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The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 nuclease system is a versatile and essential biotechnological tool in the life sciences that allows efficient genome editing. When generating gene-edited trees, T0-generation plants are often used for subsequent analysis because of the time that is required to obtain the desired mutants via crossing. However, T0-generation plants exhibit various unexpected mutations, which emphasizes the need to identify mutants with expected mutation patterns. The two critical checkpoints in this process are to confirm the expected mutation patterns in both alleles and to exclude somatic chimeric plants. In this study, we generated gene-edited Cryptomeria japonica plants and established a method to determine chimerism and mutation patterns using fragment analysis and Oxford Nanopore Technologies (ONT)-based amplicon sequencing. In the first screening, fragment analysis, i.e. indel detection via amplicon analysis, was used to predict indel mutation patterns in both alleles and to discriminate somatic chimeric plants in 188 candidate mutants. In the second screening, we precisely determined the mutation patterns and chimerism in the mutants using ONT-based amplicon sequencing, where confirmation of both alleles can be achieved using allele-specific markers flanking the single guide RNA target site. In the present study, a bioinformatic analysis procedure was developed and provided for the rapid and accurate determination of DNA mutation patterns using ONT-based amplicon sequencing. As ONT amplicon sequencing has a low running cost compared with other long-read analysis methods, such as PacBio, it is a powerful tool in plant genetics and biotechnology to select gene-edited plants with expected indel patterns in the T0-generation. [ABSTRACT FROM AUTHOR]

Cry j 1 is a major allergen present in Japanese cedar (Cryptomeria japonica) pollens. Peptides with the core sequence of KVTVAFNQF from Cry j 1 ('pCj1') bind to HLA-DP5 and activate Th2 cells. In this study, we noticed that Ser and Lys at positions −2 and −3, respectively, in the N-terminal flanking (NF) region to pCj1 are conserved well in HLA-DP5-binding allergen peptides. A competitive binding assay showed that the double mutation of Ser(–2) and Lys(–3) to Glu [S(P–2)E/K(P–3)E] in a 13-residue Cry j 1 peptide (NF-pCj1) decreased its affinity for HLA-DP5 by about 2-fold. Similarly, this double mutation reduced, by about 2-fold, the amount of NF-pCj1 presented on the surface of mouse antigen-presenting dendritic cell line 1 (mDC1) cells stably expressing HLA-DP5. We established NF-pCj1-specific and HLA-DP5-restricted CD4+ T-cell clones from HLA-DP5 positive cedar pollinosis (CP) patients, and analyzed their IL-2 production due to the activation of mouse TG40 cells expressing the cloned T-cell receptor by the NF-pCj1-presenting mDC1 cells. The T-cell activation was actually decreased by the S(P–2)E/K(P–3)E mutation, corresponding to the reduction in the peptide presentation by this mutation. In contrast, the affinity of NF-pCj1·HLA-DP5 for the T-cell receptor was not affected by the S(P–2)E/K(P–3)E mutation, as analyzed by surface plasmon resonance. Considering the positional and side-chain differences of these NF residues from previously reported T-cell activating sequences, the mechanisms of enhanced T-cell activation by Ser(–2) and Lys(–3) of NF-pCj1 may be novel. [ABSTRACT FROM AUTHOR]

The sapwood of Japanese cedar ( Cryptomeria japonica D. Don ) was decayed by the brown-rot fungus Fomitopsis palustris under bright and dark conditions. Scanning electron microscopy revealed the presence of mycelia inside the wood even after 1 week from the start of fungal exposure. Moreover, holes were observed in the torus after fungal exposure. Ruthenium red staining revealed that the pectin in pits was largely absent for 3 weeks. These events occurred before the mass loss of wood samples was confirmed at the early stage. Moreover, Fp PG28A was more highly expressed at the hyphal front on a pectin-containing medium under dark conditions compared with bright conditions. This up-regulation under dark conditions indicated that the pectin decomposition ability was promoted inside the wood where light could not reach. In conclusion, we suggest that the brown-rot fungus completed its hyphal expansion within the wood via pectin decomposition in pits before holocellulose decomposition. [ABSTRACT FROM AUTHOR]

Introductory biology lessons around the world typically teach that plants absorb water through their roots, but, unfortunately, absorption of water through leaves and subsequent transport and use of this water for biomass formation remains a field limited mostly to specialists. Recent studies have identified foliar water uptake as a significant net water source for terrestrial plants. The growing interest in the development of a new model that includes both foliar water uptake (in liquid form) and root water uptake to explain hydrogen and oxygen isotope ratios in leaf water and tree rings demands a method for distinguishing between these two water sources. Therefore, in this study, I have devised a new labelling method that utilizes two different water sources, one enriched in deuterium (HDO + D2O; δ D  = 7.0 × 10 4‰, δ18O = 4.1‰) and one enriched in oxygen-18 (H218O; δ D  = −85‰, δ18O = 1.1 × 104‰), to simultaneously label both foliar-absorbed and root-absorbed water and quantify their relative contributions to plant biomass. Using this new method, I here present evidence that, in the case of well-watered Cryptomeria japonica D. Don, hydrogen and oxygen incorporated into new leaf cellulose in the rainy season derives mostly from foliar-absorbed water (69% from foliar-absorbed water and 31% from root-absorbed water), while that of new root cellulose derives mostly from root-absorbed water (20% from foliar-absorbed water and 80% from root-absorbed water), and new branch xylem is somewhere in between (55% from foliar-absorbed water and 45% from root-absorbed water). The dual-labelling method first implemented in this study enables separate and simultaneous labelling of foliar-absorbed and root-absorbed water and offers a new tool to study the uptake, transport and assimilation processes of these waters in terrestrial plants. [ABSTRACT FROM AUTHOR]

Moso bamboo expansions into Japanese cedar forests are common. The expansion effects on soil nitrous oxide (N2O) emissions have not been thoroughly understood, and the underlying microbial mechanisms remain unclear. We studied bacterial and fungal contribution to soil N2O emissions under moso bamboo or Japanese cedar by applying bacterial or fungal inhibitors using streptomycin and iprodione, respectively. Soil N2O emissions were measured and the relative contribution of bacteria and fungi to soil N2O emissions was calculated. N2O emission from soil with moso bamboo was significantly higher than under Japanese cedar. Compared with control, bacterial or fungal inhibitor or their combination decreased N2O emissions, indicating substantial contribution of microbial activities to N2O emissions. However, the relative contribution of bacteria and fungi to N2O emissions was not affected by plants. Soil organic carbon, total and ammonium nitrogen were lower in soil under moso bamboo than Japanese cedar, suggesting faster microbial decomposition under moso bamboo. Fungal inhibitor and plants interactively affected soil pH, total phosphorus and ammonium nitrogen, while bacterial inhibitor and plants interactively affected total nitrogen, indicating substantial dependence of effects by microbial communities on plant species. Moso bamboo and Japanese cedar differed in their effects on soil N2O emissions with higher emissions under moso bamboo. Stimulation of N2O emission under moso bamboo might occur due to higher nitrogen mineralization and subsequent denitrification induced by high root exudation. These results highlight the need to consider the effect of species shifts on N2O emissions in forests. [ABSTRACT FROM AUTHOR]

The mechanisms regulating spring phenology have been extensively studied in angiosperm species. However, given that gymnosperms and angiosperms diverged 300 million years ago, phenology may be triggered by different cues in gymnosperm species. The regulatory mechanisms of phenology in subtropical regions remain largely unknown. In combination, it remains untested whether subtropical gymnosperm species have chilling requirements and are photosensitive. We conducted a climate chamber experiment with three chilling and three photoperiod treatments to investigate budburst during an 8-week forcing period. We tested whether budburst of eight gymnosperms species (Cryptomeria japonica , Cunninghamia lanceolata , Cupressus funebris , Ginkgo biloba , Metasequoia glyptostroboides , Pinus massoniana , Pseudolarix amabilis and Podocarpus macrophyllus) was photoperiod sensitive or has strong chilling requirements and whether photoperiod or chilling was more important for advancing budburst. Chilling advanced budburst and increased the percentage of budburst for gymnosperm species. Gymnosperm species required moderate chilling days to advance budburst. Interestingly, the forcing requirement for gymnosperm species was higher than that for angiosperms in the same forest, suggesting that gymnosperms may need more cumulative forcing to initiate budburst than do angiosperms. Compared with temperate gymnosperm species in Germany (194–600 °C days), the subtropical species studied here had a much higher forcing requirement (814–1150 °C days). The effects of photoperiod were minor, suggesting that chilling outweighs photoperiod in advancing budburst of gymnosperm species in this subtropical region. These results reveal that increased winter temperatures with continued global warming may impact not only angiosperms but also gymnosperms, leading to their delayed spring budburst. [ABSTRACT FROM AUTHOR]

As a method of evaluating the effect of inactivators on allergens while suppressing the effect of inactivator on the assay, we developed new dot-blot method that combines immunostaining and protein detection methods. This method is useful for evaluating whether the inactivator can inactivate allergens rather than removing them from the assay. [ABSTRACT FROM AUTHOR]

We investigated the latitudinal variation in radial growth phenology (onset, cessation, and duration of radial growth) of Cryptomeria japonica D. Don trees in Japan. The radial growth was observed using dendrometers in 16 stands at multiple latitudes throughout Japan. The onset of radial growth showed a clear latitudinal gradient: at high-latitude sites, C. japonica growth started later than at low-latitude sites. However, cessation of radial growth was independent of latitude. Consequently, the duration of radial growth (defined based on the onset and cessation dates) showed a weak but significant latitudinal gradient: at high-latitude sites, C. japonica trees grew for a shorter period than trees at low-latitude sites. The onset and duration of growth were respectively strongly and weakly affected by temperature, although growth cessation was not explained by temperature or precipitation. The latitudinal gradient for duration of radial growth weakly but significantly supports the hypothesis that regional variation in growth phenology is a key factor responsible for the regional variation in long-term growth. Furthermore, we found that growth duration was more variable at low latitudes than at high latitudes. [ABSTRACT FROM AUTHOR]

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 (PElasticm) 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 PElasticm, respectively. The contribution of PLeaf to total daily transpiration was 5-8%, while those of PSap and PElasticm 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. [ABSTRACT FROM AUTHOR]

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 microspectroscopic 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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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 (6-90 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. [ABSTRACT FROM AUTHOR]

Matrix-assisted laser desorption/ionization mass spectrometric imaging (MALDI-MSI) was employed to detect monolignol glucosides in differentiating normal and compression woods of two Japanese softwoods, Chamaecyparis obtusa and Cryptomeria japonica. Comparison of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry collision-induced dissociation fragmentation analysis and structural time-of-flight (MALDI-TOF CID-FAST) spectra between coniferin and differentiating xylem also confirmed the presence of coniferin in differentiating xylem. However, as matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and MALDI-TOF CID-FAST spectra of sucrose were similar to those of coniferin, it was difficult to distinguish the distribution of coniferin and sucrose using MALDI-MSI and collision-induced dissociation measurement only. To solve this problem, osmium tetroxide vapor was applied to sections of differentiating xylem. This vapor treatment caused peak shifts corresponding to the introduction of two hydroxyl groups to the C=C double bond in coniferin. The treatment did not cause a peak shift for sucrose, and therefore was effective in distinguishing coniferin and sucrose. Thus, it was found that MALDI-MSI combined with osmium tetroxide vapor treatment is a useful method to detect coniferin in differentiating xylem. [ABSTRACT FROM AUTHOR]

The molecular mechanisms that control male strobilus development in conifers are largely unknown because the developmental stages and related genes have not yet been characterized. The determination of male strobilus developmental stages will contribute to genetic research and reproductive biology in conifers. Our objectives in this study were to determine the developmental stages of male strobili by cytological and transcriptome analysis, and to determine the stages at which aberrant morphology is observed in a male-sterile mutant of Cryptomeria japonica D. Don to better understand the molecular mechanisms that control male strobilus and pollen development. Male strobilus development was observed for 8 months, from initiation to pollen dispersal. A set of 19,209 expressed sequence tags (ESTs) collected from a male reproductive library and a pollen library was used for microarray analysis. We divided male strobilus development into 10 stages by cytological and transcriptome analysis. Eight clusters (7324 ESTs) exhibited major changes in transcriptome profiles during male strobili and pollen development in C. japonica. Two clusters showed a gradual increase and decline in transcript abundance, respectively, while the other six clusters exhibited stage-specific changes. The stages at which the male sterility trait of Sosyun was expressed were identified using information on male strobilus and pollen developmental stages and gene expression profiles. Aberrant morphology was observed cytologically at Stage 6 (microspore stage), and differences in expression patterns compared with wild type were observed at Stage 4 (tetrad stage). [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Japanese cedar ( Cryptomeria japonica) is a major species in artificial Japanese forests. The Halomonas sp. KM-1 was recently isolated and found to grow effectively on saccharified Japanese cedar wood, resulting in the intracellular storage of poly-( R)-3-hydroxybutyric acid ( PHB) under aerobic conditions. Under microaerobic conditions, the extracellular secretion of ( R)-3-hydroxybutyric acid (( R)-3- HB) led to the degradation of intracellular PHB. In this study, the production of PHB and the secretion of ( R)-3- HB using saccharified Japanese cedar were much improved in cultures that were grown in the presence of urea. The level of intracellular PHB production after 36 h under aerobic cultivation was 23·6 g l−1; after shifting to microaerobic conditions for 24 h, the ( R)-3- HB concentration in the medium reached 21·1 g l−1. Thus, KM-1 efficiently utilizes saccharified Japanese cedar to produce PHB and secretes ( R)-3- HB, making it a practical candidate for use in the industrial production of ( R)-3- HB. Significance and Impact of the Study Japanese cedar is a major species grown in artificial Japanese forests, and its thinning is crucial for the health of artificial forests and the Japanese economy. Halomonas sp. KM-1 grew effectively on saccharified Japanese cedar wood, resulting in intracellular storage of poly-( R)-3-hydroxybutyric acid ( PHB) under aerobic conditions. Under microaerobic conditions, extracellular secretion of ( R)-3-hydroxybutyric acid (( R)-3- HB) caused intracellular PHB degradation. ( R)-3- HB is a chiral compound that is useful in the chemical, health food and pharmaceutical industries. The production of PHB and secretion of ( R)-3- HB using saccharified wood was dramatically improved, which may positively affect its future industrial production. [ABSTRACT FROM AUTHOR]

Background and Aims Distribution shifts and natural selection during past climatic changes are important factors in determining the genetic structure of forest species. In particular, climatic fluctuations during the Quaternary appear to have caused changes in the distribution ranges of plants, and thus strongly affected their genetic structure. This study was undertaken to identify the responses of the conifer Cryptomeria japonica, endemic to the Japanese Archipelago, to past climatic changes using a combination of phylogeography and species distribution modelling (SDM) methods. Specifically, this study focused on the locations of refugia during the last glacial maximum (LGM). Methods Genetic diversity and structure were examined using 20 microsatellite markers in 37 populations of C. japonica. The locations of glacial refugia were assessed using STRUCTURE analysis, and potential habitats under current and past climate conditions were predicted using SDM. The process of genetic divergence was also examined using the approximate Bayesian computation procedure (ABC) in DIY ABC to test the divergence time between the gene pools detected by the STRUCTURE analysis. Key Results STRUCTURE analysis identified four gene pools: northern Tohoku district; from Chubu to Chugoku district; from Tohoku to Shikoku district on the Pacific Ocean side of the Archipelago; and Yakushima Island. DIY ABC analysis indicated that the four gene pools diverged at the same time before the LGM. SDM also indicated potential northern cryptic refugia. Conclusions The combined evidence from microsatellites and SDM clearly indicates that climatic changes have shaped the genetic structure of C. japonica. The gene pool detected in northern Tohoku district is likely to have been established by cryptic northern refugia on the coast of the Japan Sea to the west of the Archipelago. The gene pool in Yakushima Island can probably be explained simply by long-term isolation from the other gene pools since the LGM. These results are supported by those of SDM and the predicted divergence time determined using ABC analysis. [ABSTRACT FROM PUBLISHER]

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. [ABSTRACT FROM AUTHOR]

Background and Aims Heartwood formation is a unique phenomenon of tree species. Although the accumulation of heartwood substances is a well-known feature of the process, the accumulation mechanism remains unclear. The aim of this study was to determine the accumulation process of ferruginol, a predominant heartwood substance of Cryptomeria japonica, in heartwood-forming xylem. Methods The radial accumulation pattern of ferruginol was examined from sapwood and through the intermediate wood to the heartwood by direct mapping using time-of-flight secondary ion mass spectrometry (TOF-SIMS). The data were compared with quantitative results obtained from a novel method of gas chromatography analysis using laser microdissection sampling and with water distribution obtained from cryo-scanning electron microscopy. Key Results Ferruginol initially accumulated in the middle of the intermediate wood, in the earlywood near the annual ring boundary. It accumulated throughout the entire earlywood in the inner intermediate wood, and in both the earlywood and the latewood in the heartwood. The process of ferruginol accumulation continued for more than eight annual rings. Ferruginol concentration peaked at the border between the intermediate wood and heartwood, while the concentration was less in the latewood compared wiht the earlywood in each annual ring. Ferruginol tended to accumulate around the ray parenchyma cells. In addition, at the border between the intermediate wood and heartwood, the accumulation was higher in areas without water than in areas with water. Conclusions TOF-SIMS clearly revealed ferruginol distribution at the cellular level. Ferruginol accumulation begins in the middle of intermediate wood, initially in the earlywood near the annual ring boundary, then throughout the entire earlywood, and finally across to the whole annual ring in the heartwood. The heterogeneous timing of ferruginol accumulation could be related to the distribution of ray parenchyma cells and/or water in the heartwood-forming xylem. [ABSTRACT FROM PUBLISHER]

This study analyzed wind damage caused by tropical storms from 1991 to 2007 to Japanese forests mainly consisting of Cryptomeria japonica. Statistical analyses based on logistic regression and Cox regression models were conducted in relation to conditions at the forest and stand level. Known damage to forests managed by Kyusyu Rinsan Corporation (KR forests), located on the Kyushu Island, was analyzed at the forest level, using tropical storm characteristics such as air pressure, precipitation and periods when the forests were within the storm zone as predictors. Wind damage was also examined at the stand level (150 analysis points) using Cox regression models, according to stand age, site index, terrain conditions, management practices and wind velocity indicators (horizontal and vertical velocity vectors). The results indicated that at the forest level, higher maximum hourly wind speed and longer periods of >15 m s−1 of wind speed were significantly correlated to damage occurrence. At the stand level, indicators of upward vertical velocity, thinning treatments and site index were positively associated with the probability of wind damage. For instance, stands receiving higher upward vertical velocities and thinning treatment within 2 years were more likely to have reduced stability against tropical storms. Stands with higher and lower site index than average also showed lower stability in our analysis. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM PUBLISHER]

We measured linkage disequilibrium in mostly noncoding regions of Cryptomeria japonica, a conifer belonging to Cupressaceae. Linkage disequilibrium was extensive and did not decay even at a distance of 100 kb. The average estimate of the population recombination rate per base pair was 1.55 x 10-5 and was ,1/70 of that in the coding regions. We discuss the impact of low recombination rates in a large part of the genome on association studies. [ABSTRACT FROM AUTHOR]

We estimate a suitable strip-gap arrangement in Japanese cedar (Cryptomeria japonica) plantations to promote the growth of beech (Fagus crenata) seedlings and control dwarf bamboo (Sasa kurilensis) in mountainous areas using a hemispherical model and United States Geological Survey Digital Elevation Model (DEM). Clarification was also sought on whether the gap arrangement should be altered with topography and original stand structure. Nine modelled plantations were established by combining tree inventory and DEM data from three C.japonica plantations. We simulated spatial and temporal variations in photosynthetic photon flux density in stands 5 years after strip-gap creation under six scenarios for each modelled stand and integrated the light responses of F. crenata seedlings and S. kurikensis into the model. Finer strip-gap mosaics with gap width narrower than half the tree height inhibit the growth of S. kurikensis and provide a wider area suitable for F. crenata growth. Our simulation indicated a suitable strip-gap arrangement should be based on the stand structure but not necessarily on topography. However, it indicated that the possible range of practical gap arrangements changed with topography. The decision-support model for gap arrangement used in this study combined with DEM data provides flexible gap creation options adaptable to specific plantations in mountainous areas. [ABSTRACT FROM AUTHOR]

Background and Aims Cambial reactivation in trees occurs from late winter to early spring when photosynthesis is minimal or almost non-existent. Reserve materials might be important for wood formation in trees. The localization and approximate levels of starch and lipids (as droplets) and number of starch granules in cambium and phloem were examined from cambial dormancy to the start of xylem differentiation in locally heated stems of Cryptomeria japonica trees in winter. Methods Electric heating tape was wrapped on one side of the stem of Cryptomeria japonica trees at breast height in winter. The localization and approximate levels of starch and lipids (as droplets) and number of starch granules were determined by image analysis of optical digital images obtained by confocal laser scanning microscopy. Key Results Localized heating induced earlier cambial reactivation and xylem differentiation in stems of Cryptomeria japonica, as compared with non-heated stems. There were clear changes in the respective localizations and levels of starch and lipids (as droplets) determined in terms of relative areas on images, from cambial dormancy to the start of xylem differentiation in heated stems. In heated stems, the levels and number of starch granules fell from cambial reactivation to the start of xylem differentiation. There was a significant decrease in the relative area occupied by lipid droplets in the cambium from cambial reactivation to the start of xylem differentiation in heated stems. Conclusions The results showed clearly that the levels and number of storage starch granules in cambium and phloem cells and levels of lipids (as droplets) in the cambium decreased from cambial reactivation to the start of xylem differentiation in heated stems during the winter. The observations suggest that starch and lipid droplets might be needed as sources of energy for the initiation of cambial cell division and the differentiation of xylem in Cryptomeria japonica. [ABSTRACT FROM AUTHOR]

The article discusses the discovery of a lipid transfer protein (LPT) identified as an immunoglobulin E (IgE)-binding antigen from the pollen of Cryptomeria japonica, also known as Japanese cedar. The discovered antigen, named CJP-8, along with cDNA would encode a 165-amino acid polypeptide that possess the conserved eight cysteines aspects of plant LTP family members.

The article presents a single-blind, placebo-controlled study that investigates the effect of Lactobacillus plantarum strain HSK201 on Japanese cedar pollinosis. It states that the HSK201 strain suppressed both serum Japanese cedar pollen-specific IgE elevation and T cell type ½ ratio reduction at the highest point of pollen dispersion. The study shows that ingesting HSK201 strain alleviates the symptoms of pollinosis when symptoms are mild and pollen exposure is low.

The article presents a study that describes single-chain Fv (scFv) antibodies isolation against Cryj1 and Cryj2, two allergens in the pollen of sugi (Cryptomeria japonica). It explains that recombinant phages producing scFv antibodies bound to either Cryj1 or Cryj2 were isolated through selection with immobilized antigens. It reveals that scFv antibodies against Cryj1 were bound to four non-overlapping epitopes and Cryj2-specific antibodies inhabited each other's binding to Cryj2.

One important mortality factor of Semanotus japonicus Lacordaire (Coleoptera: Cerambycidae) larvae in the trunks of Cryptomeria japonica (L.f.) D. Don (Taxodiaceae) is the presence of oleoresin secreted from traumatic resin canals (TRCs) in the phloem. The present 2-yr investigation examined whether the extent of feeding in the phloem and/or the survival of S.japonicus larvae were negatively influenced by TRCs occurred by Epinotia granitalis (Butler) (Lepidoptera: Tortricidae) larval feeding when these larvae fed simultaneously in the same trunk. The survival of S. japonicus larvae until they reach the xylem, the damage to the xylem, and emergence were investigated. The occurrence of TRCs at the E. granitalis feeding sites was significantly higher at the 1-3-yr-old annual layer compared with nonfeeding sites, suggesting that E. granitalis feeding induces the occurrence. The extent of feeding in the phloem by S. japonicus larvae that were dead there tended to be shorter with number of E. granitalis feeding sites, but the extent of some larvae was long in spite of the existence of E. granitalis feeding, suggesting that larvae feeding opposite to E. granitalis feeding sites could not be influenced by TRCs occurred by E. granitalis. The survival of S. japonicus larvae until they reached xylem did not decrease with the number of E. granitalis feeding sites because preformed TRCs considerably influenced the survival, whereas the survival until damaging the xylem and until emerging decreased. These results indicate that E. granitalis feeding induces the occurrence of TRCs in C. japonica and negatively influences the feeding behavior and survival of S.japoniens, although the survival varies with the feeding direction in respect to E. granitalis feeding sites or the existence of preformed TRCs. [ABSTRACT FROM AUTHOR]

The article discusses a diastereoselective synthesis of a bisabolane-type sesquiterpene, (1S,6R)-1-hydroxy-2,7(14),10-bisabolatrien-4-one. The synthesis of the compound, an antifeedant against Locusta migratoria and Acusta despesta, was produced from Cryptomerica japonica or Japanese cedar, from (R)-(―)-carvone through (R)-(―)-cryptomerione. It notes that the enantiomer was turned into (1S,3R,6R)-hydoxy-7(14),10-bisabola-dien-4-one by enone moiety's 1,4-selective reduction.

Sequence similarity between genes of angiosperms and conifers was analyzed to identify genes specific to conifer lineages. Using the BLAST and FASTA algorithms, we identified 156 putative conifer-specific transcripts among 5253 unique transcripts of Cryptomeria japonica D. Don. For more than half of the 156 conifer-specific transcripts obtained, the regions conserved among conifers corresponded to the predicted open reading frames, suggesting that the obtained transcripts encode functional proteins. Four of the putative conifer-specific transcripts appeared to be tissue-specific; three were specific to reproductive organs and the other was specific to wood-forming tissues. Comparisons of highly conserved nucleotide sequences in conifer lineages indicated that the nucleotide compositions of orthologous genes were similar, although no common features were found among the conifer-specific genes. [ABSTRACT FROM AUTHOR]

A decision support-based approach has been developed in order to help recommend silvicultural treatments for reducing typhoon wind damage in Japanese forests. A case study was conducted on three management scenarios (no thinning, light thinning and heavy thinning) for sugi (Cryptomeria japonica (L.f.) D.Don) stands in Himi, Toyama Prefecture, Japan. The decision support approach integrated models and tools including a mechanistic/empirical wind damage risk assessment model ForestTYPHOON, which includes a modified version of the wind damage risk model, geographical analysis of the losses and effects of storms, and an airflow model, wind atlas analysis and application program. A growth model Silve-no-Mori was linked with ForestTYPHOON to estimate wind damage risk over a SO-year period. After assessing the wind damage risk, risk stands were displayed using a geographic information system. In addition, decision tree analysis provided information on stand characteristics related to wind damage. Approximately 90 per cent agreement was found between the wind damage assessment using ForestTYPHOON and the outputs of the decision trees. The decision trees showed that top height was the most important stand characteristic and provided a critical top height at which silvicultural treatments need to be modified. If the top height exceeds the critical height, any treatments including thinning should be avoided to minimize wind damage risk. [ABSTRACT FROM AUTHOR]

The feeding preference of Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae) for 200-kGy gamma-irradiated Cryptomeria japonica D. Don (Japanese cedar) sapwood impregnated with benzoylphenylurea compounds such as hexaflumuron and noviflumuron was examined by three laboratory tests. Although termites were not deterred from feeding on gamma-irradiated wood samples that had been impregnated with hexaflumuron or noviflumuron, termite mortality was significantly higher compared with solvent controls in the no-choice test. All live termites were transferred to paper disks immediately after the no-choice test to investigate changes in mortality with time, and this test also confirmed the effects of hexaflumuron and noviflumuron on worker termites, which showed a significant feeding preference for gamma-irradiated wood. Only the 1480 ppm noviflumuron-impregnated gamma-irradiated wood specimens showed significant differences in mortality in the two-choice test. These results suggest that gamma-irradiated C. japonica wood, which is locally abundant in Japan, may have potential as a bait substrate for benzoylphenylurea compounds. [ABSTRACT FROM AUTHOR]

The article reports on results of a study of antifeedants against the insect species, Locusta migratoria from the Japanese cedar, Cryptomeria japonica II. A description of the experimental set-up and measurement methods is presented. The study showed that each compound separately showed weak activity. However, they showed intense activity against Locusta migratoria when they were combined.

We report the isolation and characterization of CjNdly, a homolog in Japanese cedar (Cryptomeria japonica D. Don) of the FLORICAULA/LEAFY (FLO/LFY) genes. We determined the entire nucleotide sequence of CjNdly, including short 5″- and 3″-untranslated regions. The deduced amino acid sequence was similar to those of the products of the FLO/LFY genes from other species. The nucleotide sequence showed the closest homology to that of the NEEDLY gene in Pinus radiata D. Don. Although no proline-rich region has been reported previously in homologous gene products from gymnosperms, we found such a region at the amino-terminal end of the deduced amino acid sequence encoded by CjNdly. We detected the expression of CjNdly in both reproductive and vegetative tissues and organs of C. japonica. Heterologous expression of CjNdly in transgenic tobacco plants induced precocious flowering of regenerating shoots on agar-solidified medium and flowers with an abnormal phenotype, namely, petal-like stamens. Our findings suggest that the CjNdly gene may have important roles in flower development in Japanese cedar, resembling those of its angiosperm homologs. [ABSTRACT FROM AUTHOR]

The article examines the effectiveness of the polysaccharide derived from black currant, named cassis polysaccharide (CAPS), for inhibiting Japanese cedar pollinosis symptoms and improving quality of life by a randomized double-blind, placebo-controlled trial in 2006. According to the authors, a smaller degree of final symptom aggravation was found in the CAPS group, although there was no significant difference between the CAPS and placebo group in the weekly mean value of any symptom.

The article discusses the identification of the antifeedants in Cryptomeria japonica against Locusta migratoria L. in a multi-component system requiring multiple components to exhibit its activity. The compounds, (1S,6R)-2,7(14),10-bisabolatrien-1-ol-4-one and (+)-7(14),10-bisaboladien-1-ol-4-one, strongly inhibited the feeding of Locusta migratoria only when they were combined, but each compound alone did not show any activity.

We determined the amount of information needed to estimate watershed-scale transpiration in a Japanese cedar (Cryptomeria japonica D. Don) forest from sap flow measurements of individual trees. Measurements of tree biometrics (diameter at breast height (DBH) and tree sapwood area (AS_tree)), and tree-to-tree and radial variations in xylem sap flux density (Fd) were made in two stand plots, an upper slope plot (UP) and a lower slope plot (LP), during a growing season characterized by significant variations in environmental factors. We then investigated how mean stand sap flux density (JS) and a tree stem allometric relationship (DBH--AS_tree) varied between the stands. Appropriate sample sizes for estimating representative JS values were determined. Both a unique and a general function allowed description of the allometric relationship along the slope, but the data for its formulation was required for both the UP and LP. Values of JS in the UP and LP were similar during the study period despite differences in tree density and size between the plots, implying that JS measured in a partial stand in a watershed is a reasonable estimate of JS in other stands in the watershed, and that stand sapwood area calculated from AS_tree is a strong determinant of water use in a forest watershed. To estimate JS in both the UP and LP, it was necessary to sample at least 10 trees in each plot. [ABSTRACT FROM AUTHOR]

Secondary metabolites called norlignans are produced in the xylem of Cryptomeria japonica D. Don. Several norlignans have roles in the defense of sapwood against microbial invasion and in the coloration of heartwood. The biosynthetic pathway of norlignans is largely unknown. Norlignans have been reported to accumulate in the sapwood during the drying of C. japonica logs. To search for genes encoding enzymes that catalyze the synthesis of norlignans, we carried out suppression subtractive hybridization using the fresh sapwood of a felled log and the drying sapwood in which a norlignan, agatharesinol, accumulated. A total of 1050 expressed sequence tags were obtained from the subtracted cDNA library, and these were assembled into 146 contigs and 361 singletons. Of these 507 unique sequences, 263 were functionally classified into 12 categories. “Metabolism” was the largest category, with 23% (61) of classified sequences. Twenty-six sequences that encode 16 enzymes were assigned to “secondary metabolism.” Expression analysis of 15 genes related to “secondary metabolism” revealed that 12 of these genes had transcripts that were induced during the sapwood drying process. Of the 12 genes, 10 encoded enzymes that use aromatic compounds as substrates. In addition, 58 sequences representing 22 defense-related proteins were found. Our subtraction library should be a useful source for isolating genes encoding proteins involved in secondary metabolism including norlignan biosynthesis and defense in C. japonica xylem. [ABSTRACT FROM PUBLISHER]

Cryptomeria japonica D. Don is one of the most important forest trees in Japan, but more than 10% of the Japanese population is allergic to its pollen. We constructed a cDNA library derived from pollen grains of C. japonica and performed an analysis of expressed sequence tags (ESTs). We obtained partial sequences from 1929 clones, which represented 1365 unique transcripts. Among the unique transcripts, 984 (72%) encoded proteins that were similar to Arabidopsis proteins with E-values of < 10−5. Analysis of funtional composition of the pollen ESTs revealed the overrepresentation of mRNAs for proteins involved in protein synthesis and post-translational modification. The most abundant transcripts were derived from novel genes (CjMP1-related genes) and encoded proteins that were not homologous to any proteins in current databases. The CjMP1-related genes formed a multi-gene family and were expressed specifically in the pollen grains of C. japonica. An analysis of homologies between ESTs from C. japonica pollen and proteins in the Structural Database of Allergenic Proteins revealed that products of 48 of the clones (2.5%) exhibited significant homology to known plant allergens. Our results provide new information about pollen-specific genes and potential allergens in C. japonica pollen. [ABSTRACT FROM PUBLISHER]

• Background and Aims The main stems of trees on forest slopes incline down the slope to various extents that are characteristic of the species. The inclination has been explained as an active response to a horizontally asymmetrical light environment, but the contributing physiological mechanisms are unknown. The present study tested the hypothesis that stem phototropism, gravitropism, or a combination of the two determines the inclination of tree stems on forest slopes.• Methods Cryptomeria japonica, Pinus densiflora, Quercus myrsinaefolia and Q. serrata were studied. Measurements were made of stem inclination of mature trees on forest slopes in uniform plantations of each species, and changes in stem inclination of potted seedlings in response to illumination treatments (unilateral or overhead) and inclination treatments (artificially inclined or erect). Indices of phototropic and gravitropic responsiveness were evaluated for each species, calculated from the change in stem inclination in response to artificial inclination with unilateral or overhead illumination.• Key Results Stem inclination on forest slopes varied significantly among species: Q. serrata inclined most in the down-slope direction, C. japonica inclined the least, and P. densiflora and Q. myrsinaefolia were intermediate. The change in stem inclination of seedlings in each treatment varied significantly among species. One-year-old stems of Q. serrata and 2-year-old stems of Q. myrsinaefolia bent toward the light source. Interspecific variation in the change in stem inclination in response to the unilateral illumination or that in the index of phototropic responsiveness was strongly correlated with the variation in stem inclination on forest slopes.• Conclusions The orientation of woody stems that have finished elongation can be actively controlled by phototropism. Interspecific variation in phototropic responsiveness of trees is a possible significant determinant of interspecific variation in stem inclination on forest slopes. [ABSTRACT FROM AUTHOR]

Thaumatin-like proteins (TLPs) are induced by a variety of phytopathogens in many plants and several TLPs are allergenic. Previously, we isolated three TLP-encoding cDNAs (Cry j 3.1, Cry j 3.2 and Cry j 3.3) from a cDNA library derived from the pollen of Cryptomeria japonica D. Don. Here, we describe three new TLP cDNAs (Cry j 3.4, Cry j 3.5 and Cry j 3.6). We compared the sequences, the genetic map location and the expression patterns of the Cry j 3 genes. The amino acid sequence predicted from Cry j 3.5 exhibits only limited similarity to those predicted from the other Cry j 3 genes. Linkage analysis showed that the Cry j 3.1 to Cry j 3.4 genes are located in the same linkage group, but Cry j 3.5 is located in a different group. Organ-specificity and induction by stresses and plant hormones differed among the Cry j 3 mRNAs. In pollen grains, the Cry j 3.5 mRNA expression level was higher than that of the other Cry j 3 genes. Exposure to UV-B and salt stress induced expression of Cry j 3.1. The ethylene-releasing compound ethephon strongly induced expression of Cry j 3.4. Salt stress and salicylic acid also induced expression of Cry j 3.4. Abscisic acid weakly induced expression of Cry j 3.5. Arachidonic acid strongly induced expression of Cry j 3.4 and Cry j 3.6, and weakly induced that of Cry j 3.3, whereas expression of Cry j 3.1 and Cry j 3.5 was unaffected. These results suggest that the roles of TLPs and the cascades that regulate their expression differ among the members of the TLP family in C. japonica. [ABSTRACT FROM PUBLISHER]

In many temperate evergreen plant species, reductions in turgor loss point of leaves (Ψtlp) and leaf osmotic potential at full turgor (πsat) occur from late summer to winter. To test the hypothesis that this seasonal change in leaf water relations is driven by root temperature, we manipulated the temperature of the roots and shoots of Cryptomeria japonica D. Don seedlings separately. Whole-plant warming diminished the seasonal changes in shoot water relations observed in the control plants, whereas shoot warming did not. Compared with the controls, root warming diminished the change in Ψtlp but not in πsat, whereas cooling accelerated the seasonal changes in shoot water relations. These results indicate that: (1) temperature responses of roots are involved in the seasonal changes in Ψtlp from late summer to winter; and (2) root temperature is partly responsible for the simultaneous changes in πsat. Whole-plant cooling caused increased root hydraulic resistance, suggesting that seasonal changes in shoot water relations represent adaptive responses to increased root hydraulic resistance at low root temperatures. [ABSTRACT FROM AUTHOR]

ent-Kaurene is a tetracyclic hydrocarbon precursor for gibberellins (GAs) in plants and fungi. To address whether fungal GA biosynthesis enzymes function in plants, we generated transgenic Arabidopsis plants overexpressing ent-kaurene synthase (GfCPS/KS) from a GA-producing fungus Gibberella fujikuroi. GfCPS/KS catalyzes a two-step reaction corresponding to ent-copalyl diphosphate synthase (CPS) and ent-kaurene synthase (KS) activities in plants. When GfCPS/KS was overexpressed and targeted to plastids, a range of GA-deficient phenotypes of the ga1-3 and ga2-1 mutants (defective in CPS and KS, respectively) were restored to wild type. Unexpectedly, the transgenic lines overproducing GfCPS/KS emitted the GA precursor ent-kaurene into the headspace besides its accumulation in the plant body. When co-cultivated with the ent-kaurene overproducers in a closed environment, the airborne ent-kaurene was able to fully complement the dwarf phenotype of ga1-3 and ga2-1 mutants, but not that of the ga3-1 mutant (defective in ent-kaurene oxidase). These results suggest that ent-kaurene may be efficiently metabolized into bioactive GAs in Arabidopsis when supplied as a volatile. We also provide evidence that ent-kaurene is released in the headspace of wild-type Chamaecyparis obtusa and Cryptomeria japonica plants, suggesting the occurrence of this hydrocarbon GA precursor as a volatile in nature. [ABSTRACT FROM AUTHOR]

Pollen from sugi (Japanese cedar, Cryptomeria japonica D. Don), a forest tree species that is widely grown in Japan, causes serious allergic disease. The major allergens from sugi pollen, Cry j 1 and Cry j 2, have been isolated and characterized. It has been reported that Cry j 1 concentration in pollen varies considerably among trees. If Cry j 1 concentration is genetically controlled, the planting of trees with low Cry j 1 concentrations would reduce pollinosis. We investigated genetic and environmental effects on Cry j 1 concentration in eight clones growing at four sites. Concentrations of Cry j 1 in pollen were measured with a monoclonal antibody-based Enzyme Linked Immunosorbent Assay (ELISA). The Cry j 1 concentrations differed significantly among clones and sites, but the site × clone interaction was not significant, suggesting that the Cry j 1 concentration is controlled primarily by genetic factors. We examined correlations between Cry j 1 concentration and temperature and precipitation from July through February. Temperature was not significantly related to Cry j 1 concentration, whereas cumulative precipitation during the 8 months and mean daily precipitation in September showed significant negative correlations with Cry j 1 concentration. [ABSTRACT FROM AUTHOR]

Phytosulfokine (PSK), which has been identified as a plant growth factor, had a dramatic stimulatory effect on the formation of somatic embryos of sugi (Cryptomeria japonica) in the presence of polyethylene glycol. The resultant somatic embryos germinated with synchronous sprouting of cotyledons, hypocotyls and roots, and most of the seedlings grew normally. A cDNA clone for the precursor to the PSK peptide of C. japonica was identified in an expressed sequence tags database. Our results support the existence of a PSK signaling pathway in C. japonica. [ABSTRACT FROM AUTHOR]

Describes the reduction of antigenecity of Cryj, major allergen of Japanese cedar pollen by the attachment of polysaccharides. Accounts on the presence and absence of N-linked carbohydrate chains; Suppression of T cell reactivity; Identification of the epitope of immunoglobulin E.

A consensus map for sugi (Cryptomeria japonica) was constructed by integrating linkage data from two unrelated third-generation pedigrees, one derived from a full-sib cross and the other by self-pollination of F[sub 1] individuals. The progeny segregation data of the first pedigree were derived from cleaved amplified polymorphic sequences, microsatellites, restriction fragment length polymorphisms, and single nucleotide polymorphisms. The data of the second pedigree were derived from cleaved amplified polymorphic sequences, isozyme markers, morphological traits, random amplified polymorphic DNA markers, and restriction fragment length polymorphisms. Linkage analyses were done for the first pedigree with JoinMap 3.0, using its parameter set for progeny derived by cross-pollination, and for the second pedigree with the parameter set for progeny derived from selfing of F[sub 1] individuals. The 11 chromosomes of C. japonica are represented in the consensus map. A total of 438 markers were assigned to 11 large linkage groups, 1 small linkage group, and 1 nonintegrated linkage group from the second pedigree; their total length was 1372.2 cM. On average, the consensus map showed 1 marker every 3.0 cM. PCR-based codominant DNA markers such as cleaved amplified polymorphic sequences and microsatellite markers were distributed in all linkage groups and occupied about half of mapped loci. These markers are very useful for integration of different linkage maps, QTL mapping, and comparative mapping for evolutional study, especially for species with a large genome size such as conifers. [ABSTRACT FROM AUTHOR]

Cell behavior in the cambium and developing xylem of 3-year-old Japanese cedar (Cryptomeria japonica D. Don.) trees, during and after an 11-day suspension of irrigation, was analyzed. Leaf xylem pressure potential and tangential strain of the stem surface were monitored throughout the experiment. Anatomical features and numbers of developing tracheids and cambial cells were observed in four trees, sampled on Days 0, 4, 8 and 11 after irrigation was suspended. Daytime xylem pressure potential decreased to –1.9 MPa on Day 7 and remained the same until irrigation was resumed on Day 11. The transverse dimensions of the tracheids, which began to form secondary walls, began to decrease on Day 4. The number of cells in the cambial zone and cell expansion zone decreased abruptly on Day 8. Tangentially aligned developing tracheids with collapsed cell walls were observed in samples harvested on Days 8 and 11. Secondary wall formation was recognized in these tracheids. After the resumption of irrigation, xylem pressure potential recovered rapidly to the same value as before the suspension of irrigation. Tangential strain increased within 30 min after the resumption of irrigation, and continued to increase until the onset of light the next day. Eighteen days after the resumption of irrigation, anatomical features of cells in the cambium and cell-expansion zone were similar to those observed before suspension of irrigation. [ABSTRACT FROM AUTHOR]

Investigates the nucleotide variation of a conifer, Cryptomeria japonica, and the divergence between this species and its closest relative, Taxodium distichum, at seven nuclear loci. Collection of samples of C. japonica from three Japanese areas Kantou-Toukai, Hokuriku and Iwate; Lack of apparent geographic differentiation among the samples; Frequency spectrum of the nucleotide polymorphism; Suggestion that the population was not panmictic and a constant size in the past.