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Abiotic elicitors play a crucial role in regulating various aspects of plant growth, development, and specialized metabolism. This study aimed to further increase the gentiopicroside content by screening elicitor types, optimizing elicitation conditions, and estimating transcriptional responses of biosynthetic genes in the adventitious roots of Gentiana scabra. The results showed that methyl jasmonate (MeJA) was the most effective inducer for biomass accumulation in the adventitious roots of G. scabra among tested elicitors, with fresh weight (FW) and dry weight (DW) of 13.26 ± 0.57 g flask−1 and 1.31 ± 0.25 g flask−1, respectively. The effects of the induction time and concentration of MeJA on the biomass and gentiopicroside content in the adventitious roots of G. scabra were investigated. The maximum FW (15.73 ± 0.41 g flask−1) and DW (1.51 ± 0.19 g flask−1) were obtained when the roots were cultured for 6 days in MS liquid medium containing 3.0 mg L−1 1-naphthlcetic acid (NAA) and 1.0 mg L−1 kinetin (KT) at MeJA concentration of 100 μM L−1. Also, the gentiopicroside content significantly increased to 62.62 ± 0.27 mg g−1 DW, and was 2.49 times higher than that for the nontreated control. The expression levels of 12 candidate gentiopicroside biosynthesis–related genes involved in the mevalonic acid (MVA), methyl erythritol phosphate (MEP), and secoiridoid pathways were estimated in the adventitious roots of G. scabra to further understand the transcriptional response to MeJA elicitation. Among these, 10 genes (ACCT1, HMGR1, MCK1, MVD1, GPPS4, G10H, IS3, DL7H1, DXS5, and ISPH5) were upregulated whereas DXR1 and IDI1 genes were downregulated in the adventitious roots of G. scabra compared with nontreated control, with significant differences having threshold P value ≤0.05. The transcriptional analyses revealed that 12 candidate genes were the key regulated genes in the gentiopicroside biosynthetic pathway. Overall, the findings provided a promising, feasible, and stable approach to utilizing MeJA elicitation to increase the production of valuable gentiopicroside. Additionally, they provided a foundation for future gentiopicroside biosynthesis through metabolic engineering strategies in the adventitious roots of G. scabra.

Non-alcoholic fatty liver disease (NAFLD) has become a progressive metabolic disease that is emerging as a global epidemic. Considering that the complex pathogenesis has not been fully elucidated, barely specific pharmacological therapy is recommended in current guidelines. Gentiana scabra (GS) is a commonly used herb in Tibetan medicine, which has received much attention in recent years due to its diverse pharmacological properties, including anti-inflammation, anti-oxidation, and anti-fibrosis. However, the therapeutic mechanisms are still unclear. Our investigation demonstrated a regulatory effect of GS on pro-inflammatory macrophages, which was extensively investigated in NAFLD that revealed intimate participation in the disease evolution, and the non-canonical IKK family member TANK-binding kinase 1 (TBK1) was involved in this process. Plasmid vectors for shTBK1 and amlexanox (AML), an inhibitor of TBK1, were used in this study to verify the mechanisms of TBK1 both in vitro and in vivo , while a co-culture system for hepatocytes and BMDMs was constructed to confirm the critical role of macrophages for inflammatory cascade. The results revealed that metabolic burden up-regulated the phosphorylation of TBK1, resulting in activation of NF-κB signaling pathway, and consequently caused an elevated expression of MCP1 to induce the macrophage recruitment and accelerate the inflammatory cascade. In contrast, GS could inhibit the TBK1 phosphorylation and the MCP1 expression to restrain the recruitment of pro-inflammatory macrophages, so as to provide curative effects on metabolic dysfunction and inflammation. Considering that GS is non-toxic and can be used as a kind of tea for long-term drinking, we propose it may be an effective option for the prevention and treatment of NAFLD, which deserves further exploration and application, and may provide new insights to improve the current standardized intervention strategy. [ABSTRACT FROM AUTHOR]

Non-alcoholic fatty liver disease (NAFLD) has become a progressive metabolic disease that is emerging as a global epidemic. Considering that the complex pathogenesis has not been fully elucidated, barely specific pharmacological therapy is recommended in current guidelines. Gentiana scabra (GS) is a commonly used herb in Tibetan medicine, which has received much attention in recent years due to its diverse pharmacological properties, including anti-inflammation, anti-oxidation, and anti-fibrosis. However, the therapeutic mechanisms are still unclear. Our investigation demonstrated a regulatory effect of GS on pro-inflammatory macrophages, which was extensively investigated in NAFLD that revealed intimate participation in the disease evolution, and the non-canonical IKK family member TANK-binding kinase 1 (TBK1) was involved in this process. Plasmid vectors for shTBK1 and amlexanox (AML), an inhibitor of TBK1, were used in this study to verify the mechanisms of TBK1 both in vitro and in vivo, while a co-culture system for hepatocytes and BMDMs was constructed to confirm the critical role of macrophages for inflammatory cascade. The results revealed that metabolic burden up-regulated the phosphorylation of TBK1, resulting in activation of NF-κB signaling pathway, and consequently caused an elevated expression of MCP1 to induce the macrophage recruitment and accelerate the inflammatory cascade. In contrast, GS could inhibit the TBK1 phosphorylation and the MCP1 expression to restrain the recruitment of pro-inflammatory macrophages, so as to provide curative effects on metabolic dysfunction and inflammation. Considering that GS is non-toxic and can be used as a kind of tea for long-term drinking, we propose it may be an effective option for the prevention and treatment of NAFLD, which deserves further exploration and application, and may provide new insights to improve the current standardized intervention strategy.

Gentiana scabra is an important medicinal herb that is used to treat inflammation, tuberculosis, rheumatism, cholecystitis, and hepatitis. The primary bioactive components in G. scabra are secoiridoids derived from the roots and rhizomes of the plant. In this study, we sought to establish a means of reliably producing these secoiridoids through G. scabra in vitro adventitious root culture. To that end, we studied the effect of phytohormones, light condition, explant type, and medium composition on adventitious root induction and secoiridoid production. Maximal induction frequency (100%) was ultimately achieved when using leaf explants cultured on Murashige and Skoog (MS) medium containing 3.0 mg L−1 NAA and 0.25 mg L−1 TDZ with a 16/8-h photoperiod, yielding 53.20 ± 3.15 roots per explant. We explored root growth curves over an 8-wk period in MS liquid medium prepared as above, and observed maximal fresh weight (FW) and dry weight (DW) values of 26.94 ± 0.38 g per flask and 1.79 ± 0.032 g per flask, respectively, in 4-wk-old cultures. The accumulation of secoiridoids (gentiopicroside, swertiamarin, and sweroside) in these adventitious root cultures was assessed via high-performance liquid chromatography (HPLC), revealing maximal gentiopicroside (25.59 ± 0.65 mg g−1 DW), swertiamarin (1.61 ± 0.04 mg g−1 DW), and sweroside (4.42 ± 0.11 mg g−1 DW) levels after 4 wk. Together, our results reveal that G. scabra adventitious root cultures represent a viable approach to the in vitro production of secoiridoids. This is the first study to our knowledge to have explored the isolation of secoiridoids following the in vitro growth of G. scabra adventitious roots. These data may represent an important step toward the development of large-scale adventitious root cultures for commercial secoiridoids production. [ABSTRACT FROM AUTHOR]

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Copyright of Journal of Shenyang Agricultural University is the property of Journal of Shenyang Agricultural University Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The first complete chloroplast (cp) genome of Gentiana scabra was determined from Illumina paired-end sequencing data in this study. The cp genome of G. scabra is 146,915 bp in length with 37.84% overall GC content, including a large single copy (LSC) region of 79,350 bp and a small single copy (SSC) region of 17,027 bp, which are separated by a pair of inverted repeats (IRs) of 25,269 bp. The cp genome is composed of 130 genes, including 85 protein-coding genes, 8 rRNA genes, and 37 tRNA genes. Phylogenetic analysis indicated that the G. scabra is more closely related to subfamily of Gentianoideae than Menyanthaceae.

Abstract Ethnopharmacological relevance The roots and rhizomes of Gentiana scabra Bunge in the family Gentianaceae comprise a major herbal medicine for skin diseases caused by wind-heat or dampness-heat in China, Japan and Korea. This treatment can clear away heat and dry dampness and purge fire from the liver and gallbladder. Aim of the study This study was designed to investigate the therapeutic potential and anti-inflammatory effects of G. scabra , roots and rhizomes. Materials and methods Thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) revealed the presence of gentiopicrin in the roots and rhizomes of G. scabra. We then investigated the effects of ethanol extract of G. scabra , roots and rhizomes (EEGS) on skin lesions and thickness, erythema and melanin index, histopathological abnormalities, and cytokine and chemokine production in mice with contact dermatitis (CD) induced by 1-fluoro-2,4-dinitrofluorobenzene (DNFB). Moreover, the effects of EEGS on body weights and spleen body weight ratio were evaluated. Results Topical application of EEGS alleviated skin lesions such as surface roughness, excoriations and scabs on the skin of CD mice, as well as prevented skin enlargement, and lowered the erythema and melanin index. In addition, EEGS prevented hyperkeratosis, epidermal hyperplasia and immune cell infiltration, and inhibited TNF-α, IFN-γ, IL-6 and MCP-1 production in inflamed tissues. EEGS did not affect changes in body weights and spleen body weight ratio in contrast to dexamethasone. Conclusions These data indicate that the roots and rhizomes of G. scabra can be used as anti-inflammatory agents for CD with relative safety and that its therapeutic mechanisms are related to regulation of pro-inflammatory cytokines such as TNF-α and IFN-γ. Graphical abstract fx1 [ABSTRACT FROM AUTHOR]

Gentianae Radix et Rhizome (Longdan in Chinese, GRR) in Chinese Pharmacopoeia is derived from the dried roots and rhizomes of Gentiana scabra and G. rigescens, that have long been used for heat-clearing and damp-drying in the medicinal history of China. However, the characterization of the chemical components of two species and the screening of chemical markers still remain unsolved. In current research, the identification and characterization of chemical components of two species was performed using ultra-high-performance liquid chromatography (UHPLC) coupled with linear ion trap-Orbitrap (LTQ-Orbitrap) mass spectrometry. Subsequently, the chemical markers of two species were screened based on metabolomics and multivariate statistical analysis. In total, 87 chemical constituents were characterized in G. scabra (65 chemical constituents) and G. rigescens (51 chemical constituents), with 29 common chemical constituents being discovered. Thereafter, 11 differential characteristic components which could differentiate the two species were designated with orthogonal partial least squares discriminant analysis (OPLS-DA) and random forest (RF) iterative modeling. Finally, seven characteristic components identified as (+)-syringaresinol, lutonarin, trifloroside, 4-O-β-d-glu-trifloroside, 4″-O-β-d-glucopyranosy1-6′-O-(4-O-β-d-glucaffeoyl)-linearroside, macrophylloside a and scabraside were selected as the chemical markers for the recognition of two Gentiana species. It was implied that the results could distinguish the GRR derived from different botanical sources, and also be beneficial in the rational clinical use of GRR.

Japanese gentians are the most important ornamental flowers in Iwate Prefecture and their breeding and cultivation have been actively conducted for half a century. With its cool climate and large hilly and mountainous area, more than 60% of gentian production in Japan occurs in Iwate Prefecture. Recent advances in gentian breeding and cultivation have facilitated the efficient breeding of new cultivars; disease control and improved cultivation conditions have led to the stable production of Japanese gentians. Molecular biology techniques have been developed and applied in gentian breeding, including the diagnosis of viral diseases and analysis of physiological disorders to improve gentian production. This review summarizes such recent approaches that will assist in the development of new cultivars and support cultivation. More recently, new plant breeding techniques, including several new biotechnological methods such as genome editing and viral vectors, have also been developed in gentian. We, therefore, present examples of their application to gentians and discuss their advantages in future studies of gentians. [ABSTRACT FROM AUTHOR]

Gentiana scabra Bunge (Gentianaceae) is an important traditional Chinese medicine commonly used as a stomachic or appetite stimulant. In this study, 21 triterpenoids ( 1- 21) were isolated from a methanol extract of the rhizomes and roots of G. scabra. Their structures were elucidated by comparing spectroscopic data with reported values. Among the isolated triterpenoids, scabanol ( 2) was firstly isolated from natural sources. All isolated compounds were evaluated for their inhibitory activity against indoleamine 2,3-dioxygenase (IDO), which catalyzes the rate limiting reaction for the conversion of tryptophan to kynurenine. Compounds 10 and 11 showed significant inhibitory activities, with IC values of 12.5 and 9.5 μM, respectively. Compound 12 showed a moderate inhibitory effect, with an IC value of 18.7 μM. Compounds 2 and 13 showed weaker inhibitory effects, with IC values of 56.8 and 60.6 μM, respectively. Kynurenine is a potent immune modulator to suppress the functions of a variety of immune cells including T cells and natural killer cells. Given that, our results that a few selected triterpenoids inhibit IDO warrant further studies on their effects on the host immune system as natural immune stimulators. [ABSTRACT FROM AUTHOR]

Three new (1–3) and 17 known (4–20) iridoid and secoiridoid glycosides were isolated from a methanol extract of the rhizomes and roots of Gentiana scabra. Their chemical structures were elucidated from 1D and 2D NMR, IR absorption, and HR-ESI-MS spectra, as well as comparisons of these data with reported values. The effects of the isolated compounds on lipopolysaccharide (LPS)-stimulated bone marrow-derived dendritic cells were investigated. Compounds 6, 10 and 20 exhibited significant inhibitory effects on LPS-induced IL-12 p40 and IL-6 production with IC50 values of 1.62–14.29 μM. Compound 10 also showed a strong inhibitory effect on the LPS-stimulated production of TNF-α with an IC50 value of 10.45 μM. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Gentiana scabra root extract (GS) is frequently prescribed as an internal remedy in traditional Korean medicine for treatment of diabetes mellitus. GS contains bitter iridoid glycosides including loganic acid, gentiopicrin, trifloroside, and rindoside. We previously reported that the intestinal bitter taste sensation stimulates GLP-1 secretion, and thereupon hypothesized that the blood glucose regulatory effect of GS is due to its GLP-1 secreting effect in enteroendocrine L cells. Materials and method We studied GLP-1 secreting effect of GS treatment and its cellular downstream mechanism in human enteroendocrine NCI-H716 cells using the G protein-coupled receptor (GPCR) pathway inhibitors. Intracellular calcium assay also demonstrated the signal transduction pathway stimulated by the GS treatment. Using db/db mice, we performed oral glucose tolerance test (OGTT) to examine the blood glucose lowering effect of GS administration. We also collected the mouse plasma during the OGTT to measure the GLP-1 and insulin levels. Result We demonstrated dose-dependent GLP-1 secreting effect of GS on the NCI-H716 cells. The GLP-1 secreting effect of GS is mediated by the G protein βγ-subunit and inositol triphosphate. Using db/db mice, we found that the effect of GS on lowering blood glucose is due to its GLP-1 secretion, and consequential insulinotropic effect. The chemical fingerprint of GS was obtained through a direct analysis in realtime mass spectrometry (DART-MS) and high-performance liquid chromatography (HPLC)/MS. Through the GLP-1 secretion study, we found that loganic acid, an iridoid glycoside, contributes to the GLP-1 secreting effect of GS. Conclusion The findings of this study highlight the potential of exploiting the antidiabetic effect of GS on type 2 diabetes mellitus patients. [ABSTRACT FROM AUTHOR]

Gentianae Scabrae Radix is a well-known traditional medicine that is used for the treatment of hepatitis, cholecystitis and inflammatory diseases. It consists mainly of secoiridoid glycosides, with representatives of gentiopicroside, sweroside and swertiamarin. In the present study, a chemical investigation of the CHCl extract of Gentianae Scabrae Radix derived from Gentiana scabra Bunge yielded seven new ( 1- 7) and ten known ( 8- 17) secoiridoid glycosides. Their structures were elucidated by extensive spectroscopic analyses and comparison with literature data. All 17 compounds were evaluated for their inhibitory effects against NO, IL-6 and TNF- α productions induced by lipopolysaccharide (LPS) in RAW264 cells. Among them, 8- epi-kingiside derivatives 1- 3; kingiside derivatives 4, 5 and 10; and a sweroside derivative 6 showed inhibition activity against IL-6 production with IC values of 51.70-61.10 μM, whereas sweroside derivatives 12 and 15- 17 and a swertiamarin derivative 13 showed inhibition effects on both NO and IL-6 productions with IC values of 64.74-94.95 and 48.91-75.45 μM, respectively. All the compounds exhibited weak inhibitory activity (IC > 100 μM) in a TNF-α bioassay. Finally, a primary structure-activity relationship of these secoiridoid glycosides is discussed. [ABSTRACT FROM AUTHOR]

The article discusses a study by Wei Li and colleagues which aimed to evaluate the inhibitory effects of each compound of the soluble epoxide hydrolase (sEH) of perennial herbaceous plant Gentiana scabra (G. scabra). Topics discussed include the role of sEH on eicosanoid epoxides metabolism and epoxyeicosatrienoic acids (EETs) hydrolisis, and the isolation of nine phenolic compounds including siringin, berchemol, and koaburaside from G. Scabra's rhizomes and roots.

Independent component (IC) analysis was applied to near-infrared spectroscopy for analysis of gentiopicroside and swertiamarin; the two bioactive components of Gentiana scabra Bunge. ICs that are highly correlated with the two bioactive components were selected for the analysis of tissue cultures, shoots and roots, which were found to distribute in three different positions within the domain [two-dimensional (2D) and 3D] constructed by the ICs. This setup could be used for quantitative determination of respective contents of gentiopicroside and swertiamarin within the plants. For gentiopicroside, the spectral calibration model based on the second derivative spectra produced the best effect in the wavelength ranges of 600--700 nm, 1600--1700 nm, and 2000--2300 nm (correlation coefficient of calibration = 0.847, standard error of calibration = 0.865%, and standard error of validation = 0.909%). For swertiamarin, a spectral calibration model based on the first derivative spectra produced the best effect in the wavelength ranges of 600--800 nm and 2200--2300 nm (correlation coefficient of calibration = 0.948, standard error of calibration = 0.168%, and standard error of validation = 0.216%). Both models showed a satisfactory predictability. This study successfully established qualitative and quantitative correlations for gentiopicroside and swertiamarin with near-infrared spectra, enabling rapid and accurate inspection on the bioactive components of G. scabra Bunge at different growth stages. [ABSTRACT FROM AUTHOR]

Background: The root of Gentiana scabra is commonly known as Longdan in Chinese herbal medicines and has been used in the treatment of inflammation, anorexia, indigestion and gastric infections for over 2000 years. High market demand had made G. scabra (GS) plants not to be the only source of Longdan in China, other Gentiana spp., G. triflora, G. manshurica and G. rigescens, were also recognized as Longdan in China now. Results: In this study, we identified three Taiwan-specific Gentiana spp., G. davidii var. formosana (GDF) and G. arisanensis (GA) and G. scabrida var. punctulata (GSP) that are phylogenetically different from GS (main source of Longdan). However, the active compounds of Longdan, gentiopicroside and swertiamari, were found in GSP and GDF showed higher antioxidant ability and free radical scavenging activities than Chinese Longdan. This discovery might explore the medicinal potential of GDF. Meanwhile, another Taiwan-specific Gentiana spp., GSP, was found to have the strongest antioxidant ability and free radical scavenging activities which might suggest a possible use of GSP as a source of natural antioxidant agents for industrial purpose. Conclusions: The finding of this study indicated that ITS analysis can be used to identify Taiwan-specific Gentiana spp. Also the Taiwan-specific Gentiana spp. which has strongest antioxidant and free radical scavenging activities among others could be a better choice for industrial purpose. [ABSTRACT FROM AUTHOR]

Background: The root of Gentiana scabra is commonly known as Longdan in Chinese herbal medicines and has been used in the treatment of inflammation, anorexia, indigestion and gastric infections for over 2000 years. High market demand had made G. scabra (GS) plants not to be the only source of Longdan in China, other Gentiana spp., G. triflora, G. manshurica and G. rigescens, were also recognized as Longdan in China now. Results: In this study, we identified three Taiwan-specific Gentiana spp., G. davidii var. formosana (GDF) and G. arisanensis (GA) and G. scabrida var. punctulata (GSP) that are phylogenetically different from GS (main source of Longdan). However, the active compounds of Longdan, gentiopicroside and swertiamari, were found in GSP and GDF showed higher antioxidant ability and free radical scavenging activities than Chinese Longdan. This discovery might explore the medicinal potential of GDF. Meanwhile, another Taiwan-specific Gentiana spp., GSP, was found to have the strongest antioxidant ability and free radical scavenging activities which might suggest a possible use of GSP as a source of natural antioxidant agents for industrial purpose. Conclusions: The finding of this study indicated that ITS analysis can be used to identify Taiwan-specific Gentiana spp. Also the Taiwan-specific Gentiana spp. which has strongest antioxidant and free radical scavenging activities among others could be a better choice for industrial purpose. [ABSTRACT FROM AUTHOR]

Abstract: Flower senescence of the potted gentian (Gentiana scabra) ‘Shinbisei’ was investigated in relation to ethylene sensitivity and production. ‘Shinbisei’ flowers were used for all experiments except for those with inflorescences. Exposure to ethylene at 0.5μLL−1 or higher concentrations for 24h markedly accelerated flower senescence, indicating that G. scabra flowers are highly sensitive to ethylene. Treatment with 0.2 or 0.5mM silver thiosulfate complex (STS) and 2μLL−1 1-methylcyclopropene (1-MCP), ethylene action inhibitors, and 50mM α-aminoisobutyric acid, an inhibitor of 1-aminocyclopropane-1-carboxylate (ACC) oxidase, did not delay flower senescence. However, treatment with 1mM l-α-(2-aminoethoxyvinyl) glycine, an inhibitor of ACC synthase, slightly delayed flower senescence. Pollination significantly accelerated petal senescence of G. scabra flowers. Ethylene production of petals, gynoecium, and stamens in unpollinated flowers slightly increased during senescence. Pollination significantly increased ethylene production of petals, gynoecium and stamens 1 day after pollination. To clarify whether 1-MCP delays senescence of cut gentian inflorescences, cut G. scabra ‘Yuki-hotaru’, G. scabra × Gentiana triflora ‘Aoi-kaze’, and G. triflora ‘Koharu’ inflorescences with various stages of flowers, including buds with colored petals, were treated with 2μLL−1 1-MCP for 24h. 1-MCP treatment delayed flower wilting of cut inflorescences of ‘Aoi-kaze’ and ‘Yuki-hotaru’ more than that of ‘Koharu’, suggesting that there is species variation in the effect of 1-MCP in delaying flower senescence of cut gentian inflorescences. [Copyright &y& Elsevier]

When the influence of host species, inoculum density, temperature, leaf wetness duration, and leaf position on the incidence of gentian brown leaf spot caused by Mycochaetophora gentianae, was examined, the fungus severely infected all seven Gentiana triflora cultivars, but failed to infect two cultivars of G. scabra and an interspecific hybrid cultivar. Inoculum density correlated closely with disease incidence, and a minimum of 10 conidia/mL was enough to cause infection. In an analysis of variance, temperature and leaf wetness duration had a significant effect upon disease incidence, which increased with higher temperature (15-25°C) and longer duration of leaf wetness (36-72 h). No disease developed at temperatures lower than 10°C or when leaf wetness lasted <24 h. At 48-h leaf wetness, disease incidence was 0, 28, 77, and 85% at 10, 15, 20, and 25°C, respectively. Middle and lower leaves on the plant were more susceptible than upper leaves. In microscopic observations of inoculated leaves, >50% of conidia germinated at temperatures >15°C after 24-h leaf wetness. More appressoria formed at higher temperatures (15-25°C) with extended duration of leaf wetness (24-72 h). At 48-h leaf wetness, appressorium formation was 0, 8, 26, and 73% at 10, 15, 20, and 25°C, respectively. These results suggest that temperature and leaf wetness duration were important factors for infection of gentian leaves. [ABSTRACT FROM AUTHOR]

The paper studies the phenology and blooming of Gentiana scabra Bunge. Thirteen species of possible agents of pollen transfer and basic insects-pollinators are revealed. [ABSTRACT FROM AUTHOR]

The first complete chloroplast (cp) genome of Gentiana scabra was determined from Illumina paired-end sequencing data in this study. The cp genome of G. scabra is 146,915 bp in length with 37.84% overall GC content, including a large single copy (LSC) region of 79,350 bp and a small single copy (SSC) region of 17,027 bp, which are separated by a pair of inverted repeats (IRs) of 25,269 bp. The cp genome is composed of 130 genes, including 85 protein-coding genes, 8 rRNA genes, and 37 tRNA genes. Phylogenetic analysis indicated that the G. scabra is more closely related to subfamily of Gentianoideae than Menyanthaceae. [ABSTRACT FROM AUTHOR]

The flavonoid 3′,5′-hydroxylase (F3′,5′H) gene is in the NADPH-Cytochrome P450 reductase family. A partial F3′,5′H cDNA was cloned by RT-PCR from corollas of Gentiana scabra. Activity of F3′,5′H was observed during flower development, and activity was highest at the stage prior to flower opening. The enzyme activity was increased with the addition of SO μM gibberellic acid (GA3) immediately prior to flower opening. Western blot analysis showed that purified F3′,5′H protein from G. scabra was cross-reacted with rabbit anti-Gentiana triflora NADPH-Cyt P450 reductase antibodies. Only one 80 to 82 kDa protein was detected by western blot analysis of fresh microsomal extracts. The enzyme activity was not distinctly changed by adding 50 μM GA3 to corollas. Therefore, we conclude that GA3 controls the expression of the F3′,5′H gene at the transcriptional level, not at the translational level. [ABSTRACT FROM AUTHOR]

In order to find out anti-platelet activating factor (PAF) from natural resources, Korean medicinal plants used for the treatments of peripheral circulation disorders were tested for their possible protective effects on PAF-induced anaphylactic shock. From the above screening, the methanol extract of Gentiana scabra showed a potent antagonistic activity against PAF. Water suspension of the extract was partitioned with CH2Cl2 and EtOAc, successively. The EtOAc fraction which showed the highest activity was chromatographed on silica gel to yield 6 fractions. From the fraction which showed higher PAF-antagonistic activity than the other fractions, compound 1 was isolated by recrystallization. On the basis of spectral data, compound 1 was identified as 2-hydroxy-3-methoxybenzoic acid glucose ester. The compound prevented the mice from the PAF-induced death at a dose of 300 μg/mouse. [ABSTRACT FROM AUTHOR]

Brown leaf spot caused by Mycochaetophora gentianae is a serious disease on gentian in Japan. Previous studies revealed that Gentiana triflora cultivars were susceptible to M. gentianae and that G. scabra cultivars and their interspecific hybrid cultivars were resistant. We subsequently analyzed the mode of inheritance of resistance to M. gentianae in gentians using several populations derived from crosses between G. scabra and G. triflora. Parental G. scabra and G. triflora lines, F and BC plants were inoculated with a conidial suspension and assessed for susceptibility to M. gentianae. All tested F plants were resistant to M. gentianae, indicating that the resistance of G. scabra was inherited dominantly. Segregation ratios in backcross progenies using G. scabra line OK fit the Mendelian ratio of 1:1, showing the involvement of a single dominant locus. These results indicate that the resistance to M. gentianae in G. scabra line OK is controlled by a single dominant allele designated as gentian brown leaf spot resistance ( GBLS- 1). Finding this allele should facilitate future breeding efforts to develop resistant gentian cultivars. [ABSTRACT FROM AUTHOR]

Gentianae Radix et Rhizome (Longdan in Chinese, GRR) in Chinese Pharmacopoeia is derived from the dried roots and rhizomes of Gentiana scabra and G. rigescens, that have long been used for heat-clearing and damp-drying in the medicinal history of China. However, the characterization of the chemical components of two species and the screening of chemical markers still remain unsolved. In current research, the identification and characterization of chemical components of two species was performed using ultra-high-performance liquid chromatography (UHPLC) coupled with linear ion trap-Orbitrap (LTQ-Orbitrap) mass spectrometry. Subsequently, the chemical markers of two species were screened based on metabolomics and multivariate statistical analysis. In total, 87 chemical constituents were characterized in G. scabra (65 chemical constituents) and G. rigescens (51 chemical constituents), with 29 common chemical constituents being discovered. Thereafter, 11 differential characteristic components which could differentiate the two species were designated with orthogonal partial least squares discriminant analysis (OPLS-DA) and random forest (RF) iterative modeling. Finally, seven characteristic components identified as (+)-syringaresinol, lutonarin, trifloroside, 4-O-β-d-glu-trifloroside, 4″-O-β-d-glucopyranosy1-6′-O-(4-O-β-d-glucaffeoyl)-linearroside, macrophylloside a and scabraside were selected as the chemical markers for the recognition of two Gentiana species. It was implied that the results could distinguish the GRR derived from different botanical sources, and also be beneficial in the rational clinical use of GRR. [ABSTRACT FROM AUTHOR]