Four glucosides ofp-hydroxyphenyl derivatives from birch leaves

The extractive of shirakamba (Betula platyphylla Sukatchev var.japonica Hara) leaves was investigated. Four glucosides ofp-hydroxyphenyl derivatives were isolated, and their structures were indentified as betuloside (I), 3,4-dihydroxy-propiophenone-3--d-glucopyranoside (II), salidroside (III), and arbutin (IV). Arbutin was newly found in the leaves of shirakamba.     

A new isoflavone glucoside from Astragalus membranaceus var. mongholicus

A new isoflavone glucoside (1) was isolated from the roots of Astragalus membranaceus var. mongholicus. Its structure was elucidated by spectroscopic methods.     

Investigation of change in tangential strain on the inner bark of the stem and root ofBetula platyphylla var.japonica andAcer mono during sap season

Although it is well known that sap exudation during early spring in temperate deciduous trees occurs in response to daytime warming and nighttime cooling, the mechanisms of the process are not yet fully understood. Previous theories suggested that changes in stress in the wood caused by daytime heating and nighttime cooling might be linked with sap flow. Consequently, a study of itaya-kaede maple (Aver mono) and shirakamba birch (Betula platyphylla var.japonica) looked at tangential strains. One-hour intervals for 3 years of the tangential strains on the inner bark of stem and root were measured in itaya-kaede maple and shirakamba birch during the sap exudation season. The measurements indicated different mechanisms of sap exudation in these two trees. During the sap exudation season in late March, when the temperature fluctuated around 0°C, the tangential strain in the root of itaya-kaede maple showed expansion in the daytime and contraction at night. Conversely, in early April the tangential strain in the root of shirakamba birch exhibited contraction in the daytime and expansion at night. The changes in tangential strains in itaya-kaede maple were attributed to conditioning, a known concept used to explain the uptake mechanism of soil water in maple and its exudation during early spring. However, because the change in tangential strain in the roots of shirakamba birch was similar to that found during the rampant season, sap exudation was not attributed to conditioning but to the plentiful supply of water from the roots. The implications of these mechanisms are that different sap harvesting techniques may be appropriate for different tree species.     

Effects of differences in forest floor and canopy vegetation on ectomycorrhizas ofBetula platyphylla var.japonica: A test using seedlings planted into soils taken from various sites

Effects of different forest floor vegetation types in secondary forest and of conversion to plantation on the quality and quantity of ectomycorrhizas are mostly unknown.Betula platyphylla var.japonica seedlings were used for bioassays of ectomycorrhizal fungal inoculum using soils from four 50-year-oldB. platyphylla var.japonica forests that had different types of forest floor vegetation: two with shrub types, one with aSasa type, and one with a grass type. Seedlings were also grown in soil from a nearby monospecific plantation ofChamaecyparis obtusa. Ectomycorrhizas formed 13 to 26% of root length of seedlings grown in soil from the five different sites. The maximum percentage of ectomycorrhizal formation was obtained from the grass-type forest. The dominant type of ectomycorrhiza in the two shrub-type forest soils was the same as that in theSasa-type forest soil. The dominant types of ectomycorrhizas in the grass-type forest soil and in theC. obtusa plantation soil were different from that in the two shrub-type forest soils and in theSasa-type forest soil. The results of this investigation suggest that the type of forest floor vegetation, accompanied with changes in thickness of the A₀ horizon, might affect the ectomycorrhizal fungi in the soils ofB. platyphylla var.japonica forests. Establishment of artificial plantations ofC. obtusa might change the ectomycorrhizal fungi that could associate withB. platyphylla var.japonica seedlings in soil.     

Essential oil from the leaves of Cryptomeria japonica acts as a silverfish (Lepisma saccharina) repellent and insecticide

This is the first article to report the evaluation of a natural product used as an antisilverfish agent. Silverfish (Lepisma saccharina), primitive wingless insects, feed on a variety of materials, including paper, cotton, starch, and cereals. They can be a problem in libraries and other places where books, documents, and papers are stored. In this pilot study, the essential oil from leaves of Cryptomeria japonica was investigated to test its properties as a silverfish repellent and insecticide. The results from a repellency bioassay show that the essential oil significantly repelled silverfish. The repellent activity was 80% at a dosage of 0.01 mg/cm³. When silverfish were exposed to a concentration of 0.16 mg/cm³ of essential oil, they were killed within 10h. The chemical composition of essential oil, the emissions from a test chamber, and the residue left on filter papers previously soaked with the essential oil in a chamber were analyzed by gas chromatography-mass spectrometry. The components of the essential oil were found to be: elemol (18.22%), 16-kaurene (11.63%), 3-carene (9.66%), sabinene (9.37%), 4-terpineol (9.06%), β-eudesmol (5.70%), α-pinene (5.62%), and limonene (5.26%). Only some constituents of the essential oil compounds collected by solid-phase microextraction were found to be emitted in the test chamber. The main constituents were: 3-carene (21.03%), p-cymene (10.95%), limonene (9.49%), β-myrcene (9.39%), γ-terpinene (9.10%), α-terpinene (8.57%), and 4-terpineol (7.97%).     

Absolute configuration of new bisaboranoids from Yaku-sugi (Cryptomeria japonica) wood

The absolute configuration of bisabolanoids isolated from Yaku-sugi wood were compared with those of dihydrocarvone and dihydrocarveol isomers based on their¹³C nuclear magnetic resonance data and optical rotations.     

Isolation of phenylisoserine methyl ester from the leaves of Taxus cuspidata var. nana

From methanolic extracts of leaves of kyaraboku, Taxus cuspidata var. nana, phenylisoserine methyl ester (3) was isolated along with taxinine (1), taxol (2), sciadopitysin (4), ginkgetin (5), isorhamnetin (6), and quercetin (7). This is the first time that phenylisoserine methyl ester has been isolated from T. cuspidata var. nana. Compound 3 was also isolated from the ethanolic extracts of leaves of T. cuspidata var. nana. Furthermore, compound 3 was identified in methanolic extracts from the bark of this tree.     

Sesquiterpenes from Ulmus davidiana var. japonica with the inhibitory effects on lipopolysaccharide-induced nitric oxide production

Investigation of antiinflammatory constituents of the stem and root barks of Ulmus davidiana var. japonica resulted in the isolation of three guaiane type sesquiterpenes, torilin, 1-hydroxytorilin, together with a new derivative, (1beta, 7beta, 8beta, 10beta)-1,8,11-trihydroxy-4-guaien-8-angeloyl-3-one named 1-hydroxytorilin A. All the three sesquiterpenes inhibited lipopolysaccharide-induced nitric oxide production in murine microglial BV2 cells.     

A new phenolic glucoside from the leaves of Heliciopsis lobata

A new phenolic glucoside ester, 6'-E-(2'-methyl-2'-butenoyl) arbutin (1), was isolated from the leaves of Heliciopsis lobata. Its structure was elucidated by spectral analysis.     

Antifungal activities of compounds isolated from the leaves of Taxus cuspidata var. nana against plant pathogenic fungi

Antifungal activities of seven compounds, taxinine (1), paclitaxel (2), phenylisoserine methyl ester (3), sciadopitysin (4), ginkgetin (5), isorhamnetin (6), and quercetin (7), isolated from the leaves of kyaraboku, Taxus cuspidata var. nana, against five plant pathogenic fungi, Gibberella fujikuroi, Cladosporium cucumeninum, Fusarium oxysporum, Colletotrichum fragariae, and Corynespora cassiicola, were investigated for utilization of extractives from trees of the genus Taxus. Also, the amounts of compounds 2 and 3 on the leaf surface was measured in relation to the antifungal activities of compounds. Taxinine (1) showed antifungal activity against G. fujikuroi, C. cucumeninum, F. oxysporum, and C. cassiicola. The minimum inhibitory concentration of taxinine for the four fungi was 0.4mol. In addition, from the results of antifungal tests, it may be concluded that paclitaxel on the leaves and stem of T. cuspidata var. nana does not play an important role as an antifungicide in the resistance of trees to plant pathogenic fungal attack.     

Environmental effects on the color of sugi (Cryptomeria japonica D. Don) heartwood

Sugi (Cryptomeria japonica D. Don) is one of the most important planted trees in Taiwan. The normal heartwood of sugi has a red to pinkish rose color. Unfortunately, this pleasant appearance is susceptible to discoloration after environmental exposures. This discoloration is a serious defect that decreases the value of sugi products. The objective of this study was to investigate the effects of environmental factors such as moisture, oxygen, and lights of different wavelengths on the discoloration of sugi. The results show that under the combined effects of light and oxygen the color of heartwood changes from red (or rosepink) to bluish green. The color further darkens and eventually turns to black with the presence of moisture. As for the effect of light wavelengths on the discoloration of sugi heartwood, it was found that the red color was enhanced after being irradiated with light of wavelengths above 600 nm. Botha ^* andb ^* values increased significantly as a result of such exposure.     

In vitro screening of angiotensin I-converting enzyme inhibitors from Japanese cedar (Cryptomeria japonica)

Screening and isolation of angiotensin I-converting enzyme (ACE) inhibitors from Japanese cedar (Cryptomeria japonica) based on the in vitro ACE inhibitory assay were attempted. The ethanol extract from outer bark showed the highest inhibitory activity (IC₅₀ is 16g/ml) among 24 extracts prepared from roots, leaves, heartwood, sapwood, inner bark, and outer bark by successive extraction with four solvents. The fractionation of the outer bark ethanol extract followed by the bioassay resulted in the isolation of two strong ACE inhibitors, catechin and dimeric procyanidin B3. The bioassay of three flavan-3-ols including (+)-catechin and six flavones revealed that most of these compounds have high ACE inhibitory activity. The results suggest that the phenolic hydroxyl group at the C7 position and heterocyclic oxygen atom of these compounds are important for expressing the inhibitory activity.     

A new steroidal saponin with antiinflammatory and antiulcerogenic properties from the bulbs of Allium ampeloprasum var. porrum

A new steroidal saponin was isolated from the bulbs of Allium ampeloprasum var. porrum L. On the basis of chemical evidence, comprehensive spectroscopic analyses and comparison of known compounds, its structure was established as (3β,5α,6β,25R)-6-[(β-d-glucopyranosyl)oxy]-spirostan-3-yl O-β-d-glucopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→3)]-β-d-galactopyranoside (1). Results of the present study indicated that the steroidal saponin showed haemolytic effects in the in vitro assays and demonstrated antiinflammatory activity and gastroprotective property using in vivo models.     

Secondary dispersal of Dioscorea japonica (Dioscoreaceae) bulbils by rodents

We examined rodent dispersal of Dioscorea japonica bulbils. Bulbils dispersed by rodents may travel farther from their parent plants than those dispersed by gravity alone. To confirm the presence of secondary dispersal and the types of vector, we used an automatic camera to photograph animals that approached bulbils. To clarify secondary dispersal distance, we set marked bulbils at eight sites and searched for the bulbils once they were removed from their original locations by a disperser. We took photographs of mice (Apodemus speciosus) approaching the bulbils. In June, 10 of 333 bulbils that were moved germinated. We also measured the primary dispersal distance at four sites by dropping bulbils from heights of 0.5, 3.0, and 12.0 m. We then compared primary and secondary dispersal distances. The mean secondary dispersal distance was significantly greater than the primary dispersal distance for dispersal heights of 0.5 and 3.0 m, but less than that for a 12.0-m dispersal height. This indicates that rodents may disperse bulbils farther than gravity alone when bulbils drop from lower heights but they contribute little to dispersal for bulbils that drop from greater heights (e.g., from the top of the canopy). Thus, we confirmed the occurrence of rodent dispersal of bulbils and rodents might to some extent contribute to the dispersal and establishment of bulbils.     

Factors shaping genetic structure within populations ofEurya japonica (Theaceae)

Starch-gel electrophoresis was used to resolve nine polymorphic enzyme loci from leaf tissue collected from 20 Korean populations ofEurya japonica in order to determine differences in allele frequencies between male and female trees. In addition, 84 adults were sampled and mapped in a population located on Naenaro Island in Korea to examine spatial genetic structure using spatial autocorrelation analysis. Allele frequencies between males and females gave few contribution to the genetic structuring within populations. Only nine (5%) of 180 cases were significantly different from both sexes in allele frequencies. On the other hand, Moran'sI was significantly different from the expected value in 31 (23.5%) of 132 cases. In the shortest distance (0<5m),I was significantly positive in 10 (22.7%) of 44 cases. The results indicate that a significant small scale genetic structure was detected in the population and patch widths were inferred to be approximately 5–7 m. A nonrandom distribution of genotypes may be indicative of restricted gene flowvia seed and pollen dispersal, and patchy establishments of genetically distinct individuals. These factors are responsible for shaping population genetic structure ofE. japonica.     

Estimation of the fine root biomass in a Japanese cedar (Cryptomeria japonica) plantation using minirhizotrons

We estimated fine root biomass in a Japanese cedar (Cryptomeria japonica) plantation using a min-irhizotron technique. Since data obtained from minirhizo-trons are limited to the length and diameter of fine roots observed on minirhizotron tubes, data conversion is necessary to determine the fine root biomass per unit soil volume or unit stand area. We first examined the regression between diameter squared and weight per unit length of fine roots in soil core samples, and calculated the fine root biomass on minirhizotron tubes from their length and diameter. Then we determined conversion factors based on the ratio of the fine root biomass in soil core samples to that on minirhizotron tubes. We examined calculation methods, using a single conversion factor for total fine root biomass in the soil for depths of 0–40cm (Cal1), or using four conversion factors for fine roots in the soil at 10-cm intervals (Cal2). Cal1 overestimated fine root biomass in the lower soil or underestimated that in the upper soil, while fine root biomass calculated using Cal2 better matched that in soil core samples. These results suggest that minirhizotron data should be converted separately for different soil depths to better estimate fine root biomass.     

Effect of simulated acid fog on membrane-bound calcium (mCa) in fir (Abies firma) and cedar (Cryptomeria japonica) mesophyll cells

The decline of virgin fir (Abies firma) forest at Mt. Oyama has been reported. Related field observations suggest that high acidity fog is linked with its decline. However, cedar (Cryptomeria japonica) in the same area shows no symptoms of decline. For assessing effects of acid fog on membrane-bound calcium (mCa) of the leaf mesophyll cells, 9-year-old seedlings of fir (Abies firma) and 8-year-old seedlings of cedar (Cryptomeria japonica) were exposed twice a week to simulated acid fog (SAF at pH 3 with pH 5 as control) for 2 h per day in a chamber during May–December 2007 (except August). Current and 1-year-old needles were collected from seedlings and analyzed at 1-month intervals. For current-year needles of fir, mCa levels in cells exposed to SAF at pH 3 were significantly lower than in cells exposed to pH 5, especially during September 2007–March 2008. In contrast, it is noteworthy that mCa levels of cedar were maintained as virtually constant irrespective of SAF acidity, indicating that fir is more sensitive to acid fog than is cedar. Based on these results, mCa loss by acid fog might also be caused in the declining virgin fir forest at Mt. Oyama.     

Histology of current shoots of Japanese cedar (Cryptomeria japonica D. don) inoculated withCercospora sequoiae Ellis et Everhart

The histological reaction of current shoots of the Japanese cedar,Cryptomeria japonica, inoculated withCercospora sequoiae was investigated. Wound periderm was formed on current shoots of the cedar on and after the 4th day after inoculation. In the cells of the cork layer, lignin and suberin were deposited, and a phenolic substance was accumulated. In other inoculated samples, plasmolysis, contraction of the cells and necrosis of the tissues were also observed. On the 32nd day, wound perioderm and phellogen were observed in samples of the tissue inside the necrotic tissues in response to hyphal invasion. Occurrence of wound periderm and phellogen in tissues of the needles was confirmed in the inoculated samples although no hyphae were observed in and around the tissues. These results suggest that this fungus causes necrosis of host tissues and induces defense responses. A part of this paper was presented at the 106th Annual Meeting of the Japanese Forestry Society (1995).     

Induction of the biosynthesis of agatharesinol, a norlignan, in sapwood sticks of Cryptomeria japonica under humidity-regulated circumstances

To establish an experimental approach for studying the biosynthetic pathway of agatharesinol, a norlignan, induction of the formation of agatharesinol in Cryptomeria japonica (Japanese cedar, sugi) was attempted. Wood sticks were prepared from the sapwood immediately after cutting a sugi tree down, and they were allowed to stand in desiccators in which the humidity was adjusted to 76% and 88%, and in a room. When the wood sticks were allowed to stand in 76% humidity and in a room, they dried rapidly and no formation of agatharesinol was demonstrated by gas chromatography-mass spectrometry. These results suggest that in the sapwood dried rapidly, the cells that were biologically active in the sapwood, probably ray parenchyma cells, die rapidly and completely before the biosyntheses of secondary metabolites including agatharesinol was wholly established. The wood sticks allowed to stand in 88% humidity, on the other hand, dried gradually compared with the sticks placed in 76% humidity and in a room, and agatharesinol was formed as one of the predominant ethyl acetate extractives. These results suggest that delaying the drying of the sapwood sticks can control the dying period to an appropriate length for the ray parenchyma cells, during which the biosynthesis of agatharesinol is induced. Part of this study was presented at the 54th Annual Meeting of the Japan Wood Research Society, Sapporo, August 2004