Uptake and accumulation of di(2-ethylhexyl) phthalate (DEHP) in a soil-ginseng system and toxicological mechanisms on ginseng (Panax ginseng C.A. Meyer).

Di(2-ethylhexyl) phthalate (DEHP) is regarded as a priority environmental pollutant. This study explored the adsorption and accumulation of DEHP within the ginseng-soil system and the mechanism of DEHP toxicity to ginseng (Panax ginseng C.A. Meyer). Under exposure to 22.10 mg/kg DEHP in soil, DEHP mainly accumulated in ginseng leaves (20.28 mg/kg), stems (4.84 mg/kg) and roots (2.00 mg/kg) after 42 days. The oxidative damage, metabolism, protein express of ginseng were comprehensively measured and analyzed. The results revealed that MDA presented an activation trend in ginseng stems and leaves after 42 days of DEHP exposure, while the opposite trend was observed for POD. Levels of ginsenoside metabolites Rg2, Rg3, Rg5, Rd, Rf and CK decreased in the ginseng rhizosphere exudates under DEHP stress. Further investigations revealed that DEHP disrupts ginsenoside synthesis by inducing glycosyltransferase (GS) and squalene synthase (SS) protein interactions. Molecular docking indicated that DEHP could stably bind to GS and SS by intermolecular forces. These findings provide new information on the ecotoxicological effect of DEHP on ginseng root.

Effects of potassium fulvic acid and potassium humate on microbial biodiversity in bulk soil and rhizosphere soil of Panax ginseng.

Potassium fulvic acid (BSFA) and potassium humate (KHM), as organic fertilizers, can improve soil structure, increase soil nutrient levels and prevent plant diseases. However, knowledge is limited regarding how BSFA and KHM influence soil microbial communities and the interrelationships between community members associated with Panax ginseng. Soil pH and nutrient content increased significantly as a result of the addition of BSFA and KHM. The pH, NH4+-N, NO3--N, AP and AK increased by 1.72 %-5.55 %, 70.09 %-108.39 %, 35.38 %-216.20 %, 1.21 %-14.19 % and 3.40 %-5.94 %, respectively, in the BSFA and KHM treatments. The soil nutrient increase may be related to Micrococcaceae and arbuscular mycorrhizal fungi (AMF). The structure of the microbial community also changed radically from that of the control group, and Chloroflexi (2.69 %-3.15 %), Actinobacteria (4.33 %-7.53 %) and Acidobacteria (9.44 %-11.62 %) were the dominant microorganisms at the phylum level in bacteria. In contrast, the dominant fungi at the phylum level were Ascomycota (77.39 %-78.08 %), Glomeromycota (0.36 %-2.68), Olpidiomycota (0.02 %-3.78 %) and Basidiomycota (0.80 %-1.17 %). Fusarium oxysporum and Ascomycota were biomarkers for BSFA and KHM, which may be related to pathogenic bacteria. Network analysis revealed that the association among members of the soil microbial community was more positive than negative following application of KHM, and more positive (62.5 %) than negative (37.5 %) correlations were observed between bacteria, whereas the fungal community exhibited more positive (97.3 %) than negative (2.7 %) correlations. PICRUST predicted the microbial function of adding KHM and BSFA to the soil, and these pathways mainly belong to the degradation and metabolism of organic matter, saprophytic organisms and plant pathogens. In summary, our study demonstrated that the addition of BSFA and KHM increased the nutrients in the ginseng soil and reshaped the microbial function in soils, providing a theoretical foundation for soil improvement and biological control of ginseng diseases. However, due to the limitations of greenhouse cultivation, additional long-term experiments on farmland with different climate changes are recommended.

Effects of ginseng soluble dietary fiber on serum antioxidant status, immune factor levels and cecal health in healthy rats.

As an important component of ginseng, the in vivo benefits of ginseng water-soluble dietary fiber (ginseng-SDF) have not been fully revealed. To explore these benefits, healthy rats were given ginseng-SDF (200, 400, and 800 mg/kg body weight/day) by gavage for 15 days. The results showed that ginseng-SDF significantly improved the rats' growth performance and serum antioxidant status. Insulin-like growth factor (IGF-1 and IGF-2) and immunoglobulin (IgA, IgM, and IgG) levels in the ginseng-SDF groups were increased. High-dose ginseng-SDF significantly increased the cecal butyric acid proportion compared with the K group. Ginseng-SDF increased the abundance of Firmicutes and promoted the proliferation of probiotics such as Lactobacillus, and cellulose decomposers such as Ruminococcus and Clostridium in cecal microflora. These altered microflora were correlated with growth performance, antioxidant status and immunoglobulin indexes. The above results suggested that ginseng-SDF might have positive effects on growth, oxidative-immune levels and cecal health in rats.

Occurrence of postharvest snow rot caused by Sclerotinia nivalis on Asian ginseng in China.

Asian ginseng (Panax ginseng) is a valuable medicinal plant that is commercially cultivated in China. A long postharvest storage period is required before ginseng is processed. From October 2019 to May 2020, snow rot was observed on the roots of 4- and 5-year-old fresh ginseng stored in three cold storage facilities located in Tonghua and Changbai cities in northeastern China, which are the most important regions for Asian ginseng production. We sampled 1,000 ginseng roots from the three cold storage facilities, and the average disease incidence was 21%. Initially, sparse hyphae and microsclerotia appeared on the root epidermis. Lesions gradually softened and the epidermis detached easily. Multiple infected sites slowly converged, resulting in the formation of a dense complex of multiple sclerotia and thick hyphae on the surface of the ginseng root as well as internal decay. The infection eventually spread to the adjacent ginseng roots (Fig. 1). Sixteen diseased ginseng roots were collected and then sclerotia were removed from the root surface, immersed in 1% NaClO for 2 min, rinsed three times with sterile water, and placed on potato dextrose agar (PDA) containing streptomycin (40 µg/mL) in Petri dishes. After a 3-day incubation at 20 °C in darkness, 22 suspected Sclerotinia isolates were obtained. Isolates SN1 and SN2 were randomly selected for identification. On PDA, fast-growing colonies produced white, sparse, powdery, and cotton-like aerial mycelia, and the reverse side showed the same color (Fig. 2). Small and white sclerotial primordia formed 3 days later and a ring of sclerotia was detected at the plate periphery. At 7 to 10 days after incubation, the mature sclerotia were black, spherical-to-subspherical, and elongated or fused to form irregular shapes. Each Petri dish produced 55-65 sclerotia, measuring 1.1 × 1.2 to 3.2 × 3.9 mm (n = 100). The sclerotia were firmly attached to the agar surface. The isolates were initially identified as Sclerotinia sp. (Saito 1997). After sequencing the nuclear ribosomal internal transcribed spacer region (MW927134 and MW927135) and the ß-tubulin gene (MW929179 and MW929180) (White et al. 1990; Glass and Donaldson 1995), BLAST searches revealed 100% homology with JX262268 and JX296007 of the published S. nivalis strain KGC-S0601, respectively. The pathogenicity of the two isolates was tested using detached ginseng roots. Briefly, healthy roots were washed, surface-disinfested with 75% alcohol, and rinsed with sterile water. Mycelial plugs (5 mm diameter) removed from the margin of actively growing colonies on PDA were placed on the ginseng roots. For each isolate, four roots were inoculated, with two plugs per root. Additionally, PDA plugs without mycelia were used as the negative control. The roots were placed in a fresh-keeping box at 20 °C in darkness and evaluated after 7 days. The pathogenicity test was repeated twice. The symptoms on the inoculated roots were the same as those observed on the roots during cold storage, whereas the control roots remained symptomless. The same fungus was reisolated consistently from all infected roots and its identity was confirmed by resequencing, thereby fulfilling Koch's postulates. To the best of our knowledge, this is the first report of S. nivalis causing postharvest snow rot on Asian ginseng in China. The occurrence of this disease threatens the postharvest storage of Asian ginseng. Hence, effective management strategies must be developed.

[Progress of studies on ATP-binding cassette transporters and transportation of secondary metabolites in medicinal plants].

ATP-binding cassette(ABC) transporters are one of the largest protein families in organisms, with important effects in regulating plant growth and development, root morphology, transportation of secondary metabolites and resistance of stress. Environmental stress promotes the biosynthesis and accumulation of secondary metabolites, which determines the quality of medicinal plants. Therefore, how to improve the accumulation of secondary metabolites has been a hotspot in studying medicinal plants. Many studies have showed that ABC transporters are extremely related to the transportation and accumulation of secondary metabolites in plants. Recently, with the great development of genomics and transcriptomic sequencing technology, the regulatory mechanisms of ABC transporters on secondary metabolites have attached great attentions in medicinal plants. This paper reviewed the mechanisms of different groups of ABC transporters in transporting secondary metabolites through cell membranes. This paper provided key theoretical basis and technical supports in studying the mechanisms of ABC transporters in medicinal plant, and promoting the accumulation of secondary metabolites, in order to improve the quality of medicinal plants.

Functional soluble dietary fiber from ginseng residue: Polysaccharide characterization, structure, antioxidant, and enzyme inhibitory activity.

Ginseng (Panax ginseng C.A. Meyer) is the most famous edible Chinese herbal medicine. In the present study, soluble dietary fiber of ginseng (ginseng-SDF, 8.98% content) was extracted from ginseng residue, and its physicochemical characterization, structure, and biological activities were studied. Ginseng-SDF was an acidic heteropolysaccharide (uronic acid, 4.42% content) rich in protein, amino acids, and mineral elements. Glucose was its main monosaccharide composition (58.03%). Ginseng-SDF had a porous microstructure, a typical cellulose I structure and a large number of hydroxyl functional groups. These chemical composition and structural characteristics gave ginseng-SDF a good water solubility (98.56%), oil-holding capacity (OHC) (3.01 g/g), and biological activities, as the antioxidant activity (13.35 µM TE/g, 105.17 µM TE/g, 54.20 µM TE/g for DPPH, ABTs, and FRAP assays, respectively), glucose diffusion retardation index (GDRI, 33.33%-7.43%), and α-amylase/α-glucosidase inhibitory activities (IC50 , 6.70 mg/ml, and 4.89 mg/ml, respectively). The results suggested that ginseng residue is a valuable source of functional dietary fiber, and the ginseng-SDF has a potential use in antioxidant and hypoglycemic foods. PRACTICAL APPLICATIONS: Ginseng has long been popular as a health food in Asia, North America, and Europe. Ginseng residue is rich in polysaccharides, dietary fiber, proteins, and other components, which is also of great research value. However, there are few studies focus on the soluble dietary fiber of ginseng at present. The research shows that ginseng residue is a valuable source of functional dietary fiber. The chemical components and structural characteristics give ginseng-SDF a noteworthy antioxidant activity and enzyme inhibitory activity in vitro. These properties and biological activities indicate that ginseng-SDF has application value in antioxidant and hypoglycemic foods.

Silicon application and related changes in soil bacterial community dynamics reduced ginseng black spot incidence in Panax ginseng in a short-term study.

BACKGROUND: This study analyzed the effect of silicon (Si) application on the occurrence of ginseng black spot caused by Alternaria panax. We explored the differences in soil physical and chemical factors and microbial community structure following Si application as well as the key factors that affected the occurrence of ginseng black spot in soil. Potted Panax ginseng plants were used to assess the effect of Si treatment on ginseng black spot. Soil physical and chemical properties were comprehensively analyzed. Bacterial communities were analyzed using Illumina HiSeq sequencing targeting the 16S rRNA gene. RESULTS: After inoculation with A. panax, the morbidity (and morbidity index) of ginseng with and without Si was 52% (46) and 83% (77), respectively. Soil physical and chemical analysis showed that under the ginseng black spot inoculation, bacterial communities were mainly affected by pH and available potassium, followed by ammonium nitrogen and available Si. NMDS and PLS-DA analyses and the heat maps of relative abundance revealed that Si application elevated the resistance of ginseng black spot as regulated by the abundance and diversity of bacterial flora in rhizosphere soils. Heatmap analysis at the genus level revealed that A. panax + Si inoculations significantly increased the soil community abundance of Sandaracinus, Polycyclovorans, Hirschia, Haliangium, Nitrospira, Saccharothrix, Aeromicrobium, Luteimonas, and Rubellimicrobium and led to a bacterial community structure with relative abundances that were significantly similar to that of untreated soil. CONCLUSIONS: Short-term Si application also significantly regulated the structural impact on soil microorganisms caused by ginseng black spot. Our findings indicated that Si applications may possibly be used in the prevention and treatment of ginseng black spot.

Effects of different substrates on low-temperature storage of fresh ginseng.

BACKGROUND: Fresh ginseng was buried in three types of sand with different moisture contents and three types of soil and then stored at 2 °C to determine the effects of these storage substrates on fresh ginseng. RESULTS: At a storage time of 200 days, ginseng stored in underforest soil softened the most slowly and had a significantly greater firmness compared to the other samples (P < 0.05). The amount of most ginsenosides changed after storage for most of the substrates. Samples stored in ginseng soil and biological fertilizer had the highest concentration of total saponin and ginseng polysaccharides, respectively. Fresh ginseng stored in medium-water content sand had a significantly lower polyphenol oxidase activity (P < 0.05). A significant difference was observed in the total concentration of nucleosides and nucleobases between the ginseng samples stored with and without substrates (P < 0.05). CONCLUSION: The data obtained in the present study suggest that the use of storage substrates is an optimal method for extending the shelf life of fresh ginseng without detrimental effects on its components. © 2019 Society of Chemical Industry.

Application of high-speed counter-current chromatography and HPLC to separate and purify of three polyacetylenes from Platycodon grandiflorum.

Three polyacetylenes were isolated and purified from Platycodon grandiflorum A. DC for the first time by high-speed counter-current chromatography using a two-phase solvent system composed of hexane/ethyl acetate/methanol/water (1:31:1:31, v/v/v/v) and high-performance liquid chromatography with an Agilent ZORBAX® SB-C18 column (4.6 mm × 150 mm, 5 µm). After separation by high-speed counter-current chromatography and high-performance liquid chromatography, we obtained 3.5 mg of platetyolin A, 4.1 mg of platetyolin B, and 18.1 mg of lobetyolin with purities of 97.2, 96.7, and 96.9%, respectively. The purity of each compound was assessed by high-performance liquid chromatography and the chemical structures were evaluated by high-resolution electrospray ionization time-of-flight mass spectrometry and one- and two-dimensional NMR spectroscopy. Among the isolated compounds, platetyolin A and platetyolin B are newly reported compounds.

Ionic Liquid-Based Ultrasonic-Assisted Extraction of Forsythosides from the Leaf of Forsythia suspensa (Thunb.) Vahl and Subsequent Separation and Purification by High-Speed Counter-Current Chromatography.

An ionic liquid-based ultrasonic-assisted extraction (ILUAE) method was developed for the extraction of the two forsythosides, namely forsythosides I and A from the leaf of Forsythia suspensa (Thunb.) Vahl. Three kinds of l-alkyl-3-methylimidazolium ionic liquids with different alkyl chain and anion were investigated. The results indicated that ionic liquids showed remarkable effects on the extraction yields of forsythosides. In addition, several ILUAE ultrasonic parameters, such as the solvent concentration, solvent to solid ratio and extraction time have been optimized. Under these optimal conditions (e.g., with 0.6 M [C6MIM]Br, solvent to solid ratio of 15 mL/g and extraction time of 10 min), this approach gained the highest extraction yields of forsythoside I (0.89%) and forsythoside A (10.74%). Meanwhile, forsythosides in the ILUAE extract were separated and purified successfully through the high-speed counter-current chromatography with a two-phase solvent system consisting of ethyl acetate-ethanol-acetic acid-water (4 : 1 : 0.25 : 6, v/v). 5.4 mg of forsythoside I and 59.7 mg of forsythoside A were obtained from 120 mg of the prepurified sample in one-step separation, with the purity of 96.1 and 97.9%, respectively, as determined by high-performance liquid chromatography. Their structures were identified by (1)H nuclear magnetic resonance (NMR) and (13)C NMR.

Preparative isolation, quantification and antioxidant activity of dihydrochalcones from Sweet Tea (Lithocarpus polystachyus Rehd.).

Dihydrochalcones are the main active components of Lithocarpus polystachyus Rehd. (Sweet Tea), they are directly related to the sweet tonic beverage and traditional herb. In this work, two runs of preparative high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hexane/ethyl acetate/ethanol/water (1:4:3:4, v/v) were employed to separate three dihydrochalcones (phloridzin, trilobatin and phloretin) from Sweet Tea. About 6.4mg of phloridzin, 48.4mg of trilobatin, and 4.7mg of phloretin with purities of 96.7%, 98.4% and 98.1% were obtained from 130mg of the crude Sweet Tea extract. Phloridzin, trilobatin, and phloretin had effective radical scavenging activities, with IC50 values of 866.80, 20.16 and 179.47µg/mL, respectively, in a 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical method. The contents of phloridzin, trilobatin and phloretin in dried old leaves and tender leaves of tea were in the range of 10.1-18.0, 113.7-128.8, 3.6-4.3mg/g and 9.3-9.8, 82.9-103.1, 1.9-2.5mg/g, respectively. The results indicated that the HPLC had good precision, accuracy and repeatability for the determination of three dihydrochalcones in samples.

Aqueous ionic liquid based ultrasonic assisted extraction of four acetophenones from the Chinese medicinal plant Cynanchum bungei Decne.

In this study, an aqueous ionic liquid based ultrasonic assisted extraction (ILUAE) method for the extraction of the four acetophenones, namely 4-hydroxyacetophenone (1), 2,5-dihydroxyacetophenone (2), baishouwubenzophenone (3) and 2,4-dihydroxyacetophenone (4) from the Chinese medicinal plant Cynanchum bungei was developed. Three kinds of aqueous l-alkyl-3-methylimidazolium ionic liquids with different anion and alkyl chain were investigated. The results indicated that ionic liquids (ILs) showed remarkable effects on the extraction efficiency of acetophenones. In addition, the ILUAE, including several ultrasonic parameters, such as the ILs concentration, solvent to solid ratio, power, particle size, temperature, and extraction time have been optimized. Under these optimal conditions (e.g., with 0.6M [C(4)MIM]BF(4), solvent to solid ratio of 35:1, power of 175 W, particle size of 60-80 mesh, temperature of 25 ° C and time of 50 min), this approach gained the highest extraction yields of four acetophenones 286.15, 21.65, 632.58 and 205.38 µg/g, respectively. The proposed approach has been evaluated by comparison with the conventional heat-reflux extraction (HRE) and regular UAE. The results indicated that ILUAE is an alternative method for extracting acetophenones from C. bungei.

An application of high-speed counter-current chromatography for separation and purification of bungeiside-A, bungeiside-B and baishouwubenzophenone from Cynanchum bungei Decne.

INTRODUCTION: Cynanchum bungei Decne (Baishouwu in China), is a famous traditional Chinese medicine that has been widely used as a tonic medicine or health food for centuries. Bungeiside-A, bungeiside-B and baishouwubenzophenone, as the major bioactive constituents in C. bungei, are challenging to separate and purify since bungeiside-A and -B are present in very low concentrations and have similar structures and high polarity. OBJECTIVE: To develop a method of isolation and purification of bungeiside-A and -B and baishouwubenzophenone from the Chinese medicinal plant Cynanchum bungei Decne by high-speed counter-current chromatography (HSCCC). METHODOLOGY: The roots of C. bungei were extracted with light-petroleum (60-90 °C) and chloroform to remove the lipid substance. Then the residuals were extracted with methanol. The methanol extract was prepared for the subsequent HSCCC separation. The simple HSCCC method of separation and purification of bungeiside-A and -B and baishouwubenzophenone was established according to the solvent system, which was selected according to the measurement of partition coefficient (K). The purities of target compounds were test by HPLC and the structure was identified by ¹H NMR and ¹³C NMR. RESULTS: Bungeiside-A (9.4 mg), bungeiside-B (8.6 mg) and baishouwubenzophenone (5.7 mg) were obtained from 1.5 g of the methanol extract with purities of 93.2, 98.7 and 95.4%, respectively. CONCLUSION: These results clearly demonstrate that HSCCC is a powerful tool for isolating and purifying components with similar structures, low concentration and high polarity from medicinal plant, such as bungeiside-A and -B and baishouwubenzophenone.

[Determination of four acetophenones in Radix Cynanchi bungei by high performance liquid chromatography-photodiode array detection].

A high performance liquid chromatographic (HPLC) method was developed for the determination of four acetophenones, namely 4-hydroxyacetophenone (I), 2,5-dihydroxyacetophenone (II), baishouwubenzophenone (III) and 2,4-dihydroxyacetophenone (IV), in Radix Cynanchi bungei. The HPLC with photodiode array (PDA) detection was carried out on a Symmetry-C18 (4.6 mm x 250 mm, 5 microm) column with CH3OH-H2O (26 : 74, v/v) as the mobile phase at the flow rate of 1.0 mL/min and 30 degrees C. The compounds I , III, IV were monitored at 280 nm, and the compound II was at 224 nm. Four acetophenones (I - IV) had good linearities (r = 0.999 6 - 0.999 9) at the ranges of 0.080 - 0.560 microg, 0.080 -0.560 microg, 0.100 - 0.700 microg and 0.092 -0.644 microg, respectively. The average recoveries were from 98.0% to 104.0% with relative standard deviations (RSD) ranging from 0.8% to 2.6%. The method is of quick, simple and accurate. The method can be used for the quality control of this product.