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Transformation of Ophiostoma picea and Trichoderma harzianum with green fluorescent protein (GFP)
2003 - IRG/WP 03-10477
While microbial colonization of wood is presumed to be characterized by a myriad of interactions between numerous organisms, studying these processes is often difficult owing to the opaque nature of the wood and the inability to readily distinguish among the many species colonizing the material. One method for enhancing the ability to distinguish organisms is to induce specific proteins in one or more organisms that can be detected using fluorescence or other light microscopic techniques. The insertion of genes for the production of green fluorescent proteins produced by the jellyfish, Aequora victoria, has been widely used to visualize a variety of organisms. In this report, we describe transformation of two fungi, Ophiostoma picea and Trichoderma harzianum using a green fluorescent protein (SGFP) gene under the control of the ToxA promoter of Pyrenophora tritici-repentis. The growth and wood colonization by two transformed fungi were compared to their non-transformed strains. The expression of gfp was particularly useful for studying the spatial distribution of young hyphae in wood.
Ying Xiao, L M Ciuffetti, J J Morrell

Effects of Prior Establishment of Trichoderma harzianum on Ophiostoma picea Growth in Freshly Sawn Douglas-fir Sapwood
2003 - IRG/WP 03-10476
Trichoderma harzianum has been shown to be an effective biocontrol agent against a number of wood inhabiting fungi under laboratory conditions, but this fungus has performed poorly in field trials. Understanding the interactions between biocontrol agents and their intended targets in wood may provide important clues for developing improved approaches to biocontrol, potentially reducing our reliance on pesticides. One particularly difficult problem with studying biocontrol in wood is the inability to accurately resolve individual organisms as they interact. The opaque nature of wood makes it difficult to observe interactions more than a few cells from the surface. Wood can be cut into thin sections, but the structural similarity of many fungal hyphae makes it difficult to determine which fungus is present. Transformation using genes for the synthesis of green fluorescent protein (GFP) has proven useful for visualizing specific microorganisms in their hosts and could be useful for biocontrol studies. In this study, interactions between gfp transformed Ophiostoma picea and non-transformed T. harzianum were studied on freshly sawn Douglas-fir sapwood blocks. Growth of O. picea was significantly retarded when applied to wood blocks where T. harzianum was well-established. Interestingly the Graphium state of O. picea was still observed on surfaces of the blocks, but the fungus was unable to penetrate deeply into the wood. The results suggest that failure of the biocontrol may not represent an inability to protect the wood beneath the surface and implies that a more detailed study of the causes of previous failures would be useful.
Ying Xiao, J J Morrell, L M Ciuffetti

Physical properties of ß-1,4-Xylanase produced by Postia (=Poria) placenta: Implications for the control of brown rot
1987 - IRG/WP 1318
The degradation of hemicelluloses is an early event in wood decay by brown-rot fungi. An understanding of the physical properties of hemicellulases may suggest target mechanisms for the development of new control agents. Endo-b-1,4-xylanase was partially purified by column chromatography from wood decayed by Postia (= Poria) placenta. The enzyme was extremely resistant to denaturing conditions; no loss of activity was detected after 2 h in 9 M urea or 6 M guanidine-HCl. Boiling the enzyme for 5 min in 2.5% SDS + 0.5% b-mercaptoethanol reduced its activity by 65%, as measured by the production of reducing sugars. The activity of a-D-galactosidase, another enzyme detected in large quantities in the decayed wood, was reduced by 98% under these conditions. Optimum pH and temperature ranges were pH 2-6 and 50-60°C, respectively. The enzyme appears to be a glycoprotein containing 50-60% carbohydrate (w/w); the carbohydrate moiety may protect the enzyme from adverse environmental conditions. The control of brown rot by in situ inactivation of xylanase may not be feasible because of the enzyme's extreme stability.
J A Micales, F Green III, C A Clausen, T L Highley

Report of Section 1 2003
2003 - IRG/WP 03-10502
F Green III

Treatment of fresh green round bamboos culms (Dendrocalamus strictus) by sap-displacement (wick) method
2005 - IRG/WP 05-40311
Sap displacement method has great potential for treating short length bamboos as it does not require any technical equipment. The process is simple and large nos. of bamboos can be simultaneously treated in relatively short period. There is no wastage of chemicals as the remnant solution reused. Bottom ends and middle portion had better treatment in compared to lop end portion. With increasing length of the flow of preservatives is impaired. The method culms can conveniently 3m length.
R Lal, C N Vani

Potentialities of protein borates as low-toxic, long-term wood preservatives - Preliminary trials
1999 - IRG/WP 99-30212
Boron compounds are efficient wood preservatives, as well as safe for the mammals and environmentally acceptable. Their natural solubility allows them to treat almost any wood species, but is also the cause of their high depletion from treated timber in outside exposure. In order to reduce this leachability, potentialities of proteinic polymer networks retaining boron within the wood have been investigated. Several mixtures of boric acid and proteins (including ovalbumin, collagen, casein, soya flour) have been used to treat pine sapwood miniblocks. The insoluble networks were obtained by protein gelation or coagulation, induced by a physical and/or a chemical factor. These systems appeared to retard boron leaching, the decrease of the leachability rate depending on the protein and the denaturing agent involved in the network creation. The best results have generally been observed for the irreversible heat-induced protein gels. These associations are also able to conserve some boron mobility and activity. Accelerated biological tests of leached wood samples showed good durability performances against Basidiomycetes. The use of protein borates seems to be an interesting basis for low-toxic wood preservatives. Furthermore, in some cases, proteins could add their anti-nutritional factors to boron efficacy to enhance wood protection.
M-F Thévenon, A Pizzi, J P Haluk

Molecular studies on isolates of Serpula lacrymans
1989 - IRG/WP 1421
The major protein species present in detergent extracts of 14 different Serpula lacrymans isolates have been compared, by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), with a standard strain, viz. Serpula lacrymans FPRL 12C. Following silver staining of SDS gels the major protein species identified in 12 isolates were similar to those found in the standard strain. However differences were found when the final two isolates were compared with FPRL 12C, both isolates had extra molecular species not present in 12C and both were lacking some present in the standard strain. Comparison of the protein species identified in Serpula lacrymans isolates with those identified in extracts of other fungal organisms, viz. brown and white rot causing basidiomycetes and non-basidiomycetes indicated that the Serpula isolates were more similar to each other than to other organisms. Some molecular differences could be identified when individual isolates were cultured on different media, i.e. liquid culture or agar, only minor differences were seen when individual isolates were subcultured. These results indicate that whilst care must be taken to ensure as near identical conditions as possible for culture of organisms if their molecular species are to be compared by SDS-PAGE and silver staining, consistent results can be obtained using this technique. The technique may therefore offer a method of distinguishing between isolates, strains and species of wood decay basidiomycetes, and identifying new isolates.
A Vigrow, D Button, J W Palfreyman, B King, B M Hegarty

Observations on the penetration of preservatives into green timber
1985 - IRG/WP 3335
Differences in the rate and extent of diffusion of MBT and NaPCP-based preservatives following a short dip treatment were observed. The importance of preservative distribution as well as toxicity to the target organisms in governing the ultimate performance of any anti-sapstain compound is discussed.
G R Williams, R A Eaton, D A Lewis

Protection of Ochroma pyramidale from fungal decay with N,N-napthaloylhyroxylamine
1998 - IRG/WP 98-30182
Fungal decay of wood in service results in billions of dollars (U.S.) in losses annually. Recent environmental restrictions, both U.S. and international, are limiting and eliminating the use of broad-spectrum, heavy metal biocides for wood preservation. Restrictions result primarily from problems with disposal. New wood preservatives need to be developed and tested which specifically target key elements in the sequence of fungal decay mechanisms. Our laboratory has been experimenting with chemicals which inhibit pectin hydrolysis during incipient brown-rot and white-rot decay in southern pine sapwood (Inter. Biodeter. Biodegrad. 39:103). In the present paper these results are extended to include the tropical hardwood Ochroma pyramidale (balsa). Balsa blocks (24x18x12mm) were exposed to two brown-rot fungi and one white-rot fungus in ASTM soil block tests for 10 weeks. CCA (6.4 km/m3 ) was compared with the calcium binding agent N,N-napthaloylhydroxylamine (NHA; 1.6, 3.2 & 6.4 km/m3 ) in leached and unleached blocks. CCA protected balsa with minimal weight loss (> 7.4%) with no leaching effects. NHA (6.4 km/m3 ) protected balsa (0.3-1.2%) weight loss but leaching raised the weight losses to 25% with the brown-rot fungus Tyromyces palustris. We conclude that NHA can protect balsa against G. trabeum and T. versicolor with comparable efficiency to CCA (leached and unleached) but not T. palustris.
F Green III, T L Highley

Use of fluorescent-coupled lectins as probes for studying fungal degradation of wood
1986 - IRG/WP 1288
The ability of the fluorescent-coupled lectins wheat germ agglutin (WGA) and Concanavalin A (Con A) to react with selected Basidiomycetes, Ascomycetes, and Fungi Imperfecti was evaluated using pure cultures of 35 fungi grown on malt extract agar. WGA, which binds specifically to the n-acetylglucosamine residues found in fungal chitin, reacted with nearly all hyaline fungal structures but did not react with dematiaceous (dark) structures. Several reasons are suggested for this variation. Con A, which is specific for a-D-mannosyl and a-D-glucosyl residues, reacted with about one half of the fungi that reacted with WGA. This variation in reactivity may be useful for studying simultaneous degradation by morphologically similar fungi having different lectin specificities. The results indicate that WGA is a useful probe for studying fungal degradation by non-dematiaceous fungi particularly at the early stages of decay.
J J Morrell, R L Krahmer, L C Lin

Oxalic acid production of fifteen brown-rot fungi in copper citrate- treated southern yellow pine
2001 - IRG/WP 01-10388
Non-arsenical copper-based wood preservatives have grown in number since the 1980's as a response to environmental concerns posed by arsenicals. Interest in copper tolerant decay fungi has increased accordingly. Oxalic acid (OA) production by brown-rot fungi has been proposed as one mechanism of copper tolerance. Fifteen brown-rot fungi representing the genera Postia, Wolfiporia, Serpula, Gloeophyllum, Laetiporus, Coniophora, Antrodia, and Tyromyces were evaluated for OA production bi- weekly in southern yellow pine (SYP) blocks treated with 1.2% AI copper citrate (CC). Ten fungi were designated copper tolerant and produced 2 to 17 times more OA in the CC-treated blocks than in untreated SYP after 2 weeks. Weight losses ranged from 20 to 55% in CC-treated SYP after 10 wks. Five fungi were copper-sensitive, producing low levels of OA and low weight losses on CC-treated blocks. In this study, early induction of OA appears to closely correlate with copper tolerance. We conclude that brown-rot fungi able to exceed and maintain an OA concentration of >400 mM in this study effectively decayed SYP treated with copper citrate.
F Green III, C A Clausen

Inhibition of wood decay and termite damage by calcium precipitation
1996 - IRG/WP 96-30111
Fungal decay of wood in service, especially brown rot, results in billions of dollars (US) of losses annually. Recent environmental restrictions, both U.S. and international, are limiting or eliminating the use of broad spectrum biocides for wood preservation, primarily due to problems with disposal. In order to design new, environmentally benign methods for control of wood decay fungi, it is essential to understand the precise sequence of biochemical events as wood is colonized. The production of polygalacturonase (PG) and hydrolysis of bordered pit membranes during incipient brown-rot decay has recently been described by our laboratory. One key to pectin hydrolysis by plant pathogens has been shown to be fungal production of oxalic acid which lowers the pH of the substrate and chelates calcium ions. Production of oxalic acid may serve a similar role during incipient wood decay as calcium oxalate has been visualized by scanning electron microscopy during both brown-rot and white-rot decay. Therefore, it was hypothesized that in situ precipitation of existing calcium ions in wood may prevent the cascade of biochemical events involved in colonization of wood by brown-rot fungi, expecially hydrolysis of bordered pit membranes. Preliminary experiments in our laboratory have shown that brown-rot fungi, white-rot fungi, and termites are inhibited from effecting weight loss of wood following pretreatment of wood blocks with the selective water soluble calcium precipitating agent N,N-naphthaloylhydroxylamine (NHA).
F Green III, T A Kuster, L Ferge, T L Highley

Effect of light and ventilation condition on the rate of wood decay by the brown rot basidiomycete, Tyromyces palustris
1991 - IRG/WP 1517
Effect of light and the ventilation conditions of incubation jars on the wood decay by Tyromyces palustris (Berk. et Curt.) Murr. FFPRI 0507 was investigated. Under no irradiation of light, the ventilation conditions gave extensive effect on mass loss of the test pieces when the culturing was performed with culture medium designated in Japanese Industrial Standard (JIS) A 9302 (Medium A; glucose 4.0%, malt extract 1.5%, peptone 0.3%). On the other hand, no such kind of effect for ventilation conditions was recognized under light conditions, and the wood decay by the fungus accelerated by additional light irradiation of a considerably small intensity. Next, we investigated the relationship between light and the culture medium composition during the wood decay by the fungus. It was found that almost equivalent mass loss occurred after 60-90 days of cultures when the culturings were performed under light-shield conditions with medium A, the culture medium designated in Japan Wood Preserving Association (JWPA) standard No.1 (medium B; malt extract 2.0%, peptone 1.0%), and another culture medium diluted medium B by two times (medium C). Under the irradiation of light, the mass loss on the cultures in medium B and C was markedly less than that in the same media under no irradiation conditions. These results suggested that effect of light on the wood decay by Tyromyces palustris depended on the concentration of glucose in the culture medium. Further, we also investigated the activities of several extracellular and cell wall bound enzymes in wood meal medium contained medium A. From our experimental results, the activities of cellulase (b-1,4-glucan 4-glucanohydrolase, E.C. and mannanase (b-1,4-mannan mannanohydrolase) depended on light irradiation during the wood decay and these enzyme activities may give extensive effect on the mass loss by Tyromyces palustris.
T Suzuki, M Higaki

Inhibition of termite damage by N'N-napthaloylhydroxyamine (NHA): Reticulotermes flavipes (Kollar) vs. Coptotermes formosanus Shiraki
2000 - IRG/WP 00-10354
The calcium precipitating agent NHA has been shown to protect southern yellow pine (SYP) from wood decay and termite damage comparable to CCA in field tests (Gulfport, MS) for two years (Crawford and Green, 1999). In a collaborative study, SYP wood blocks were vacuum treated with three concentrations of aqueous NHA and exposed in a no-choice test to Eastern subterranean termites (FPL, USA) and Formosan subterranean termites (WRI, Japan) to determine protection against termite damage. Individual blocks (leached and unleached) were exposed to R. flavipes (AWPA) or C. formosanus (JWPA) for 3-4 weeks. Mean weight loss of wood blocks after termite exposure ranged from 0.0 to 18.0% for R. flavipes and 6.0 to 20% for C. formosanus. Wood blocks exposed to R. flavipes were completely protected by 0.5 and 1.0% NHA, but weight loss in similar blocks challenged by C. formosanus were 6.0% and 6.2% respectively at the same concentrations. NHA acted as an effective termiticide for R. flavipes with 100% mortality after 3 weeks, but only soldiers were preferentially killed in C. formosanus. Formosan subterranean termite workers showed enhanced resistance to NHA treatment when compared to Eastern subterranean termites.
F Green III, S T Lebow, T Yoshimura

Laboratory tests on light organic solvent preservatives for use in Australia. - Part 6: Soft rot resistance of three fully formulated preservatives on different timber substrates
2000 - IRG/WP 00-30245
The above-ground soft rot resistance of substrates treated with three fully formulated light organic solvent preservatives (Cuprivac Green WR, Impresol WR 205 and Vacsol) was studied using a modified vermiculite burial method. The substrates were sapwood of Pinus elliottii and P. radiata and heartwood of Eucalyptus regnans, Pseudotsuga menziesii, Shorea sp. (a lower and a higher density source) and Thuja plicata. Following artificial weathering, replicate test blocks were exposed to either Chaetomium globosum or Lecythophora mutabilis. C. globosum caused 3% or more mass loss of the water and solvent (white spirit) impregnated controls of all three hardwoods and two of the four softwoods, whereas L. mutabilis caused similar attack in only E. regnans and P. radiata. The P. menziesii and T. plicata heartwoods were naturally durable to both soft rot fungi and, hence, no further conclusions can be drawn. None of the preservatives, at the highest retention tested, protected E. regnans from attack by C. globosum, whereas the highest retentions of both the Cuprivac Green WR and Impresol WR 205 protected all other timbers from this fungus. At the highest retention, the latter preservative was the only one to protect E. regnans from L. mutabilis.
G C Johnson, M A Tighe, J D Thornton

Treatability problems - Relationships between anatomy, chemical composition and treatability
2001 - IRG/WP 01-40213
This report documents the results of phase 1 and 2 of a 3-phase research program. In phase 1, two hundred and fifty-six (256) Southern pine (pinus spp.) nominal 2 x 6's (38 mm x 140 mm) from a single mill in Georgia (southeastern US) were evaluated for treatability with CCA preservative. After treatment, 128 pieces representing a broad range of treatment characteristics were selected and more fully evaluated for anatomical and chemical composition. No direct correlation was noted between CCA treatability and any of the anatomical characteristics evaluated in this study. Neither did a direct correlation between chemical composition and treatability seem to exist. The pit tori of all pits evaluated in this study were aspirated against the pit wall for this difficult to treat Southern pine lumber. This pit aspiration might have resulted from pre-treatment kiln drying. Later reductions in kiln temperature and increased kiln humidity (i.e., lowered wet-bulb depression temperature) reportedly resulted in improved treatability. In Phase 2, a methodology was developed to assess treatability after 3 treating schedules and then compare those results to permeability measurements and anatomical characteristics of matched material from 7 growth regions.
J E Winandy, F Green III, D Keefe

Evaluating the natural durability of native and tropical wood species against Reticulitermes flavip
2004 - IRG/WP 04-10539
Environmental pressures to eliminate arsenate from wood preservatives has resulted in voluntary removal of CCA for residential applications in the United States. A new generation of copper organic preservatives has been formulated to replace CCA for decking and in-ground applications but there is no guarantee that these preservatives represent a permanent solution to all related problems. Therefore, it is still necessary to evaluate alternative treatments, as well as naturally durable wood species, in order to be prepared for future changes in the field. In this study, six hardwoods and six softwoods have been evaluated for their ability to resist termite damage by Reticulitermes flavipes in a 4-week laboratory no-choice test. In addition, moderately resistant Douglas-fir and southern pine wood blocks were evaluated after treatment with copper borate, copper naphthanate, and N,N-naphthaloylhydroxylamine (NHA). Erisma, juniper, ipe and white-cedar were shown to be highly resistant. NHA protected Douglas-fir and southern pine as effectively as copper borate or copper naphthanate. These results suggest that some naturally durable wood species, both tropical and native, can inhibit R. flavipes as effectively as preservative treatment.
R A Arango, F Green III, K Hintz, R B Miller

Commercial potential of the four-cycle method for the impregnation of green beech sleepers in Yugoslavia
1975 - IRG/WP 347
This paper was presented to the Conference on Wood Protection held in Sarajevo (Yugoslavia) in 1973. The paper was based partially on the report prepared by J. Struhar and G. F. Franciosi, who were appointed in 1972 as FAO consultants to demonstrate the new impregnation process for green beech sleepers in Yugoslavia. The so-called 4-cycle method was developed at the State Forest Research Institute in Bratislava (Czechoslovakia). The demonstration treatment was carried out at the Kolasin plant, where the double Rueping method is currently used. Comparing the double Rueping method with the 4-cycle method, the author discusses the possibility of introducing the new method into existing impregnation plants in Yugoslavia. Analysis has shown that the production costs of impregnated beech sleepers under the new method would cost 44 Dinars per m³ more for sleepers, or 530,000 Dinars annually (based on the Kolasin plant capacity). On the other hand, formation of tyloses and doat in "white" sleepers during seasoning, estimated to be 10%, would be completely avoided, saving 972,000 Dinars annually at the Kolasin plant. Besides the stock of untreated sleepers would be considerably reduced which would enable the saving of working capital at the Kolasin plant of about 2,5 mill of Dinars. It is also expected that due to higher creosote oil consumption and its even spreading through the wood, the service life of sleepers would be extended by 10 years (from 30 to 40). In such a way the Yugoslavian Railways would decrease the purchasing and installation costs by about 10 Dinars per m³ of sleepers or 1,270,000 Dinars annually (in 1972 USA$ 1 equalled 15 Dinars).
N Vidovic

Application of roller-pressing method to the novel liquid impregnation treatment of green timber
2001 - IRG/WP 01-40198
A transverse compression technique enabled us to impregnate liquid into green timber through replacement of the free water in the timber with a treatment liquid without any critical physical damage to the timber by the roller-pressing method. The roller-pressing is supposed to remove free water the cell cavities and to concurrently introduce the treatment solution into green timber in the treatment vessel. This process seems applicable to the preservative treatment of timber without drying prior to the treatment, and definitely contributes to both the reduction of production costs and the greater yield of the products due to the occurrence of the fewer undesirable checks. When Cryptomeria japonica D. Don was treated with an aqueous solution of phenolic-resin according to this process, weight gain exceeded 40% and anti-swelling efficiency went up to over 60% even for 1 min immersion period after pressing. Further trials are planned to evaluate biological resistance of the treated timber when fungicide(s) and/or termiticide(s) are incorporated into the treatment solution(s).
M Inoue, K Adachi, K Tsunoda, Y Imamura, S Kawai

Serpula lacrymans, The Dry Rot Fungus and its Tolerance towards Copper-based Wood Preservatives
2005 - IRG/WP 05-10555
Serpula lacrymans (Wulfen : Fries) Schröter, the dry rot fungus, is considered the most economically important wood decay fungus in temperate regions of the world i.e. northern Europe, Japan and Australia. Previously copper based wood preservatives were the most commonly used preservatives for pressure treatment of wood for building constructions. Because of a suspicion about tolerance toward copper components, a soil block test was undertaken to clarify the effect of two copper based preservatives, copper citrate and ACQ-D, on the dry rot fungus, Serpula lacrymans compared to an alternative non-copper containing wood preservative. The extensive use of copper-based wood preservatives has hastened the need for understanding why some fungi are able to attack copper-treated wood. The copper tolerance of S. lacrymans and other brown-rot fungi is thought to be due in part to oxalic acid production and accumulation. Oxalic acid has been implicated in copper tolerance by the formation of copper oxalate crystals. Twelve isolates of the dry rot fungus, S. lacrymans and four other brown rot species were evaluated for weight loss on wood treated with 1.2% copper citrate, 0.5% ACQ-D and 0.5% N’N-naphthaloylhydroxylamine (NHA). Eleven out of 12 S. lacrymans were shown to be tolerant towards copper citrate. ACQ-D and NHA, on the other hand, were both effective against the dry rot isolates. These wood preservatives are less toxic toward the environment than traditional copper based preservatives.
A C Steenkjær Hastrup, F Green III, C A Clausen, B Jensen

Gas chromatographic determination of 1,8-naphthalimide, N-Hydroxy-1,8-naphthalimide (N,N-naphthaloylhydroxylamine) and the sodium salt of N-Hydroxy-1,8-naphthalimide
2003 - IRG/WP 03-20259
A number of naphthalimide (NI) derivatives are used as efficient laser dyes, in medicine or in scanning electron microscopy. Only N,N-naphthaloylhydroxylamine (NHA) has been shown to be an effective wood preservative against wood decay fungi and termite damage. However, limited information is available concerning the analytical detection of NI-derivatives in treated timber. There is a clear need for the analytical characterisation; e.g. with regard to the penetration depth or the assessment of retention after leaching. This paper describes the development of a gas chromatographic method for determination of NI and their derivatives in timber. These investigations were carried out by means of direct thermal desorption-gas chromatography-mass spectrometry (TD-GS-MS) using the pure substances in solution, as well as direct analysis of treated southern yellow pine (SYP). It was shown that the identification of NHA in treated SYP is possible using this analytical technique. Furthermore first evidence is given for determining quantitative data. Surprisingly, the chromatograms and especially mass spectra obtained for NHA and the sodium salt of NHA are identical to the mass spectra of NI. The first results show that TD-GC-MS can be an option in determining the retention levels of NI and their derivatives in wood.
E Melcher, F Green III

Proposed model for the penetration and decay of wood by the hyphal sheath of the brown-rot fungus Postia placenta
1989 - IRG/WP 1391
Scanning electron microscopy (SEM) of Pinus sp. decayed by the brown-rot fungus Postia placenta confirmed the existence of extracellular membranous structures previously described by transmission electron microscopy (TEM). These structures appear to be an integral part of the hyphal sheath and assume a variety of forms including lamellar sheets, fibrils, and vesicles. These structures were observed (a) on the surface of hyphae, (b) extending from hyphae into the wood surface and covering the S3 layer, (c) embedded in the hyphal sheath and, (d) penetrating into the wood cell wall layers from S3 - S3. We conclude that penetration of the wood cell wall by the complex, multistructured hyphal sheath facilitates the decay process in an orderly and linear progression. The directionality of the decay process appears to alter the fiber orientation of individual wood cell wall layers. This model offers an alternative hypothesis to simple diffusion of wood degrading agents during wood decay by Postia placenta.
F Green III, M J Larsen, L L Murmanis, T L Highley

Early detection of brown-rot decay in southern yellow pine using immunodiagnostic procedures
1990 - IRG/WP 2356
Immunodiagnostic procedures have been used to detect incipient decay of southern yellow pine by six common brown-rot fungi. Rabbit polyclonal antibodies were raised to liquid culture fractions of the six fungi. The antibodies, after preadsorption to sawdust, were tested in particle agglutination assays, immuno-dot blot, and ELISA for their·ability to detect the decay organisms when they were grown in southern yellow pine. Results were correlated to wood block weight loss. Each method specifically detected five of the six test organisms at very low wood block weight loss. Agglutination assays were the least effective and lacked the sensitivity of the other assays, even though the latex particles were more sensitive and reproducible than charcoal particles. Cross-reactivity was only noted for one of the control organisms and only in the charcoal agglutination assay. Both dot blot and ELISA were sensitive test methods, but ELISA had the advantage of quantification. We feel that two of these assays, latex for presumptive results and ELISA for definitive results, could effectively detect incipient decay of common brown-rot fungi.
C A Clausen, F Green III, T L Highley

Steam/hold/APM boron treatment - Treatability trials with green gauged radiata pine
1987 - IRG/WP 3439
Freshly sawn 100 x 50 mm radiata pine was green gauged, steam conditioned and preservative treated with a mixture of borax and boric acid using a modified Alternating Pressure Method (APM) treatment schedule. A 12 hour holding period between steam conditioning and treatment resulted in the necessary moisture loss and moisture re-distribution to facilitate treatment to NZ Timber Preservation Authority preservative retention and distribution requirements.
P Vinden

Detection of a brown-rot fungus using serological assays
1986 - IRG/WP 1305
Polyclonal antisera produced to Poria placenta (Fr.) Cooke were used in two tests to qualitatively assay for the fungus. Fungal hyphae were fixed to slides and fluorescent antibody (FA) techniques used to visualize the hyphae under the microscope. Fluorescence of non-Poria fungi, when present, could be reduced but not eliminated by cross-absorbing the sera with these fungi. The antisera was also used in an enzyme-linked immunosorbent assay (ELISA) to distinguish between Poria placenta and a non-Poria control fungus Rhizoctonia solani.
B Goodell, J Jellison

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