Search Results (21)

Background: The GRAS transcription factor family plays pivotal regulatory roles in plant growth and development, hormone signaling, and responses to environmental stresses. Taxus cuspidata, an endangered and nationally protected conifer species endemic to China, is recognized as the sole natural source of paclitaxel—an anti-cancer compound of considerable pharmaceutical importance. While GRAS family genes have been systematically characterized in various plant species, a comprehensive investigation in T. cuspidata has yet to be conducted. Methods: In this study, 26 TcGRAS genes were identified and characterized through genome-wide analysis. Results: Phylogenetic analysis grouped these genes into seven subfamilies, indicating a high degree of evolutionary conservation. Gene structure analysis revealed that most TcGRAS genes lacked introns, while promoter regions were enriched with cis-acting elements related to light responsiveness, hormone signaling (e.g., MeJA, ABA, SA), and abiotic stress responses. Tissue-specific expression profiling and qRT-PCR analyses showed that several TcGRAS genes were highly expressed in paclitaxel-accumulating organs such as roots, stems, and bark—particularly TcGRAS6, TcGRAS13, and TcGRAS24—suggesting their putative involvement in paclitaxel biosynthesis through hormone-mediated regulatory pathways. Co-expression network analysis further identified TcGRAS13 and TcGRAS14 as central nodes within the transcriptional regulatory network. Conclusions: This study provides the first genome-wide identification and comprehensive characterization of the GRAS gene family in T. cuspidata, thereby establishing a theoretical framework and providing valuable candidate gene resources for elucidating their biological functions and regulatory roles in secondary metabolism, particularly in paclitaxel biosynthesis. Full article

Understanding ecological niche evolution patterns is crucial for elucidating biogeographic history and guiding biodiversity conservation. Taxus is a Tertiary relict gymnosperm with 11 lineages mainly distributed across East Asia, spanning from tropical to subarctic regions. However, the spatiotemporal dynamics of its ecological niche evolution and the roles of ecological and geographical factors in lineage diversification, remain unclear. Using occurrence records, environmental data, and reconstructed phylogenies, we employed ensemble ecological niche models (eENMs), environmental principle components analysis (PCA-env), and phyloclimatic modeling to analyze niche similarity and evolution among 11 Taxus lineages. Based on reconstructed Bayesian trees and geographical distribution characteristics, we classified the eleven lineages into four clades: Northern (T. cuspidata), Central (T. chinensis, T. qinlingensis, and the Emei type), Western (T. wallichiana, T. florinii, and T. contorta), and Southern (T. calcicola, T. phytonii, T. mairei, and the Huangshan type). Orogenic activities and climate changes in the Tibetan Plateau since the Late Miocene likely facilitated the local adaptation of ancestral populations in Central China, the Hengduan Mountains, and the Yunnan–Guizhou Plateau, driving their expansion and diversification towards the west and south. Key environmental variables, including extreme temperature, temperature and precipitation variability, light, and altitude, were identified as major drivers of current niche divergence. Both niche conservatism and divergence were observed, with early conservatism followed by recent divergence. The Southern clade exhibits high heat and moisture tolerance, suggesting an adaptive shift, while the Central and Western clades retain ancestral drought and cold tolerance, displaying significant phylogenetic niche conservatism (PNC). We recommend prioritizing the conservation of T. qinlingensis, which exhibits the highest PNC level, particularly in the Qinling, Daba, and Taihang Mountains, which are highly degraded and vulnerable to future climate fluctuations. Full article

Climate change is becoming an important driver of biodiversity loss by altering the habitat, distribution and interspecific relationships of species. Japanese yew (Taxus cuspidata) is a first class protected plant in China, which has important ecological significance and occupies a certain position in the feeding habit of wapiti (Cervus elaphus) and Siberian roe deer (Capreolus pygargus). Due to human and animal damage, the number of Japanese yew has gradually decreased. Therefore, understanding the potential distribution of Japanese yew and the suitable areas for deer to browse on it under climate change will help to further protect these three species in Northeast China, especially migrate to more suitable areas in different scenarios in the future. From July 2021 to July 2024, we collected the information of species distribution and the variables associated with the species’ ecological limits in Muling National Nature Reserve to cross-reflect the current and future distribution and feeding area of the two species to assess each other’s impacts with Maximum entropy model (MaxEnt). The results showed that under the SSP2-4.5 and SSP5-8.5 scenarios, feeding pressure, driest quarter precipitation (BIO17) and seasonal temperature variation coefficient (BIO4) were the main variables affecting the distribution of Japanese yew, and the driest quarter precipitation (BIO17) and annual precipitation (BIO12) were the main variables affecting wapiti and Siberian roe deer foraging them. Under SSP2-4.5 and SSP5-8.5 scenarios, the suitable area of Japanese yew and the feeding area of the two species of deer gradually decreased from 2041 to 2100. Compared with wapiti, Siberian roe deer has a greater impact on the distribution range of Japanese yew, and the suitable feeding area is wider. It is expected that the potential centroid of Japanese yew, wapiti and Siberian roe deer will migrate to higher latitudes in the future. These findings provide a scientific basis for the reserve to develop relevant measures and plans and effectively protect the three species. Full article

Taxus × media, belonging to the genus Taxus of the Taxaceae family, is a unique hybrid plant derived from a natural crossbreeding between Taxus cuspidata and Taxus baccata. This distinctive hybrid variety inherits the superior traits of its parental species, exhibiting significant biological and medicinal values. This paper comprehensively analyzes Taxus × media from multiple dimensions, including its cultivation overview, chemical composition, and multifaceted applications in the medical field. In terms of chemical constituents, this study delves into the bioactive components abundant in Taxus × media and their pharmacological activities, highlighting the importance and value of these components, including paclitaxel, as the lead compounds in traditional medicine and modern drug development. Regarding its medicinal value, the article primarily discusses the potential applications of Taxus × media in combating tumors, antibacterial, anti-inflammatory, and antioxidant activities, and treating diabetes. By synthesizing clinical research and experimental data, the paper elucidates the potential and mechanisms of its primary active components in preventing and treating these diseases. In conclusion, Taxus × media demonstrates its unique value in biological research and tremendous potential in drug development. Full article

Taxanes are the best-known compounds in Taxus cuspidata owing to their strong anticancer effects. However, the traditional taxanes extraction method is the solid–liquid extraction method, which is limited by a large energy consumption and low yield. Therefore, it is urgent to find an efficient method for taxanes extraction. The ultrasonic microwave synergistic extraction (UME) method integrates the cavitation effect of ultrasound and the intensifying heat transfer (ionic conduction and dipole rotation of molecules) effect of microwave to accelerate the release of intracellular compounds and is used in active ingredient extractions. This study aimed to evaluate the performance of UME in extracting taxanes from T. cuspidata needles (dichloromethane-ethanol as extractant). A single-factor experiment, Plackett–Burman design, and the response surface method showed that the optimal UME parameters for taxanes extraction were an ultrasonic power of 300 W, a microwave power of 215 W, and 130 sieve meshes. Under these conditions, the taxanes yield was 570.32 μg/g, which increased by 13.41% and 41.63% compared with the ultrasound (US) and microwave (MW) treatments, respectively. The reasons for the differences in the taxanes yield were revealed by comparing the physicochemical properties of T. cuspidata residues after the UME, US, and MW treatments. The cell structures were significantly damaged after the UME treatment, and numerous tiny holes were observed on the surface. The absorption peaks of cellulose, hemicellulose, and lignin increased significantly in intensity, and the lowest peak temperature (307.40 °C), with a melting enthalpy of −5.19 J/g, was found after the UME treatment compared with the US and MW treatments. These results demonstrate that UME is an effective method (570.32 μg/g) to extract taxanes from T. cuspidata needles by destroying cellular structures. Full article

In this research, the cell growth, physiological, and biochemical reactions, as well as the paclitaxel production, of Taxus cuspidata suspension cells after treatment with polyethylene glycol (PEG), cyclodextrin (CD), or salicylic acid (SA) (alone or in combination) were investigated. To reveal the paclitaxel synthesis mechanism of T. cuspidata suspension cells under elicitor treatment, the transcriptomics of the Control group and P + C + S group (PEG + CD + SA) were compared. The results show that there were no significant differences in cell biomass after 5 days of elicitor treatments. However, the content of hydrogen peroxide (H₂O₂) and malondialdehyde (MDA), and the activities of phenylalanine ammonia-lyase (PAL) and polyphenol oxidase (PPO) after elicitor combination treatments were decreased compared with the single-elicitor treatment. Meanwhile, the antioxidant enzyme activity (superoxide dismutase (SOD), catalase (CAT), and peroxidase (PO)) and the contents of soluble sugar and soluble protein were increased after combination elicitor treatments. Additionally, the paclitaxel yield after treatment with the combination of all three elicitors (P + C + S) was 6.02 times higher than that of the Control group, thus indicating that the combination elicitor treatments had a significant effect on paclitaxel production in T. cuspidata cell suspension culture. Transcriptomics analysis revealed 13,623 differentially expressed genes (DEGs) between the Control and P + C + S treatment groups. Both GO and KEGG analyses showed that the DEGs mainly affected metabolic processes. DEGs associated with antioxidant enzymes, paclitaxel biosynthesis enzymes, and transcription factors were identified. It can be hypothesized that the oxidative stress of suspension cells occurred with elicitor stimulation, thereby leading to a defense response and an up-regulation of the gene expression associated with antioxidant enzymes, paclitaxel synthesis enzymes, and paclitaxel synthesis transcription factors; this ultimately increased the production of paclitaxel. Full article

Taxus cuspidata is a rare and endangered plant species with an extremely small population which is endemic to China. This study focused on the natural T. cuspidata population in Jilin Province in China. Conventional population ecology survey methods were used to describe its population structure characteristics. Then, we chose diameter structure instead of temporal structure to establish a static population life table, draw a population survival curve, and quantify the future development trend by using population dynamic analysis and time sequence prediction. The results showed that: (1) the static life table suggested that the population of T. cuspidata was stable overall. The population survival curve tended to be Deevey Ⅱ, with a high early seedling mortality, and the later population growth tended to be stable; (2) the survival curve suggested that the population initially experienced higher mortality rates in the early stage. However, as time progressed and the population aged, the mortality rates decreased, resulting in a more stable population in the middle and late stages; (3) The diameter class structure of T. cuspidata was stable overall, and the dynamic indices showed that the population was fluctuating. The population was influenced by external disturbances and showed some resistance to human disturbance; (4) time sequence prediction analysis showed that the mortality rate of young individuals was high, natural renewal could be maintained, and the population size would remain at a certain amount in the future. The result shows that the Jilin region is a highly suitable area for the growth of Taxus cuspidata’s population in Northeast China. We recommend in situ conservation of remaining wild populations, relocation of germplasm resources, and reduction of human activities; these actions will be beneficial to Taxus cuspidata’s long-term survival. Full article

Our research group successfully designed the F. carica and Taxus cuspidata Sieb. et Zucc mixed forest, and confirmed that their interspecific relationship was stable and F. carica has a promoting effect on neighboring T. cuspidata growth. However, the promoting mechanism has not been elucidated. In this study, F. carica was used as the donor plant and T. cuspidata was used as the recipient plant. T. cuspidata seedlings were irrigated with F. carica root extracts of different concentrations (10.0, 20.0, 40.0 g·L⁻¹), and the plant height, base diameter, photosynthetic parameters, photosynthetic pigments, MDA contents, and antioxidant enzyme activities were measured. Soil physical and chemical properties, enzyme activities, and microbial diversity were measured. The results showed that the abundance of growth-promoting bacteria increased and the number of pathogenic bacteria decreased in the rhizosphere of T. cuspidata soil. It was speculated that the chemicals secreted by F. carica roots interacted with soil microorganisms of T. cuspidata soil after enrichment, changed soil microbial diversity, and indirectly promoted the growth of T. cuspidata. UPLC-QTOF-MS/MS was used to analyze F. carica root water extract and F. carica root exudates, respectively, and it was found that the main components were similar. Therefore, the promoting effect of F. carica on T. cuspidata is mainly caused by the accumulation of potential chemicals in F. carica root exudates in the soil through interaction with soil microorganisms. Furthermore, the allelopathic-promoting mechanism of F. carica on T. cuspidata was discussed from various aspects, to provide a theoretical basis for the protection, breeding, and sustainable management of T. cuspidata resources. Full article

In the present study, an effective method of preparative high-performance liquid chromatography (Prep-HPLC) was established to purify two taxanes in Taxus cuspidata. During the experimental operation, the effects of flow rate, injection volume, and column temperature on the purity of 10-deacetyltaxol (10-DAT) and paclitaxel (PTX) were investigated, and the optimized conditions were as follows: flow rate of 10 mL/min, injection volume of 0.5 mL, and column temperature of 30 °C. Under these conditions, the purity of 10-DAT and PTX reached 95.33% and 99.15%, respectively. The purified products were characterized by scanning electron microscopy (SEM), high-performance liquid chromatography (HPLC), and electrospray ionization-high resolution mass spectrometry (ESI-HRMS). The results demonstrated that preparative HPLC can effectively purify 10-DAT and PTX from Taxus cuspidata with a purity of >95%, which was suitable for the large-scale preparation of 10-DAT and PTX. Full article

High concentrations of airborne particulate matter (PM) in urban areas are of great concern to human health. Urban greening has been shown to be an effective and eco-friendly way to alleviate particle pollution, and attention to its role in mitigating particle pollution has increased worldwide. The species-specific PM-capturing capacity of ten urban-greening species in Seoul was evaluated by leaf functional traits (average leaf area (ALA), specific leaf area (SLA), and leaf width-to-length ratio (W/L)), microstructures (roughness, stomata, and trichomes), and physicochemical traits (contact angle (θ_w), surface free energy (r_s), the work of adhesion for water (W_a), and epicuticular wax loads (EWL)). The relationships between leaf traits and PM adsorption by leaves were revealed by Pearson’s correlations and principal component analysis (PCA). A gravimetric method was used to quantify, by particle size, the PM adsorbed on leaf surfaces or embedded in leaf epicuticular wax layers. The key factors for PM adsorption on leaf surfaces were the SLA, the mean roughness value (R_a), and stomatal size. The SLA and R_a of adaxial leaf surfaces were negatively correlated with PM accumulation on leaf surfaces, while stomatal length and width were positively correlated with surface PM load. The r_s and EWL positively affected the in-wax PM load. Species-specific PM deposition was the result of complicated mechanisms of various leaf traits. Three evergreen shrub species, Buxus sinica (Rehder & E.H. Wilson) M.Cheng var. insularis (Nakai) M.Cheng, Taxus cuspidata Siebold & Zucc., and Euonymus japonicus Thunb., were efficient in capturing both surface PM and in-wax PM. The PCA revealed that the high PM accumulation efficiency of these three species might be attributable to the interaction between stomatal size and EWL. Aesculus turbinata Blume, Chionanthus retusus Lindl. & Paxton, and Rhododendron schlippenbachii Maxim. had intermediate PM adsorption ability, which might be a result of interactions among stomatal density, the W_a of adaxial surfaces, and ALA. Magnolia denudata Desr., Styphnolobium japonicum (L.) Schott, Liriodendron tulipifera L., and Ginkgo biloba L. had low PM accumulation efficiency. These four species exhibited correlations among SLA, the R_a of adaxial leaf surfaces, and W/L, which had negative effects on PM adsorption. Full article

Taxanes are natural compounds with strong antitumor activity. In this study, we first enriched taxanes by ultrasonic extraction and liquid–liquid extraction from Taxus cuspidata, then purified these taxanes by the antisolvent recrystallization method, and discussed the effects of four recrystallization conditions on the purity of eight target compounds. The most promising purification results were obtained using methanol as a solvent and water as an antisolvent. Response surface methodology (RSM) was used to further optimize the optimal purification conditions: when the crude extraction concentration was 555.28 mg/mL, an antisolvent to solvent volume ratio was 28.16 times, the deposition temperature was 22.91 °C, and the deposition time was 1.76 min, the purity of the taxanes reached its maximum. The scanning electron microscopy (SEM) results showed that recrystallization could effectively reduce the particle size of crude Taxus cuspidata and control the particle morphology. X-ray diffraction (XRD) and Raman spectrum experiments demonstrated that the amorphous state of the crude Taxus cuspidata did not change during the recrystallization process, and always remained amorphous. This recrystallization method can effectively improve the purity of taxanes in Taxus cuspidata, and is suitable for the preliminary purification of taxanes. Full article

Taxanes are natural compounds with strong antitumor activity. In this study, we first extracted taxanes from the needles of Taxus cuspidata using ultrasonic (US) extraction, and then assessed the effects of different extraction conditions on the yields of eight target compounds. Response surface methodology (RSM) was further used to optimize the extraction conditions: when the liquid-to-solid ratio was 20.88 times, ultrasonic power was 140.00 W, ultrasonic time was 47.63 min, and ethanol content in solvent was 83.50%, taxane yields reached the maximum value of 354.28 μg/g. Under these conditions, the actual extraction rate of taxanes from the needles was 342.27 μg/g. The scanning electron microscopy (SEM) results indicated that the morphology of the needles, suspension cells, and callus of Taxus cuspidata extracted by ultrasonic wave had changed, the pores of the sections of the needles extracted by ultrasonic wave had become relatively loose, and the pore diameter had obviously increased. The callus and overall structure of the suspension cells extracted by ultrasonic wave were destroyed, forming cell fragments. The components of Taxus cuspidata are complex; the high-performance liquid chromatography (HPLC) method established in this paper is suitable for the rapid and effective separation of taxanes in Taxus cuspidata. We systematically and comprehensively compared the yields of taxanes in needles, callus, and suspension cells of Taxus cuspidata, and the taxane yields were increased by the suspension cell culture. Full article

Taxanes are a series of natural compounds with great application potential in antitumor therapy, whereas the lack of efficient taxanes extraction methods significantly hinders the development of taxanes. The high-intensity pulsed electric field (PEF) is a novel technology used to extract bioactive ingredients from food and other natural products. However, the prospect of using PEF for taxanes extraction remains to be elucidated. Herein, we extracted taxanes from Taxus cuspidata via PEF and explored the effects of seven extraction conditions on the yields of target compounds. The Placket–Burman design (PBD) assay revealed that electric field strength, pulse number, and particle size are key factors for taxanes extraction. The response surface methodology (RSM) and back-propagation neural network conjugated with genetic algorithm (GA-BP) were further used to model and predict the optimal extraction conditions, and GA-BP exerted higher reliability, leading to a maximum extraction yield of 672.13 μg/g under electric field strength of 16 kV/cm, pulse number of 8, particle size of 160 meshes, solid–liquid ratio of 1:60, a single extraction, centrifugal speed of 8000 r/min, and flow rate of 7 mL/min, which was 1.07–1.84 folds that of control, solid–liquid extraction (SL), and ultrasonic extraction (US) groups. Additionally, the scanning electron microscopy (SEM) results indicated that the sample particles extracted by PEF method exhibited a coarser surface morphology. Thus, we present for the first time that PEF is feasible for the extraction of taxanes from Taxus cuspidata and highlight the application value of the PBD, RSM, and GA-BP models in parameters optimization during extraction process. Full article

Medicinal-agroforestry systems are one of the multi-functional medicinal plant production systems, gaining attention as a sustainable alternative to traditional monoculture systems. In this study, three planting patterns were established which included: (1) monoculture F. carica (MF); (2) monoculture T. cuspidata (MT); and (3) interplanting F. carica with T. cuspidata (IFT). The differences of growth biomass, photosynthesis, soil nutrients, soil enzyme activities, soil microorganisms, and main secondary metabolites of F. carica and T. cuspidata under the above three models were investigated. Compared with the MF and MT patterns, IFT pattern for 5 months significantly increased the plant growth biomass, photosynthesis, soil organic carbon, total nitrogen, and secondary metabolites content. The activities of acid phosphatase, sucrase, protease, polyphenol oxidase, urease, dehydrogenase, and catalase in soil of IFT were significantly higher than MF and MT patterns. Results showed that IFT pattern is preferred compared to the MF and MT patterns. Our result will help to provide a feasible theoretical basis for the large-scale establishment of F. carica and T. cuspidata mixed forests and obtain high-quality medicine sources for extracting important active ingredients, psoralen and paclitaxel, which are crucial to the long-term sustainable development and production of medicinal plants. Full article

Endophytic fungi inside a plant can degrade a portion of plant lignin and cellulose. Endophytic Penicillium is one of the industrial microorganisms with the advantage of producing enzymes with a complete enzyme system that can be secreted into the extracellular space. The natural evolution of ancient tree species from special natural geographic environments to screen out cellulase-producing strains with excellent characteristics provides a promising direction for future industrial enzymes. The present study successfully isolated and screened a novel fungal endophyte, Penicillium oxalicum R4, with higher cellulase activity from Taxus cuspidata. Under the optimized culture conditions obtained by a Box–Behnken design (BBD) and an artificial neural network–genetic algorithm (ANN–GA), yields of Filter Paperase (FPase), Carboxymethyl Cellulase (CMCase) and β-glucosidase (βGLase) produced by P. oxalicum R4 were 1.45, 5.27 and 6.35 U/mL, which were approximately 1.60-fold, 1.59-fold and 2.16-fold higher than those of the non-optimized culture, respectively. The discovery of cellulase-producing strains of endophytic fungi located in special natural geographic environments, such as Taxus cuspidata, which is known as a living plant fossil, provides new research directions for future industrial enzymes. Full article