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The antinociceptive activity of the flower extracts of Styrax japonicus was confirmed in our previous study. However, the key compound for analgesia has not been distinguished, and the corresponding mechanism is obscure. In this study, the active compound was isolated from the flower by multiple chromatographic techniques and structurally illustrated using spectroscopic methods and referring to the related literature. The antinociceptive activity of the compound and the underlying mechanisms were investigated using animal tests. The active compound was determined to be jegosaponin A (JA), which showed significant antinociceptive responses. JA was also shown to possess sedative and anxiolytic activities but no anti-inflammatory effect, implying the association of the antinociceptive effects with the sedative and anxiolytic activities. Further antagonists and calcium ionophore tests showed that the antinociceptive effect of JA was blocked by flumazenil (FM, antagonist for GABA-A receptor) and reversed by WAY100635 (WAY, antagonist for 5-HT1A receptor). Contents of 5-HT and its metabolite (5-HIAA) increased significantly in the hippocampus and striatum tissues after JA administration. The results indicated that the antinociceptive effect of JA was regulated by the neurotransmitter system, especially GABAergic and serotonergic systems.

Styrax japonicus is a small tree or shrub with flowers of great ornamental value. This experiment was conducted to study the metabolites of S. japonicus petals in two colors to improve their value of ornamental and economic. Two kinds of petals were sampled at the pre-bloom stage (S1), the full-bloom stage (S2), and the post-peak stage (S3), respectively. Extensive non-targeted metabolomic assays were performed on the petals using LC-MS. A total of 552 distinct metabolites were found. Additionally, the relative contents of organooxygen compounds, carboxylic acids and derivatives, and fatty acyls in each developmental stage of both petals were the top 3. Notably, a high relative content of kaempferol-3- O -rutinoside was found in each developmental stage of both petals. The differential metabolites of the two types of petals varied considerably at different developmental stages. A portion of the differential metabolites in the S1 was enriched for amino acid and protein synthesis, while a portion of the differential metabolites in the S2 and S3 were enriched for aromatic compound degradation. Furthermore, compared to white flowers, pink flowers might have more metabolic activity. At the same time, in all developmental stages, there is an enrichment of some differential metabolites into the phenylpropanoid synthesis pathway.

Global climate change has led to an increased frequency of extreme weather events, with flooding caused by heavy rainfall posing a significant threat to plant growth and survival. Styrax japonicus, a species of ecological and economic importance, exhibits stronger flooding tolerance compared to its congener Styrax tonkinensis. Endogenous hormonal systems in plants are indispensable for integrating growth dynamics, developmental transitions, and ecological stress perception-transduction pathways. To investigate the response of S. japonicus to flooding stress at both hormonal and molecular levels, this study utilized annual seedlings of S. japonicus as experimental material. Two levels of flooding stress, waterlogging and submergence, were applied to examine the variations in endogenous hormone levels in S. japonicus roots under different stress conditions and durations. Combined with transcriptome sequencing, critical genes associated with hormone-mediated signaling and biosynthetic processes were identified. The results showed that the content of the ethylene precursor ACC exhibited a trend of “increase–decrease–increase”, with an earlier decline under submergence compared to waterlogging stress by approximately 10 days. Abscisic acid content sharply decreased at 5 d, followed by an initial increase and subsequent decrease, with higher ABA levels observed under waterlogging stress than under submergence. GA content significantly decreased after 10 d in both stress conditions. KEGG enrichment analysis revealed that the most prominently enriched pathway for DEGs was plant hormone signal transduction under both waterlogging and submergence stress, with 314 and 370 DEGs identified, respectively. Analysis of common genes indicated their association with ethylene, ABA, auxin, and BRs. After further investigation of DEGs in the ethylene and ABA biosynthesis process, we identified key enzyme genes encoding ACS, ACO, and NCED, which are critical for their biosynthesis.

Styrax japonicus is a medicinal and ornamental shrub belonging to the Styracaceae family. To explore the diversity and characteristics of the chloroplast genome of S. japonicus, we conducted sequencing and comparison of the chloroplast genomes of four naturally distributed S. japonicus. The results demonstrated that the four chloroplast genomes (157,914–157,962 bp) exhibited a typical quadripartite structure consisting of a large single copy (LSC) region, a small single copy (SSC) region, and a pair of reverse repeats (IRa and IRb), and the structure was highly conserved. DNA polymorphism analysis revealed that three coding genes (infA, psbK, and rpl33) and five intergene regions (petA-psbJ, trnC-petN, trnD-trnY, trnE-trnT, and trnY-trnE) were identified as mutation hotspots. These genetic fragments have the potential to be utilized as DNA barcodes for future identification purposes. When comparing the boundary genes, a small contraction was observed in the IR region of four S. japonicus. Selection pressure analysis indicated positive selection for ycf1 and ndhD. These findings collectively suggest the adaptive evolution of S. japonicus. The phylogenetic structure revealed conflicting relationships among several S. japonicus, indicating divergent evolutionary paths within this species. Our study concludes by uncovering the genetic traits of the chloroplast genome in the differentiation of S. japonicus variety, offering fresh perspectives on the evolutionary lineage of this species.

To prepare and manage urban greenspace for a hotter future, we must select trees that can tolerate or acclimate to high temperatures. Here, we compared tolerance and acclimation to high temperatures among nine urban tree species in Japan. Saplings were exposed to heat from different times (early and mid-summer) during the growing season in a greenhouse. With the exception of Ginko, heating in early summer did not affect whole-tree health, suggesting that most urban tree species may be able to acclimate to higher temperatures during the early growing season. However, continued exposure to higher temperatures, as well as heating from mid-summer, had negative effects on tree health, leading to wilting/browning, especially for evergreen broadleaved species whose leaves mature slowly. Cornus florida, Styrax japonicus and Morella rubra were the most vulnerable to heating, such that all heated saplings had died by the end of summer. At the leaf level, leaf maturation of the deciduous species and Morella was negatively affected by heating, whereas that of Eurya emarginata and Euonymus japonicas were not affected. These two species also showed heat tolerance, having a higher T50 (temperature where leaf quantum yield declined to 50% of maximum value due to heat stress) compared to other species, as well as heat acclimation, where T50 was higher for the heated saplings compared to the control. Our results indicate that, while some species that cannot recover from heat damage in early summer could die, others can acclimate to sustained high temperatures, as well as to late summer heat. As heatwaves are expected to become more frequent and severe due to global warming, tree species need to be screened individually to assess their ability to tolerate or acclimate to high temperature.

Styrax japonicus sieb. et Zucc. is widely distributed in China with ornamental and medicinal values. However, the transcriptome of S. japonicus has not yet been reported. In this study, we carried out the first transcriptome analysis of S. japonicus and developed a set of expressed sequence tag–simple sequence repeats (EST–SSRs). We obtained 338,570,222 clean reads in total, of which the mean GC content was 41.58%. In total, 136,071 unigenes were obtained having an average length of 611 bp and 71,226 unigenes were favorably annotated in the database. In total, we identified 55,977 potential EST–SSRs from 38,611 unigenes, of which there was 1 SSR per 6.73 kb. The di-nucleotide repeats (40.40%) were the most identified SSRs. One set of 60 primer pairs was randomly selected, and the amplified products in S. japonicus were validated; 28 primer pairs successfully produced clear amplicons. A total of 21 (35%) polymorphic genic SSR markers were identified between two populations. In total, 15 alleles were detected and the average number was 6. The average of observed heterozygosity and expected heterozygosity was 0.614 and 0.552, respectively. The polymorphism information content (PIC) value fluctuated between 0.074 and 0.855, with a mean value of 0.504, which was also the middle level. This study provides useful information for diversity studies and resource assessments of S. japonicus.

Styrax japonicus is a small ornamental tree with medicinal value. An S. japonicus variety with purplish red calyxes and white petals has higher ornamental value. The mechanism underlying calyx pigmentation in S. japonicus is still unclear. In this study, metabolome data combined with transcriptome profiling were used to explore the molecular mechanisms underlying the difference in the color of calyx in two varieties of S. japonicus, namely, Red Linglong (RA; purplish red calyx) and Green Linglong (GA; green calyx). The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis indicated that the levels of delphinidin 3,5-diglucoside, delphinidin 3-O-glucoside, and malvidin 3-O-glucoside when enriched in delphinidin biosynthesis were significantly higher in the RA calyx than in the GA calyx. These key differentially accumulated metabolites were highly correlated with five late biosynthetic genes that were enriched in the anthocyanin biosynthesis pathway. MYB1, MYB82, and MYB113 were the three probable transcription factors responsible for anthocyanin accumulation. This study provides novel insights into secondary metabolism pathways, their regulators, and the changes in the transcription and metabolite levels in the calyx of S. japonicus regulating sepal color. The results provide a theoretical basis for exploring the mechanism of calyx color formation in S. japonicus and provide genetic material and a reference for molecular breeding to obtain desired flower colors in the future.

Germination of Styrax japonicus seeds is promoted by warm stratification (WS) at 18–20°C followed by cold stratification (CS) at 4–5°. The objective of this work was to analyze the state and mobility of water molecules measured by 1H-NMR and endogenous gibberellins (GAs) and abscisic acid (ABA) by ultra performance liquid chromatography/mass spectrometry/mass spectrometry (UPLC-MS/MS) as influenced by WS and CS treatments had not previously been investigated. Styrax seeds that received 35 days of WS (35D WS) followed by 63 days of CS (63D CS) (35D WS + 63D CS) germinated. Seeds that received only 35D WS failed to germinate. Endogenous GA1, GA8, GA19, GA20, and GA53 were identified as well as GA17, GA23, GA28, GA29, and GA97 by gas chromatography/MS (GC/MS) and UPLC-MS/MS in seeds that were treated with warm and cold stratification (WS + CS). This suggests that the early C-13 hydroxylation pathway [-GA53-(GA44)-GA19-GA20-GA1-GA8] of GAs is a major biosynthetic pathway in the seeds. The concentration of GA53 and GA19 increased following WS and that of GA53 increased after WS + CS. The concentration of GA19 increased only slightly after WS + CS. The concentration of GA1 increased only after WS + CS. ABA concentration significantly decreased following the WS treatment. It is concluded that the mobility of water molecules and water content in cotyledons and endosperm is increased following WS + CS treatments. The occurrence of C-13 hydroxylated GAs suggests that the early C-13 hydroxylation pathway, → GA53 → GA44 → GA19 → GA20 → GA1 → GA8, is a major biosynthetic pathway in Styrax seeds.

Styrax japonicus is a small ornamental tree with medicinal values, although its flowering period is short. To date, information about the morphological and physiological characteristics of the petals during the flowering period is limited. In this study, we observed the structure of the petals at the full flowering stage with a scanning electron microscope and detected the contents of nutrients, minerals, and endogenous hormones and the activities of enzymes at different flowering stages. The results showed that the content of soluble sugar exhibited an ‘increase-decrease’ trend, whereas the contents of soluble protein, nitrogen (N), phosphorus (P), and abscisic acid (ABA) showed a ‘decrease-increase’ pattern. The content of starch descended continuously, but the contents of potassium (K), gibberellic acid (GA3), indoleacetic acid (IAA), and malondialdehyde (MDA) ascended continuously. The activities of peroxidase (POD) and superoxide dismutase (SOD) first rose and then declined during the flowering period. Higher contents of soluble sugar, N, K, and IAA promoted S. japonicus flowering; meanwhile, lower contents of starch, soluble protein, P, and GA3 in addition to the lower activity of SOD might be some of the causes of the short flowering period. This work will serve as the foundation for a scientific technique to utilize the flowers and extend the flowering period in S. japonicus.

This study was carried out to clarify the vegetation decline due to air pollutants emitted in the process of industrial activities and the passive restoration of the vegetation due to socioeconomic changes after economic growth. To achieve this goal, we investigated the spatial distribution of vegetation, differences in species composition and diversity among vegetation types different in damage degree, vegetation dynamics, the age structure and annual ring growth of two dominant plant species, and the landscape change that occurred in this area over the last 50 years. Plant communities tended to be spatially distributed in the order of grassland, shrubland (dominated by Styrax japonicus Siebold and Zucc. community), and forests (dominated by Pinus thunbergii Parl. and Pinus densiflora Siebold and Zucc. communities), with increasing distance from the pollution source. The result of stand ordination based on vegetation data reflected the trend of such a spatial distribution. Species richness evaluated based on the species rank dominance curve was the highest in shrubland and the lowest in grassland; species richness in forests was intermediate. The size class distribution of woody plant species in four plant communities composing three vegetation types showed the possibility of them being replaced by forest in the late successional stage. However, the density of successor trees was relatively low, whereas the density of shrubby plants, which are resilient to air pollution, was very high. The age class distribution of a dominant species forming shrubland and pine forest showed that most of them were recruited after industrialization in this area. The period when young individuals in both vegetation types were recruited corresponded to the period when the annual ring growth of the pine trees that survived air pollution was reduced. An analysis of the landscape change in this area indicated that coniferous forest and agricultural field decreased greatly, whereas industrial area, residential area, mixed forest, and broadleaved forest showed increasing trends since construction of the industrial complex. As a result, the decrease in coniferous forest is usually due to vegetation decline and partially to succession, as the pine trees dominating the forest are not only sensitive to air pollution but are also shade-intolerant. The increase in mixed and broadleaved forests reflects vegetation decline or succession. Vegetation decline progressed for about 30 years after the construction of the industrial complex; it has begun to be restored passively since then, although the change has been slow. These results are in line with the environmental Kuznets curve hypothesis that environmental degradation increases in the early stages of economic growth to a certain point, and, after a turning point, economic development leads to environmental improvements—thus, there is an inverted U-shaped relationship between economic growth and environmental degradation.