Antifungal activity of Liriodenine from Michelia formosana heartwood against wood-rotting fungi

Wood-rotting fungi lead to great economic losses of lignocellulosic materials. The influence of extractives on the decay resistance of Michelia formosana wood and the antifungal activities of heartwood extract and its constituent against wood-rotting fungi were evaluated. Results revealed that extractives had a strong influence on the decay resistance of M. formosana heartwood. The unextracted heartwood block was resistant to fungi, while the extracted heartwood block was not. Antifungal activities of ethanolic extract from heartwood were assessed; results revealed that the ethanolic extract possessed good antifungal activity against wood-rotting fungi. The ethanolic extract was further separated into four fractions including n-hexane-soluble fraction, ethyl acetate-soluble fraction, n-butanol-soluble fraction, and water-soluble fraction. Among these fractions, the n-hexane-soluble fraction exhibited the best antifungal ability. An alkaloid, liriodenine, was isolated from the active n-hexane-soluble fraction. IC₅₀ values of liriodenine against the white-rot fungi Lenzites betulina and Trametes versicolor were 0.76 and 3.50?μg/mL, respectively. IC₅₀ values of liriodenine against the brown-rot fungi Laetiporus sulphureus, Gloeophyllum trabeum, and Fomitopsis pinicola were all lower than 2.0?μg/mL. Results indicated that liriodenine exhibited a broad spectrum of antifungal activity and great potential to develop as an environmentally benign fungicide for the preservation of lignocellulosic materials.     

Quantitative comparisons of antitermite extractives in heartwood from the same clones of <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> planted at two different sites

The quantities of some extractives in the heartwood of 25 plus tree clones of Japanese cedar (Cryptomeria japonica) from two different sites were investigated to clarify the differences in antitermite properties in relation to clones and environmental factors. The measured compounds were cubebol, epicubebol, sandaracopimarinol, and ferruginol. The total amounts of extractives were calculated from gas chromatogram peak areas. The heartwoods from Tano (Tano Forest Research Station, Miyazaki University; 31° N, 131° E, 130 m asl) tended to contain more of the individual extractives and total extractives than those from Komenono (Komenono Forest Research Station, Ehime University; 33° N, 132° E, 700 m asl). There was a significant difference (calculated by analysis of variance) at the 1% level among clones at both sites in quantities of total and individual extractives. This result suggests that the qualities and quantities of heartwood extractives are largely affected by genetic factors. Spearman’s rank correlation with the average of the extractive quantities at the sites was investigated. The correlation coefficients of total extractives, cubebol, epicubebol, sandaracopimarinol, and ferruginol were 0.61, 0.85, 0.76, 0.67, and 0.74, respectively. This result means that the order of the amounts of these compounds among the cultivars could be maintained at different sites. It is apparent that both environmental and genetic factors affected the quantities of these extractives in the heartwoods of Japanese cedar.     

Extractives relating to heartwood color changes in sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) by a combination of smoke-heating and UV radiation exposure

 Sugi green logs with red or black heartwood were smoke-heated, and the changes in the color of the heartwood after ultraviolet (UV) (λ = 365 nm) radiation exposure were then observed. After UV radiation exposure, the redness and yellowness increased in both the red and black heartwoods, whereas the brightness decreased. In the black heartwood, the resulting color turned from yellowish white to reddish brown. Reddening in black heartwood after exposure to a combination of smoke heating and UV radiation is thought to be due to a decrease in brightness and an increase in both redness and yellowness. However, the degree of change in heartwood color by UV radiation exposure was not greatly affected by smoke-heating treatments of various lengths. When methanol extracts were fractionated and exposed to UV radiation, the yellowness increased in the n-hexane-soluble portion and the redness increased in the acetone-soluble fractions from the n-hexane-insoluble portion. These results suggest that the n-hexane-soluble fraction contains the substances that allow heartwood color to change to yellow after UV radiation exposure, and the acetone-soluble-fraction from the n-hexane-insoluble portion contains the substances that allow it to change to red. Received: November 14, 2001 / Accepted: June 3, 2002 Acknowledgment This research was supported in part by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science. This study was presented in part at the 51st Annual Meeting of the Japan Wood Research Society, Tokyo, April 2001 Correspondence to:N. Yoshizawa     

Antifungal agents from heartwood extract of <Emphasis Type="Italic">Taiwania cryptomerioides</Emphasis> against brown root rot fungus <Emphasis Type="Italic">Phellinus noxius</Emphasis>

Taiwania (Taiwania cryptomerioides Hayata) is a native and plantation species in Taiwan, and its heartwood has demonstrated excellent decay resistance. Brown root rot fungus Phellinus noxius has caused severe damage to more than 100 tree species in Taiwan. The aims of this study were to investigate the antifungal activities of Taiwania heartwood extracts against P. noxius and to identify the antifungal compounds. Antifungal activities were examined using the agar dilution method (a direct contact assay) and the inverted lid method (a vapor contact assay). Active compounds were purified using liquid–liquid partition, open-column chromatography and HPLC. Among four soluble fractions (n-hexane, ethyl acetate, n-butanol and water soluble fractions), results showed that n-hexane soluble fraction (HSF) can significantly inhibit the growth of P. noxius via the agar dilution method. Moreover, active compounds present in HSF were obtained using antifungal assay-guided fractionation. Results showed that ferruginol, T-cadinol, α-cadinol and T-muurolol were found to exhibit excellent antifungal activities against P. noxius, and their IC₅₀ values were 16.9, 25.8, 33.8 and 50.6 μg/mL, respectively. According to the current findings, phytochemicals (ferruginol, T-cadinol, α-cadinol and T-muurolol) of Taiwania heartwood had the potential to be used as environmentally benign antifungal agents against P. noxius in place of synthetic or inorganic fungicides.     

Bioactivity and characterization of exudates from Cryptomeria japonica bark

Tree exudates play an important role in the defense system of trees. In this study, the composition of exudates secreted from the wound site of Cryptomeria japonica was examined. Seven diterpenoids, including ferruginol (1), cryptojaponol (2), isopimaric acid (3), 7β-hydroxydeoxocryptojaponol (4), isopimarol (5), sandaracopimarinol (6) and phyllocladan-16α-ol (7), were identified through chromatographic separation and spectroscopic analyses. Among them, ferruginol was the most abundant compound, accounting for 76.6 % of the total content in exudates. Exudates, ferruginol and isopimaric acid possessed the strongest antibacterial activity and had MIC values of 25–250, 12.5–100 and 6.25–100 μg/mL, respectively, against one strain of Gram-negative bacteria (Escherichia coli) and four strains of Gram-positive bacteria (Staphylococcus aureus, methicillin-resistant S. aureus, S. epidermidis and Enterococcus faecalis). In addition, exudates and ferruginol presented 79 and 53 % at 100 μg/mL, respectively, of antioxidant activities against 1,1-dipheny1-2-picrylhydrazyl radical. C. japonica bark exudates and their diterpenes exhibit the strongest antioxidant activity and can inhibit bacterial growth and thus have the potential to be developed into natural antioxidants and bactericides.     

Antimicrobial activity of the Maniltoa schefferi extracts

All fractions of Maniltoa schefferi exhibited a broad spectrum of antibacterial activity. Good activity was demonstrated by the petrol and dichloromethane fractions of seed pods, seeds with coat and by methanol fractions of stem heartwood, root bark, root heartwood and stem bark whose butanol fraction was equally good. None of the fractions showed any antifungal activity.     

Inhibition activity of essential oils obtained from Japanese trees against <Emphasis Type="Italic">Skeletonema costatum</Emphasis>

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.     

Termite repellent sesquiterpenoids from Callitris glaucophylla heartwood

Fractions of methanol and ethanol extracts from the heartwood of white cypress pine (Callitris glaucophylla Thompson et Johnson) were investigated for their repellent activity against subterranean termite Coptotermes formosanus Shiraki worker using a two-choice semicircular filter paper test at 0.5% (w/w) concentration. Fraction CY-E2 composed of (−)-citronellic acid, guaiol, α-, β-, and γ-eudesmol isomers as well as an unknown compound, showed the highest statistically significant repellency (97.8% ± 2.2 SEM) of all fractions tested. Bioactivity-guided fractionations using high-performance liquid chromatography led to the isolation of two, oxygenated eudesmane-type sesquiterpenes with α-methylene moieties, both termite-repellent compounds. These compounds were subsequently identified as ilicic acid methyl ester (IAME) and costic acid by means of spectroscopic analyses, electron impact mass spectrometry, and nuclear magnetic resonance spectroscopy. We report the isolation of both IAME and costic acid from C. glaucophylla heartwood for the first time. Part of this paper was presented at the 47th Symposium on the Chemistry of Terpenes, Essential Oils, and Aromatics, Tokyo, November 2003     

Antibacterial and antifungal activities of Myracrodruon urundeuva heartwood

The aim of this work was to isolate a lectin from Myracrodruon urundeuva heartwood and to evaluate its antimicrobial activity against bacteria and fungi that attack plants, including woods. The lectin was isolated from heartwood through affinity chromatography on a chitin column monitored by hemagglutination assay. The lectin inhibited Gram-negative and Gram-positive bacteria and was more effective than antifungal Cercobin in growth inhibition of phytopathogenic fungi. The detected antimicrobial activity reveals the possible role of the lectin in the resistance of M. urundeuva heartwood against deteriorative biological agents. The M. urundeuva lectin is the first bioactive peptide found in heartwood, probably stored as a chemical protection against biodegradation.     

Abietane-type and labdane-type diterpenoids from the cones of Chamaecyparis obtusa

A novel compound, 8(17),12E,14-labdatrien-19-al (trans-communal), was isolated from ethyl acetate extract of young cones of hinoki (Chamaecyparis obtusa Endl.). The chemical structure of the compound was determined mainly with various nuclear magnetic resonance spectral techniques. Its stereochemistry was determined by derivation to a known compound,trans-communol. In addition to this compound, four known compounds — ferruginol, chamaecydin, 12-hydroxy-6,7-seco-abieta-8,l 1,13-triene-6,7-dial, and frans-communic acid — were isolated. All isolated compounds were subjected to an antifeedant bioassay against the pest insectSpodoptera litura. Results of the bioassay showed that chamaecydin and 12-hydroxy-6,7-seco-abieta-8,11,13-triene-6,7-dial had antifeedant activity.Part of this report was presented at 45th Annual Meeting of the Japan Wood Research Society, Tokyo, April 1995     

Antibacterial activities of some mosses including Hylocomium splendens from South Western British Columbia

The antibacterial activity of methanol extracts of ten moss species and fractions prepared from 80% methanol extract of Hylocomium splendens were evaluated by disk diffusion method. Nine moss species showed antibacterial activity against Gram (+) bacteria, in particular H. splendens and its ethyl acetate fractions showed stronger activity. Enhancement of antibacterial activity against Staphylococci by UV-A light irradiation was demonstrated in the extracts of Bartramia pomiformis, Ceratodon purpureus and Neckera douglasii.     

In vitro characterization of Inocutis jamaicensis and experimental inoculation of Eucalyptus globulus standing trees

Lesions of variable size, associated with the hymenomycete Inocutis jamaicensis, a white‐rot fungus, have been observed on the stems of Eucalyptus globulus trees in Uruguay. The aim of this study was to evaluate some ecophysiological characteristics of I. jamaicensis and assess its ability to colonize E. globulus trees of two different seed origins (Geeveston and Jeeralang) and the clone, 334‐1‐AR, obtained by micropropagation (ENCE, Spain). The growth of an I. jamaicensis isolate (MVHC11379) was evaluated at 25°C in a medium with a water potential of 0 (Ψ = 0). The growth rate did not vary significantly with a growth medium pH of between 4 and 7. I. jamaicensis showed no growth at either 5 or 37°C at any pH or Ψ tested. Weight loss of heartwood and sapwood of different plant provenances inoculated with I. jamaicensis under laboratory conditions was evaluated, and significant differences observed. Lignin‐modifying enzyme activity was evaluated in culture medium with or without E. globulus sawdust as substrate or inducer. Laccase activity was observed with sawdust and manganese peroxidase activity with and without sawdust. Only slight activity of aryl‐alcohol oxidase and lignin peroxidase was detected without sawdust. Experimental inoculation with I. jamaicensis of 3‐year‐old Geeveston and Jeeralang, and of 4‐year‐old 334‐1‐AR stems, resulted in successful fungal colonization of 56% of the 334‐1‐AR, 50% of Geeveston and 25% of Jeeralang trees. Only the heartwood was decayed. In 334‐1‐AR, the rotted wood was delimited by a reaction zone. Wood characteristics and the ability of I. jamaicensis to overcome the chemical reactions in the tree could partially explain differences in susceptibility to the fungus among provenances observed under natural and laboratory conditions.     

Antioxidant activity of extracts from <Emphasis Type="Italic">Calocedrus formosana</Emphasis> leaf,bark, and heartwood

Calocedrus formosana Florin (Cupressaceae) is an endemic tree to Taiwan. To evaluate the antioxidant activity of the plant extracts from heartwood, bark, and leaf of C. formosana, assays for 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and superoxide anion scavenging activities, as well as prevention of DNA strand cleavage were performed in this study. Similar IC₅₀ values against the DPPH radical were found for the heartwood and bark extracts at approximately 23µg/ml. Moreover, the heartwood extract exhibited the highest inhibitory activity against superoxide radicals among the test samples; a 2.3-fold lower value of IC₅₀ for superoxide radical inhibition was found in the heartwood extract relative to that of (+)-catechin. Much less effect on inhibition of DPPH and superoxide radicals was found from the leaf extract of C. formosana. More than 70% of superoxide radicals were inhibited in the presence of 10µg/ml heartwood extract, whereas only 15% inhibition was obtained from the leaf extract. The heartwood extract, at a dose of approximately 0.5mg/ml, apparently completely prevented the X174 supercoiled DNA cleavage induced by ultraviolet photolysis of H₂O₂, as judged by agarose gel electrophoresis. This report also suggests that the antioxidant activities of the plant extracts of C. formosana are in good correlation with their phenolic contents.     

Identification and evaluation of antioxidant activities of bamboo extracts

The antioxidant activity of solvent extracts from two main bamboo species, moso bamboo (Phyllostachys pubescens) and madake bamboo (P bambusoides) in Japan, was first evaluated by scavenging free radical of 1,1-diphenyl-2-picrylhydrazyl (DPPH), the inhibition activity for peroxidation of linoleic acid, and the reduction power. The methanol-extracts of moso bamboo culms and madake bamboo leaves presented stronger antioxidant activity compared with DPPH scavenging activity. Methanol-extract of moso bamboo culms was further fractionated by different solvents and n-butanol soluble fraction exhibited the most significant activity in the DPPH scavenging assay. The fractionation of n-butanol soluble extract was isolated by silica gel column with gradient mixture solvent of chloroform and methanol. The isolated fractions were directed by the antioxidant activity measured by scavenging the stable DPPH free radical. It was observed that most of the eluted fractions showed the antioxidative activity. Fractions acquired from elution with the mixture solvent of chloroform and methanol (10:1-5:1) showed stronger antioxidant activity than the other fractions.     

Antioxidant activity and constituents of extracts from the root of Garcinia multiflora

The methanolic extract of the root of Garcinia multiflora and its derived soluble fractions, namely those soluble in ethyl acetate (EtOAc), n-butanol, and water, were screened for their antioxidant activities. Among them, the EtOAc-soluble fraction exhibited the highest scavenging activity against the 1,1-dipheny1,2-picrylhydrazyl radical, the highest superoxide radical scavenging activity, and the strongest reducing power. In addition, according to the bioactivity-guided isolation, 26 specific phytocompounds, including 3 aromatics, 3 benzophenones, 3 flavonoids, 3 isocoumarins, 1 phloroglucinol, 6 steroids, and 7 xanthones, were isolated from the EtOAc-soluble fraction and identified. Of these, 2,4,3′,4′-tetrahydroxy-6-methoxybenzophenone and 1,3,6,7-tetrahydroxyxanthone were found to be the major bioactive constituents, present in the crude extract in concentrations of 3.9 and 15.6 mg/g, respectively. These two compounds had similar antioxidant activities to (+)-catechin, a well-known antioxidant.     

Antioxidant activity of heartwood extracts of Papua New Guinean woods

Antioxidant effects of methanol extracts from the heartwood of 23 Papua New Guinea (PNG) wood species were examined. The extract ofAmoora sp. (Meliaceae) showed the strongest antioxidant activity against lipid peroxidation in rabbit erythrocyte membrane and linoleic acid autoxidation. Also, the extract ofAmoora sp. showed potent 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicalscavenging activity. These results showed that the heartwood ofAmoora sp. is a possible source of antioxidative agents. The result of antioxidant activity-guided fractionation suggested that gallic acid, protocatechuic acid, and hydrolyzable tannins in the extract ofAmoora sp. caused the potent antioxidant activity.Part of this report was presented at the 51st Annual Meeting of the Japan Wood Research Society, Tokyo, 2001     

Chemical constituents from Gmelina arborea bark and their antioxidant activity

Gmelina arborea Roxb. is a fast-growing species and is known to have been used in traditional Indian medicine. Chemical constituents from the bark have not been reported, although some chemical constituents from part of this plant (heartwood, leaf, and root) are known. In this study, the bark meal was successively extracted with acetone and methanol. Fractionation of the acetone extract with n-hexane, diethyl ether, and ethyl acetate and subsequent chromatographic separation of the fractions led to the isolation of four compounds. The diethyl ether-soluble fraction yielded tyrosol [2-(4-hydroxyphenyl)ethanol] (1); (+)-balanophonin (2), an 8-5′ neolignan, with opposite optical rotation to known (−)-balanophonin; and gmelinol (3), a known lignan. The ethyl acetate-soluble fraction afforded a new phenylethanoid glycoside to the best of our knowledge, which was identified as (−)-p-hydroxyphenylethyl[5′″-O-(3,4-dimethoxycinnamoyl)-β-d-apiofuranosyl(1′" → 6′)]-β-d-glucopyranoside (4). From the methanol extract, two known compounds, 2,6-dimethoxy-p-benzoquinone (5) and 3,4,5-trimethoxyphenol (6), were isolated and identified. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay of the identifi ed compounds indicated that 3,4,5-trimethoxyphenol (6) exhibited moderate activity. Part of this report was presented at the 57th and 58th Annual Meetings of the Japan Wood Research Society, Hiroshima and Tsukuba, August 2007 and March 2008, respectively     

Evaluation of biological activities of extracts from the fruiting body of <Emphasis Type="Italic">Pleurotus citrinopileatus</Emphasis> for skin cosmetics

Pleurotus citrinopileatus Singer has recently become a popular delicacy in East Asian countries. We prepared a methanol extract, soluble fractions from the methanol extract, and a hot water extract of the fruiting bodies of P. citrinopileatus. The biological activities such as melanin biosynthesis inhibition, tyrosinase inhibition, antioxidant activities [1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, oxygen radical absorbance capacity (ORAC), and superoxide dismutase (SOD)-like activity], antibacterial activities, and antihyaluronidase activities of these extracts were evaluated. We found that the n-hexanesoluble, diethyl ether-soluble, and ethyl acetate-soluble fractions exhibited melanin biosynthesis inhibition in B16 melanoma cells, as well as antioxidant (ORAC) and antibacterial activities. However, the n-butanol-soluble and water-soluble fractions and the methanol and hot water extracts exhibited antioxidant (DPPH radical scavenging, SOD-like activity) and antihyaluronidase activities. These results indicate that the fruiting bodies of P. citrinopileatus have the potential to be used as an ingredient in skin cosmetics.     

Antimicrobial activity of Micrechites novoguineensis

The methanol extracts of Micrechites novoguineensis, leaves and stem bark were partitioned (petrol, ethyl acetate, methanol). All obtained fractions showed a broad spectrum of antibacterial activity, while only the leaf ethyl acetate fraction showed activity against moulds.     

The influence of heartwood on the pulping properties of Acacia melanoxylon wood

The pulping wood quality of Acacia melanoxylon was evaluated in relation to the presence of heartwood. The sapwood and heartwood from 20 trees from four sites in Portugal were evaluated separately at 5% stem height level in terms of chemical composition and kraft pulping aptitude. Heartwood had more extractives than sapwood ranging from 7.4% to 9.5% and from 4.0% to 4.2%, respectively, and with a heartwood-to-sapwood ratio for extractives ranging from 1.9 to 2.3. The major component of heartwood extractives was made up of ethanol-soluble compounds (70% of total extractives). Lignin content was similar in sapwood and heartwood (21.5% and 20.7%, respectively) as well as the sugar composition. Site did not influence the chemical composition. Pulping heartwood differed from sapwood in chemical and optical terms: lower values of pulp yield (53% vs 56% respectively), higher kappa number (11 vs. 7), and lower brightness (28% vs 49%). Acacia melanoxylon wood showed an overall good pulping aptitude, but the presence of heartwood should be taken into account because it decreases the raw-material quality for pulping. Heartwood content should therefore be considered as a quality variable when using A. melanoxylon wood in pulp industries