Search Results (12)

Angelica dahurica var. formosana (ADF), which belongs to the Umbelliferae family, is one of the original plants of herbal raw material Angelicae Dahuricae Radix. ADF roots represent an enormous biomass resource convertible for disease treatment and bioproducts. But, early bolting of ADF resulted in lignification and a decrease in the coumarin content in the root, and roots lignification restricts its coumarin for commercial utility. Although there have been attempts to regulate the synthesis ratio of lignin and coumarin through biotechnology to increase the coumarin content in ADF and further enhance its commercial value, optimizing the biosynthesis of lignin and coumarin remains challenging. Based on gene expression analysis and phylogenetic tree profiling, AdNAC20 as the target for genetic engineering of lignin and coumarin biosynthesis in ADF was selected in this study. Early-bolting ADF had significantly greater degrees of root lignification and lower coumarin contents than that of the normal plants. In this study, overexpression of AdNAC20 gene plants were created using transgenic technology, while independent homozygous transgenic lines with precise site mutation of AdNAC20 were created using CRISPR/Cas9 technology. The overexpressing transgenic ADF plants showed a 9.28% decrease in total coumarin content and a significant 12.28% increase in lignin content, while knockout mutant plants showed a 16.3% increase in total coumarin content and a 33.48% decrease in lignin content. Furthermore, 29,671 differentially expressed genes (DEGs) were obtained by comparative transcriptomics of OE-NAC20, KO-NAC20, and WT of ADF. A schematic diagram of the gene network interacting with AdNAC20 during the early-bolting process of ADF was constructed by DEG analysis. AdNAC20 was predicted to directly regulate the transcription of several genes with SNBE-like motifs in their promoter, such as MYB46, C3H, and CCoAOMT. In this study, AdNAC20 was shown to play a dual pathway function that positively enhanced lignin formation but negatively controlled coumarin formation. And the heterologous expression of the AdNAC20 gene at Arabidopsis thaliana proved that the AdNAC20 gene also plays an important role in the process of bolting and flowering. Full article

Objective: To clarify the accumulation and mutual transformation patterns of the chemical components in Angelica dahurica (A. dahurica) and predict the quality markers (Q-Markers) of its antioxidant activity. Method: The types of and content changes in the chemical components in various parts of A. dahurica during different periods were analyzed by using gas chromatography-mass spectrometry technology (GC-MS). The antioxidant effect of the Q-Markers was predicted using network pharmacological networks, and molecular docking was used to verify the biological activity of the Q-Markers. Result: The differences in the content changes in the coumarin compounds in different parts were found by using GC-MS technology, with the relative content being the best in the root, followed by the leaves, and the least in the stems. The common components were used as potential Q-Markers for a network pharmacology analysis. The component-target-pathway-disease network was constructed. In the molecular docking, the Q-Markers had a good binding ability with the core target, reflecting better biological activity. Conclusions: The accumulation and mutual transformation patterns of the chemical components in different parts of A. dahurica were clarified. The predicted Q-Markers lay a material foundation for the establishment of quality standards and a quality evaluation. Full article

Pine wood nematode (PWN), Bursaphelenchus xylophilus, is a major pathogen of pine wilt disease (PWD), which is a devastating disease affecting pine trees. Eco-friendly plant-derived nematicides against PWN have been considered as promising alternatives to control PWD. In this study, the ethyl acetate extracts of Cnidium monnieri fruits and Angelica dahurica roots were confirmed to have significant nematicidal activity against PWN. Through bioassay-guided fractionations, eight nematicidal coumarins against PWN were separately isolated from the ethyl acetate extracts of C. monnieri fruits and A. dahurica roots, and they were identified to be osthol (Compound 1), xanthotoxin (Compound 2), cindimine (Compound 3), isopimpinellin (Compound 4), marmesin (Compound 5), isoimperatorin (Compound 6), imperatorin (Compound 7), and bergapten (Compound 8) by mass and nuclear magnetic resonance (NMR) spectral data analysis. Coumarins 1–8 were all determined to have inhibitory effects on the egg hatching, feeding ability, and reproduction of PWN. Moreover, all eight nematicidal coumarins could inhibit the acetylcholinesterase (AChE) and Ca²⁺ ATPase of PWN. Cindimine 3 from C. monnieri fruits showed the strongest nematicidal activity against PWN, with an LC₅₀ value of 64 μM at 72 h, and the highest inhibitory effect on PWN vitality. In addition, bioassays on PWN pathogenicity demonstrated that the eight nematicidal coumarins could effectively relieve the wilt symptoms of black pine seedlings infected by PWN. The research identified several potent botanical nematicidal coumarins for use against PWN, which could contribute to the development of greener nematicides for PWD control. Full article

Plants are affected by changes in light and adaptation mechanisms can affect secondary metabolite synthesis. In this study, the physiological response and regulation of the coumarin biosynthetic pathway of Angelica dahurica to different light intensities (natural light (CK), shade rate 50% (L1), shade rate 70% (L2), and shade rate 90% (L3)) were examined. The chlorophyll content, level of the enzymes of the antioxidant system, extent of lipid peroxidation, and concentrations of the osmoregulatory solute levels were determined in potted plants. Root transcriptome under different light intensities was sequenced using high-throughput technology, and differentially expressed genes (DEGs) related to coumarin biosynthesis were analyzed by quantitative real-time PCR (qRT-PCR). With increasing shade, Chl a, Chl b, Chl a + b, and Chl a/b content increased, while the Chl a/b ratio decreased. The antioxidant enzyme system activity and extent of membrane lipid peroxidation increased. The soluble protein (SP) and proline (Pro) content decreased with the reduction in the light intensity, and soluble sugar (SS) content was found to be highest at 50% shade. The RNA-seq analysis showed that 9388 genes were differentially expressed in the L3 group (7561 were upregulated and 1827 were downregulated). In both the L1 and L2 groups, DEGs were significantly enriched in “Ribosome biosynthesis”; meanwhile, in the L3 group, the DEGs were significantly enriched in “Amino and ribonucleotide sugar metabolism” in KEGG metabolic pathway analysis. Additionally, 4CL (TRINITY_DN40230_c0_g2) and COMT (TRINITY_DN21272_c0_g1) of the phenylpropanoid metabolic pathway were significantly downregulated in the L3 group. In conclusion, A. dahurica grew best under 50% shade and the secondary-metabolite coumarin biosynthetic pathway was inhibited by 90% shade, affecting the yield and quality of medicinal compounds. Full article

Endophytic fungi are recognized for their many potential applications in agriculture, such as supporting cropland expansion and increasing the yield and resistance of plants by creating antibiotics that inhibit the growth of pathogenic microorganisms. In addition, they can produce enzymes that break down hard-to-solubilize substances within soil, dissolve phosphates, fix nitrogen, reduce metals, and produce hormones that promote plant growth (auxin, cytokinin, and gibberellins) to keep crops healthy. In this report, three strains of endophytic fungi, namely, N1, N2, and N3, were isolated from the roots of Stevia rebaudiana (Bert.) Hemsl., Polyscias fruticosa, and Angelica dahurica in some localities in Vietnam. Through a screening process, it was found that they can produce high levels of indole acetic acid (IAA), resolve phosphates, and resist disease, and they were selected to as an alternative to chemical fertilizers to make probiotics in order to increase medicinal plant yields. The results show that the three strains of fungi have the ability to degrade phosphate to 341.90, 1498.46, and 390.79 ppm; the content of IAA produced in the culture medium reached 49.00, 52.35, and 33.34 ppm. Based on some morphological characteristics and an internal transcribed spacer gene sequence analysis of the fungal strains, N1, N2, and N3 were named Penicillium simplicissimum CN7, Talaromyces flavus BC1, and Trichoderma konilangbra DL3, respectively, which have the ability to inhibit the growth of pathogenic fungal strains, such as fungus C. gloeosporioides (CD1), fungus F. oxysporum, fungus L. theobromae N13, and N. dimidiatum. They grow significantly over a period of 5 to 6 days. Full article

The present review comprehensively gathered phytochemical, bioactivity, and pharmacokinetic reports on a linear furanocoumarin, namely oxypeucedanin. Oxypeucedanin (OP), which structurally contains an epoxide ring, has been majorly isolated from ethyl acetate-soluble partitions of several genera, particularly Angelica, Ferulago, and Prangos of the Apiaceae family; and Citrus, belonging to the Rutaceae family. The methanolic extract of Angelica dahurica roots has been analytically characterized as the richest natural OP source. This naturally occurring secondary metabolite has been described to possess potent antiproliferative, cytotoxic, anti-influenza, and antiallergic activities, as assessed in preclinical studies. In order to explore potential drug candidates, oxypeucedanin, its derivatives, and semi-synthetically optimized analogues can be considered for the complementary assessments of biological assays. Full article

Ionic liquids (ILs) have sparked much interest as alternative solvents for plant materials as they provide distinctive properties. Therefore, in this study, the capacity of ILs to extract oxypeucedanin hydrate and byakangelicin from the roots of Angelica dahurica (A. dahurica) was investigated. The back-extraction method was examined to recover target components from the IL solution as well. Herein, [Bmim]Tf₂N demonstrated outstanding performance for extracting oxypeucedanin hydrate and byakangelicin. Moreover, factors including solvent/solid ratio, extraction temperature and time were investigated and optimized using a statistical approach. Under optimum extraction conditions (solvent/solid ratio 8:1, temperature 60 °C and time 180 min), the yields of oxypeucedanin hydrate and byakangelicin were 98.06% and 99.52%, respectively. In addition, 0.01 N HCl showed the most significant ability to back-extract target components from the [Bmim]Tf₂N solution. The total content of both oxypeucedanin hydrate (36.99%) and byakangelicin (45.12%) in the final product exceeded 80%. Based on the data, the proposed approach demonstrated satisfactory extraction ability, recovery and enrichment of target compounds in record time. Therefore, the developed approach is assumed essential to considerably reduce drawbacks encountered during the separation of oxypeucedanin hydrate and byakangelicin from the roots of A. dahurica. Full article

Solid-phase microextraction (SPME), purge and trap (P&T), stir bar sportive extraction (SBSE), and dynamic headspace sampling (DHS) were applied to extract, separate and analyze the volatile compounds in the roots of Hangbaizhi, Qibaizhi, and Bobaizhi and the GC-O-MS/MS (AEDA) was utilized for the quantification of key aroma compounds. Totals of 52, 54, and 43 aroma-active compounds extracted from the three samples by the four extraction methods were identified. Among these methods, the SPME effectively extracted the aroma compounds from the A. dahurica. Thus, using the SPME methods for quantitative analysis based on external standards and subsequent dilution analyses, totals of 20, 21, and 17 aroma compounds were detected in the three samples by the sniffing test, and sensory evaluations indicated that the aromas of A. dahurica included herb, spice, and woody. Finally, principal component analysis (PCA) showed that the three kinds A. dahurica formed three separate groups, and partial least squares discriminant analysis (PLS-DA) showed that caryophyllene, (−)-β-elemene, nonanal, and β-pinene played an important role in the classification of A. dahurica. Full article

Osthenol is a prenylated coumarin isolated from the root of Angelica koreana and Angelica dahurica, and is an O-demethylated metabolite of osthole in vivo. Its various pharmacological effects have been reported previously. The metabolic pathway of osthenol was partially confirmed in rat osthole studies, and 11 metabolic products were identified in rat urine. However, the metabolic pathway of osthenol in human liver microsomes (HLM) has not been reported. In this study, we elucidated the structure of generated metabolites using a high-resolution quadrupole-orbitrap mass spectrometer (HR-MS/MS) and characterized the major human cytochrome P450 (CYP) and uridine 5′-diphospho-glucuronosyltransferase (UGT) isozymes involved in osthenol metabolism in human liver microsomes (HLMs). We identified seven metabolites (M1-M7) in HLMs after incubation in the presence of nicotinamide adenine dinucleotide phosphate (NADPH) and uridine 5′-diphosphoglucuronic acid (UDPGA). As a result, we demonstrated that osthenol is metabolized to five mono-hydroxyl metabolites (M1-M5) by CYP2D6, 1A2, and 3A4, respectively, a 7-O-glucuronide conjugate (M6) by UGT1A9, and a hydroxyl-glucuronide (M7) from M5 by UGT1A3 in HLMs. We also found that glucuronidation is the dominant metabolic pathway of osthenol in HLMs. Full article

The accurate identification of plant species is of great concern for the quality control of herbal medicines. The Korean Pharmacopoeia and the Pharmacopoeia of the People’s Republic of China define Angelicae Dahuricae Radix (Baek-Ji in Korean and Bai-zhi in Chinese) as the dried roots of Angelica dahurica or A. dahurica var. formosana belonging to the family Apiaceae. Discrimination among Angelica species on the basis of morphological characteristics is difficult due to their extremely polymorphic traits and controversial taxonomic history. Furthermore, dried roots processed for medicinal applications are indistinguishable using conventional methods. DNA barcoding is a useful and reliable method for the identification of species. In this study, we sequenced the internal transcribed spacer (ITS) region of nuclear ribosomal RNA genes in A. dahurica, A. dahurica var. formosana, and the related species A. anomala and A. japonica. Using these sequences, we designed species-specific primers, and developed and optimized a multiplex sequence-characterized amplified region (SCAR) assay that can simply and rapidly identify respective species, and verify the contamination of adulterant depending on the polymerase chain reaction (PCR) amplification without sequencing analysis in a single PCR reaction. This assay successfully identified commercial samples of Angelicae Dahuricae Radix collected from Korean and Chinese herbal markets, and distinguished them from adulterants. This multiplex SCAR assay shows a great potential in reducing the time and cost involved in the identification of genuine Angelicae Dahuricae Radix and adulterant contamination. Full article

DA-9805 is a botanical anti-Parkinson’s drug candidate formulated from ethanol extracts of the root of Bupleurum falcatum, the root cortex of Paeonia suffruticosa, and the root of Angelica dahurica. The pharmacokinetics (PKs) and brain distribution of active/representative ingredients of DA-9805, Saikosaponin a (SSa; 1.1–4.6 mg/kg), Paeonol (PA; 14.8–59.2 mg/kg), and Imperatorin (IMP; 1.4–11.5 mg/kg) were evaluated following the intravenous or oral administration of each pure component and the equivalent dose of DA-9805 in rats. All three components had greater dose-normalized areas under the plasma concentration-time curve (AUC) and slower clearance with higher doses, following intravenous administration. By contrast, dose-proportional AUC values of SSa, PA, and IMP were observed following the oral administration of each pure component (with the exception of IMP at the highest dose) or DA-9805. Compared to oral administration of each pure compound, DA-9805 administration showed an increase in the AUC of SSa (by 96.1–163%) and PA (by 155–164%), possibly due to inhibition of their metabolism by IMP or other component(s) in DA-9805. A delay in the absorption of PA and IMP was observed when they were administered as DA-9805. All three components of DA-9805 showed greater binding values in brain homogenates than in plasma, possibly explaining why the brain-to-plasma ratios were greater than unity following multiple oral administrations of DA-9805. By contrast, their levels in cerebrospinal fluid were negligible. Our results further our understanding of the comprehensive PK characteristics of SSa, PA, and IMP in rats and the comparative PKs between each pure component and DA-9805. Full article

A series of new furanocoumarins with long-chain hydrophobic groups, namely andafocoumarins A–H and J, have been isolated from the dried roots of Angelica dahurica cv. Hangbaizhi (Angelicae Dahuricae radix) in our previous study, among which andafocoumarins A and B were demonstrated to have better anti-inflammatory activity than the positive controls. In this work, a sensitive, accurate, and efficient ultra-fast liquid chromatography coupled with triple quadrupole mass spectrometer (UFLC-MS/MS) method was developed and validated for simultaneous quantification of above-mentioned nine compounds in four cultivars of Angelicae Dahuricae Radix. Chromatographic separation was performed on a Kinetex 2.6u C18 100 Å column (100 × 2.1 mm, 2.6 µm). The mobile phases were comprised of acetonitrile and water with a flow rate of 0.5 mL/min. Using the established method, all components could be easily separated within 12 min. With the multiple reaction monitor mode, all components were detected in positive electrospray ionization. The method was validated with injection precision, linearity, lower limit of detection, lower limit of quantification, precision, recovery, and stability, respectively. The final results demonstrated that the method was accurate and efficient, which could be used to simultaneously quantify the nine andafocoumarins in Angelicae Dahuricae Radix. The results also indicated that in different batches of Angelicae Dahuricae Radix, some of the andafocoumarins were significantly different in terms of content. Full article