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The influence of crystalline and amorphous cellulose on extracellular hydrogen peroxide production by brown-rot fungi
1991 - IRG/WP 1482
The production of hydrogen peroxide (H2O2) has been suggested to play a key role in the degradation of wood by wood-rotting fungi. The production of extracellular hydrogen peroxide was studied by a quantitative method which detects the oxidation of the 2,2-azinobis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) by H2O2 and horseradish peroxidase (HRP) in liquid culture medium. The carbon sources used were crystalline and amorphous cellulose. Two brown-rotters, Serpula lacrymans and Poria placenta, were able to produce clearly detectable amounts of extracellular hydrogen peroxide in liquid medium which contained crystalline cellulose as carbon source. No detectable H2O2 was produced in conditions where amorphous medium was used as carbon source. This result suggests that the conformational structure of the substrate may induce H2O2 production by brown-rot fungi.
A-C Ritschkoff, L Viikari
Cytochemical localization of hydrogen peroxide in brown rot fungus Tyromyces palustris by cerium chloride technique
1999 - IRG/WP 99-10299
Cerium chloride (CeCl3) was used to localize H2O2 cytochemically for studying relationship between ultrastructural and functional characteristics of cellulose degradation by brown rot fungi. This technique proved very useful in localizing discrete electron-densereactionproducts at high resolution with minimal nonspecific deposition. The cytochemical localization of extracellular H2O2 by CeCl3 using TEM demonstrated the presence of H2O2 within the fungal hyphae. Furthermore, our results give an indication of the diffusion of extarcellular H2O2 from brown-rot decay fungi into the intact wood cell walls in the early stages of decay.
Yoon Soo Kim, Seung-Gon Wi
Effect of fatty acid removal on treatability of Douglas-fir
1993 - IRG/WP 93-40008
Treatment of Douglas-fir with chromated-copper-arsenate (CCA) poses a major challenge. Several hypotheses based on the anatomical aspects as well as chemical reactivity of the preservative formulations with cell wall constituents and deposits have been proposed. Techniques to prevent pit aspiration or slow fixation reactions have, however, not significantly improved treatment. The presence of high molecular weight fatty acids have been reported to be responsible for higher hydrophobicity in some wood species. These acids can react with Cu+2/Cr+3 ions to form insoluble metallic soaps, thereby immobilizing Cu/Cr and increasing wood hydrophobicity by a mechanism similar to that employed in paper sizing. The effect of fatty acids on treatability was explored by removing these components via several extraction methods. In general, extracted wood had higher gross solution absorptions and chemical retentions, but preservative penetration was largely unaffected. The results suggest that removal or disruption of fatty acids can improve treatability of Douglas-fir heartwood.
S Kumar, J J Morrell
Involvement of hydrogen peroxide in wood decay by brown-rot and white-rot fungi
1985 - IRG/WP 1256
To gain further understanding of the role of H2O2 in wood degradation by brown- and white-rot fungi, we studied the following: (a) extracellular H2O2 production, (b) effect of various hydroxyl radical (·OH) and singlet O2 (1O2) quenching agents on wood and cellulose degradation, (c) intracellular H2O2 production and catalase activity, and (d) cytochemical localization of H2O2 with diaminobenzidine (DAB) during wood degradation. Extracellular H2O2 detection varied with the growth media and chromogen. The chromogen 2,2'-azino-di-(3-ethyl benzthiazoline-6-sulphonic acid) (ABTS) was more sensitive than o-dianisidine. Extracellular H2O2 was not detected in half of the brown-rot fungi. One white-rot fungus did not produce detectable amounts of H2O2. ·OH and 1O2 quenching agents generally did not inhibit decay of wood or decomposition of cellulose by either brown- or white-rot fungi. DAB did not detect the presence of H2O2 within or outside cells of the brown-rot fungus, Poria placenta. Nor was H2O2 -generating activity detected in sonicated extracts of this fungus. With the white-rot fungus, Coriolus versicolor, H2O2 occurred predominantly in the periplasmic space, but also in the cytoplasm and hyphal sheath. Sonicates of Coriolus versicolor contained H2O2 -generating activity. These observations provide further support for H2O2 involvement in degradation of wood by white-rot fungi, but raise doubts concerning its participation in wood degradation by brown-rot fungi.
T L Highley, L L Murmanis
Iron in stone wool - one reason for the increased growth and decay capacity of Serpula lacrymans
1992 - IRG/WP 92-1537
The chemical compositions of stone wool and glass wool were analysed. There was more iron in the stone wool than in the glass wool. It was found that iron present in stone wool was easily dissolved by oxalic acid that Serpula lacrymans is able to produce. The stone wool promoted the decay of pine wood by Serpula lacrymans. The glass wool had no effect on the decay capacity of Serpula Iacrymans. The iron derived from the stone wool may be one reason for the increased growth and decay capacity of Serpula lacrymans. Transition metals (Fe2+, Mn2+, etc.) combined with hydrogen peroxide (H2O2) are believed to be necessary for the oxidative breakdown of polysaccharides.
L Paajanen, A-C Ritschkoff
Screening of lignin peroxidase from white-rot fungi
1998 - IRG/WP 98-10274
Since lignin peroxidase was first found in the ligninolytic culture of Phanerochaete chrysosporium, it has been regarded as an enzyme responsible for the lignin biodegradation and studied comprehensively. As the results of these studies its unique characteristics have been revealed. On the other hand, the features of lignin peroxidase produced by other white-rot fungi have not been studied so much. In this study, over 80 white-rot fungi were examined for their ability to produce lignin peroxidase. Five fungi produced lignin peroxidase, and they were subjected to further investigations. These lignin peroxidases showed higher activity at pH 3.5 than pH 4.5 in the oxidation of veratryl alcohol. It could oxidize guaiacol slowly. These features are similar to those from P. chrysosporium. Newly isolated lignin peroxidase were inactivated by the reaction with hydrogen peroxide. There was a difference in the rates of the inactivation by the hydrogen peroxide treatment. The rate observed in the inactvation of lignin peroxidase from P. chrysosporium was moderate.
Controlling the sapstain fungus Ceratocystis coerulescens by metabolites obtained from Bjerkandera adusta and Talaromyces flavus
1993 - IRG/WP 93-10024
Sapstain causes severe damage to wood and wood products, posing a major economic problem for the wood industry. The purpose of this study was to determine if metabolites from Bjerkandera adusta and Talaromyces flavus would (1) decolorize stain in wood caused by Ceratocystis coerulescens and (2) prevent sapstain by Ceratocystis coerulescens. We studied the interaction of the sapstain fungus Ceratocystis coerulescens against the test fungi Bjerkandera adusta and Talaromyces flavus in dual cultures on agar medium. The metabolites obtained from test fungi were examined on pine veener disks stained by Ceratocystis coerulescens. Our results indicate that the test fungi were antagonistic to the sapstain fungus Ceratocystis coerulescens. The combination of metabolites from the antagonists decolorize the sapstained pine veener disks and killed the existing growth of Ceratocystis coerulescens.
S C Croan, T L Highley
Production of exraxellular hydrogen peroxide and oxalic acid by brown-rot fungus Poria placenta
1995 - IRG/WP 95-10112
Hydrogen peroxide and oxalic acid have been suggested to be essential in the degradation of wood carbohydrates by brown-rot fungi. Hydrogen peroxide has been suggested to be one of the diffusible low molecular weight agents produced by brown-rot fungi for the degradation of wood carbohydrates by generating highly active radicals. The production of hydrogen peroxide and oxalic acid by Poria placenta was studied on crystalline cellulose, amorphous cellulose and glucose media. Hydrogen peroxide and oxalic acid were clearly produced on culture media containing either amorphous or crystalline cellulose. Hydrogen peroxide and oxalic acid were formed simultaneously and highest amounts of these compounds were obtained on amorphous cellulose. The production of hydrogen peroxide and oxalic acid was surprisingly low on glucose medium.
A-C Ritschkoff, M Rättö, J Buchert, L Viikari
Extracellular hydrogen peroxide producing and hydrogen peroxide reducing compounds of wood decay fungi
1991 - IRG/WP 1516
Extracellular H2O2-producing and H2O2-reducing compounds were isolated from wood-containing cultures of all the white-rot and brown-rot fungi and Ascomycetes which well degraded wood, but were not detected in the culture of the fungi which degraded little wood. The compounds are glycopeptides with a low molecular weight, require H2O2 for one-electron oxidation, catalyze the redox reaction between an electron donor such as NADH or ascorbic acid and O2 to produce H2O2 via O2·-, and produce ·OH by Fenton's reaction between the ferrous iron bound to the ligands and H2O2. The compounds show no phenol-oxidase activity and catalyze the hydroxylation of phenol to catechol and hydroquinone in the presence of H2O2.
A Enoki, G Fuse, H Tanaka
Extracellular substance from the white rot basidiomycete Irpex lacteus involved in wood degradation
1992 - IRG/WP 92-1571
Cellulases, phenol oxidase, and a substance that both produced and reduced H2O2 were isolated from cultures containing wood or glucose on which the white-rot fungus Irpex lacteus was growing. The rate of wood degradation by the fungus with different amounts of glucose in the medium was measured. More of the substance that produced and reduced H2O2 was found extracellularly in cultures containing wood than in cultures containing glucose. The extent of one-electron oxidation activity of the extracellular substance was correlated with that of the wood-degrading activity, but the extent of phenol oxidase activity was not. The extracellular substance catalyzed the hydroxylation of phenol to hydroquinone and catechol in the presence of H2O2 or of NADH and O2. The substance seemed to be a glycopeptide containing Fe(II) and its molecular weight was very low.
H Tanaka, T Hirano, G Fuse, A Enoki
The role of oxidation in wood degradation by brown-rot fung
1992 - IRG/WP 92-1562
Brown-rot fungi are suggested to degrade cellulose by oxidation with hydrogen radicalsformed eg. in the conversion of hydrogen peroxide in the Fenton type reactions. The stuctural changes in the holocellulose in wood induced by Fenton's reaction on wood components are very similar to those caused by brown-rot fungi. In this work the effect of the Fenton reaction on wood components was studied. The brown-rot fungi produce extracellular hydrogen peroxide while growing on sawdust medium and on crystalline cellulose. Hydrogen peroxide is apparently formed by the action of an oxidase enzyme. In this work the role of enzymatic oxidation in cellulose degradation by the brown-rot fungus, Poria placenta was studied.
A-C Ritschkoff, J Pere, J Buchert, L Viikari
Mechanistic aspects of the reactions of copper complexes with lignin in the presence of hydrogen peroxide and lipid hydroperoxide model compounds as proposed for white rot fungi
2001 - IRG/WP 01-10399
The structural principles and the oxidative reactions of the catalytic system consisting of copper, hydroperoxides, and copper coordination compounds that contain aromatic nitrogen atoms are similar to those of catalytic centers of oxidative enzymes involved in delignification reactions of white rot fungi. The system is capable of selectively degrading lignin and its low molecular weight would allow to react oxidatively also at sites of the wood cell wall, where enzymes cannot penetrate. Understanding the mechanisms of white rot would offer perspectives to develop targeted wood preservatives to inhibit these enzymatic and non-enzymatic reactions of wood decay. Studies on the mechanism of the copper/pyridine and 4-aminopyridine resp./hydrogen peroxide system that were carried out with 14C-labeled lignin and cellulose model compounds demonstrated the selectivity of the copper system for lignin structures and the ability of the system to oxidise even non-phenolic lignin. Product profiles of the reactions supported by ESR measurements suggested a non-radical reaction pathway involved in that case. However, when a lipid hydroperoxide model (cumene hydroperoxide, CHP) was used as peroxide source, a variety of free radicals derived from this peroxide could be detected in ESR. Selective staining of treated wood sections with safranin and astra blue showed degradation of softwood and hardwood lignin in the middle lamella and secondary cell wall. Besides its importance for developing targeted wood preservatives, the coordinated copper system has great potential for application for selective lignin degradation in the pulp and paper industry and could therefore replace environmentally problematic chlorine and chlorine dioxide based pulp bleaching stages.
K Fackler, P Lamaipis, E Srebotnik, M Humar, C Tavzes, F Pohleven, M Sentjurc, T Watanabe, K Messner
Extracellular hydrogen peroxide-producing and one-electron oxidation system of brown-rot fungi
1990 - IRG/WP 1445
Wood-component-degrading compounds involved in the initial degradation of the cellulose and lignin in wood were isolated from wood-containing culture of brown-rot fungi, Gloeophyllum trabeum and Tyromyces palustris and partially purified by gel filtration on Sephadex G-25 and DEAE-Sepharose ion-exchange chromatography. The compounds were glycoproteins. The molecular weights of the glycoproteins as determined by gel filtration were very small and about 1,600-2,000. The one-electron oxidation activity of the peptides was determined by measuring ethylene production from 2-keto-4-thiomethylbutyric acid (KTBA). The peptides contained ferrous iron,required H2O2 for KTBA oxidation, were capable of catalyzing the oxidation of NADH to produce H2O2 in the presence of 02 and showed little phenol-oxidase activity under conditions giving high activity against KTBA. The ferrous iron combined with the glycopeptides was oxidized to the ferric state by H2O2.
A Enoki, S Yoshioka, H Tanaka, G Fuse
Experiments on the degradation of tributyltin oxide: A progress report
1984 - IRG/WP 3287
A variety of experiments designed to assess the chemical and physical factors affecting the degradation of tributyltin oxide in treated timber are described. Simple procedures in which temperature and oxygen availability were increased in the presence of wood and water suggest that the wood itself was of prime importance. Attempts to decrease degradation with antioxidants were unsuccessful but led to the idea that free radicals may be instrumental in the degradative mechanism. Subsequent work in which a range of antioxidants and free radical producing systems were used confirmed the susceptibility of tributyltin oxide to the action of free radicals. It is suggested that the presence of free radicals in the painted and treated wood system may be an important factor in the eventual degradation of tributyltin oxide.
R J Orsler, G E Holland
In-house accelerated method for testing decay resistance of treated wood
2004 - IRG/WP 04-20286
Fungicidal compounds often change their effectiveness when they are incorporated into candidate formulations. For this reason fungistatic effectiveness needs to be re-evaluated as many times as the formula has been modified for better performance. To avoid multiple expenses, in-house fungistatic tests are essential. Our goal was to develop in-house usable, simple but reliable and reproductive procedure for testing decay resistance of the Sansin priming formulae without a need for special testing equipment. Our preliminary trials have proved that commercially available, heat processed wood pellet fuel can be used as an excellent substrate for growing certain wood rotting fungi. In contact with water wood pellets quickly disintegrate and convert into sawdust, gaining up to three times increase in volume. When wood pellets are wetted with water containing hydrogen peroxide, the vapors of this antiseptic kill non-desirable air-borne contaminants and protect the substrate from further contamination for as long period as peroxide remains in the substrate at sufficient concentration. When peroxide protected substrate is spawned with well-organized fungal organisms (for example true decay/test fungi), they will rapidly colonize the substrate and develop a powerful mycelial network capable of decomposing most of the natural fiber based materials. Treated and non-treated wood specimens were buried in spawned, hydrogen peroxide protected wood pellet fuel-based substrate and placed in transparent, perforated plastic containers to determine the effect of this procedure on rapid colonization and accelerated wood degradation. Parallel test was set up with specimens inserted in 150mm diameter dishes containing fungal cultures developed on the beer based agar medium. The results after 45 and 90 days of exposure to the brown rot causing fungus Gloeophyllum trabeum indicate that the weight loss in control blocks in containers was higher than in the dishes for 7 and 11 percent respectively.
The production of extracellular hydrogen peroxide by some brown-rot fungi
1990 - IRG/WP 1446
The role of hydrogen peroxide (H2O2) has been discussed in the degradation of wood by wood-rotting fungi. The production of extracellular hydrogen peroxide was studied by detecting the oxidation of the chromogen 2,2'-azinobis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) by H2O2 and horse radish peroxidase (HPR). ABTS and HPR were added to a solid wood based culture media. In this study two brown-rotters, Poria placenta and Serpula lacrymans, produced detectable extracellular hydrogen peroxide.
A-C Ritschkoff, L Paajanen, L Viikari
Micromorphology of oak wood degraded by brown rot fungus Coniophora puteana
2000 - IRG/WP 00-10356
To characterize the degradation of hardwood by brown rot fungus, Coniophora puteana was incubated on the wood blocks of Quercus accutissima. For comparison, the same fungus was also incubated on the pine wood blocks of Pinus densiflora. In softwood, the shape of the wood cells and thickness of the cell appeared unchanged even the loss of birefringence in the affected areas. In contrast, oak wood decayed by brown rot fungus showed the erosion of wood cell walls from lumen towards the middle lamella. Complete degradation of cell walls including the lignin- rich middle lamella occurred in some areas of the oak wood cells. Degradation modes of brown rot fungus C. puteana in oak wood followed a typical white rot. The production of hydrogen peroxide was confirmed cytochemically by the deposition of cerium chloride in the degraded wood cell walls. However, Bavendamm test was negative to C. puteana. The present work suggested that hydrogen peroxide produced by brown rot fungus might be associated with not only the depolymerization of cellulose in softwood but also the degradation of lignin in hardwood.
Yoon Soo Kim, Seung-Gon Wi, Kwang-Ho Lee
Effects of bleaching process on the roughness values of wood surfaces of Lebanon cedar (Cedrus libani A. Rich.) and Black poplar (Populus nigra L.) using NaOH (sodium hydroxide), H2O2 (hydrogen peroxide) and Ca(OH)2 (calcium hydroxide)
2008 - IRG/WP 08-40403
Technical progress in the wood industry has been rapid in recent times. In this case, the quality assurance of the consumer products aligned with aesthetics value appears as one of the most important parameters. Because of the outer appearance of goods exert an effect on customers, interest in production of high quality surfaces of wooden commodities has increased essentially based on the surface smoothness (and/or the surface roughness of wood) aiming to reach the customer-oriented quality criteria. An aesthetics behaviour is being more influenced than the functional situation of the merchandise when the customers making the decision to buy wood products. It has been well estabilished that some of the properties of wood material (i.e. density, porosity, moisture content, fiber directions), and the wood machining process and its conditions (i.e. kinematics of the cutting process, wood sanding process) make the surface smoothness of wood problematic. There is a lack of information about the effects of bleaching process (i.e. one of the special technical ways to increase the aesthetics of wood products) on the smoothness of wood surfaces despite numerous reports published on the machining tools and the cellular structure of wood. In this study, therefore, effects of bleaching process on the surface roughness of wood was investigated for Lebanon cedar (Cedrus libani A. Rich.) and Black poplar (Populus nigra L.) using the bleaching chemicals NaOH (sodium hydroxide), H2O2 (hydrogen peroxide) and Ca(OH)2 (calcium hydroxide) by the two prescriptions with or without calcium hydroxide.
I Usta, E Aydinlar
Degradation of wood veneers by Fenton’s reagents: effects of low molecular weight phenolic compounds on hydrogen peroxide decay and tensile strength loss
2009 - IRG/WP 09-20400
Pine wood (Pinus sylvestris) veneers strips were incubated in acetate buffer containing hydrogen peroxide and iron to mimic mechanisms of brown rot decay and assess the degradation of cellulose through analysis of the tensile properties of the decayed wood. The tensile properties of thin wood strips treated with Fenton system reagents or precursors were determined and correlated to weight loss as reaction conditions were varied. The effect of 2,3‑DHBA concentration on the decay rate of H2O2 in the reaction mixture and the mechanisms of H2O2 decomposition will be discussed. The concentration and ratio of the low molecular weight, phenolic compound 2,3-dihydroxybenzoic acid (2,3‑DHBA) were also studied with regard to the generation of hydroxyl radicals from H2O2 and the resulting weight and strength loss. The pathway and rate of H2O2 decay depended on the pH and the medium (water, buffer) of the reaction mixture. As the concentrations of 2,3-DHBA above an optimal ration with iron, strength loss of the wood strips was reduced. This is likely to have occurred because concentrations of catechols that exceed a 1:1 ratio sequester iron via hexadentate and quadradentate ligand formation as has been previously reported Wood constituents were also shown to reduce Fe(III) to Fe(II) and to drive the Fenton reaction.
Yanjun Xie, R Well, Zefang Xiao, B Goodell, J Jellison, H Militz, C Mai
Chemical mediated depolymerization of cotton cellulose for the understanding of non-enzymatic fungal decay
2010 - IRG/WP 10-10731
Small, low molecular weight non-enzymatic compounds have been linked to the early stages of brown rot decay as the enzymes involved with holocellulose degradation are found to be too large to penetrate the S3 layer of intact wood cells. The most pronounced of these which were analyzed in this study are hydrogen peroxide, iron, and oxalic. The compounds related to the Fenton reaction: the combination of hydrogen peroxide and iron caused marked lowering of the degree of polymerization in the cotton cellulose after treatment. This was the case for both iron ions; Fe3+ and Fe2+. A 10mM solution of oxalic acid also showed significant depolymerizing effect on cotton cellulose, whereas diluting the oxalic acid with sodium oxalate to obtain a pH gradient, showed that this decreased the effect reducing of oxalic acid. In addition an organic iron chelator, EDTA, was tested but was found to inhibit depolymerization when in solution with chemicals related to Fenton chemistry. Manganese was tested to see if other metals than iron could generate an significant impact on the degree of polymerization of cotton cellulose and the metal showed good depolymerizing properties as a substitute for iron. We conclude that low molecular weight metabolites are capable of effectively depolymerizing cellulose during incipient decay by brown-rot fungi.
A C Steenkjær Hastrup, B Jensen, F Green III
Health aspects concerning the use of bifluorides in wood
1981 - IRG/WP 3173
An attempt has been made to find a correlation between the quantities F¯ and HF present among other ions in an 'Improsol' solution consisting of NH4F.HF and KF.HF, the quantity absorbed by the wood from this after immersion and the toxicological effects of this treated wood when it is used in rooms destined for the residence of people or animals or for the storage of foodstuffs.
H F M Nijman
Studies on the infestation behaviour of the powder-post beetle Lyctus brunneus (Steph.) and its physical control in the wood yards of the Caspian forests of Iran
1985 - IRG/WP 1271
Lyctus brunneus (Steph.) is a pest which has not been previously thoroughly studied in Iran. It severely attacks Iranian hardwoods, especially those used in wooden houses and that have not been treated. Research work was necessary to determine the natural resistance of the most important timber species in Iran against this insect.
The use of bifluorides-diffusion in remedial treatments
1983 - IRG/WP 3256
A description is given about the experience to date of a 14 years' development to bring bifluorides-diffusion from laboratory evaluation to commercial service application. The described treatment is an adjunct to good design and not a replacement for it. The process is also used with an injection method for the remedial treatment of premature decayed external joinery.
H F M Nijman
Diffusion of bifluorides and borates from preservative rods in laminated beams
1991 - IRG/WP 3644
Laminated beams from spruce, pine and larch were treated with two preservative rods, based on bifluorides and octaborate respectively. After 3 and 6 months placed in different climates (65% r.h., 100% r.h., water) the diffusion of the active ingredient was measured. After the 6 months period, the diffusion of the bifluorides was, in all species, ca. 5 times larger than the diffusion from octaborate rods. The bifluorides were not hindered by the glue-lines. The diffusion of the borate was poor, mainly perpendicular to the grain and was hindered by the glue-line.
Remedial treatment of joinery. Comparison of distribution of organic solvent preservative and ammonium bifluoride
1978 - IRG/WP 3115
It is some years since it was recognised in the United Kingdom that there was a problem of premature decay in untreated external joinery. Consequently, recommendations for preservative treatment were made by the Building Research Establishment, Princes Risborough Laboratory. In addition, the National House-Building Council has also included the preservative treatment of external joinery in its requirements. Thus, a large proportion of external joinery manufactured in the U.K. is now preservative treated, mainly in double vacuum impregnation plants. There still remain, however, relatively new buildings where preservative treatment has not been specified and where fungal decay is a problem. Decay occurs most frequently in the lower horizontal members such as cills, and the lower rails of opening lights. The damage is often most severe at the joints, where entry of water is facilitated by exposure of end grain. In some circumstances such as in Local Authority buildings where some of the more exposed joinery has already decayed, it has been considered economic to treat the remaining sound or near-sound joinery in situ in order to prevent future attack and to arrest any slight decay that may already be present. This treatment is normally applied by injecting organic solvent preservative under pressure in the region of the joints and where considered necessary along the lower horizontal member. As an alternative method of treatment, we have investigated the possible use of ammonium bifluoride, since diffusion of gaseous hydrogen fluoride may be expected to penetrate more extensively into wet timber than organic solvent preservative. The work described below was carried out in order to assess the comparative penetration of fluoride and organic solvent preservative in redwood (Pinus sylvestris).
J M Taylor