Divergent mechanisms of reduced growth performance in Betula ermanii saplings from high-altitude and low-latitude range edges.

The reduced growth performance of individuals from range edges is a common phenomenon in various taxa, and considered to be an evolutionary factor that limits the species' range. However, most studies did not distinguish between two mechanisms that can lead to this reduction: genetic load and adaptive selection to harsh conditions. To address this lack of understanding, we investigated the climatic and genetic factors underlying the growth performance of Betula ermanii saplings transplanted from 11 populations including high-altitude edge and low-latitude edge population. We estimated the climatic position of the populations within the overall B. ermanii's distribution, and the genetic composition and diversity using restriction-site associated DNA sequencing, and measured survival, growth rates and individual size of the saplings. The high-altitude edge population (APW) was located below the 95% significance interval for the mean annual temperature range, but did not show any distinctive genetic characteristics. In contrast, the low-latitude edge population (SHK) exhibited a high level of linkage disequilibrium, low genetic diversity, a distinct genetic composition from the other populations, and a high relatedness coefficient. Both APW and SHK saplings displayed lower survival rates, heights and diameters, while SHK saplings also exhibited lower growth rates than the other populations' saplings. The low heights and diameters of APW saplings was likely the result of adaptive selection to harsh conditions, while the low survival and growth rates of SHK saplings was likely the result of genetic load. Our findings shed light on the mechanisms underlying the reduced growth performance of range-edge populations.

Gross primary production of dwarf bamboo, Sasa senanensis, in a mature beech forest with a substantial gap-mosaic structure.

Forest understory plays an important role in the gross primary production (GPP) of some forest ecosystems. However, differences in understory GPP caused by obviously different overstory canopy structure have not been taken into consideration in previous studies, thus potentially over- or underestimating understory GPP. To estimate the understory GPP more accurately, we separated a forest into "canopy area", with closed-overstory canopy, and "gap area", with open-overstory canopy. The study was conducted in a mature deciduous forest dominated by beech and with an understory dominated by dwarf bamboo, Sasa senanensis. We measured S. senanensis GPP at the community scale (GPPSasa-community) using a static chamber system that covered the aboveground part of the plants and then upscaled it to the ecosystem scale (GPPSasa-ecosystem) by considering the proportions of canopy and gap areas within the forest. GPPSasa-community was 192 g C m-2 year-1 in the canopy area and 699 g C m-2 year-1 in the gap area. The large difference likely occurred because the photosynthetic ability and biomass of the S. senanensis community differed strongly between the two areas. The seasonal dynamics of GPPSasa-community also differed between the areas. The 10-day cumulative GPPSasa-community peaked from July to August in the gap area, whereas there was no clear peak of GPPSasa-community in the canopy area. Multiple linear regressions showed that light intensity and biomass were significant predictors of GPPSasa-community in the canopy area, whereas air temperature and biomass were significant predictors of GPPSasa-community in the gap area. GPPSasa-ecosystem during growing season in 2019 was 3.74 t C ha-1 year-1, which contributed between 16.37 and 19.85% of the entire forest ecosystem GPP. This study highlights the need to consider differences in overstory structure for the accurate estimation of understory GPP.

New xanthine oxidase inhibitors from the fruiting bodies of Tyromyces fissilis.

Excessive uric acid production, which causes gout and hyperuricemia, can be blocked by inhibiting xanthine oxidase (XO). However, some agents to block on XO often cause side effects, thereby necessitating the identification of new inhibitors. During the screening of XO inhibitors from various mushroom extracts, we found that a methanolic extract of the fruiting bodies of Tyromyces fissilis, an inedible and non-toxic fungus, showed inhibitory activity. Both n-hexane and ethyl acetate layers, obtained by partitioning this extract exhibited XO inhibitory activity. Subsequently, using an activity-guided separation method, eight active compounds (1-8) were isolated. The structures of five of the new compounds, 2-4, 6, and 7, were elucidated by spectral analysis and chemical derivatization. All compounds had a salicylic acid moiety with an aliphatic group at the C-6 position. Notably, 2-hydroxy-6-pentadecylbenzoic acid (1) showed the highest level of XO noncompetitive inhibition (58.9 ± 2.2% at 25 µM).

Ergosterol and its derivatives from Grifola frondosa inhibit antigen-induced degranulation of RBL-2H3 cells by suppressing the aggregation of high affinity IgE receptors.

Grifola frondosa is an edible mushroom consumed as a health food and/or traditional medicine in Asia. However, the anti-allergic effects of G. frondosa are not yet understood. In this study, we demonstrated the effects of G. frondosa extract (GFE) on IgE-mediated allergic responses, using antigen-stimulated RBL-2H3 cells. Three active compounds: ergosterol, 6ß-methoxyergosta-7,22-dien-3ß,5α-diol (MEDD), and 6-oxoergosta-7,22-dien-3ß-ol (6-OXO) were isolated from GFE and shown to inhibit the antigen-induced release of ß-hexosaminidase and histamine. Among the three active components, we focused on ergosterol because of its high content in GFE. Ergosterol inhibited the aggregation of high-affinity IgE receptor (FcεRI), which is the first step in the activation of mast cells and antigen-induced tyrosine phosphorylation. Furthermore, ergosterol suppressed antigen-increased IL-4 and TNF-α mRNA. Taken together, our findings suggest that G. frondosa, including ergosterol and its derivatives as active components, has the potential to be a novel functional food that prevents type I allergies.

Response of stomatal density and size in Betula ermanii to contrasting climate conditions: The contributions of genetic and environmental factors.

As plant distribution and performance are determined by both environmental and genetic factors, clarifying the contribution of these two factors is a key for understanding plant adaptation and predicting their distribution under ongoing global warming. Betula ermanii is an ideal species for such research because of its wide distribution across diverse environments. Stomatal density and size are crucial traits that plants undergo changes in to adapt to different environments as these traits directly influence plant photosynthesis and transpiration. In this study, we conducted a multi-location common garden experiment using B. ermanii to (1) clarify the contribution of both environmental and genetic factors to the variation in stomatal density and size of B. ermanii, (2) demonstrate the differences in the plasticity of stomatal density and size among B. ermanii populations, and (3) understand how stomatal density and size of B. ermanii would respond to increased temperature and changing precipitation patterns. Genetic factors played a more significant role in stomatal size than environmental factors, suggesting that B. ermanii struggles to adjust its stomatal size in response to a changing environment. Our results also revealed a positive correlation between stomatal size plasticity and original habitat suitability, indicating that in B. ermanii populations in harsh environments exhibit lower adaptability to environmental shifts. Although stomatal density and size of B. ermanii showed the significant responses to increased temperature and shifting precipitation patterns, the response ranges of stomatal density and size to the environmental factors varied among populations. Our findings highlighted the interplay between genetic and environmental factors in determining the intraspecific variation in stomatal density and size in B. ermanii. This indicated that certain populations of B. ermanii exhibit limited stomatal plasticity and adaptability, which could directly affect photosynthesis and transpiration, suggesting potential population-specific fitness implications for B. ermanii under future climate change.

Spatial variation in forest soil respiration: A systematic review of field observations at the global scale.

As it is responsible for the second largest CO2 flux in the terrestrial ecosystem, the accurate estimation and prediction of soil respiration (SR) are necessary, especially for forest ecosystems, which are a major contributor to the total terrestrial SR. Spatial variation is one of the challenges affecting the accurate estimation and prediction of forest SR in ecosystems. Although a number of studies have examined spatial variation in SR within individual forests, the magnitude and patterns of spatial variation in SR within forest ecosystems (CV of SR [%]) remain unexplored at the global scale. In this study, we collected 94 field observation studies to demonstrate the range and pattern of the CV of SR, and to clarify the controlling factors. Through our analysis, the CV of SR was found to range from 1.8 % to 89.3 % on the global scale; it was highest in the equatorial zone (39.0 % ± 13.8 %) and followed by the warm temperate zone (32.6 ± 14.5 %) and the snow zone (30.0 % ± 16.3 %). There was a significant negative correlation between the CV of SR and soil water content, bulk density, fine root biomass, and elevation at both the global scale and in each climatic zone (P < 0.01). Other factors such as total nitrogen content, mean of diameter at breast height, slope, etc., were also significantly correlated with the CV of SR, but the correlation was different among climatic zones. This study provides an overall perspective of the CV of SR by clarifying the range, patterns, and controlling factors at both the global scale and in each climatic zone. However, further research is needed, especially regarding the mechanisms between the CV of SR and its controlling factors.

Diversity and Distribution of Aspergillus fumigatus and Its Related Species in Izu and Ogasawara Islands, Japan.

The taxon Aspergillus section Fumigati comprises several causative agents of aspergillosis. Here, the distribution of Aspergillus sect. Fumigati in outdoor environments of Izu and Ogasawara Islands was investigated. Different strains were isolated from soil samples collected from 68 sites on 9 islands (Izu-oshima, Toshima, Shikinejima, Kozushima, Miyakejima, Hachijojima, Mukojima, Chichijima, and Hahajima), including different landscapes, and identified using morphological characteristics and calmodulin (CaM) sequences. Seven Aspergillus sect. Fumigati species were identified. The occurrence frequency of Aspergillus fumigatus was higher in forest sites on the islands, except for Ogasawara Islands, whereas that of species other than A. fumigatus was higher in bare land and grassland sites on all islands. The occurrence frequency of A. fumigatus was more than 50% on islands between Izu-oshima and Toshima, decreased on islands between Shikinejima and Hachijojima, and was zero on Ogasawara Islands. Considering other Aspergillus species, Aspergillus felis showed high occurrence frequency on islands between Izu-oshima and Shikinejima, Aspergillus pseudoviridinutans on islands between Kozushima and Hachijojima, and Aspergillus udagawae on Ogasawara Islands. At two study sites (grassland and forest sites), the soil was sampled throughout the year to evaluate whether the occurrence frequency of each fungal species was affected by sampling season. At the grassland site, A. pseudoviridinutans was isolated at more than 90% frequency, regardless of the sampling season. A. fumigatus occurrence frequency at the forest site ranged from 0% to 60% and greatly varied among sampling seasons. Thus, differences in island location and landscape affected the distribution of Aspergillus sect. Fumigati.

Photosynthetic Properties of Co-Occurring Pioneer Species on Volcanically Devastated Sites in Miyake-jima Island, Japan.

Pioneer species differing in their inherent ecological characteristics (e.g., N-fixing ability, photosynthetic pathway) can have a large impact on local ecosystems in the early stages of volcanic succession. However, it remains unclear as to how these pioneer species adapt to the extreme environment of volcanically devastated sites in terms of ecophysiological leaf traits. In this study, we compared the leaf traits (including morphological, physiological) of three co-occurring pioneer species, including a C4 non-N-fixing grass, a C3 N-fixing tree, and a C3 non-N-fixing herb from a newly created (18 years after eruption) volcanically devastated site in Miyake-jima, Japan. Our results showed that three pioneer species have different sets of leaf traits that are associated with their ecophysiological growth advantages, respectively. Miscanthus condensatus shows the highest light-saturated photosynthetic rate (Amax). The higher Amax were partially the result of higher water use efficiency (WUE) and photosynthetic N-use efficiency (PNUE). The PNUE in M. condensatus appears to be high, even for a C4 grass. Alnus sieboldiana rely on its N-fixing ability, has a higher leaf N content (Narea) that compensates for its photosynthetic machinery (Rubisco), and further ensures its photosynthetic capacity. Fallopia japonica var. hachidyoensis has a higher leaf mass per area (LMA), chlorophyll content (Chl), and maximum quantum yield of PSII (Fv/Fm), demonstrating its higher light capturing ability. These results make it possible to predict certain ecological processes that take place in the early stages of volcanic succession resulting from ecological characteristics and from some key leaf traits of pioneer species. It also provides a theoretical basis for species selection and species combination for volcanic ecological restoration.

Synthesis of procyanidin B3 and its anti-inflammatory activity. the effect of 4-alkoxy group of catechin electrophile in the Yb(OTf)(3)-catalyzed condensation with catechin nucleophile.

Yb(OTf)(3)-catalyzed equimolar condensation of the benzylated catechin with various 4-alkoxy catechin derivatives was studied. In particular, the reaction using 4-(2''-ethoxyethoxy)flavan derivative gave good yield with excellent stereoselectivity. The condensed product was successfully converted to procyanidin B3 (1). The anti-inflammatory effect of procyanidin B3 (1) on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation of mouse ears was examined. The anti-inflammatory activity of 1 was stronger than that of indomethacin and glycyrrhetinic acid, the normally used anti-inflammatory agents.

Asymmetric syntheses of daedalin A and quercinol and their tyrosinase inhibitory activity.

Stereoselective syntheses of daedalin A and quercinol, an enantiomer of daedalin A, is described. The tyrosinase inhibitory activities of daedalin A and quercinol were examined. The activity of quercinol was weaker than that of daedalin A at high concentration.

Small-scale variation in ecosystem CO2 fluxes in an alpine meadow depends on plant biomass and species richness.

Characterizing the spatial variation in the CO2 flux at both large and small scales is essential for precise estimation of an ecosystem's CO2 sink strength. However, little is known about small-scale CO2 flux variations in an ecosystem. We explored these variations in a Kobresia meadow ecosystem on the Qinghai-Tibetan plateau in relation to spatial variability in species composition and biomass. We established 14 points and measured net ecosystem production (NEP), gross primary production (GPP), and ecosystem respiration (Re) in relation to vegetation biomass, species richness, and environmental variables at each point, using an automated chamber system during the 2005 growing season. Mean light-saturated NEP and GPP were 30.3 and 40.5 micromol CO2 m(-2) s(-1) [coefficient of variation (CV), 42.7 and 29.4], respectively. Mean Re at 20 degrees C soil temperature, Re(20), was -10.9 micromol CO2 m(-2) s(-1) (CV, 27.3). Re(20) was positively correlated with vegetation biomass. GPP(max) was positively correlated with species richness, but 2 of the 14 points were outliers. Vegetation biomass was the main determinant of spatial variation of Re, whereas species richness mainly affected that of GPP, probably reflecting the complexity of canopy structure and light partitioning in this small grassland patch.

Photosynthetic Properties of Miscanthus condensatus at Volcanically Devastated Sites on Miyake-jima Island.

How photosynthetic-related leaf traits of non-nitrogen (N)-fixing pioneer species respond to extreme habitat conditions of primary succession is still not well-elucidated, especially in volcanically N-deplete habitats. The effect of N-deplete soil on photosynthetic-related leaf traits can provide a basis for predicting how plants adjust their strategies to adapt to such habitats. To examine the responses of leaf traits to extreme conditions, we investigated Miscanthus condensatus (a non-N-fixing C4 pioneer grass) which grows on a volcanically devastated area on Miyake-jima Island, Japan, in which the volcanic ash has been deposited for 17-18 years since the 2000-year eruption. Leaf N content (Narea), light-saturated photosynthetic rate (Amax), and photosynthetic N use efficiency (PNUE) in three contrasting study sites: bare land (BL), shrub land gap (SLG), and shrub land under canopy (SLUC) were determined. Results indicated that compared to previous studies and internal comparison of Miyake Island, M. condensatus in BL was able to maintain a relatively high Amax, Narea and PNUE. The higher Amax was in part a result of the higher PNUE. This is a characteristic necessary for its successful growth in N-deplete soils. These results suggest that M. condensatus has photosynthetic-related advantages for adaptation to volcanically N-deplete habitats.

Anti-proliferative and apoptosis-inducible activity of Sarcodonin G from Sarcodon scabrosus in HeLa cells.

There is an ongoing search for plant-derived diterpenes, especially for diterpenes with anti-inflammatory activity that also have anti-proliferative effects on human cancer cells. A cyathane-type diterpene, Sarcodonin G (SG), isolated from the mushroom Sarcodon scabrosus and already reported to have anti-inflammatory activity, inhibited proliferation of HeLa cells to the greatest extent among 4 cyathane diterpenes tested. SG showed an IC50 (50% inhibition concentration) of 20 microM, estimated by MTT assay 2 days after culture of cells with the chemical. SG treatment of HeLa cells resulted in dose-dependent generation of apoptotic events such as DNA-laddering (< or =100 microM). Moreover, SG-treated HeLa cells showed activation of caspase-3 and caspase-9 and increase of Bax/Bcl-2 ratios, as analyzed by Western blot analysis. The anti-proliferative effects of SG treatment on HeLa cells were lessened by a caspase inhibitor, Z-VAD-FMK. SG also showed anti-proliferative effects toward 5 other human cancer cell lines with IC50 values of 20-40 microM. Because of these anti-proliferative effects via possible caspase activation, SG holds promise of being a novel anti-proliferative agent deserving further investigation.

Daedalin A, a metabolite of Daedalea dickinsii, inhibits melanin synthesis in an in vitro human skin model.

The culture broth of Daedalea dickinsii was found to predominantly contain the tyrosinase inhibitor, (2R)-6-hydroxy-2-hydroxymethyl-2-methyl-2H-chromene, daedalin A (1). Ongoing research into bioactive metabolites resulted in the identification of two new 2H-chromenes, 6-hydroxy-5,7-dimethoxy-2,2-dimethyl-2H-chromene (3) and 6-hydroxy-2-hydroxymethyl-5-methoxy-2-methyl-2H-chromene (4), together with 6-hydroxy-2,2-dimethyl-2H-chromene (2). Comparative studies of isolated compounds 1-4 and related compounds (+/-)-1 and 1a-1c showed 1 to have the strongest tyrosinase inhibitory activity. These results suggest that the hydroxyl groups at positions 6 and 9 of 1 were important for the potent activity. A Lineweaver-Burk plot for a kinetic analysis indicates that 1 competed with L-tyrosine for tyrosinase. Compound 1 also suppressed melanogenesis in a human skin model (up to 49% at 2.8 micromol/tissue application) without affecting the cell viability. Compounds 1, 1b and 1c also showed 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity comparable to that of alpha-tocopherol.

Synthesis of (-)-epicatechin 3-(3-O-methylgallate) and (+)-catechin 3-(3-O-methylgallate), and their anti-inflammatory activity.

A concise synthesis of (-)-epicatechin 3-(3-O-methylgallate) (1; ECG3''Me), which is a minor constituent of tea, and (+)-catechin 3-(3-O-methylgallate) (2; CG3''Me) via condensation of equimolar amount of catechin and gallate derivatives has been achieved. The anti-inflammatory effect of the synthetic compounds on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation of mouse ears was examined. Compounds 1 and 2 suppressed the TPA-induced inflammation of mouse ears by 50 and 43%, respectively, at a dose of 200 microg. Their activities are stronger than those of indomethacin and glycyrrhetinic acid, the normally used anti-inflammatory agents.

Limited distribution of natural cyanamide in higher plants: occurrence in Vicia villosa subsp. varia, V. cracca, and Robinia pseudo-acacia.

Cyanamide (NH2CN) has recently been proven to be a natural product, although it has been synthesized for over 100 years for agricultural and industrial purposes. The distribution of natural cyanamide appears to be limited, as indicated by our previous investigation of 101 weed species. In the present study, to investigate the distribution of natural cyanamide in Vicia species, we monitored the cyanamide contents in V. villosa subsp. varia, V. cracca, and V. amoena during their pre-flowering and flowering seasons. It was confirmed that V. cracca was superior to V. villosa subsp. varia in accumulating natural cyanamide, and that V. amoena was unable to biosynthesize this compound under laboratory condition examined. The localization of cyanamide in the leaves of V. villosa subsp. varia seedlings was also clarified. In a screening study to find cyanamide-biosynthesizing plants, only Robinia pseudo-acacia was found to contain cyanamide among 452 species of higher plants. We have investigated 553 species to date, but have so far found the ability to biosynthesize cyanamide in only three species, V. villosa subsp. varia, V. cracca and R. pseudo-acacia.

A tyrosinase inhibitor, Daedalin A, from mycelial culture of Daedalea dickinsii.

A chromene-type compound, daedalin A (1), was isolated from mycelial culture broth of Daedalea dickinsii. Based on spectroscopic data, the structure of 1 was found to be (2R)-6-hydroxy-2-hydroxymethyl-2-methyl-2H-chromene. Daedalin A (1) strongly inhibited the activity of tyrosinase (IC(50): 194 muM). In addition, 1 also showed 1,1-diphenyl-2-picrylhydrazyl scavenging activity (IC(50): 16.9 microM) and superoxide anion scavenging activity (IC(50): 28.5 microM).

Fluxes of carbon dioxide, methane and nitrous oxide in two contrastive fringing zones of coastal lagoon, Lake Nakaumi, Japan.

We measured fluxes of carbon dioxide (CO(2)), methane (CH(4)), and nitrous oxide (N(2)O) simultaneously in two typical fringing zones, sandy shore and salt marsh, of coastal lagoon, Lake Nakaumi, Japan, in mid-summer 2003. Our aim was to quantify net the greenhouse gases (GHGs) fluxes and examine key factors, which control variation of the GHGs fluxes in the two sites. Net CO(2) and CH(4) fluxes were markedly different between the two sites; magnitudes and variations of the both fluxes in sandy shore were lower than those of salt marsh. Meanwhile, magnitude and variation of net N(2)O flux in the two sites were similar. In sandy shore, temporal and spatial variation of the three GHGs fluxes were highly controlled by water level fluctuation derived from astronomic tide. In salt marsh, spatial variation of the three GHGs fluxes were correlated with aboveground biomass, and temporal variation of CO(2) and CH(4) fluxes were correlated with soil temperature. The sum of global warming potential, which was roughly estimated using the observed GHGs fluxes, was ca. 174-fold higher in salt marsh than in sandy shore.

Evidence of cyanamide production in hairy vetch Vicia villosa.

Cyanamide (NH(2)CN) has recently been isolated as a plant growth inhibitor from Vicia villosa, which is the first discovery of cyanamide from natural sources. To reveal the presence of the biosynthesized cyanamide in plants, 3.4 mM potassium ((15)N)nitrate was administered to 15- to 35-day-old plants of V. villosa, from which the cyanamide was purified and subjected to GC/MS analysis. The isotopic ratio (15)N/((14)N + (15)N) of the cyanamide was calculated to be 0.143, while that of the cyanamide extracted from V. villosa grown in the presence of a natural N source was 0.0065. The (15)N-enrichment proved de novo biosynthesis of cyanamide.