Specific degradation ofβ-aryl ether linkage in synthetic lignin (dehydrogenative polymerizate) by bacterial enzymes ofSphingomonas paucimobilis SYK-6 produced in recombinantEscherichia coli

Sphingomonas paucimobilis SYK-6 produces unique and specific enzymes, such as-etherases,O-demethylases, and ring fission dioxygenases, for lignin degradation. Cleavage of arylglycerol--aryl ether linkage is the most important process in the lignin metabolic pathway ofS. paucimobilis SYK-6. We reported the genes (ligD, ligE, ligF) for enzymes that cleaved-aryl ether linkage of dimeric compounds in previous studies. In this study we synthesized the fluorescent high-molecular-weight lignin (UBE-DHP) by dehydrogenative polymerization. We investigated the-aryl ether cleavage ability of these enzymes produced in recombinantEscherichia coli. When UBE-DHP was incubated with LigF, 4-methylumbeliferone was released as a result of-aryl ether cleavage of O-methylumbelliferyl--hydroxypropiovanillone (compound III) incorporated in UBE-DHP. Here, we report that-etherase ofS. paucimobilis SYK-6 can be expressed inE. coli and is able to cleave the-aryl ether linkage in synthetic high-molecular-weight lignin.     

Effects of stilbenes from bark of <Emphasis Type="Italic">Picea glehnii</Emphasis> (Sieb. et Zucc) and their related compounds against feeding behaviour of <Emphasis Type="Italic">Reticulitermes speratus</Emphasis> (Kolbe)

The effects of stilbene glucosides and related compounds on termite feeding behavior were investigated using paper disc methods against the subterranean termite Reticulitermes speratus. The stilbene-rich fraction and isorhapontin (3-methoxy-3,4, 5-trihydroxystilbene-3--d-glucoside) from bark extracts of Picea glehnii showed avoidance by termites in choice tests. In the no-choice tests using compounds purified from the stilbene-rich fraction, the largest feeding deterrent effect was observed for piceid (3,4,5-trihydroxystilbene-3--d-glucoside), followed by isorhapontin, and astringin (3,3,4,5-tetrahydroxystilbene-3--d-glucoside), at the concentrations from 0.63 to 2.5µmol/disc. No change in activity was observed at retentions of more than 5.0µmol/disc. When the activities of isorhapontin and its aglycone derivative (isorhapontigenin: 3-methoxy-3,4,5-trihydroxystilbene) were compared with that of taxifolin (3,3,4,5,7-pentahydroxyflavanone) in the no-choice test, the stilbenes exhibited a larger antifeedant potential. Methylation of isorhapontigenin increased its termiticidal activity.Part of this study was presented at the 32nd Annual Meeting of the International Research Group on Wood Preservation, 2001     

Effect of rhizobial inoculation methods and host plant provenances on nodulation and growth of <Emphasis Type="Italic">Acacia senegal</Emphasis> and <Emphasis Type="Italic">Acacia nilotica</Emphasis>

The purpose of this work was to determine the most efficient methods of inoculation to significantly improve nodulation and growth of Acacia senegal and Acacia nilotica, grown under greenhouse conditions. Our results showed that inoculation using dissolved alginate beads containing rhizobia significantly improved the growth of both acacia species better than the growth of plants in other treatments. The experiment with A. nilotica was conducted in two unsterilised soils from different areas. Plants grown in soil from Bel Air were well-nodulated and showed better growth than plants grown in soil from Sangalkam. However, no difference between these soils was shown between the several methods of inoculation and their effect on the nodulation and growth of plants. An interaction between A. senegal and A. nilotica provenances and the effect of inoculation with rhizobia was also demonstrated. Bel Air provenance of A. senegal, Dahra and RIM provenances of A. nilotica grew best of several provenances tested. These results suggest that (1) it may be possible to improve growth and yield of A. senegal and A. nilotica by careful selection of each symbiotic partner; and (2) nursery-grown seedlings of A. senegal and A. nilotica should be inoculated, just after sowing, with dissolved alginate beads containing a mixture of selected rhizobia. Résumé. Lobject de notre travail était de déterminer la méthodologie dinoculation la plus efficace pour améliorer significativement la nodulation et la croissance dAcacia senegal et dAcacia nilotica poussant dans des conditions de serre. Nos résultats ont montré que linoculation utilisant des billes dalginate dissoutes améliorait significativement la croissance des deux espèces dAcacia par rapport à celle des plantes des autres traitements. Lexpérience avec A. nilotica a été faite dans deux types de sol non stériles. Nous avons observé que les plants poussant dans le sol de Bel Air étaient plus nodulés et mieux développés que ceux poussant dans le sol de Sangalkam. Cependant, aucune différence significative entre les deux sols na été mise en évidence en termes deffet des différentes méthodologies dinoculation sur la nodulation et la croissance des plantes. Il a aussi été démontré lexistence dune interaction entre les provenances dA. senegal et dA. nilotica et leffet de linoculation avec rhizobium. Cest ainsi que les provenances Bel Air dA. senegal, Dahra et RIM dA. nilotica ont le mieux poussé parmi les différentes provenances testées. Lensemble de ces résultats suggèrent: ⁽¹⁾ il devrait être possible daméliorer la croissance et la production dA. senegal et dA. nilotica par une sélection des deux partenaires symbiotiques; ⁽²⁾ Les jeunes plantules dA. senegal et dA. nilotica produites en pépinière devraient être inoculées juste après le semis avec des billes dalginate dissoutes contenant un cocktail de souches sélectionnées.     

5α-Reductase inhibitory component from leaves ofArtocarpus altilis

The inhibitory effects of 17 samples prepared from Thailand plants on 5-reductase activity were examined. The acetone extract of leaves ofArtocarpus altilis showed potent 5-reductase inhibitory activity. Fractionation guided by 5-reductase inhibition led to the isolation of 2-geranyl-2,3,4,4-tetrahydroxydihydrochalcone from the acetone extract of leaves ofA. altilis. This compound showed more potent inhibitory effect (IC₅₀=38M) than-linolenic acid known as a naturally occurring potent inhibitor.     

Fogging can control<Emphasis Type="Italic"> Tetranychus urticae</Emphasis> on greenhouse cucumbers

The effect of a fogging system on interactions between Tetranychus urticae and Phytoseiulus persimilis was studied in two cucumber greenhouses in northern Italy. The greenhouses were divided into two compartments, one (referred to as fog area) was humidified using a fogging system and the other was not humidified (no fog area). Four cultivars were investigated: Akito and Frontera during 2000, Dinero and Jazzer during 2001. Tetranychus urticae was released on a number of plants in both compartments. The release of P. persimilis was planned in both compartments at definite T. urticae thresholds. Phytoseiid releases failed, and the colonization by other predators was scarce.Thus, the effect of the fogging system was evaluated only on T. urticae populations. During the first experimental year, spider mite populations reached low densities in the fog area of both cultivars. Higher densities were found in the no fog area, especially in late June and early July. One year later, spider mites were more abundant in the no fog area of the Dinero cultivar. On the Jazzer cultivar, where most plants were infected by powdery mildew, spider mites were less abundant and differences between the two areas were less marked. Since relative humidity values exceeding 90% (considered as effective on T. urticae) were recorded during the night when the fogging system was not active, we suggest that contact with misty water was the main factor responsible for the reduction of T. urticae populations in the fog area.     

Compounds inhibitory to rat liver 5α-reductase from tropical commercial wood species: resveratrol trimers from melapi (Shorea sp.) heartwood

5-Reductase inhibitory activity of methanol extracts of the heartwood of 13 tropical wood species were examined. Strong 5-reductase inhibitory activity was observed withShorea species. From melapi (Shorea sp.), two known resveratrol trimers, vaticanol A and ampelopsin C, and two novel trimers were isolated as active compounds. The structures of the two novel resveratrol trimers were elucidated by one- and two-dimensional nuclear magnetic resonance spectroscopic analyses, including¹H-¹HCOSY, HMQC, and HMBC. The compounds were named melapinol A and melapinol B. There were no significant differences among the 5-reductase inhibitory activities of the four resveratrol trimers, which were significantly stronger than those of-linolenic acid and epigallocatechin gallate, known 5-reductase inhibitors.     

Role of caffeic glucoside esters in defense-repair processing of trees II: Synthesis of 2-(3,4-dihydroxyphenyl)-ethyl 3-α-L-rhamnopyranosylβ-D-glucopyranoside

The reference compound (9), with a partial structure of acteoside, was synthesized to elucidate the relation between structural features and the precipitation or solubility of the oxidation products of acteoside: 2-(3,4-dihydroxyphenyl)-ethyl 3-O-(-L-rhamnopyranosyl)--D-glucopyranoside (9). The glycosyl acceptor 2-O-benzoyl-3-O-(2,3,4-tri-O-acetyl--L-rhamnopyranosyl)-4,6-O-benzylidene--D-glucopyranosyl trichloroacetimidate (7) was prepared from allyl 2-O-benzoyl-3-O-(2,3,4-tri-O-acetyl--L-rhamnopyranosyl)-4,6-O-benzylidene--D-glucopyranoside (4) via isomerization of the allyl group with an iridium complex to the 1-propenyl group and its hydrolysis with HgCl₂lHgO, followed by treatment with CCl₃CN and DBU in a 65.5% overall yield. The glycosyl acceptor 3,4-diacetoxyphenethyl alcohol (16) was prepared from homoveratric acid via demethylation with 57% HI and its acetylation with Ac₂O and 85% H₃PO₄, followed by selective reduction of the carboxyl group to the alcohol with a borane-tetrahydrofurane complex in a 61% overall yield. The glycosylation of7 with16 in dichloromethane promoted by BF₃-Et₂O gave a 74.3% yield of8. Hydrolysis of8 with 90% CF₃COOH gave the debenzylidenated product, which was treated with NaOMe to afford a 32% overall yield of the desired compound9. This compound9 was identical with the natural specimen.This study was presented in part at the 46th annual meeting of the Japan Wood Research Society, Kumamoto, April 3–5, 1996     

Absolute configuration of arylglycerol-β-aryl ethers obtained by asymmetric reduction of the correspondingα-ketonic compound with intactFusarium solani cells

When (±)--oxo-guaiacylglycerol--(vanillic acid) ether (1) is degraded byFusarium solani M-13-1, the-ketone is initially reduced to giveerythro andthreo guaiacylglycerol--(vanillic acid) ethers (2), arylglycerol--aryl ethers, both of which are enantiomerically pure. The absolute configuration in each2 was determined by Mosher's method; the products were converted to,-di-(R)--methoxy--trifluoromethylphenylacetates (MTPA esters) (3) oferythro (-)- andthreo (+)-veratrylglycerol--(methyl vanillate) ethers (3), whose¹H nuclear magnetic resonance (NMR) spectra were examined and compared with those of four di-(R)-MTPA ester (3) diastereomers from chemically synthesizederythro (±)-3 andthreo (±)-3. To assign the- and-MTPA-OCH₃ peaks, the¹H NMR scans of several compounds that have substructures of 3 and their 3,4,5-trimethoxyphenyl analogues were examined. When a racemic alcohol reacts with (R)-MTPA to give a pair of (R)-MTPA ester diastereomers, the value was defined as the absolute value of the difference in the¹H chemical shifts of the peak between the diastereomers. It was found that the values of-MTPA-OCH₃ were larger than those of-MTPA-OCH₃ owing to a shielding effect of the veratryl ring located on the-MTPA-OCH₃, and that the-MTPA-OCH₃ peaks in the 3,4,5-trimethoxyphenyl compounds shifted downfield relative to those in the veratryl compounds. On the basis of the¹h NMR data of (R)-MTPA esters, the absolute configuration of the four chemically prepared diastereomers (3) were determined. The catabolicerythro 3 [fromerythro (-)-3] andthreo 3 [fromthreo (+)-3] were identical to (R, S, R)-erythro 3 and (R, S, S)- threo 3, respectively. An hydrogen species in the fungal reduction would attack the-ketone fromre-face of both (R)-1 and (S)-1, givingerythro (S, R)-2 andthreo (S, S)-2, respectively.Part of this paper was presented at the 33rd Lignin Symposium, Tsukuba, November 1988     

Studies on the synthesis and properties of terpene-phenol-aldehyde resin with a high softening point

Synthesis conditions of terpene-phenol-aldehyde resin with a high softening point were studied based on the reactions among-pinene, phenol, and formaldehyde. A suitable catalyst system and the best processing conditions were obtained by laboratory experiments. The influence of the raw materials ratios, reaction temperature, reaction time, and catalyst dose on the softening point, yield, and color of the synthesized resin was examined. The physical and chemical characteristics of the resin were determined: softening point 140°C (ball and ring method); color value 7 (Gardner); acid value (KOH mg/g) <1; bromine value (Br₂ mg/100g) <64; saponification value (KOH mg/g) <1; average molecular weight 830. The yield of resin was more than 85% (based on the total raw material).     

Effect of cutting position on rooting and shoot growth of two poplar clones

One-year-old shoots from stooling beds of Populus × deltoides Walker and Populus jackii Northwest were collected in late fall to determine the effect of cutting position on rooting and growth of hardwood poplar cuttings. Cuttings with a minimum diameter of 9.0 mm were obtained from basal, middle and distal locations on the poplar shoot. Rooting and shoot growth were assessed in the greenhouse and under field conditions. Walker poplar cuttings collected from basal portions of the shoot and planted in the field had 87.6% rooting and 103.8 cm growth compared to 78.8% and 103.4 cm and 67.6% and 88.8 cm for middle and distal locations respectively. For Northwest rooting and growth under field conditions was 84.4% and 94.7 cm for basal, 78.4% and 90.5 cm for mid and 64.4% and 84.3 cm for distal locations. Rooting in the greenhouse was superior to the field. Walker cuttings had fewer roots and buds per cutting than Northwest, however, growth of Walker in the field was superior.     

Interactions between<Emphasis Type="Italic"> Orthotomicus erosus</Emphasis> (Woll.) (Col., Scolytidae) and the Argentine ant<Emphasis Type="Italic"> Linepithema humile</Emphasis> (Mayr) (Hym., Formicidae)

Interactions occurring between Orthotomicus erosus, an economically important pine bark beetle in Portugal, and the aggressive Argentine ant Linepithema humile were investigated. Tri-trophic experimental enclosures consisting of Pinus pinaster logs, O. erosus adults and L. humile workers were set up in the field and in the laboratory. Interactions between ants and scolytids were observed and quantified in terms of the bark beetles success in colonizing the logs. The presence of L. humile did not influence the ability of O. erosus to colonize the logs, apparently due to the difficulties met by the ants in handling the bark beetles and to their inability to injure them. Thus, although abundant in many pine stands in Portugal and in spite of its aggressiveness, L. humile does not seem to affect populations of O. erosus.     

In vitro propagation of <Emphasis Type="Italic">Oroxylum indicum</Emphasis> Vent. a medicinally important forest tree

In vitro propagation of Oroxylum indicum Vent. was carried out using cotyledonary node explants. Among the different types of cytokinins used for culture establishment, 6-benzyladenine exhibited the best response with higher concentrations (8.87µM or above) for inducing multiple shoots. Inclusion of indole-3-acetic acid (2.85µM) into 6-benzyladenine-supplemented medium triggered a high frequency of response as well as a proliferation of shoots. The best medium for proliferation was Murashige-Skoog (MS) medium with 6-benzyladenine (8.87µM) and indole-3-acetic acid (2.85µM). However, incorporation of gibberellic acid (1.44µM) was mandatory to enhance shoot elongation. Repeated subculturing of cotyledonary node and in vitro developed nodal segments in MS medium with 6-benzyladenine (4.44µM) at 4-week intervals resulted in continuous mass multiplication of shoots without any evidence of decline. Root induction was best (91.6%) when MS strength was reduced to one-quarter and combined with -naphthalene acetic acid (2.69µM) and indole-3-acetic acid (5.71µM), with a high survival rate (70–72%) of plantlets hardened in either soil rite or soil:sand:soil rite (1:1:2).     

Five years of Paulownia field trials in North Carolina

Performance of Paulownia elongata, Paulownia fortunei, and Paulownia×Henan 1 was monitored at three sites in North Carolina between 1996 and 2000, and P. elongata clones were tested at an additional two sites. The primary objective was to define the potential for growing Paulownia in the state. Species and clones within species differed in survival and height and diameter growth whenever genotype was included as a treatment. P. elongata was superior at two and P. fortunei was superior at one of the three sites that included P. elongata, P. fortunei, and P.×Henan 1. Trees produced through vegetative means were more likely to survive and were taller and of greater DBH than seed produced trees. Mortality in trees produced from seed was greater for P. fortunei and P. ×Henan 1 than for P. elongata. Application of animal waste or different rates of nitrogen as inorganic fertilizer did not have a pronounced effect on tree survival and growth.     

Paraiso (Melia azedarach var. “Gigante”) woodlots: an agroforestry alternative for the small farmer in Paraguay

This paper describes the salient aspects and analyses the potential of the Paraiso Woodlot System, which is becoming popular as an agroforestry alternative to land use in the degraded acid sandy soils of the humid sub-tropical Guayaybi area of Paraguay. The system consists of a combination of paraiso (Melia azedarach var gigante) with other trees, especially Leucaena leucocephala, and annual crops.The fast growth habit, deep root system, addition of large quantities of organic matter through leaf and litter fall, compatibility with agricultural crops, high value of the sawlogs and production of substantial quantities of poles and firewood make paraiso an excellent species for agroforestry combination. Preliminary results of the trials indicate that the woodlots are successful and with increasing efforts of the extension agencies, they are being accepted as a viable alternative to traditional agricultural systems.Besides discussing the potential of these woodlots based on initial results, the paper identifies the constraints of the system and highlights the priority research areas.     

Antioxidative activity of tree phenolic constituents 1: Radical-capturing reaction of flavon-3-ols with radical initiator

The present work was undertaken from the standpoint of radical-capturing ability with regard to the antioxidative ability of flavonoids, especially flavonols distributed widely in woody plants. In regard to the flavonols, six methyl derivatives were initially prepared from quercetin and its litinoside. Their radical-capturing constants were determined strictly by the stopped-flow spectroscopic method. It was proved that the radical-capturing ability of quercetin mainly involves the vicinal C₃. and C₄, hydroxyl groups and the C₃ hydroxyl group. To clarify the reaction mechanism begun at the C₃ hydroxyl group of quercetin, 5,7,3,4-tetramethylquercetin (TMQ), flavon-3-ol (F30) and so on were treated with 2,2-azo-bis-(2,4-dimethylvaleronitrile) (AMVN). Six products (1–6) containing one depside and its two hydrolytic products, two valeronitrile adducts, and others were isolated from the reaction mixture of TMQ and their structures determined by instrumental analyses. Similarly, F30 gave four products, 7–10, which corresponded to the above products 1–3 and 5 (one depside, its two hydrolytic products, and one adduct), respectively. 3,5,7,3,4-Pentamethylquercetin (PMQ) and flavon-3-O-methylate (F3M) gave no products. The quantitative change of the products with reaction time was determined spectroscopically. An initial reaction pathway for the radical-capturing reaction of flavon-3-ols with AMVN was proposed based on the products and their amounts. The main route — formation of depside and its hydrolytic products via ketohydroperoxide (3) or ketohydroperoxy radical (4) - was similar to that of the oxidation reaction of quercetin with quercetinase and light.Part of this paper was presented at the 46th and 47th annual meetings of the Japan Wood Research Society, Kumamoto and Kouchi, April 1996 and 1997     

Effect of collar insertion on soil respiration in a larch forest measured with a LI-6400 soil CO2 flux system

Little information is available on the effect of root cutting by the collar pre-insertion technique on soil respiration. In this study, we found that soil respiration rates decreased with increasing depth of collar insertion in both the with live roots intact and with live roots severed treatments, but the rate of decrease was substantially higher in the former. The cutting of roots, especially fine roots, may be responsible for this result.     

Vertikalbewegungen Rickettsiose-kranker Larven vonAmphimallon solstitiale (LINNAEUS),Anomala dubia aenea (DE GEER) undMaladera brunnea (LINNAEUS)(Col., Lamellicornia)

Zusammenfassung DurchRickettsiella melolonthae (Krieg) PHILIP infizierte Maikäfer-Engerlinge wandern während des Spätherbstes und Winters im Freiland absterbend auf die Erdoberfläche. Als Begleitarten inMelolontha Larvenpopulationen kamen im Gebiet vor:Amphimallon solstitiale (L.),Anomala dubia aenea (DeG.) undMaladera brunnea (L.). MitR. melolonthae künstlich infiziert, zeigten sie im Orientierungsversuch die gleiche Aufwanderungstendenz wie kranke Maikäferengerlinge, am deutlichsten die vonM. brunnea. Auf dem Waldboden fanden sich kranke Larven dieser Art in einigen Fällen.

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Summary Larvae ofMelolontha spp. infested withRickettsiella melolonthae (Krieg) PHILIP move during October– January to the top of the soil where they eventually die. Larvae ofAmphimallon solstitiale (L.),Anomala dubia aenea (DeG.) andMaladera brunnea (L.) are susceptible forR. melolonthae, too. In experiments, artificially infested individuals showed a similar tendency to move upward. This was most pronounced inM. brunnea, of which some infested larvae were found on the forest floor.

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Résumé Larves du hanneton, infectées parRickettsiella melolonthae (Krieg) PHILIP, se rendent à la surface de la terre au fin de l'automme et dans l'hivers. Comme espèces accompagnant les populations larvaires deMelolontha nous avons trouvés:Amphimallon solstitiale (L.),Anomala dubia aenea (DeG.) etMaladera brunnea (L.). Ils out démonstré, après une infection artificielle avecR. melolonthae, la mème tendence dans les essais d'orientation comme les vers blancs malades deMelolontha. Le phé nomène etait le plus prononcé parmi les larves deMaladera. De cette espèce nous avons trouvés plusieurs larves malades à la surface de la terre en plein champs.

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Rickettsiella melolonthae (Krieg) Philip Melolontha- . . Melolontha- :Amphimallon solstitiale (L.),Anomala dubia aenea (De G.) Maladera brunnea (L.). R. melolonthae , Melolontha-, M. brunnea. .

     

5<Emphasis Type="Italic">Α</Emphasis>-Reductase inhibitory tannin-related compounds isolated from <Emphasis Type="Italic">Shorea laeviforia</Emphasis>

Five tannin-related compounds – gallic acid, flavogallonic acid dilactone, valoneic acid dilactone, gallagyldilactone, ellagic acid – were isolated from the heartwood of Shorea laeviforia, and the inhibitory activity of each against rat liver 5-reductase was evaluated. Valoneic acid dilactone and gallagyldilactone exhibited positive inhibitory activity, but gallic acid and ellagic acid did not. Flavogallonic acid dilactone stimulated 5-reductase activity, even though this compound is structurally similar to valoneic acid dilactone. The kinetic study of valoneic acid dilactone and gallagyldilactone indicated that the inhibitory behavior of 5-reductase were not competitive against the substrate (testosterone) and were partially competitive against the cofactor (NADPH). Additionally, double inhibition analysis of valoneic acid dilactone and NADP⁺ showed synergetic inhibition. These results suggested that neither valoneic acid dilactone nor gallagyldilactone can affect the binding of testosterone but that either compound could interact with an enzyme–NADP⁺ complex to inhibit 5-reductase.     

Lignans ofLinum flavum var.compactum

A new dibenzylbutyrolactone lignan 7.6-dihydroxybursehernin, together with six known lignans (pinoresinol, lariciresinol, secoisolariciresinol,-peltatin,-peltatin, 5-methoxypodophyllotoxin) were isolated from the methanol extracts ofLinum flavum var.compactum. The enantiomeric analysis of pinoresinol and lariciresinol isolated from the species, which are upstream lignans in the lignan biosynthetic pathway, indicated that they are not optically pure, which is in accordance with our recent findings on lignans occurring in other plant species.Parts of this report were presented at the 42nd Lignin Symposium, Sapporo, October 1997; and the 43rd Lignin Symposium, Fuchu, October 1998     

A new diterpene dimer from the bark ofChamaecyparis obtusa

This paper describes characterization of diterpenes of the bark of Japanese cypress,Chamaecyparis obtusa (S. and Z.) Endl, without the resinous stem canker to learn the difference between the cypress bark affected with the canker. A diterpene dimer and two diterpenes, 6,12-dihydroxyabieta-8,11,13-trien-7-one and 6,12-dihydroxyabieta-5,8,11, 13-tetraen-7-one, were firstly isolated from Japanese cypress. The dimer, 6-(abieta-6,8,11, 13-tetraenyl-12-oxy)-7-methoxyabieta-8,11,13-trien-12-ol, was a new compound. It is a terpene indicative ofC. obtusa not infected with the resinous stem canker. Five known diterpenes were also isolated.