Effects of chronic acid fog exposure with soil acidification or nitrogen loading on nutrient status in Abies firma seedlings

To clarify the nutrient status in momi fir (Abies firma Sieb. et Zucc.) seedlings under complex stress conditions of acid fog with soil acidification or nitrogen loading, we exposed seedling shoots to simulated acid fog (pH 3) and simultaneously loaded the soil with acid or excess nitrogen for 17 months. Proton and nitrogen loading reduced K concentration in soil, but these treatments had little effect on the nutrient status of fine roots in momi fir seedlings. Acid fog exposure resulted in reduced concentrations of Mg, Al, and Fe in current-year needles and Mg, Al, Cu, Fe, and Zn in 1-year-old needles. The complex effects of acid fog exposure with proton or nitrogen loading on nutrient status were relatively slight. However, elements such as Al and Cu notably reflect the effects of proton loading. These results show that the nutrient status of trace elements, rather than major elements, in needles of momi fir seedlings is sensitive to exposure to acid fog or proton loading.     

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.     

Mitigation of aluminum toxicity by calcium and magnesium in Japanese cedar (Cryptomeria japonica)

We investigated the short-term and long-term mitigation of Al toxicity by Ca and Mg in pot trials of Japanese cedar (Cryptomeria japonica D. Don). We found that in the initial stages of treatment, Al toxicity at high Al concentration (5mM) was mitigated by Ca and Mg through the stimulation of antioxidant enzyme activities in needles. However, growth reduction occurred after 11 months exposure to Al despite the coexistence of Ca and Mg. Growth reduction was related to Al³⁺ activity in solution rather than the concentration of Ca and Mg. Therefore, when considering the influence of soil acidification on Al toxicity in forest ecosystems, it is important to consider not only the potential for mitigation of Al toxicity by base cations, but also the potential for factors in the soil solution to change the chemical form of Al.     

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.     

Response of gas exchange rates in <Emphasis Type="Italic">Abies firma</Emphasis> seedlings to various additional stresses under chronic acid fog stress

In order to clarify the chronic influence of acid fog on the gas exchange rates of momi fir (Abies firma Sieb. et Zucc.) trees, we exposed them to simulated acid fog (pH 3) for 3 years. The composition of the acid fog was similar to that observed in a region where momi fir trees have been declining, and it contained organic acids. We then treated the firs with various additional stresses, such as drought, low temperature, fine root cutting, ozone exposure, soil acidification, nitrogen load, and rhizosphere aluminum stress. Under chronic exposure to acid fog, the momi fir seedlings exhibited a pattern of stomatal behavior whereby they excessively opened in summer and closed in winter. Furthermore, the stomata of these seedlings tended to open during drought stress, and their needles were visibly injured after ozone exposure. The net photosynthesis rates of the seedlings exposed to acid fog were regulated by their stomatal aperture, rather than directly by acid fog. These results suggest that acid fog exposure disturbs the control of stomatal function in the momi fir seedlings. In addition, we found that chronic acid fog exposure suppressed the decrease in net photosynthesis rate, due to its nitrogen load.     

Effects of chronic nitrogen application on the growth and nutrient status of a Japanese cedar (Cryptomeria japonica) stand

To investigate the potential effects of nitrogen (N) deposition on Japanese forests, a chronic N-addition experiment that included three treatments (HNO₃, NH₄NO₃, and control) was carried out in a 20-year-old Japanese cedar (Cryptomeria japonica D. Don) stand in eastern Japan over 7 years. The amount of N applied was 168 kg N ha⁻¹ year⁻¹ on the HNO₃ plots and 336 kg N ha⁻¹ year⁻¹ on the NH₄NO₃ plots. Tree growth, current needle N concentration, and soil solution chemistry were measured. Nitrogen application decreased the pH and increased NO₃ ⁻, Ca²⁺, Mg²⁺, and Al concentrations in the soil solution. The needle N concentration increased in both of the N plots during the first 3 years. Nevertheless, the annual increments in height and in the diameter at breast height of the Japanese cedars were not affected by N application, and no visible signs of stress were detected in the crowns. Our results suggest that young Japanese cedar trees are not deleteriously affected by an excess N load.     

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).     

Effects of the absorption of artificial acidic solutions on the root systems of Japanese red cedar (Cryptomeria japonica D. Don) cutting and saplings

The effects of the absorption of artificial acidic solutions only from below-ground parts on root systems were examined for 60 days in summer and 72 days in fall using cuttings and saplings of Japanese red cedar (Cryptomeria japonica D. Don). Cuttings and saplings absorbed the solutions through the potted soils from the reservoir of an autoirrigator and/or a simplified-autoirrigator. Nitric acid solutions of pH 2.0, 4.0 and 6.0 and distilled water (control) were used in summer, mixtures of H₂SO₄ and HNO₃ solutions of pH 2.0, 3.0 and 4.0, HNO₃ solution of pH 3.0, H₂SO₄ solution of pH 3.0 and distilled water (control) were used in fall. Although no detrimental effects of acidic solutions on saplings were observed, the rooting rate of cuttings treated at pH 2.0 in summer was significantly lower than that of the control. Root lengths and root dry weight per cutting at pH 2.0 in summer and fall were smaller than those of the control, and the difference in fall was significant. The present study revealed that the acidic treatment only from below-ground parts had detrimental effects on the root systems of cuttings, suggesting the importance of quantification of the effects on below-ground parts, as well as on above-ground parts, to evaluate the effects of acidic precipitation on trees.     

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.     

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.     

Comparison of organic matter dynamics in soil between Japanese cedar (Cryptomeria japonica) forest and adjacent Japanese red pine (Pinus densiflora) forest established on flatland

In order to clarify the effects of tree species on organic matter dynamics in soil, we investigated the amount of forest floor material, leaf litter decomposition rate, soil chemical characteristics, soil respiration rate and cellulose decomposition rate in a Japanese cedar forest (cedar plot) and an adjacent Japanese red pine forest (pine plot) established on a flatland. The amount of forest floor material in the cedar plot was 34.5 Mg ha⁻¹ which was greater than that in the pine plot. Because the leaf litter decomposition rate was higher in the pine plot than in the cedar plot, it is likely that the difference in the amount of forest floor material between the plots is caused by the difference in the leaf litter decomposition rate. The C concentrations of soil in the cedar plot were 1.2–2.1 times higher than those in the pine plot. Soil pH(H₂O)s in the cedar plot were significantly higher than those in the pine plot. The soil respiration rates and the rates of mineralized C in the cedar plot byin vitro incubation were higher than those in the pine plot. From this result, it is assumed that soil organic matter in the cedar plot was decomposed relatively faster compared with the pine plot. Furthermore, microbial activities, which were reflected as cellulose decomposition rates in the cedar plot, were higher than those in the pine plot. A part of this paper was presented at the 109th Annual Meeting of the Japanese Forestry Society (1998).     

Treatment of sugi （Cryptomeria japonica D.） sapwood with aqueous solution of acetic acid

Sugi sapwood samples were processed with aqueous solution of acetic acid in order to find the response of the weight of sugi sapwood and the treatment of aqueous solution of acetic acid. The result showed that loss of weight for the treated sugisapwood was about equal to yield of extracts from sugi sapwood, and increased with the increment of the concentration of aqueous solution of acetic acid. Fourier transform infrared spectroscopy spectra changes of the treated sugi wood and extracts from sugi sapwood were analyzed by FT-IR spectroscopic technique. Increasing tendency of absorption intensities of the stretching vibration at 3 400 cm^-1 of hydroxyl group (OH) and C=C in lignin stretching vibration at 1510 cm^-1 of benzene ring inlignin were observed from FT-IR of the treated sugi sapwood. From FT-IR spectra of extracts from sugi sapwood by aqueoussolution of acetic acid, the dissolution of lignin was observed during the treatment with 30% acetic acid solution aqueous.     

Antimicrobial activity of heartwood components of sugi (Cryptomeria japonica) against several fungi and bacteria

Methanol extract of sawdust of sugi (Cryptomeria japonica) heartwood was fractionated with toluene and n-hexane to give solvent-soluble and solvent-insoluble fractions. The n-hexane-soluble fraction showed the most inhibition activity among the fractions against phytopathogenic microorganisms, namely Fusarium oxysporum, Phytophthora capsici, Pythium splendens, and Ralstonia solanacearum. Sandaracopimarinol and ferruginol, isolated from the n-hexane-soluble fraction, showed moderate antifungal activity against the three fungi and strong antibacterial activity against R. solanacearum. The content of sandaracopimarinol (7.07 g/kg based on the dried sawdust) in the heartwood was about twice that of ferruginol. Sandaracopimarinol and ferruginol strongly inhibited the growth of Gram-positive bacteria but did not show inhibitory action against Gram-negative bacteria except for R. solanacearum. The antibacterial effect of sandaracopimarinol was first found in the present study and was stronger than that of ferruginol.     

Spatiotemporal distribution of aboveground litter in a Cryptomeria japonica plantation

We assessed the vertical distribution of litter and its seasonal patterns in the canopy and on the forest floor (soil), as well as litterfall (the flux of litter from the canopy to the soil) in a 33-year-old plantation of Japanese cedar (Cryptomeria japonica D. Don). The masses of total litter, dead leaves, and dead branches in the canopy of C. japonica trees averaged 34.09, 19.53, and 14.56 t dry wt ha⁻¹, respectively, and were almost constant during the study period. The total masses of the annual litterfall were 4.17 and 5.88 t dry wt ha⁻¹ year⁻¹ in the two consecutive years of the study. The mass of the soil litter averaged 7.95 t dry wt ha⁻¹ during the same period. All relationships between the mass of canopy litter and tree-size parameters (diameters at breast height and at the lowest living branch) were linear in a log-linear regression. Compared with the results for this plantation at a younger stage (16 years old), our results suggest that the total mass of dead leaves attached to each tree increases markedly with increasing age, but that the trajectory of this increase as a function of tree size may change from an exponential to a saturation curve with increasing stand age.     

The composition and abundance of microarthropod communities on arboreal litter in the canopy of Cryptomeria japonica trees

In a Cryptomeria japonica plantation, we examined the composition and seasonal abundance of microarthropods in communities associated with habitat substrates in the canopy (defined as dead leaves, dead branches, and living leaves) and compared them with those in soil communities. Habitat substrates and microarthropods were periodically collected by the branch-clipping and washing method from the canopy and by the Tullgren method from the soil. Oribatida, Collembola, and larvae of the Chironomidae, most of which are detritivorous or fungivorous, were dominant in the canopy. The dominant oribatid and collembolan families differed markedly between the canopy and the soil. Numbers of all microarthropods per unit dry weight of leaf or per unit area of branch ranged from 4.2 to 11.7g^–1 dry wt on dead leaves, 0.13–0.48cm^–2 on dead branches, and 1.3–6.4g^–1 dry wt on living leaves. In the soil, the number of individuals per unit ground area ranged from 24000 to 220000m^–2. The total abundances of microarthropods on dead leaves and dead branches were almost constant throughout the year. These results suggest that the arboreal litter characteristic of C. japonica canopies is utilized consistently by large numbers of detritivorous and fungivorous microarthropods, and that the decomposition of dead foliage and branches is initiated in the canopy.     

Study of sugi (Cryptomeria japonica D. Don) with male-sterility gene using CAPS markers

The frequency of open-pollinated male-sterile seedlings from a male-sterile mother tree in Cryptomeria japonica D. Don was determined in 765 open-pollinated seedlings. Pollen was not formed by 29 out of the 765 seedlings. To find pollen donor candidates for 29 male-sterile seedlings, a male-sterile tree was crossed with five individuals in the neighborhood of the mother tree. Analysis of microspores in male flowers of F₁ seedlings revealed that one of the five individuals had a heterozygous male-sterility gene. Paternity tests were conducted on the 29 male-sterile seedlings using eight kinds of CAPS markers in C. japonica. The pollen parents of 29 male-sterile seedlings were not only a tree in the neighborhood of the mother tree, but also other trees. Since it was found that the pollen parents of four of the male-sterile seedlings were not in the study plot, it was inferred that C. japonica trees that possess the male-sterility gene grow in other places and may be widely distributed.     

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%).     

Effects of complex stress of chronic acid fog exposure with rhizosphere aluminum on nutrient status in Abies firma seedlings

We exposed momi fir (Abies firma Sieb. et Zucc.) seedlings to simulated acid fog of pH 3.0 for about 3 years (from July 1999). During the last year of acid fog exposure, half of the seedlings were subjected to rhizosphere Al stress (complex stress) and the nutrient status of seedlings was determined. Chronic acid fog exposure decreased Fe and Zn concentrations in current-year and 1-year needles, and Al in 1-year needles, but had little effect on major element concentrations. Aluminum treatment had a broad impact on nutrient status in fine roots and needles. In fine roots, increases in Al and Cu concentrations and decreases in B, Mn, and Zn concentrations were observed. In 1-year needles, Al treatment increased Al, B, and Mn concentrations and decreased Cu concentration. The complex effect of acid fog with aluminum on nutrient status was relatively slight. These results show that changes in nutrient status under chronic acid fog exposure and/or Al stress are induced before the decline of photosynthesis in momi fir seedlings, mainly due to Al stress.     

Contribution of active defense responses in the limitation of fungal spread in the sapwood of living sugi (Cryptomeria japonica) tree

The participation of active defense responses in the limitation of fungal spread in the sapwood of living sugi tree was determined. Isolates ofGuignardia cryptomeriae were inoculated to living trees, fresh logs, frozen and thawed logs and autoclaved logs, and the spread of wood discoloration and fungal hyphae was assessed. In autoclaved logs and freeze-treated logs, wood discoloration and reaction zone formation were not observed, and fungal hyphae of both of virulent and avirulent isolates grew rapidly in the wood. In fresh logs, wood discoloration and the formation of a pale reaction zone and transition zone were observed. Virulent isolates spread rapidly, but the spread of avirulent isolates was inhibited. In living trees, wood discoloration was observed and reaction zones were formed at lesion margins. These results indicated that constitutive defense mechanisms had no effect on the spread of both of virulent and avirulent isolates, and that active defense responses were important in the limitation of spread of fungal hyphae, hence in the limitation of wood discoloration. Further, it appeared that avirulent, but not virulent, races ofG. cryptomeriae were easily inhibited even by weak host responses. This study was presented at 105th and 106th Annual Meeting of the Japanese Forestry Society.