective than TCI in detecting the green attack stage and provides a more accurate picture of beetle spread patterns. C 2011 Society of Photo-Optical Instrumentation Engineers

Evaluation of the interspecific competitive ability of the bioincising fungus Physisporinus vitreus

A hierarchical set of assays were used to evaluate the interspecific competitive ability of the bioincising fungus Physiporinus vitreus (response species). The competitiveness and growth mode of P. vitreus against a range of blue stain fungi and Trichoderma species (challenge species) was investigated in dual culture tests (a) and a spatially heterogeneous system of tessellation agar (b) on different media. In addition, the robustness of the bioincising process against biotic influences was determined by controlled interaction tests with different inocula of the challenge species (colony-forming units: 10(2), 10(4), 10(6) ml(-1)) on Norway spruce (Picea abies Karst.) heartwood. The results of the dual culture and tessellation agar tests revealed a significant (P ≤ 0.001) antagonistic potential (lethal effect) of the T. atroviride strain (15603.1), whereas no or only a slight effect of the blue stain fungi on P. vitreus was detectable. The interaction tests on Norway spruce heartwood confirmed that an increase in inoculum size of the challenge fungi correlated with an increased negative effect (Spearman's rho coefficient (ρ) = -0.521). Among the challenge species, T-15603.1 revealed the most negative effect on the homogeneous development of P. vitreus on wood and thus on the outcome of the bioincising process. To improve the uniformity of colonization and to reduce the rate of contamination, the incubation of wood in a sealed system (bioreactor) after sterilization is suggested.

Resistance of bioincised wood treated with wood preservatives to blue-stain and wood-decay fungi

Bioincising is a biotechnological process that aims at the improvement of wood preservative uptake in wood species with a low permeability, such as Norway spruce (Picea abies (L.) Karst). The process is based on a short-term pre-treatment with white-rot fungus Physisporinus vitreus. During incubation the membranes of bordered and half bordered pits are supposed to be degraded by fungal activity resulting in a better treatability of the wood structure for wood preservatives. In the present study, first of all the resistance of bioincised Norway spruce heartwood and untreated controls against blue-stain and wood-decay fungi (white- and brown-rot) was determined. Then, bioincised and untreated specimens were dipped or vacuum impregnated with six wood preservatives and substance uptake was assessed gravimetrically. Additionally, the penetration of 3-iodo-2-propynyl butylcarbamate (IPBC) into the wood was analyzed by high-pressure liquid chromatography (HPLC). Finally, wood resistance was assessed according to the European standards EN 152 and EN 113. Results showed no difference between bioincised wood without preservatives and the untreated wood against blue-stain discolouration. However, a significant (P < 0.05) increase in susceptibility against wood decay was recorded. In the bioincised wood samples a significantly higher uptake of all the different preservatives was determined and the HPLC-method revealed that IPBC penetrated deeper into bioincised wood than into control samples. The improved uptake of preservatives into bioincised wood resulted in a significantly higher resistance against white- and brown-rot fungi. However, only a slight protection against wood discolouration by blue-stain fungi was recorded. The results of this study show for the first time that the biotechnological process with P. vitreus can be used to improve wood durability by increasing the uptake and penetration of wood preservatives.

The influence of Ceratocystis polonica inoculation and methyl jasmonate application on terpene chemistry of Norway spruce, Picea abies

Constitutive and inducible terpene production is involved in conifer resistance against bark beetles and their associated fungi. In this study 72 Norway spruce ( Picea abies) were randomly assigned to methyl jasmonate (MJ) application, inoculation with the bluestain fungus Ceratocystis polonica, or no-treatment control. We investigated terpene levels in the stem bark of the trees before treatment, 30 days and one year after treatment using GC–MS and two-dimensional GC (2D-GC) with a chiral column, and monitored landing and attack rates of the spruce bark beetle, Ips typographus, on the trees by sticky traps and visual inspection. Thirty days after fungal inoculation the absolute amount and relative proportion of (+)-3-carene, sabinene, and terpinolene increased and (+)-α-pinene decreased. Spraying the stems with MJ tended to generally increase the concentration of most major terpenes with minor alteration to their relative proportions, but significant increases were only observed for (−)-β-pinene and (−)-limonene. Fungal inoculation significantly increased the enantiomeric ratio of (−)-α-pinene and (−)-limonene 1 month after treatment, whereas MJ only increased that of (−)-limonene. One year after treatment, both MJ and fungal inoculation increased the concentration of most terpenes relative to undisturbed control trees, with significant changes in (−)-β-pinene, (−)-β-phellandrene and some other compounds. Terpene levels did not change in untreated stem sections after treatment, and chemical induction by MJ and C. polonica thus seemed to be restricted to the treated stem section. The enantiomeric ratio of (−)-α-pinene was significantly higher and the relative proportions of (−)-limonene were significantly lower in trees that were attractive to bark beetles compared to unattractive trees. One month after fungal inoculation, the total amount of diterpenes was significantly higher in putative resistant trees with shorter lesion lengths than in putative susceptible trees with longer lesions. Thus, terpene composition in the stem bark may be related to resistance of Norway spruce against I. typographus and C. polonica.

Penetration of 3-iodo-2-propynyl butylcarbamate (IPBC) in coniferous wood pre-treated with Physisporinus vitreus

The permeability of Norway spruce wood can be reduced to 1–5% of that of green timber when dried, resulting in only slight radial penetration of chemical solutions. We investigated the penetration of the biocide 3-iodo-2-propynyl butylcarbamate, (IPBC) in Norway spruce and Scots pine sapwood when applied in a water-based coating system. Penetration of both untreated wood and specimens pretreated with the white-rot fungus Physisporinus vitreus was analyzed by the absence or presence of blue staining after incubation with Aureobasidiumpullulans and Sclerophoma pithyophila. The qualitative results were compared with quantitative data obtained by chemical analysis. It was assumed a pre-treatment with Physisporinus vitreus could make the surface of the substrate more porous and improve the penetration of fluid substances. All analytical investigations of the samples were done after an outdoor weathering period of 6 months. From a practical point of view the performance of the coated substrate is more relevant after a certain aging procedure then right after the application. The chemical measured uptake of IPBC for Scots pine was similar for all samples at the surface. The penetration depth was higher for those samples with a pre-treatment with Physisporinus vitreus. The colonization pattern of the samples by blue-stain fungi depended upon the pre-treatment but did not show a clear correlation between IPBC-content and staining. The cross-section of the samples with pre-treatment was completely stained, the cross-section of the samples without pre-treatment did show a distinctive blue stain-free zone. In Norway spruce, the chemical measured uptake of IPBC was similar for all samples at the surface independent of the pre-treatment. The penetration depth was higher for those samples with a pre-treatment with Physisporinus vitreus. But a clear visible colonization by blue-stain fungi was only observed for the samples with a pre-treatment with Physisporinus vitreus. The samples without pre-treatment were locally stained across the entire cross-section. As with the Scots pine, a clear correlation between IPBC-content and blue staining was not visible, following comparison of the samples with and without pre-treatment with Physisporinus vitreus. The results suggest that not only the presence of IPBC influences colonization by A.pullulans and S.pithyophila, but also a range of other factors in wood pre-treated with P. vitreus.