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Lucidophyllous (evergreen broad-leaved) forests are the dominant forests in human-dominated subtropical/warm-temperate regions in East Asia. Biometric-based estimates of net primary production (NPP) were conducted in a secondary lucidophyllous forest on Mt. Kinka (35°26′ N, 136°47′ E) near the northern limit of their distribution in central Japan for three years, including the masting event. The forest stand mainly consists of Castanopsis cuspidata (Thunb.) Schottky and Cleyera japonica Thunb. in the canopy and subtree layers, respectively. In 2018, the total NPP of the masting year was 14.53 ± 2.03 ton ha−1 yr−1, including woody NPP (above: 2.63 ± 0.35 ton ha−1 yr−1; below: 0.57 ± 0.08 ton ha−1 yr−1), foliage NPP (4.07 ± 0.23 ton ha−1 yr−1), reproductive NPP (4.81 ± 0.77 ton ha−1 yr−1), and fine root production (Pfr) (2.46 ± 1.84 ton ha−1 yr−1). Pfr and belowground production comprised 16.9% and 20.9%, respectively, of the total NPP. The nut production of C. cuspidata in 2018 (4.31 ± 0.75 ton ha−1 yr−1) was significantly higher than that in 2017 (0.77 ± 0.13 ton ha−1 yr−1) and 2019 (0.23 ± 0.06 ton ha−1 yr−1). No significant change was observed for the three years of foliage NPP and total NPP without Pfr. However, the woody NPP in 2018 (3.20 ± 0.43) was lower than in 2017 (5.37 ± 0.33 ton ha−1 yr−1) and 2019 (4.71 ± 0.38 ton ha−1 yr−1). This suggests that nut production in the masting years compensated by decreasing woody production in the Castanopsis forest.

The Castanopsis cuspidata var. sieboldii (CCS) plant grows predominantly in temperate regions of Asian countries, such as South Korea. Research on CCS has so far concentrated on the nutritional analysis, antioxidant activity, and anti-inflammation properties of its branches. However, the isolation of compounds and structural elucidation of effective single molecules remain unexplored, necessitating further exploration of CCS branches. Therefore, this study demonstrates the antioxidant and antimelanogenic activity of a single substance of ethyl gallate (EG) isolated from CCS branch extracts. Notably, the antimelanogenic (whitening) activity of EG extracted from CCS branches remains unexplored. Tyrosinase inhibition, kinetic enzyme assays, and molecular docking studies were conducted using mushroom tyrosinase in order to examine the antioxidant mechanism and antimelanin activity of EG in B16F10 melanoma cells. Nontoxic EG concentrations were found to be below 5 µg/mL. While EG significantly reduced the levels of whitening-associated proteins, p-CREB, and p-PKA, it dose-dependently inhibited the expression of TYR, TRP-1, TRP-2, and transcription factor (MITF). In addition, EG downregulated melanogenetic gene expression and activated autophagy signals. Therefore, EG extracted from CCS branches could serve as a novel functional cosmetic material with antimelanogenic and autophagy-enhancing activity.

Secondary lucidophyllous forest is one of the dominant forests in human-dominated subtropical/warm-temperate regions in East Asia. There were few direct monitoring techniques to elucidate the following hypotheses: (a) self-thinning may govern the stand development process and (b) wood production decline can be observed during secondary succession in a lucidophyllous forest. We conducted a long-term study at a permanent plot in central Japan, since 1989. The forest consists mainly of Castanopsis cuspidata in a canopy layer, Cleyera japonica, and Eurya japonica in a subtree layer. During the 28-year period, the basal area of the stand significantly increased due to the growth of C. cuspidata, from 29.18 ± 1.84 (87.8% of total) to 38.71 ± 2.22 m2 ha−1 (91.9%), while the stem density of C. cuspidata significantly decreased from 666 ± 13 to 404 ± 10 stems ha−1 in proportion to accumulating biomass (117.8 to 166.6 ton ha−1). The annual woody net primary production ranged from 2.40 ± 0.13 to 3.93 ± 0.33 ton ha−1 year−1 as a nearly 70-year-old forest. There was no age-related decline of woody net primary production (NPP) was found during secondary succession, and the growth of individual tree still increased when the self-thinning process governed the stand.

Among 276 herbal extracts, a methanol extract of Castanopsis cuspidata var. sieboldii stems was selected as an experimental source for novel acetylcholinesterase (AChE) inhibitors. Five compounds were isolated from the extract by activity-guided screening, and their inhibitory activities against butyrylcholinesterase (BChE), monoamine oxidases (MAOs), and β-site amyloid precursor protein cleaving enzyme 1 (BACE-1) were also evaluated. Of these compounds, 4′- O -(α- l -rhamnopyranosyl)-3,3′,4-tri- O -methylellagic acid ( 3 ) and 3,3′,4-tri- O -methylellagic acid ( 4 ) effectively inhibited AChE with IC 50 values of 10.1 and 10.7 µM, respectively. Ellagic acid ( 5 ) inhibited AChE (IC 50  = 41.7 µM) less than 3 and 4 . In addition, 3 effectively inhibited MAO-B (IC 50  = 7.27 µM) followed by 5 (IC 50  = 9.21 µM). All five compounds weakly inhibited BChE and BACE-1. Compounds 3 , 4 , and 5 reversibly and competitively inhibited AChE, and were slightly or non-toxic to MDCK cells. The binding energies of 3 and 4 (− 8.5 and − 9.2 kcal/mol, respectively) for AChE were greater than that of 5 (− 8.3 kcal/mol), and 3 and 4 formed a hydrogen bond with Tyr124 in AChE. These results suggest 3 is a dual-targeting inhibitor of AChE and MAO-B, and that these compounds should be viewed as potential therapeutics for the treatment of Alzheimer’s disease.

Castanopsis argentea is an endangered tree naturally distributed in Java and Sumatera Island and considered a keystone species in the submontane forest ecosystem. A laboratory study was conducted to test the cross-amplification potential of seven pairs of Castanopsis cuspidata var. sieboldii microsatellite markers to be used for Castanopsis argentea. The rate of cross-amplification success, average number of alleles per locus (N A ), observed heterozygosity (H O ), expected heterozygosity (H E ), and fixation index (F IS ) were determined, and cross-amplification potential was investigated. The rate of amplification success was 85.71% (6 out of 7 markers were successful amplified), N A was 7.67, H O ranged from 0.467 to 0.923, H E ranged from 0.441 to 0.745, and the fixation index (F IS ) ranged from −0.413 to 0.278. Results indicate the high transferability of C. cuspidata var. sieboldii microsatellite markers to C. argentea. Six markers were suitable for genetic diversity and population genetic studies on C. argentea.

The growth trajectories of trees are not fully understood due to their long life span. We characterized the population dynamics of the canopy tree Castanopsis cuspidata (Thunb.) Schottky in a Japanese warm‐temperate forest over 49 years (1966–2015). Our study was initiated approximately 50 years after our study site was clear‐cut. The forest had a closed canopy for the first 23 years of the study. Strong typhoons in 1991 and 1993 seriously damaged the forest, and since then the forest has been recovering from these disturbances. The diameter distribution of this species was bell‐shaped in 1966, suggesting that the trees emerged simultaneously after the clear‐cut in the 1910s, and the recruitment of trees has remained unchanged since then. The lack of recruitment of C. cuspidata before the typhoon disturbance supports this conclusion. Assuming that the C. cuspidata trees in 1966 were cohorts that were established soon after the clear‐cut, the size differences reflect differences in growth rate, with small trees corresponding to slow growers and large trees corresponding to fast growers. Before the typhoon, slow growers had low survival, and the mortality rate of fast growers was low. Many fast growers were uprooted or snapped by strong winds by the typhoons. However, their mortality rate did not differ from that of slow growers because many slow growers were killed by large fallen trees. The growth of some slow‐growing survivors increased after the typhoon, which allowed them to rapidly reach the canopy. Therefore, the typhoon altered the distribution of canopy trees among slow and fast growers. Survivors experienced faster growth than trees that died during the census period, suggesting that growth rate provides a robust indicator of future survival. Before the typhoon, the survival of fast growers was higher than that of slow growers. This suggests that fast growers disproportionally contribute to reproduction compared with slow growers. However, no recruited tree was observed in this subperiod, suggesting that fast growers made no contribution to reproduction. Fast growers might not play a more significant demographic role than slow growers in this species.

The broadleaved evergreen forests of the East Asian warm temperate zone are characterised by their high biodiversity and endemism, and there is therefore a need to extend our understanding of its genetic diversity and phylogeographic patterns. Castanopsis (Fagaceae) is one of the dominant tree species in the broadleaved evergreen forests of Japan. In this study we investigate the genetic diversity, genetic structure and leaf epidermal morphology of 63 natural populations of C. sieboldii and C. cuspidata, using 32 Expressed Sequence Tag associated microsatellites. The overall genetic differentiation between populations was low (GST = 0.069 in C. sieboldii and GST = 0.057 in C. cuspidata). Neighbor-joining tree and Bayesian clustering analyses revealed that the populations of C. sieboldii and C. cuspidata were genetically clearly differentiated, a result which is consistent with the morphology of their epidermal cell layers. This suggests that C. sieboldii and C. cuspidata should be treated as independent species, although intermediate morphologies are often observed, especially at sites where the two species coexist. The higher level of genetic diversity observed in the Kyushu region (for both species) and the Ryukyu Islands (for C. sieboldii) is consistent with the available fossil pollen data for Castanopsis-type broadleaved evergreen trees during the Last Glacial Maximum and suggests the existence of refugia for Castanopsis forests in southern Japan. Within the C. sieboldii populations, Bayesian clustering analyses detected three clusters, in the western and eastern parts of the main islands and in the Ryukyu Islands. The west-east genetic differentiation observed for this species in the main islands, a pattern which is also found in several plant and animal species inhabiting Castanopsis forests in Japan, suggests that they have been isolated from each other in the western and eastern populations for an extended period of time, and may imply the existence of eastern refugia.

The role of plant hormones in tension wood (TW) formation has been studied but is still unclear. IAA, ABA, ACC, tZ, tZR, iP, and iPR in cambial region tissues were identified and quantified by liquid chromatography/mass spectrometry (LC/MS). We examined the distribution of plant hormones and their precursors in the stems of Quercus myrsinifolia Blume and Castanopsis cuspidata var.sieboldii Nakai after bending the stems or applying an ethylene precursor (ACC). After 3 weeks of bending, though not after 1 week of bending, the auxin (IAA) and abscisic acid (ABA) amounts were larger on the TW side than on the opposite wood (OW) side and in upright trees. After 2 weeks of bending, the peak concentrations of IAA in cambium on the TW side were obviously higher than those on the OW side. After 1 week of bending, the ACC amounts on both sides were larger than in upright trees, but after 3 weeks of bending, they were smaller than in upright trees. Applied ACC did not enhance TW formation but induced axical parenchyma and phloem formation in C. cuspidata var.sieboldii. These results indicated that the distribution patterns of IAA and ABA might have important roles in TW formation in these two species. The role of ACC might be limited in the early stages of TW formation.

The growth dynamics and mode of competition between adult trees > = 5.0 cm in diameter at breast height (DBH) of nine abundant tree species occupying ca. 85% of the total basal area were investigated in a 4 ha study plot (200 m x 200 m) of a warm-temperate old-growth evergreen broad-leaved forest in the Tatera Forest Reserve of Tsushima Island, southwestern Japan. In the plot, adult trees > = 5.0 cm DBH co-occurred with 35 woody plant species (except for woody vine species). The most dominant and largest species, Castanopsis cuspidata var. sieboldii exhibited a bimodal DBH distribution; it was found in both the upper and lower vertical layers. Other tree species had unimodal DBH distributions corresponding mostly to the lower vertical layer. We developed a model for individual growth incorporating both intra- and interspecific competition and degree of competitive asymmetry. One-sided interspecific competition was detected in 17 cases out of the 66 possible combinations on the scale of the 4 ha study plot. The direction of interspecific competition was generally one-sided from layer-I species to layer-II and III ones. The effects of two-sided competition were detected only in layer-II and III species. Only Distylium racemosum exhibited one-sided intraspecific competition. We also found 11 cases of positive interspecific relationships. Generally, competitive relationships prevailed over positive relationships between adult trees in this warm-temperate evergreen broad-leaved forest. Competition between adult trees > = 5.0 cm in DBH did not occur in the same vertical layer, but occurred only between trees in different vertical layers. This suggests that competition between adult trees > = 5.0 cm in DBH plays a key role in the variation in species coexistence between different vertical layers on the 4 ha scale of the warm-temperate evergreen broad-leaved forests. Moreover, it was found by comparing with three different forest types that interspecific competition is more intense in warm-temperate forests than in cool-temperate or sub-boreal forests. We conclude that, compared to cool-temperate or sub-boreal forests (which have little interspecific competition), warm-temperate forests support more complex interspecific relationships and species-specific habitat preferences that result in higher species diversity.

Non-ectomycorrhizal fungi that associate with typical ectomycorrhizae often remain hidden, and their localization inside ectomycorrhizal (ECM) roots has remained uncharacterized. In this study, the fungal community associated with the ectomycorrhizae of Castanopsis cuspidata was investigated using a culture-dependent isolation technique. Additionally, the species composition and localization were determined using molecular techniques. The results of the isolation and identification of fungal species revealed the predominance of a few species belonging to the order Helotiales. Furthermore, the fungal community structures were significantly different depending on the taxa of the ectomycorrhiza-forming fungi. A taxon-specific probe was developed to analyze the localization of one dominant Hyaloscyphaceae (Helotiales) species in ECM tissues by in situ hybridization. Hybridization signals were detected on the surface of the fungal mantle and around the ECM fungal cells within the mantle. Hyphal penetration into ECM hyphal cells of fungal mantles was also observed. Signals were not detected in the Hartig net or plant tissues inside the mantle in healthy ectomycorrhizae. These findings suggest that the analyzed species interact not only with host plant as root endophyte but also directly with the ECM fungi.