Tatsuya Ashitani

Antitermitic Activities of Abietane-type Diterpenes from Taxodium distichum Cones

Eight known abietane-type diterpenes were isolated from the weak acidic fraction of the n-hexane extract from cones of Taxodium distichum, one of the extant, living fossil conifers. They were identified as 6,7-dehydroroyleanone (1), taxodal (2), taxodione (3), salvinolone (4), 14-deoxycoleon U (5), 5,6-dehydrosugiol (6), sandaracopimaric acid (7), and xanthoperol (8). The structures of these compounds were determined by comparison of NMR spectral data with published data. The antitermitic (termicidal and antifeedant) activities of the compounds 1–8 against the subterranean termite, Reticulitermes speratus Kolbe, were evaluated. Compounds 1 and 3 showed potent termicidal activity, and 5 and 8 showed potent antifeedant activity. Compound 1 was found to be one of the representative bioactive compounds in the n-hexane extract of T. distichum cones. Compounds 1–8, with the exception of 7, were oxides of ferruginol (9). Therefore, the presence of various oxidation forms of the abietane-type structure reflects their various bioactivities.

Antifungal Abietane-Type Diterpenes from the Cones of Taxodium distichum Rich

The chemical composition of Taxodium distichum cones and the antifungal activities of twelve diterpenoids against two wood decay fungi, Trametes versicolor (white-rot) and Fomitopsis palustris (brown-rot) were examined. The chemical composition of the major extractive fraction, the n-C6H14 extract, was evaluated and its antifungal properties were identified. Twelve diterpenoids including ten abietane-type components were isolated from the n-C6H14 extract: 6,7-dehydroferruginol (1), ferruginol (2), 6,7-dehydroroyleanone (3), sandaracopimaric acid (4), taxodione (5), taxodal (6), taxodone (7), sugiol (8), xanthoperol (9), salvinolone (10), 5,6-dehydrosugiol (11), and 14-deoxycoleon U (12). Compounds 5 and 12 were highly active against both wood-decay fungi. In particular, the activities of these compounds against F. palustris were potent. The results suggest that the position and the number of hydroxyl groups on abietane-type structures may be related to antifungal activities against T. versicolor and F. palustris.

Volatile and non-volatile monoterpenes produced by elicitor-stimulated Cupressus lusitanica cultured cells

Elicitor treatment initiates defense responses in cultured Cupressus lusitanica cells. In order to investigate the defense mechanism with a yeast extract elicitor, we carried out solid-phase microextraction coupled with gas chromatography for monoterpene analysis. Ten hydrocarbon monoterpenes, including high amounts of sabinene and limonene, were detected in the gas phase of the elicitor-treated cell cultures. Six oxidized monoterpenes including beta-thujaplicin were also detected in the ether extract of the cells and the medium. Time-course profiles of volatile monoterpenes showed that one group of hydrocarbon monoterpenes was maximized on the second day after elicitation, while the other group was maximized on the third day. There were no oxidized monoterpenes that are structurally related to sabinene and limonene in the gas phase or cell extracts, suggesting that these compounds are produced exclusively for emission. Other monoterpenes, which are produced during later stages of elicitation, are metabolized into more complex compounds such as oxidized monoterpenes, including beta-thujaplicin. Although terpinolene synthase was the principal monoterpene synthase in these cell cultures, terpinolene was detected only as a minor compound in the gas phase. The time course for terpinolene synthase activity coincided with beta-thujaplicin biosynthesis. Thus, most of the terpinolene is metabolized rapidly to oxidized terpenes such as beta-thujaplicin rather than emitted.

Antioxidant activity and mechanism of the abietane-type diterpene ferruginol

The antioxidant activity of the abietane-type diterpene ferruginol was evaluated by comparison with that of carnosic acid, ( ± )-α-tocopherol and dibutylhydroxytoluene using 2,2-diphenyl-1-picrylhydrazyl, β-carotene bleaching and linoleic acid assays. Ferruginol had the lowest antioxidant activity of this group using the 2,2-diphenyl-1-picrylhydrazyl and β-carotene methods in polar solvent buffer. However, ferruginol exhibited stronger activity than carnosic acid and α-tocopherol for linoleic acid oxidation under non-solvent conditions. Five peaks corresponding to ferruginol derivatives were detected through GC-MS analysis of the reaction between ferruginol and methyl linoleate. The three reaction products were identified as dehydroferruginol, 7β-hydroxyferruginol and sugiol, and the other two peaks were assumed to be 7α-hydroxyferruginol and the quinone methide derivative of ferruginol. The time course of the reaction suggests that the quinone methide was produced early in the reaction and reacted further to produce dehydroferruginol, 7-hydroxyferruginol and sugiol. Thus, we inferred that quinone methide formation was a key step in the antioxidant reaction of ferruginol.

Color and chemical characterization of partially black-streaked heart-wood in teak (Tectona grandis)

The heartwood of teak in certain areas of Java Island displays irregular black streaks along the annual rings. We investigated the color and chemical characteristics in a radial direction of partially black-streaked heartwood samples. Color properties (pH value, inorganic element, extractive content and extractive characterization) were measured in the color co-ordinates CIELAB system. The results show that the black streak part was 12–15 brightness (L*) value units less than the normal heartwood. Furthermore, the black streak part had more red (a*) but less yellow (b*), hue (h) and chroma (C*) than the normal wood. The pH value, ash content and calcium contents of the black streak part were slightly higher than those obtained for the normal wood. The content of the low-polar extractive (n-hexane and ethyl acetate) of the black streak part was considerably higher than that for the normal wood. The blackening process was speculated to be a kind of defense mechanism indicated by remarkable amount of bioactive compound called tectoquinone.

Inhibition activity of essential oils obtained from Japanese trees against Skeletonema costatum

The growth inhibition activities of essential oils obtained from Cryptomeria japonica, Chamaecyparis obtusa, and Pinus thunbergii were examined against the bacillariophyceae Skeletonema costatum, also known as red tide plankton. The essential oils were extracted from the heartwood, leaves, and bark of these typical indigenous Japanese conifers. The essential oils from C. japonica bark and P. thunbergii heartwood possessed strong growth inhibition activity. The chemical compositions of these essential oils were analyzed by gas chromatography/flame ionization detection (GC-FID) and gas chromatography/mass spectrometry (GC-MS). α-Terpineol and longifolene were the main components of the essential oil from P. thunbergii heartwood. The C. japonica bark essential oil was mainly composed of α-terpineol, δ-cadinene, isophyllocladene, and ferruginol. Ferruginol and longifolene showed more potent growth inhibition against S. costatum than hinokitiol (β-thujaplicine), which is known to be a strong antifungal compound among wood components. Ferruginol and longifolene were important factors for the growth inhibition activity of the essential oils from C. japonica bark and P. thunbergii heartwood, respectively. These results suggest the possibility of using C. japonica bark and P. thunbergii heartwood for the control of red tide plankton.

Monoterpenes produced by Cupressus lusitanica cultured cells including a novel monoterpene (1S, 2S, 6S)-(+)-1,6-epoxy-4(8)-p-menthen-2-ol

A novel monoterpene, (1S, 2S, 6S)-(+)-1,6-epoxy-4(8)-p-menthen-2-ol, was isolated from an elicitor-treated cell culture of Cupressus lusitanica (Mexican cypress). Ten known monoterpenes - limonene, myrcene, β-ocimene, sabinene, terpinolene, 4-terpineol, α-terpineol, 4(8)-p-menthen-1,2-diol, 4-hydroxyphellandric acid methylester and β-thujaplicin methylether - were also identified. Regioselective metabolisms of monoterpenes were observed in this culture.

Direct Episulfidation of Caryophyllene and Humulene

Abstract Caryophyllene-6,7-episulfide (3), humulene-9,10- (4) and -6, 7-episulfide (5) found in hop oil, were prepared by heating caryophyllene or humulene with sulfur at 120°C. Examination of various types of olefins showed that this peculiar reaction occurred only to the medium ring olefins.

New method to determine the hydroxyl value in liquefied bark as polyurethane material

ConclusionsWhen the hydroxyl values of materials containing phenolic 3 or sterically hindered hydroxyl groups were determined by the esterification method, the values were considerably smaller than the theoretical values. In contrast, the NCO method provided hydroxyl values much closer to the theoretical values for such materials, though the values varied depending on the kinds and amounts of catalysts.The quantity of hydroxyl groups reacting with the NCO group could be directly determined by the NCO method using a catalyst mixture consisting of DABCO and DBTDL in a molar ratio of 2∶1. Therefore, this method can be considered useful for determining the hydroxyl value before preparing polyurethane from liquefied wood or bark.

Induced monoterpene and lignin production in mechanically stressed and fungal elicited cultured Cupressus lusitanica cells

Cultured Cupressus lusitanica cells induced by various stresses are thought to produce different complexes of defense chemicals to optimize defense. To compare the induced products of two stimulations, we investigated the emission of monoterpenes, biosynthesis of β-thujaplicin, and accumulation of lignin in mechanically stressed and fungal elicited cultured C. lusitanica cells. Both mechanical stress and fungal elicitor caused emission of qualitatively similar monoterpene blends indicating de novo biosynthesis of these compounds after stimulation, while mechanical stress alone is sufficient to induce fungal elicitor-related monoterpene emission. Sabinene and limonene were the dominant compounds over the time course in both volatile blends. Although the emitted volatile blends were qualitatively similar, the time course and the relative ratios of the constituents of the volatile blends differed with the type of stimulation. While fungal elicited cells produced significant amounts of β-thujaplicin over the 5-day time course, no β-thujaplicin was observed in the mechanically stressed cells. The production of β-thujaplicin was the main dissimilarity of the induced products of these two treatments, suggesting that synthesis of β-thujaplicin is not a general response to all types of stresses, but is a specific response and serves as a strong toxic compound against already invaded fungus. Significantly higher amounts of lignin accumulations were observed in the fungal elicited and mechanically stressed cells on the 5th day after induction. Based on these results, we suggest the composition of induced products was dependent on the method of stimulation.