Effects of forest thinning on sap flow dynamics and transpiration in a Japanese cedar forest.

Historically, forest thinning in Japan was conducted to obtain high-quality timber from plantations. Today, in contrast, thinning is also motivated by forest water balance and climate change considerations. It is in this context that the present study examines the effects of thinning on the ecophysiological responses of remaining trees, which are inadequately understood, especially in relation to changes in the magnitude and duration of transpiration. Sap flux densities were measured in both outer and inner sapwood to obtain stand-scale transpiration for two years in the pre-thinning state and three years post-thinning. The effects of thinning on transpiration were quantitatively evaluated based on canopy conductance models. The larger increases in outer sap flux density were found in the first year after the treatment, while those in inner sap flux density were detected in the second and third years. The remaining trees required a few of years to adjust to improved light conditions of the lower crown, resulting in a delayed response of inner sap flux density. As a result of this lag, transpiration was reduced to 71 % of the pre-thinning condition in the first year, but transpiration recovered to the pre-thinning levels in the second and third years due to compensating contributions from inner sap flow. In terms of more accurately chronicling the thinning effect, the distribution of sap flux density with respect to its radial pattern, is necessary. Such measurements are key to more comprehensively examining the ecophysiological response of forest plantations to thinning and, ultimately, its effect on the forest water balance.

Changes in annual transcriptome dynamics of a clone of Japanese cedar (Cryptomeria japonica D. Don) planted under different climate conditions.

Environmental responses are critical for plant growth and survival under different climate conditions. To elucidate the underlying biological mechanisms of environmental responses in Japanese cedar (Cryptomeria japonica D. Don), the annual transcriptome dynamics of common clonal trees (Godai1) planted at three different climate sites (Yamagata, Ibaraki, and Kumamoto Prefectures) were analyzed using microarrays. Both principal component analysis (PCA) and hierarchical clustering of the microarray data indicated the transition to dormant transcriptome status occurred earlier and the transition to active growth status later in the colder region. Interestingly, PCA also indicated that the transcriptomes of trees grown under three different conditions were similar during the growth period (June to September), whereas the transcriptomes differed between sites during the dormant period (January to March). In between-site comparisons, analyses of the annual expression profiles of genes for sites 'Yamagata vs. Kumamoto', 'Yamagata vs. Ibaraki', and 'Ibaraki vs. Kumamoto' identified 1,473, 1,137, and 925 targets exhibiting significantly different expression patterns, respectively. The total of 2,505 targets that exhibited significantly different expression patterns in all three comparisons may play important roles in enabling cuttings to adapt to local environmental conditions. Partial least-squares regression analysis and Pearson correlation coefficient analysis revealed that air temperature and day length were the dominant factors controlling the expression levels of these targets. GO and Pfam enrichment analyses indicated that these targets include genes that may contribute to environmental adaptation, such as genes related to stress and abiotic stimulus responses. This study provided fundamental information regarding transcripts that may play an important role in adaptation to environmental conditions at different planting sites.

Verification of our empirical understanding of the physiology and ecology of two contrasting plantation species using a trait database.

The effects of climate change on forest ecosystems take on increasing importance more than ever. Information on plant traits is a powerful predictor of ecosystem dynamics and functioning. We reviewed the major ecological traits, such as foliar gas exchange and nutrients, xylem morphology and drought tolerance, of Cryptomeria japonica and Chamaecyparis obtusa, which are major timber species in East Asia, especially in Japan, by using a recently developed functional trait database for both species (SugiHinokiDB). Empirically, C. obtusa has been planted under drier conditions, whereas C. japonica, which grows faster but thought to be less drought tolerant, has been planted under wetter conditions. Our analysis generally support the empirical knowledge: The maximum photosynthetic rate, stomatal conductance, foliar nutrient content and soil-to-foliage hydraulic conductance were higher in C. japonica than in C. obtusa. In contrast, the foliar turgor loss point and xylem pressure corresponding to 50% conductivity, which indicate drought tolerance, were lower in C. obtusa and are consistent with the drier habitat of C. obtusa. Ontogenetic shifts were also observed; as the age and height of the trees increased, foliar nutrient concentrations, foliar minimum midday water potential and specific leaf area decreased in C. japonica, suggesting that nutrient and water limitation occurs with the growth. In C. obtusa, the ontogenetic shits of these foliar traits were less pronounced. Among the Cupressaceae worldwide, the drought tolerance of C. obtusa, as well as C. japonica, was not as high. This may be related to the fact that the Japanese archipelago has historically not been subjected to strong dryness. The maximum photosynthetic rate showed intermediate values within the family, indicating that C. japonica and C. obtusa exhibit relatively high growth rates in the Cupressaceae family, and this is thought to be the reason why they have been selected as economically suitable timber species in Japanese forestry. This study clearly demonstrated that the plant trait database provides us a promising opportunity to verify out empirical knowledge of plantation management and helps us to understand effect of climate change on plantation forests by using trait-based modelling.

Estimating spatial variation in the effects of climate change on the net primary production of Japanese cedar plantations based on modeled carbon dynamics.

Spatiotemporal prediction of the response of planted forests to a changing climate is increasingly important for the sustainable management of forest ecosystems. In this study, we present a methodology for estimating spatially varying productivity in a planted forest and changes in productivity with a changing climate in Japan, with a focus on Japanese cedar (Cryptomeria japonica D. Don) as a representative tree species of this region. The process-based model Biome-BGC was parameterized using a plant trait database for Japanese cedar and a Bayesian optimization scheme. To compare productivity under historical (1996-2000) and future (2096-2100) climatic conditions, the climate scenarios of two representative concentration pathways (i.e., RCP2.6 and RCP8.5) were used in five global climate models (GCMs) with approximately 1-km resolution. The seasonality of modeled fluxes, namely gross primary production, ecosystem respiration, net ecosystem exchange, and soil respiration, improved after two steps of parameterization. The estimated net primary production (NPP) of stands aged 36-40 years under the historical climatic conditions of the five GCMs was 0.77 ± 0.10 kgC m-2 year-1 (mean ± standard deviation), in accordance with the geographical distribution of forest NPP estimated in previous studies. Under the RCP2.6 and RCP8.5 scenarios, the mean NPP of the five GCMs increased by 0.04 ± 0.07 and 0.14 ± 0.11 kgC m-2 year-1, respectively. The increases in annual NPP were small in the southwestern region because of the decreases in summer NPP and the small increases in winter NPP under the RCP2.6 and RCP8.5 scenarios, respectively. Under the RCP2.6 scenario, Japanese cedar was at risk in the southwestern region, in accordance with previous studies, and monitoring and silvicultural practices should be modified accordingly.

Transcriptome analysis of heat stressed seedlings with or without pre-heat treatment in Cryptomeria japonica.

With global warming as a major environment concern over the coming years, heat tolerance is an important trait for forest tree survival during the predicted future warmer weather conditions. Cryptomeria japonica is a coniferous species widely distributed throughout Japan, and thus, can adapt to a wide range of air temperatures. To elucidate genes involved in heat response in Cryptomeria japonica, transcriptome analysis was conducted for seedlings under heat shock conditions. To test whether heat acclimation affects levels of gene expression, half of the seedlings were pretreated with moderately high temperatures prior to heat shock. De novo assembly of the transcriptome generated 107,924 unigenes and the analysis of differentially expressed genes was conducted using these unigenes. A total of 5217 differentially expressed genes were identified. Most genes upregulated by heat shock, regardless of pre-heat treatment, were conserved to heat response genes of angiosperm species, such as heat shock factors (Hsf) and heat shock proteins (Hsp). Pre-heating of seedlings affected expression levels of several Hsfs and their induction was lower in pre-heated seedlings than in seedlings without pre-heat treatment. This suggests a conserved role of Hsfs in heat response and heat acclimation in seed plants. On the other hand, many unknown genes were upregulated in only seedlings without pre-heat treatment after heat exposure. Notably, expression of gypsy/Ty3 type retrotransposons was dramatically induced. These findings provide valuable information to develop a better understanding of the molecular mechanisms of heat response and acclimation in C. japonica.

Climate sensitivity of Cryptomeria japonica in two contrasting environments: Perspectives from QTL mapping.

Long-lived forest tree species experience a wide range of environmental conditions throughout their lifespan. Evaluation of the underlying growth and development mechanisms of these species is essential to predict tree growth under climate change. This study investigated climate sensitivity to temperature, precipitation, dry periods, and the associated genomic regions in Cryptomeria japonica, Japan's most commercially important tree. We used tree rings and common garden experiments with three clonal replicates planted in two contrasting environments in Kyushu (Kumamoto site) and Honshu (Chiba site), Japan. Tree growth showed a significant negative correlation with the dry period (>4 days) in March of the year of tree-ring formation at the Chiba site. In contrast, temperature and precipitation had little influence on tree growth. Quantitative trait locus (QTL) analysis was performed to investigate climate sensitivity to dry periods at the Chiba site, revealing 13 significant QTLs. One QTL showed a substantially large contribution to the overall climate sensitivity, accounting for 12.4% of the total phenotypic variation. The phenotypic variance explained (PVE) by other QTLs ranged from 0.9% to 2.9%, and the total PVE by all QTLs was 35.6%. These findings indicate that the tree population at the Chiba site could be vulnerable to drought in early spring and that the QTL showing the greatest impact on climate sensitivity may be closely related to genes associated with tolerance or adaptation to drought stress. The QTLs identified in this study could be useful for molecular breeding, forest management, and predicting the growth of C. japonica under a changing climate.

Dispersal of allergenic pollen from Cryptomeria japonica and Chamaecyparis obtusa: characteristic annual fluctuation patterns caused by intermittent phase synchronisations.

Trees produce pollen during specific times of the year. Pollen can induce pollinosis, a type of allergic rhinitis, in humans. In Japan, allergenic pollen is mainly dispersed from February to May. Using data collected at 120 observation sites managed by the Japanese Ministry of the Environment, we studied the annual patterns of airborne allergenic pollen. The allergenic pollen showed an alternating ON-OFF cycle, but the length of the cycle differed among regions. We used an in-phase/out-of-phase analysis to quantify two characteristic features of the synchronisation. The degrees of phase synchronisation were strong in eastern and weak in western Japan. The pattern of allergenic pollen dispersal throughout Japan is typical intermittent synchronisation. This is the first study to evaluate allergenic pollen's distribution from a phase synchronisation viewpoint.

Identification of novel putative causative genes and genetic marker for male sterility in Japanese cedar (Cryptomeria japonica D.Don).

BACKGROUND: Japanese cedar (Cryptomeria japonica) is an important tree for Japanese forestry. Male-sterile marker development in Japanese cedar would facilitate selection of male-sterile plus trees, addressing the widespread social problem of pollinosis and facilitating the identification of heterozygotes, which are useful for breeding. RESULTS: This study used next-generation sequencing for single-nucleotide polymorphism discovery in libraries constructed from several organs, including male-sterile and male-fertile strobili. The single-nucleotide polymorphisms obtained were used to construct a high-density linkage map, which enabled identification of a locus on linkage group 9 strongly correlated with male-sterile trait. Expressed sequence tags corresponding to 11 marker loci from 5 isotigs were associated with this locus within 33.4-34.5 cM. These marker loci explained 100% of the phenotypic variation. Several homologs of these sequences are associated with male sterility in rice or Arabidopsis, including a pre-mRNA splicing factor, a DEAD-box protein, a glycosyl hydrolase, and a galactosyltransferase. These proteins are thus candidates for the causal male-sterile gene at the ms-1 locus. After we used a SNaPshot assay to develop markers for marker-assisted selection (MAS), we tested F2 progeny between male-sterile and wild-type plus trees to validate the markers and extrapolated the testing to a larger plus-tree population. We found that two developed from one of the candidates for the causal gene were suitable for MAS. CONCLUSIONS: More than half of the ESTs and SNPs we collected were new, enlarging the genomic basis for genetic research on Japanese cedar. We developed two SNP markers aimed at MAS that distinguished individuals carrying the male-sterile trait with 100% accuracy, as well as individuals heterozygous at the male-sterile locus, even outside the mapping population. These markers should enable practical MAS for conifer breeding.

Effects of soil water content and elevated CO2 concentration on the monoterpene emission rate of Cryptomeria japonica.

Monoterpenes emitted from plants contribute to the formation of secondary pollution and affect the climate system. Monoterpene emission rates may be affected by environmental changes such as increasing CO2 concentration caused by fossil fuel burning and drought stress induced by climate change. We measured monoterpene emissions from Cryptomeria japonica clone saplings grown under different CO2 concentrations (control: ambient CO2 level, elevated CO2: 1000µmolmol-1). The saplings were planted in the ground and we did not artificially control the SWC. The relationship between the monoterpene emissions and naturally varying SWC was investigated. The dominant monoterpene was α-pinene, followed by sabinene. The monoterpene emission rates were exponentially correlated with temperature for all measurements and normalized (35°C) for each measurement day. The daily normalized monoterpene emission rates (Es0.10) were positively and linearly correlated with SWC under both control and elevated CO2 conditions (control: r2=0.55, elevated CO2: r2=0.89). The slope of the regression line of Es0.10 against SWC was significantly higher under elevated CO2 than under control conditions (ANCOVA: P<0.01), indicating that the effect of CO2 concentration on monoterpene emission rates differed by soil water status. The monoterpene emission rates estimated by considering temperature and SWC (Improved G93 algorithm) better agreed with the measured monoterpene emission rates, when compared with the emission rates estimated by considering temperature alone (G93 algorithm). Our results demonstrated that the combined effects of SWC and CO2 concentration are important for controlling the monoterpene emissions from C. japonica clone saplings. If these relationships can be applied to the other coniferous tree species, our results may be useful to improve accuracy of monoterpene emission estimates from the coniferous forests as affected by climate change in the present and foreseeable future.

Water retained in tall Cryptomeria japonica leaves as studied by infrared micro-spectroscopy.

Recent studies in the tallest tree species suggest that physiological and anatomical traits of tree-top leaves are adapted to water-limited conditions. In order to examine water retention mechanism of leaves in a tall tree, infrared (IR) micro-spectroscopy was conducted on mature leaf cross-sections of tall Cryptomeria japonica D. Don from four different heights (51, 43, 31 and 19 m). We measured IR transmission spectra and mainly analyzed OH (3700-3000 cm-1) and C-O (1190-845 cm-1) absorption bands, indicating water molecules and sugar groups, respectively. The changes in IR spectra of leaf sections from different heights were compared with bulk-leaf hydraulics. Both average OH band area of the leaf sections and leaf water content were larger in the upper-crown, while osmotic potential at saturation did not vary with height, suggesting higher dissolved sugar contents of upper-crown leaves. As cell-wall is the main cellular structure of leaves, we inferred that larger average C-O band area of upper-crown leaves reflected higher content of structural polysaccharides such as cellulose, hemicellulose and pectin. Infrared micro-spectroscopic imaging showed that the OH and C-O band areas are large in the vascular bundle, transfusion tissue and epidermis. Infrared spectra of individual tissue showed that much more water is retained in vascular bundle and transfusion tissue than mesophyll. These results demonstrate that IR micro-spectroscopy is a powerful tool for visualizing detailed, quantitative information on the spatial distribution of chemical substances within plant tissues, which cannot be done using conventional methods like histochemical staining. The OH band could be well reproduced by four Gaussian OH components around 3530 (free water: long H bond), 3410 (pectin-like OH species), 3310 (cellulose-like OH species) and 3210 (bound water: short H bond) cm-1, and all of these OH components were higher in the upper crown while their relative proportions did not vary with height. Based on the spectral analyses, we inferred that polysaccharides play a key role in biomolecular retention of water in leaves of tall C. japonica.

Leaf water maintains daytime transpiration in young Cryptomeria japonica trees.

Compared with stem water storage, leaf water storage is understudied although it may be important for alleviating water stress by contributing quickly and directly to transpiration demand. To quantify the relative contribution of stem versus leaf water storage to daily water deficit, we measured diurnal changes in transpiration rate, sap-flow velocity and stem radius of 10-year-old Cryptomeria japonica D. Don trees. We assumed that the duration of time lags between transpiration rate and sap-flow velocity reflected stored water in the stem and leaf, and that stem volume change represented water content of elastic tissue. The relationship between fresh mass and water potential of the whole tree indicated that the study trees had capacity to store, on average, 91.4 ml of water per kg fresh mass at turgor loss. Leaves, sapwood and elastic tissue contributed around 51%, 29% and 20% of stored water, respectively. During morning, transpiration rates were higher than sap-flow velocity suggesting depletion of stored water. During the first 2 h after onset of transpiration, stored water contributed more than 100% of whole-tree transpiration. Depletion of leaf water (PLeaf) and sapwood water (PSap) coincided with the onset of transpiration and became maximum around 15:00 h. Depletion of elastic tissue water (PElastic) lagged behind that of PLeaf and PSap by 1-2 h, indicating that replenishment of stored water occurs late in the day when low leaf water potentials resulting from daytime transpiration drive water uptake. Maximum depletion of PLeaf was about 1-3 times and 5-10 times that of PSap and PElastic, respectively. The contribution of PLeaf to total daily transpiration was 5-8%, while those of PSap and PElastic were 3-4% and 0.7-1%, respectively. Our results suggest the importance of leaf water storage in maintaining daily transpiration in young C. japonica trees.

Species characteristics and intraspecific variation in growth and photosynthesis of Cryptomeria japonica under elevated O3 and CO2.

In order to predict the effects of future atmospheric conditions on forest productivity, it is necessary to clarify the physiological responses of major forest tree species to high concentrations of ozone (O3) and carbon dioxide (CO2). Furthermore, intraspecific variation of these responses should also be examined in order to predict productivity gains through tree improvements in the future. We investigated intraspecific variation in growth and photosynthesis of Cryptomeria japonica D. Don, a major silviculture species in Japan, in response to elevated concentrations of O3 (eO3) and CO2 (eCO2), separately and in combination. Cuttings of C. japonica were grown and exposed to two levels of O3 (ambient and twice-ambient levels) in combination with two levels of CO2 (ambient and 550 µmol mol-1 in the daytime) for two growing seasons in a free-air CO2 enrichment experiment. There was no obvious negative effect of eO3 on growth or photosynthetic traits of the C. japonica clones, but a positive effect was observed for annual height increments in the first growing season. Dry mass production and the photosynthetic rate increased under eCO2 conditions, while the maximum carboxylation rate decreased. Significant interaction effects of eO3 and eCO2 on growth and photosynthetic traits were not observed. Clonal effects on growth and photosynthetic traits were significant, but the interactions between clones and O3 and/or CO2 treatments were not. Spearman's rank correlation coefficients between growth traits under ambient conditions and for each treatment were significantly positive, implying that clonal ranking in growth abilities might not be affected by either eO3 or eCO2. The knowledge obtained from this study will be helpful for species selection in afforestation programs, to continue and to improve current programs involving this species, and to accurately predict the CO2 fixation capacity of Japanese forests.

Changes in the anatomy, morphology and mycorrhizal infection of fine root systems of Cryptomeria japonica in relation to stand ageing.

Biomass allocation to fine roots often increases under soil nutrient deficiency, but the fine root biomass does not often increase in old stands, even under nutrient limitation. Therefore, in old stands, the morphology, anatomy, branching architecture and mycorrhization of fine roots may compensate efficiently for nutrient acquisition by the low fine root biomass. In this study, changes in the morphology, anatomy and arbuscular mycorrhizal infection at each branching position of fine root clusters were evaluated in relation to stand age. A chronosequence (6­90 years of age) of stands in a Cryptomeria japonica D. Don plantation was used for these analyses. The fine root size parameters, such as length, weight and tip numbers of fine root clusters, increased with stand age. The specific root tip length (SRTL) decreased with increasing stand age, suggesting that the allocation to root active portions decreased with stand age. From the anatomical observation, the ephemeral root tips increased with stand age, suggesting that root tip turnover within a root cluster was high in old stands. The proportions of proto-xylem groups among branching positions indicated that the life cycles in branching hierarchy should be clearer in old stands than that in younger stands. The increasing in the mycorrhizal infection of root tips in old stands should enhance the root tip absorptive functions. The SRTL was correlated with the wood/needle ratio, suggesting that carbon limitation as the stand ages may result in decline of carbon allocation to maintain active root tips. However, increasing of the ephemeral tips and mycorrhizal infection rates may compensate the declines of tip allocation in old stands.

Terrestrial-aquatic linkage in stream food webs along a forest chronosequence: multi-isotopic evidence.

Long-term monitoring of ecosystem succession provides baseline data for conservation and management, as well as for understanding the dynamics of underlying biogeochemical processes. We examined the effects of deforestation and subsequent afforestation of a riparian forest of Japanese cedar (Cryptomeria japonica) on stable isotope ratios of carbon (δ¹³C) and nitrogen (δ¹5N) and natural abundances of radiocarbon (Δ¹4C) in stream biota in the Mt. Gomadan Experimental Forest and the Wakayama Forest Research Station, Kyoto University, central Japan. Macroinvertebrates, periphytic algae attached to rock surfaces (periphyton), and leaf litter of terrestrial plants were collected from six headwater streams with similar climate, topography, and bedrock geology, except for the stand ages of riparian forests (from 3 to 49 yr old in five stands and > 90 yr old in one reference stand). Light intensity and δ¹³C values of both periphyton and macroinvertebrates decreased synchronously with forest age in winter. A Bayesian mixing model indicates that periphyton contributions to the stream food webs are maximized in 23-yr-old forests. Except for grazers, most macroinvertebrates showed Δ¹4C values similar to those of terrestrial leaf litter, reflecting the influence of modern atmospheric CO2 Δ¹4C values. On the other hand, the Δ¹4C values of both periphyton and grazers (i.e., aquatic primary consumers) were significantly lower than that of modern atmospheric CO2, and were lowest in 23-yr-old forest stands. Previous studies show that root biomass of C. japonica peaks at 15-30 yr after planting. These evidences suggest that soil CO2 released by root respiration and dispersed by groundwater weathers carbonate substrata, and that dissolved inorganic carbon (DIC) with low Δ¹4C is incorporated into stream periphyton and some macroinvertebrates. The ecological response in the studied streams to clear-cutting and replanting of Japanese cedar is much slower (~20 yr) than the chemical response (< 5 yr). More than 50 yr is required for the food web structure to completely recover from clear-cutting. The ecological delay is attributed to several biogeochemical factors, the understanding of which is critical to integrated management of forest-stream continuum and the prediction of ecosystem resilience in response to environmental change.

Localized cooling of stems induces latewood formation and cambial dormancy during seasons of active cambium in conifers.

BACKGROUND AND AIMS: In temperate regions, trees undergo annual cycles of cambial growth, with periods of cambial activity and dormancy. Environmental factors might regulate the cambial growth, as well as the development of cambial derivatives. We investigated the effects of low temperature by localized cooling on cambial activity and latewood formation in two conifers, Chamaecyparis obtusa and Cryptomeria japonica. METHODS: A plastic rubber tube that contained cooled water was wrapped around a 30-cm-wide portion of the main stem of Chamaecyparis obtusa and Cryptomeria japonica trees during seasons of active cambium. Small blocks were collected from both cooled and non-cooled control portions of the stems for sequential observations of cambial activity and for anatomical measurements of cell morphology by light microscopy and image analysis. KEY RESULTS: The effect of localized cooling was first observed on differentiating tracheids. Tracheids narrow in diameter and with significantly decreased cambial activity were evident 5 weeks after the start of cooling in these stems. Eight weeks after the start of cooling, tracheids with clearly diminished diameters and thickened cell walls were observed in these stems. Thus, localized low temperature induced narrow diameters and obvious thickening of secondary cell walls of tracheids, which were identified as latewood tracheids. Two months after the cessation of cooling, a false annual ring was observed and cambium became active again and produced new tracheids. In Cryptomeria japonica, cambial activity ceased earlier in locally cooled portions of stems than in non-cooled stems, indicating that the cambium had entered dormancy sooner in the cooled stems. CONCLUSIONS: Artificial cooling of stems induced latewood formation and cessation of cambial activity, indicating that cambium and its derivatives can respond directly to changes in temperature. A decrease in the temperature of the stem is a critical factor in the control of cambial activity and xylem differentiation in trees.

Equation to predict the (137)Cs leaching dynamic from evergreen canopies after a radio-cesium deposit.

The Fukushima Daiishi nuclear power plant (FDNPP) accident led to a massive radionuclide deposition mainly onto Japanese forest canopies. In our previous study, an improved double exponential (IDE) equation including rainfall intensity was proposed to estimate the (137)Cs hydrological transport from evergreen canopies to the ground. This equation used two types of parameters, kinetic (k1 and k2) and leachable stock (A1 and A2). Those parameters have been estimated by adjusting them in the IDE equation in order to accurately describe the measured cumulative leached (137)Cs from canopies (k1 = 4.2E-04-5.0E-04 d(-1), k2 = 1.2E-02-1.7E-02 d(-1), A1 = 62-99 kBq/m(2), A2 = 25-61 kBq/m(2)). In this study, we linked the total leachable stock (Aleachable, a parameter of the IDE equation corresponding to A1 + A2) to a physiological criteria (the canopy closure CC, which can be measured with a simple camera equipped with a fish-eye objective). Furthermore, the kinetic parameters measured for Japanese cedar (k1 = 5.0E-04 d(-1), k2 = 1.2E-02 d(-1), and r12 = 0.22 (r12 = A1/A2) could also be used for two other coniferous species: Japanese cypress and spruce. This suggests that these parameters could be constants for coniferous forests.

Seasonal and clonal variation in cellulose microfibril orientation during cell wall formation of tracheids in Cryptomeria japonica.

To investigate the biological mechanism by which trees control the changes in microfibril (MF) orientation among secondary cell wall layers of conifer tracheids, we studied seasonal variation in the orientation of newly deposited MFs during tracheid cell wall development in Japanese cedar (Cryptomeria japonica D. Don) trees growing in Central Japan (36°36'N, 140°39'E). Sample blocks were repeatedly collected from four 16-year-old clones of different origins during the growing season of 2010 to investigate the hypotheses that changes in cellulose MF orientation between wall layers exhibited seasonal and clonal differences. The progressive change in the orientation of newly deposited MFs on the primary and secondary cell wall layers of tracheids was detected by field-emission-scanning electron microscopy. Tracheid production and differentiation was studied by light microscopy. We observed a decreasing trend in the orientation of deposited MFs from earlywood to latewood in the S2 and S1 layers, where MFs appeared in a Z-helix. In contrast, no seasonal pattern in the orientation of the MFs in the S-helix was observed. Minor clonal variation was observed in the phenology of tracheid production and differentiation. We concluded that a seasonal decreasing trend in the orientation of the MFs in the Z-helix in S1 and S2 was present, whereas the MFs in other layers exhibited minor random variations. Thus, the orientation of the MFs in S2 was affected by seasonal factors, whereas the MFs in other layers were more intrinsically controlled. The within-ring variations in the MF orientation and thus the resulting average MF angle might also be related to genotypic differences in the tracheid production and differentiation rate. However, our results do not exclude other intrinsic and environmental regulations in the change in MF orientation, which remains a topic for future studies.

Azimuthal and radial variations in sap flux density and effects on stand-scale transpiration estimates in a Japanese cedar forest.

Understanding radial and azimuthal variation, and tree-to-tree variation, in sap flux density (Fd) as sources of uncertainty is important for estimating transpiration using sap flow techniques. In a Japanese cedar (Cryptomeria japonica D. Don.) forest, Fd was measured at several depths and aspects for 18 trees, using heat dissipation (Granier-type) sensors. We observed considerable azimuthal variation in Fd. The coefficient of variation (CV) calculated from Fd at a depth of 0-20 mm (Fd1) and Fd at a depth of 20-40 mm (Fd2) ranged from 6.7 to 37.6% (mean = 28.3%) and from 19.6 to 62.5% (mean = 34.6%) for the -azimuthal directions. Fd at the north aspect averaged for nine trees, for which azimuthal measurements were made, was -obviously smaller than Fd at the other three aspects (i.e., west, south and east) averaged for the nine trees. Fd1 averaged for the nine trees was significantly larger than Fd2 averaged for the nine trees. The error for stand-scale transpiration (E) estimates caused by ignoring the azimuthal variation was larger than that caused by ignoring the radial variation. The error caused by ignoring tree-to-tree variation was larger than that caused by ignoring both radial and azimuthal variations. Thus, tree-to-tree variation in Fd would be more important than both radial and azimuthal variations in Fd for E estimation. However, Fd for each tree should not be measured at a consistent aspect but should be measured at various aspects to make accurate E estimates and to avoid a risk of error caused by the relationship of Fd to aspect.

A novel approach of preventing Japanese cedar pollen dispersal that is the cause of Japanese cedar pollinosis (JCP) using pollen-specific fungal infection.

In Japan, Japanese cedar pollen dispersal is one of the major causes of pollinosis. Sydowia japonica is an ascomycetous fungus that grows exclusively on the male strobili of Japanese cedar, suggesting a possible mechanism for controlling pollen dispersal. To evaluate this possibility, eleven isolates of S. japonica were collected from around Japan and used as an inoculum to male strobili of Japanese cedar. The treatment demonstrated that the fungus infected only the pollen and prevented pollen dispersal. The fungus did not cause any additional symptoms to other parts of Japanese cedar, such as needles, stems, and buds. All S. japonica isolates collected around Japan could serve to control pollen dispersal. Periodic observation of the fungal pathogenesis with stereomicroscope and scanning electron microscope showed that hyphal fragments and conidia of S. japonica germinated on the surface of male strobili, and the germ tube entered pollen sacs through opening microsporophylls. Within the pollen sacs, the hyphae penetrated pollen gradually, such that all pollen was infected by the fungus by approximately one month before the pollen dispersal season. The infected pollen was destroyed due to the fungal infection and was never released. Our data suggests a novel approach of preventing pollen dispersal using pollen-specific fungal infection.

Inflammatory and degranulation effect of yellow sand on RBL-2H3 cells in relation to chemical and biological constituents.

Recent studie pointed out that allergic diseases have increased during the Asian dust storm event (ADSE) in Japan. Daily observations and the atmospheric concentrations of yellow sand (YS) aerosol have been increasing. In this study, YS samples collected from three sites of Japan during ADSE in 2009-2010 were used. The particles were analyzed by X-ray photoelectron spectroscopy (XPS) and X-ray fluorescence-energy dispersive spectrometer (XRF-EDS). We investigate ability of YS extract on enhancing the chemical mediator release and cytokine production from rat basophilic leukemia (RBL-2H3) cells. The dust particles at Fukuoka and Tsukuba were abundant in aluminum (Al), iron (Fe), potassium (K) and titan (Ti) than those at Naha. Concentration of the trace endotoxin and Cryptomeria japonica pollen allergen (Cry j 1) were measured in YS extract. After exposure of RBL-2H3 cells to YS extract, the ß-hexosaminidase (ß-hex) release, tumor necrosis factor-alpha (TNF-α) production were enhanced in RBL-2H3 cells. This process depends on endotoxin, Cry j 1 and other allergen present in the YS extract. YS water extract also show a strong cytotoxic effect on the cells. This data suggest that low levels of endotoxin and Cry j 1 in YS may cause allergy during the ADSE.