Coumarin Content Research Articles

Aerospace Mutagenesis Increases the Content of Characteristic Metabolites of Tea Tree Leaves and Enhances Taste Characteristics of Tea Leaves

Aerospace mutagenesis can alter the physiological metabolism and growth of tea trees and affect tea leaf quality. In this study, the effect of aerospace mutagenesis on the metabolite content and taste characteristics of Dahongpao fresh leaves and raw tea were analyzed. The results showed that aerospace mutagenesis had little effect on the composition and total amount of metabolites in fresh leaves and raw tea, but significantly changed the content of different metabolites. Aerospace mutagenesis improved the content of lipids, lignans and coumarins, and amino acids and derivatives, which in turn enhanced the aroma and fresh and brisk taste of fresh leaves. Aerospace mutagenesis increased the content of alkaloids, tannins, flavonoids, lignans and coumarins, amino acids and derivatives, and organic acids in raw tea, and enhanced the bitterness, mellowness, and fresh and brisk taste of raw tea. This study provides a basis for the development of aerospace mutagenesis Dahongpao tea products and the establishment of processing techniques.

Genes to specialized metabolites: accumulation of scopoletin, umbelliferone and their glycosides in natural populations of Arabidopsis thaliana

BackgroundScopoletin and umbelliferone belong to coumarins, which are plant specialized metabolites with potent and wide biological activities, the accumulation of which is induced by various environmental stresses. Coumarins have been detected in various plant species, including medicinal plants and the model organism Arabidopsis thaliana. In recent years, key role of coumarins in maintaining iron (Fe) homeostasis in plants has been demonstrated, as well as their significant impact on the rhizosphere microbiome through exudates secreted into the soil environment. Several mechanisms underlying these processes require clarification. Previously, we demonstrated that Arabidopsis is an excellent model for studying genetic variation and molecular basis of coumarin accumulation in plants.ResultsHere, through targeted metabolic profiling and gene expression analysis, the gene-metabolite network of scopoletin and umbelliferone accumulation was examined in more detail in selected Arabidopsis accessions (Col-0, Est-1, Tsu-1) undergoing different culture conditions and characterized by variation in coumarin content. The highest accumulation of coumarins was detected in roots grown in vitro liquid culture. The expression of 10 phenylpropanoid genes (4CL1, 4CL2, 4CL3, CCoAOMT1, C3’H, HCT, F6’H1, F6’H2,CCR1 and CCR2) was assessed by qPCR in three genetic backgrounds, cultured in vitro and in soil, and in two types of tissues (leaves and roots). We not only detected the expected variability in gene expression and coumarin accumulation among Arabidopsis accessions, but also found interesting polymorphisms in the coding sequences of the selected genes through in silico analysis and resequencing.ConclusionsTo the best of our knowledge, this is the first study comparing accumulation of simple coumarins and expression of phenylpropanoid-related genes in Arabidopsis accessions grown in soil and in liquid cultures. The large variations we detected in the content of coumarins and gene expression are genetically determined, but also tissue and culture dependent. It is particularly important considering that growing plants in liquid media is a widely used technology that provides a large amount of root tissue suitable for metabolomics. Research on differential accumulation of coumarins and related gene expression will be useful in future studies aimed at better understanding the physiological role of coumarins in roots and the surrounding environments.

AdNAC20 Regulates Lignin and Coumarin Biosynthesis in the Roots of Angelica dahurica var. Formosana.

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.

Integrative analyses of metabolome and transcriptome reveal the dynamic accumulation and regulatory network in rhizomes and fruits of Polygonatum cyrtonema Hua

BackgroundAccording to Chinese ancient books, both fruits and rhizomes of Polygonatum cyrtonema Hua have medicinal and edible values. Up to now, there is no report about the metabolite profiles and regulatory network in fruits and different year-old rhizomes of P. cyrtonema.ResultsIn this study, we performed integrative analyses of metabolome and transcriptome to reveal the dynamic accumulation and regulatory network of fruits and different year-old rhizomes in P. cyrtonema. The relative content of phenolic acids, lignans and coumarins, flavonoids and alkaloids increased with growth years, while steroids and lipids decreased with it. In addition, the relative content of nucleotides and derivatives, flavonoids, organic acids, steroids and lipids in fruits were higher than rhizomes. Genes that might relate to the biosynthesis of polysaccharides, flavonoids, triterpene saponins and alkaloids biosynthesis were further analyzed by transcriptome analysis, including sacA, GMPP, PMM, CCoAOMT, CHI, ANR, CHS, DXS, GGPS, ZEP, CYP72A219 and so on, for their expressions were positively correlated with the relative content of the metabolites. Additionally, the correlation network in sugar and aromatic amino acids metabolites were constructed to further illustrate the biosynthesis of polysaccharides, flavonoids and alkaloids in P. cyrtonema, and some transcription factors (TFs) were screened, such as C2C2, MYB, bZIP, GRAS and NAC.ConclusionsThis study can deepen our understanding of the accumulation patterns and molecular mechanism of the main compounds in P. cyrtonema, and provide reference for the standardize production of P. cyrtonema.

Quantitative Analysis of Isopimpinellin from Ammi majus L. Fruits and Evaluation of Its Biological Effect on Selected Human Tumor Cells.

Ammi majus L. (Apiaceae) is a medicinal plant with a well-documented history in phytotherapy. The aim of the present work was to isolate isopimpinellin (5,8-methoxypsoralen; IsoP) from the fruit of this plant and evaluate its biological activity against selected tumor cell lines. The methanol extract obtained with the use of an accelerated solvent extraction (ASE) method was the most suitable for the quantitative analysis of coumarins in the A. majus fruit matrix. The coumarin content was estimated by RP-HPLC/DAD, and the amount of IsoP was found to be 404.14 mg/100 g dry wt., constituting 24.56% of the total coumarin fraction (1.65 g/100 g). This, along with the presence of xanthotoxin (368.04 mg/100 g, 22.36%) and bergapten (253.05 mg/100 g, 15.38%), confirmed A. majus fruits as an excellent source of these compounds. IsoP was isolated (99.8% purity) by combined liquid chromatography/centrifugal partition chromatography (LC/CPC) and tested for the first time on its antiproliferative activity against human colorectal adenocarcinoma (HT29, SW620), osteosarcoma (Saos-2, HOS), and multiple myeloma (RPMI8226, U266) cell lines. MTT assay results (96 h incubation) demonstrated a dose- and cell line-dependent decrease in cell proliferation/viability, with the strongest effect of IsoP against the Saos-2 cell line (IC50; 42.59 µM), medium effect against U266, HT-29, and RPMI8226 (IC50 = 84.14, 95.53, and 105.0 µM, respectively), and very weak activity against invasive HOS (IC50; 321.6 µM) and SW620 (IC50; 711.30 µM) cells, as well as normal human skin fibroblasts (HSFs), with IC50; 410.7 µM. The mechanistic study on the Saos-2 cell line showed that IsoP was able to reduce DNA synthesis and trigger apoptosis via caspase-3 activation. In general, IsoP was found to have more potency towards cancerous cells (except for HOS and SW620) than against healthy cells. The Selective Index (SI) was determined, underlining the higher selectivity of IsoP towards cancer cells compared to healthy cells (SI = 9.62 against Saos-2). All these results suggest that IsoP might be a promising molecule in the chemo-prevention and treatment of primary osteosarcoma.

Novel coumarins from green sweet bell pepper seeds: Their isolation, characterization, oxidative stress-mitigating, anticancer, anti-inflammatory, and antidiabetic properties

In an attempt to develop a potent, eco-friendly, and selective technique for isolating natural coumarins, this study introduced a novel extraction technique called kinetic thermomagnetic. It involved the application of four different variables: temperature, time interval, magnetic, and rotation speed. Phytochemical analysis was performed on the eighty one crude chloroform extracts of green sweet bell pepper seeds collected, identifying the presence of many botanical categories. Nine extracts were found to have the same two extraction variables—a 90-minute extraction period and a 55 °C extraction temperature—and to have the coumarin chemical class. Two of these extracts with coumarin were selected so that their coumarin contents could be chemically isolated and chromatographically purified. The molecular frameworks of four novel phytocoumarins have been discovered and differentiated by the analysis of their mass, 1H NMR, 13C NMR, and FTIR spectra. These phytocoumarins were found to have anticancer properties against eight malignant populations by MTT probing and to mitigate oxidative stress in human SH-SY5Y populations. Likewise, three and two related enzymes were used to specify the anti-inflammatory and antidiabetic capabilities, respectively. Comparing the isolated phytocoumarins to a positive control, the results showed that they had outstanding oxidative stress-mitigating properties, ranging from 71.51 to 81.48 %. Additionally, they demonstrated excellent cytotoxicity (IC50 values of 46.76–81.45 μg/ml) against the test malignant populations. The isolates also demonstrated an intriguing selective anti-inflammatory action, making them worthy of consideration as dual COX-2/5-LOX antagonists. Ultimately, when they submitted to the test against two blood-controlling enzymes, they showed a moderate antidiabetic effect. The authors concluded that by adjusting its factors, the approach used is effective and adaptable for extraction. Furthermore, natural-B4 in particular and other isolated phytocoumarins in general offer naturally derived remedies for alleviating oxidative stress and its related pathologies.

Unveiling the Biomedical Applications of Novel Coumarins Isolated From Capsicum Annuum L. Seeds by a Multivariate Extraction Technique.

For the first time, kinetic thermomagnetic extraction is a novel approach presented in this work. It required the application of four distinct variables: rotation speed (50, 75, and 100 rpm), magnetic field (0.8, 1.2, and 1.6 T), time interval (30, 60, and 90 min), and temperature (45, 55, and 65 °C). Numerous phytochemical categories were detected in the 81 crude chloroform extracts of green sweet bell pepper seeds that were collected, according to phytochemical analysis. Nine extracts were discovered to be linked to the coumarin chemical class and to have the same two extraction parameters: a 90-minute extraction duration and a 55 °C extraction temperature. To enable their coumarin contents to be chemically separated and chromatographically purified, two of these extracts containing coumarin were chosen. Four new phytocoumarins have been identified and their molecular structures distinguished using FTIR spectra, 1H-NMR, 13C-NMR, and mass analysis. By using MTT probing, it was discovered that these phytocoumarins exhibited anticancer activities against eight malignant populations and reduced oxidative stress in human SH-SY5Y populations. Similarly, the anti-inflammatory and antidiabetic properties were determined using three and two associated enzymes, respectively. The results demonstrated that the extracted phytocoumarins have exceptional oxidative stress-mitigating characteristics, ranging from 71.51 to 81.48 %, when compared to a positive control. Furthermore, they showed excellent cytotoxicity against the test malignant populations (IC50 values of 46.76-81.45 μg/ml). The isolates need to be taken into account as dual COX-2/5-LOX antagonists because they also showed a fascinating selective anti-inflammatory effect. The phytocoumarins under investigation have selectivity indices that are higher than those of the standards used, suggesting that they may have the ability to selectively block the COX2 enzyme that induces harmful inflammation. Compared to the standards, the phytocoumarins have a higher ability to block the catalytic activity of 5-LOX. This observation suggests that the phytocoumarins are powerful 5-LOX agents. Finally, they had a modest antidiabetic impact when tested against two blood-controlling enzymes. The authors came to the conclusion that the technique adopted is flexible and successful for extraction after modifying its components. Moreover, isolated phytocoumarins in general and natural-B1 in particular provide naturally derived solutions for oxidative stress and its associated diseases.

Root uptake of umbelliferone enhances pea's resistance against root-knot nematodes

Coumarins are secondary plant metabolites which play a key role in plant–plant and plant–microbe interactions. In particular, they are highly involved in environmental stress responses. Coumarins can transfer from plants producing coumarins to non-coumarin-producing neighbouring plants. As in vitro studies have shown that coumarins are nematicidal, we hypothesized that this transfer may also result in enhanced resistance against plant parasitic nematodes in coumarin-receiving plants. To test if the uptake of coumarins in a non-coumarin- producing plant protects the plant against nematode attack, we incubated non-coumarin- producing pea seedlings in growth media with the coumarin umbelliferone for three weeks, after which the plants were transplanted into soil and inoculated with root-knot nematodes (Meloidogyne incognita). We quantified the coumarin content in pea organs and nematode root invasion 2, 4, and 6 weeks after transplantation. Umbelliferone was taken up by the pea roots and translocated to the shoots. As a result of metabolization, umbelliferone and its derived metabolites coumarin, scopoletin, and scopolin were detected in the plants. The root uptake of umbelliferone reduced root-knot nematode invasion significantly up to 4 weeks after the root exposure. Our results suggest that the root uptake and the transfer of bioactive compounds between plants can expand the understanding of plant–plant interactions.

Phytochemical and Pharmaceutical Characterization of Seseli L. Species From Georgia

The Apiaceae family, formerly known as Umbelliferae, is one of the largest plant families, composed of 466 genera and nearly 3800 species worldwide. According to the Royal Kew Garden, it is also one of 12 plant families with a higher than-normal percentage of medicinal species, ranking 6th with 15% of its plants counted as medicinal. Seseli species, known for their high coumarin content as well as terpenoids, flavonoids, essential oils, and more, have been extensively investigated and found to possess various pharmacological properties such as antimicrobial, antitumor, antioxidant, cytotoxic, anti-nociceptive, anti-inflammatory, etc. Some Seseli species have also been used in treating central nervous system disorders such as anxiety. From 135 species 12 are distributed in Georgia (original name of the country is Sakhartvelo), from which 3 are endemics. Despite numerous studies on the chemical composition and pharmacological properties of various Seseli species, some remain unexplored, especially those found in Georgia, where data is limited and out of date. Therefore, further extensive research is necessary to improve understanding of the pharmacological activities, chemical constituents, and efficacy of plants from the Georgian Seseli species. The presented review draws attention to Seseli species distributed in Georgia, their phytochemistry, botanical characterisation and traditional use. However, as the information on biological activity of Georgian Seseli spp. is limited, we discussed potential pharmacological properties expected based on their chemical composition, also in relation with the same species from other countries or botanical gardens.

Pharmacological and phytochemical insights on the pancreatic β-cell modulation by Angelica L. roots

Ethnopharmacological relevanceAngelica roots are a significant source of traditional medicines for various cultures around the northern hemisphere, from indigenous communities in North America to Japan. Among its many applications, the roots are used to treat type 2 diabetes mellitus; however, this application is not mentioned often. Ethnopharmacological studies have reported the use of A. japonica var. hirsutiflora, A. furcijuga, A. shikokiana, and A. keiskei to treat diabetes symptoms, and further reports have demonstrated the three angelica roots, i.e., A. japonica var. hirsutiflora, A. reflexa, and A. dahurica, exhibit insulin secretagogue activity. Aim of the studyThis study aimed to phytochemically characterize and compare angelica roots monographed in the European Pharmacopeia 11th, isolate major plant metabolites, and assess extracts and isolates' capability to modulate pancreatic β-cell function. Materials and methodsRoot extracts of Angelica archangelica, Angelica dahurica, Angelica biserrata, and Angelica sinensis were phytochemically profiled using liquid chromatography method coupled with mass spectrometry. Based on this analysis, simple and furanocoumarins were isolated using chromatography techniques. Extracts (1.6–50 μg/mL) and isolated compounds (5–40 μmol/L) were studied for their ability to modulate insulin secretion in the rat insulinoma INS-1 pancreatic β-cell model. Insulin was quantified by the homogeneous time-resolved fluorescence method. ResultsForty-one secondary metabolites, mostly coumarins, were identified in angelica root extracts. A. archangelica, A. dahurica, and A. biserrata root extracts at concentration of 12.5–50 μg/mL potentiated glucose-induced insulin secretion, which correlated with their high coumarin content. Subsequently, 23 coumarins were isolated from these roots and screened using the same protocol. Coumarins substituted with the isoprenyl group were found to be responsible for the extracts' insulinotropic effect. ConclusionsInsulinotropic effects of three pharmacopeial angelica roots were found, the metabolite profiles and pharmacological activities of the roots were correlated, and key structures responsible for the modulation of pancreatic β-cell function were identified. These findings may have implications for the traditional use of angelica roots in treating diabetes. Active plant metabolites may also become lead structures in the search for new antidiabetic treatments.

Genome-wide analysis of the cytochrome P450 superfamily suggests its roles in coumarin biosynthesis and salt stress response in Melilotus albus

Coumarin, a secondary metabolite from the plant phenylpropane pathway, was certified as an important natural product for plants in response to abiotic stresses. Cytochrome P450s represent the largest monooxygenase superfamily in kingdoms, participating in various biochemical processes including coumarin biosynthesis. Here, we reported a total of 290 P450 genes identified from the Melilotus albus genome, and these MaP450s were grouped into 9 clans. These genes were unevenly distributed in eight chromosomes and all of them contain highly conserved feature motifs of P450s. MaCYP82L1 was differentially expressed in JiMa46 and JiMa49, meanwhile up-regulated in response to NaCl treatment. The heterologous expression in yeast showed that the MaCYP82L1 transformed yeast cells exhibited obvious resistance under drought and salt stresses. Furthermore, overexpression of MaCYP82L1 increased coumarin content in transgenic hairy roots. This study not only identified MaCYP82L1 as an important regulator that adjusts drought and salt resistance, but also provided a link between coumarin metabolism and abiotic stresses.

Transcriptomic and targeted metabolome analyses revealed the regulatory mechanisms of the synthesis of bioactive compounds in Citrus grandis 'tomentosa'.

Exocarpium Citri Grandis is a popular Chinese herbal medicine prepared from Citrus grandis 'tomentosa', and it is rich in several bioactive compounds, including flavonoids, coumarins, and volatile oils. However, studies are yet to elucidate the mechanisms of synthesis and regulation of these active components. Therefore, the present study examined the profiles of flavonoids and volatile oil bioactive compounds in plant petals, fruits, and tender leaves, and then performed RNA sequencing on different tissues to identify putative genes involved in the synthesis of bioactive compounds. The results show that the naringin, naringenin, and coumarin contents of the fruitlets were significantly higher than those of the tender leaves and petals, whereas the tender leaves had significantly higher levels of rhoifolin and apigenin. A total of 49 volatile oils, of which 10 were mainly found in flowers, 15 were mainly found in fruits, and 18 were mainly found in leaves, were identified. RNA sequencing identified 9,942 genes that were differentially expressed in different tissues. Further analysis showed that 20, 15, and 74 differentially expressed genes were involved in regulating flavonoid synthesis, regulating coumarin synthesis, and synthesis and regulation of terpenoids, respectively. CHI1 (Cg7g005600) and 1,2Rhat gene (Cg1g023820) may be involved in the regulation of naringin synthesis in C. grandis fruits. The HDR (Cg8g006150) gene, HMGS gene (Cg5g009630) and GGPS (Cg1g003650) may be involved in the regulation and synthesis of volatile oils in C. grandis petals. Overall, the findings of the present study enhance our understanding of the regulatory mechanisms of secondary metabolites in C. grandis, which could promote the breeding of C. grandis with desired characteristics.

Metabolome and Transcriptome Analysis Revealed the Basis of the Difference in Antioxidant Capacity in Different Tissues of Citrus reticulata 'Ponkan'.

Citrus is an important type of fruit, with antioxidant bioactivity. However, the variations in the antioxidant ability of different tissues in citrus and its metabolic and molecular basis remain unclear. Here, we assessed the antioxidant capacities of 12 tissues from Citrus reticulata 'Ponkan', finding that young leaves and root exhibited the strongest antioxidant capacity. Secondary metabolites accumulated differentially in parts of the citrus plant, of which flavonoids were enriched in stem, leaf, and flavedo; phenolic acids were enriched in the albedo, while coumarins were enriched in the root, potentially explaining the higher antioxidant capacities of these tissues. The spatially specific accumulation of metabolites was related to the expression levels of biosynthesis-related genes such as chalcone synthase (CHS), chalcone isomerase (CHI), flavone synthase (FNS), O-methyltransferase (OMT), flavonoid-3'-hydroxylase (F3'H), flavonoid-6/8-hydroxylase (F6/8H), p-coumaroyl CoA 2'-hydroxylase (C2'H), and prenyltransferase (PT), among others, in the phenylpropane pathway. Weighted gene co-expression network analysis (WGCNA) identified modules associated with flavonoids and coumarin content, among which we identified an OMT involved in coumarin O-methylation, and related transcription factors were predicted. Our study identifies key genes and metabolites influencing the antioxidant capacity of citrus, which could contribute to the enhanced understanding and utilization of bioactive citrus components.

QUANTITATIVE ANALYSIS OF KHELLACTONE AND OTHER COUMARINS IN PHLOJODICARPUS SIBIRICUS (APIACEAE)

Phlojodicarpus sibiricus (Fisch.) Koso-Pol. is a medicinal plant of the Apiaceae family, the underground organs of which accumulate various groups of coumarins, including simple coumarins, furocoumarins and angular dihydropyranocoumarins in the form of esters of kellactone (3′,4′-dihydroxy-3′,4′-dihydroseselin), which are the dominant group of secondary metabolites. Known methods for the analysis of rhizomes and roots of P. sibiricus determine the content of individual compounds (visnadin, dihydrosamidin), which does not allow obtaining complete information about the presence of coumarin compounds in the plant. In the present study, a method of quantitative analysis of coumarins was developed. It includes preliminary hydrolysis of raw materials in aqueous potassium hydroxide, which leads to deacylation of esterified derivatives of kellactone and lomatin, without affecting glycosides of simple coumarins, followed by analysis of hydrolysis products (sum of kellactones, lomatin, 6′-O-apiosyl skimmin) by HPLC. The developed technique is characterized by high analysis speed, satisfactory validation characteristics, and accuracy. Approbation of the technique was carried out on samples of rhizomes and roots of P. sibiricus collected in three regions of Siberia (Zabaikalsky Krai, Buryatia Republic, Sakha (Yakutia) Republic). The total content of coumarins in plant materials from the studied populations was 12.70–74.03 mg/g. The method can be used for quality analysis and standardization of P. sibiricus rhizomes and roots.

Effect of fertilizers and growth stimulants on the content of coumarin and its derivatives in the vegetative mass of melilot.

The paper presents the results of determining the content of coumarin and its derivatives in the melilot using nuclear magnetic resonance spectroscopy, including the study of the effect of fertilizers on the quantitative content of coumarin and its derivatives in the composition of the melilot treated by phases using the method of 1H and 13C nuclear magnetic resonance spectroscopy. Based on the results of the nuclear magnetic resonance spectroscopy, the possibility of qualitative and quantitative determination of the content of coumarin and its derivatives isolated from extracts of fertilized melilot was evaluated. The effect of eight preparations on the plant and its concentration in solution on the content of coumarin substances in the Altynbas yellow melilot variety was studied. It was shown that the preparations have a slight effect on the coumarin content in the melilot and, accordingly, on the quality of feed based on it. In the control variant (variant 9), the coumarin proton content was 9.85%, and the maximum coumarin proton content was observed in the variant where BioEnergy was used as fertilizer and amounted to 22.44%.

Analysis of coumarin content and key enzyme genes expression involved in coumarin biosynthesis from Peucedanum praeruptorum Dunn at different stages

Analysis of coumarin content and key enzyme genes expression involved in coumarin biosynthesis from Peucedanum praeruptorum Dunn at different stages

Phytochemical screening and in vitro evaluation of antioxidant and antimicrobial efficacies of Pteropyum scoparium (Jaub. & Spach) Sidaf crude extracts

ObjectivePteropyrum scoparium Jaub. & Spach locally known as “Sidaf” is a meal known to the ancient Omani people with many health benefits. It is traditionally used in Oman to treat high cholesterol, hypertension, indigestion problems, wound healing, and diabetes. However, these claims are yet to be scientifically proven. Hence, this study aimed to perform phytochemical, antioxidant, and antimicrobial analysis of P. scoparium leaves aqueous and alcoholic extracts to confirm its medicinal potential. MethodsA detailed phytochemical analysis of ethanol and aqueous extracts of leaves was carried out to confirm the presence of bioactive substances. DPPH (2,2′-diphenyl-1-picrylhydrazyl), agar-well diffusion and disc diffusion methods were used to evaluate antioxidant and antimicrobial potential, respectively. The extracts were tested against four microorganisms viz. E. coli (ATCC 25922), S. aureus (ATCC 23235), Penicillium sp. (ATCC 11597) and Rhizopus stolonifer (ATCC 14037). ResultsThe ethanol extract exhibited higher DPPH scavenging activity than aqueous extract that was confirmed with IC50 values of both extracts. However, the aqueous extract was found to be significantly more effective as an antimicrobial agent than the ethanol extract. This could be due to higher coumarins content that is thrice as much as in ethanol extract. One-way repeated measure RM ANOVA showed that there was a statistically significant difference in the antimicrobial susceptibility of all four organisms for the aqueous and ethanol well diffusion extracts (DF = 7; SS = 56.350, MS = 8.050; F = 5.865; P < 0.001). The highest mean zone of inhibition was recorded for S. aureus (12 ± 3.851 mm) well diffusion aqueous extract followed by R. stolonifer (11.750 ± 4.250 mm) well diffusion aqueous extract, and S. aureus (10.625 ± 3.771 mm) well diffusion ethanol extract. ConclusionsPhytochemical screening of ethanol and aqueous extracts revealed the presence of alkaloids, glycosides, carbohydrates, amino acid, fats & fixed oils, phenolic compounds & tannins, proteins, phytosterols, saponins, gum & mucilage, terpenoids, coumarins and anthocyanins. The findings from this study will be useful in evaluating the phytochemical constituents present in the extract and developing commercial drugs as antioxidant and antimicrobial agents based on this plant.

Effect of different degrees of withering on gene expression and metabolite content of Wuyi rock tea leaves

The degree of withering significantly affected the formation of tea quality. This study analyzed the effects of light withering (LW), moderate withering (MW) and heavy withering (HW) on tea quality jointly by transcriptome and metabolomics. It was found that LW was more beneficial in increasing gene expression of the ko00500, ko00511, ko00940, ko00944, ko00941, ko00740 and ko02010 pathways in tea leaves, thereby increasing the content of quinones, phenolic acids, lignans and coumarins, and flavonoids; MW was more beneficial in increasing gene expression of ko00500, ko00511, ko00130, ko00380 and ko00520 pathways, thereby increasing the content of terpenoids, organic acids and lipids. The aroma intensity was highest in moderately withering tea leaves and bitterness intensity was highest in lightly withering tea leaves, while the intensity of mellowness, fresh and brisk taste was significantly higher in MW and LW than in HW. This study provides an important basis for guiding the processing of Wuyi rock tea with different flavors.

Help sought for gout sufferers: A historical look at the treatment of gout in the New Kingdom of Granada

IntroductionDuring the colonial period, despite scarce knowledge and resources, doctors and apothecaries developed medical prescriptions mainly of vegetable origin for the management of multiple diseases, including gout. ObjectiveTo contextualise and describe the use of medical prescription in the early 19th century in the New Kingdom of Granada for the treatment of gout. Material and methodsA documentary search of medical prescription was carried out in the Cipriano Rodríguez Santamaría Historical Archive of the Octavio Arizmendi Posada Library of the University of La Sabana. Subsequently, an open literature review was carried out in the ScienceDirect, ClinicalKey, and Scielo databases in English and Spanish. ResultsPistacia lentiscus, the basis of the recipe described, has a high content of terpenes, tannins, flavonoids, and coumarins, which generate anti-inflammatory, antimicrobial, antioxidant, antibiotic, antiviral, and anti-atherogenic effects, among others. ConclusionMastic, one of the components of these recipes, possesses anti-inflammatory and antioxidant properties, which could explain its relative efficacy in the treatment of gout in the 19th century.

Genome sequencing-based transcriptomic analysis reveals novel genes in Peucedanum praeruptorum

BackgroundPeucedanum praeruptorum Dunn, a traditional Chinese herbal medicine, contains coumarin and volatile oil components that have clinical application value. However, early bolting often occurs in the medicinal materials of Apiaceae plants. The rhizomes of the medicinal parts are gradually lignified after bolting, resulting in a sharp decrease in the content of coumarins. At present, the link between coumarin biosynthesis and early bolting in P. praeruptorum has not been elucidated.ResultsCombining the genome sequencing and the previous transcriptome sequencing results, we reanalyzed the differential transcripts of P. praeruptorum before and after bolting. A total of 62,088 new transcripts were identified, of which 31,500 were unknown transcripts. Functional classification and annotation showed that many genes were involved in the regulation of transcription, defense response, and carbohydrate metabolic processes. The main domains are the pentatricopeptide repeat, protein kinase, RNA recognition motif, leucine-rich repeat, and ankyrin repeat domains, indicating their pivotal roles in protein modification and signal transduction. Gene structure analysis showed that skipped exon (SE) was the most dominant alternative splicing, followed by the alternative 3’ splice site (A3SS) and the alternative 5’ splice site (A5SS). Functional enrichment of differentially expressed genes showed that these differentially expressed genes mainly include transmembrane transporters, channel proteins, DNA-binding proteins, polysaccharide-binding proteins, etc. In addition, genes involved in peroxisome, hexose phosphate pathway, phosphatidylinositol signaling system, and inositol phosphate metabolism pathway were greatly enriched. A protein-protein interaction network analysis discoverd 1,457 pairs of proteins that interact with each other. The expression levels of six UbiA genes, three UGT genes, and four OMT genes were higher during the bolting stage. This observation suggests their potential involvement in the catalytic processes of prenylation, glycosylation, and methylation of coumarins, respectively. A total of 100 peroxidase (PRX) genes were identified being involved in lignin polymerization, but only nine PRX genes were highly expressed at the bolting stage. It is worth noting that 73 autophagy-related genes (ATGs) were first identified from the KEGG pathway-enriched genes. Some ATGs, such as BHQH00009837, BHQH00013830, and novel8944, had higher expression levels after bolting.ConclusionsComparative transcriptome analysis and large-scale genome screening provide guidance and new opinions for the identification of bolting-related genes in P. praeruptorum.