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ObjectiveRoot rot is one of the most serious diseases in the cultivation and production of Atractylodes lancea. Trichoderma spp. are effective in the biocontrol of root rot without causing environmental pollution. This study aims to isolate and study a Trichoderma strain capable preventing and controlling root rot from the rhizosphere of A. lancea and to solve the problem of disease prevention and control in the planting and production of A. lancea. MethodTrichoderma T2204 was isolated by the dilution-coating method and identified by ITS sequencing. The inhibitory activities of T2204 and its volatiles against two pathogenic fungal strains were examined by dual-culture and co-culture experiments. The biocontrol potential of T2204 on root rot of A. lancea and the effect of T2204 on the accumulation of medicinal compounds in the rhizosphere of A. lancea were investigated by pot experiments and GC-MS, respectively. In addition, the optimal medium, photoperiod, temperature, pH, and carbon and nitrogen sources for the culture of T2204 were explored. ResultThe Trichoderma isolate T2204 was identified as T. citrinoviride and had direct inhibitory effects on two highly pathogenic strains causing root rot. In the dual-culture experiments with the two pathogenic strains, T2204 showcased the inhibition rates of 77.90% and 76.80%, respectively. In the co-culture experiments with the two pathogenic strains, the volatile organic compounds produced by T2204 showed the inhibition rates of 57.11% and 81.11%, respectively. The pot experiments showed that the survival rate of A. lancea seedlings infected by root rot reached 100% after inoculation with T2204 and was only 50% in the case without inoculation of T2204. After 150 days of cultivation, the dry weight and atractylodin content of the rhizome of A. lancea plants treated with T2204 increased by 32% (P<0.05) and 11%, respectively, compared with the untreated group. The optimal conditions for the growth of T2204 were PDA or PSA medium, photoperiod of 12 h dark/12 h light, 25-30 °C, pH 5-6, carbon sources of glucose, D-fructose, soluble starch, and maltose, and the nitrogen sources of ammonium sulfate and ammonium dihydrogen phosphate. The optimal conditions for the sporulation of T2204 were PSA or CMA medium, photoperiod of 12 h dark/12 h light, 20-30 °C, pH 8, carbon source of sucrose, and nitrogen source of sodium nitrate. ConclusionT2204 could improve the growth and root rot resistance of A. lancea and promote the accumulation of medicinal compounds. The findings laid a foundation for the industrialized production and application of T2204 in the production of A. lancea in the future.
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ObjectiveTo study the differences in volatile oil content of bran-processed Atractylodes lancea and its standard decoction concentrate and freeze-dried powder, as well as the differences in the types and contents of chemical components in volatile oil, and to clarify the quality value transmitting. MethodTen batches of A. lancea rhizoma were collected and prepared into raw products and bran-processed products of A. lancea, standard decoction concentrate and freeze-dried powder of bran-processed A. lancea in order to extract the volatile oil, and the transfer rate of volatile oil in each sample was calculated. Quantitative analysis of the main chemical components(β-eudesmol, atractylon, atractylodin) in each volatile oil was performed by gas chromatography(GC) on the HP-5 quartz capillary column(0.32 mm×30 m, 0.25 μm) with a flame ionization detector(FID), a split ratio of 10∶1 and a temperature program(initial temperature at 80 ℃, hold for 1 min, rise to 150 ℃ at 10 ℃·min-1, hold for 10 min, rise to 155 ℃ at 0.5 ℃·min-1, hold for 5 min, rise to 240 ℃ at 8.5 ℃·min-1, hold for 8 min). Cluster analysis and principal component analysis(PCA) were used to explore the overall differences in types and contents of chemical components between the standard decoction concentrate and freeze-dried powder. ResultThe transfer rates of volatile oil in the bran-processed products, standard decoction concentrate and freeze-dried powder were 70.51%, 1.57% and 40.90%, respectively. The average transfer rates of β-eudesmol, atractylon and atractylodin in the volatile oil of bran-processed A. lancea were 58.45%, 48.49% and 55.64%, respectively. In the standard decoction concentrate, only β-eudesmol and atractylodin were detected, and their average transfer rates were 0.22% and 0.10%, respectively. And only β-eudesmol was detected in the freeze-dried powder with the average transfer rate of 8.37%. The results of cluster analysis and PCA showed that there are obvious differences in the types and contents of chemical components between the standard decoction concentrate and freeze-dried powder. ConclusionThe quality value transmitting between bran-processed A. lancea and its standard decoction concentrate and freeze-dried powder is stable, and if the freeze-dried powder is selected as the reference material of dispensing granules, appropriate amount of volatile oil should be added back to make it consistent with the quality of the standard decoction concentrate.
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This study investigated the choroplast genome sequence of wild Atractylodes lancea from Yuexi in Anhui province by high-throughput sequencing, followed by characterization of the genome structure, which laid a foundation for the species identification, analysis of genetic diversity, and resource conservation of A. lancea. To be specific, the total genomic DNA was extracted from the leaves of A. lancea with the improved CTAB method. The chloroplast genome of A. lancea was sequenced by the high-throughput sequencing technology, followed by assembling by metaSPAdes and annotation by CPGAVAS2. Bioiformatics methods were employed for the analysis of simple sequence repeats(SSRs), inverted repeat(IR) border, codon bias, and phylogeny. The results showed that the whole chloroplast genome of A. lancea was 153 178 bp, with an 84 226 bp large single copy(LSC) and a 18 658 bp small single copy(SSC) separated by a pair of IRs(25 147 bp). The genome had the GC content of 37.7% and 124 genes: 87 protein-coding genes, 8 rRNA genes, and 29 tRNA genes. It had 26 287 codons and encoded 20 amino acids. Phylogenetic analysis showed that Atractylodes species clustered into one clade and that A. lancea had close genetic relationship with A. koreana. This study established a method for sequencing the chloroplast genome of A. lancea and enriched the genetic resources of Compositae. The findings are expected to lay a foundation for species identification, analysis of genetic diversity, and resource conservation of A. lancea.
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Phylogeny , Atractylodes/genetics , Genome, Chloroplast , Whole Genome Sequencing , Microsatellite Repeats , LamialesABSTRACT
We explored the correlations between the color difference values [ΔL~*(lightness), Δa~*(red-green), Δb~*(yellow-blue)] and the content of four active components(including sesquiterpenoids and polyacetylenes) in the powder of Atractylodes lancea and A. chinensis, aiming to provide reference for the quality evaluation of Atractylodis Rhizoma and establish a qualitative model that can distinguish between A. lancea and A. chinensis based on the chromatic values. The tristimulus values(L~*, a~*, and b~*) of 23 batches of A. lancea and A. chinensis were measured by a color difference meter. The content of atractylenolide Ⅱ, β-eudesmol, atractylodin, and atractylone in the 23 batches of samples were measured by high performance liquid chromatography(HPLC). Principal component analysis(PCA) and partial least squares-discriminant analysis(PLS-DA) were performed to establish the qualitative models for distinguishing between A. lancea and A. chinensis. SPSS was employed to analyze the correlations between the tristimulus values and the content of the four index components. The results showed that the established PCA and PLS-DA models can divide the A. lancea and A. chinensis samples into two regions, and the tristimulus values of A. lancea and A. chinensis were positively correlated with the content of β-eudesmol and atractylodin. Therefore, the PCA and PLS-DA models can successfully identify A. lancea and A. chinensis, and the appearance color can be used to quickly predict the internal quality of Atractylodis Rhizoma. This study provides a reference for the quality evaluation of Atractylodis Rhizoma and the modern research on the color of Chinese medicinal materials.
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Atractylodes , Sesquiterpenes, Eudesmane , Drugs, Chinese Herbal , Rhizome , ExcipientsABSTRACT
4-(Cytidine 5′-diphospho)-2-C-methyl-D-erythritol kinase (CMK) was one of the key enzymes in the methylerythritol-4-phosphate (MEP) pathway to generate terpenoids. In this study, based on the transcriptome data of Atractylodes lancea, the sequence of the CMK gene was cloned, named AlCMK (GenBank accession number OM283293). The results showed that AlCMK contains a 1 230 bp open reading frame (ORF) encoding 409 amino acids. The deduced protein had a theoretical molecular weight of 44 752.53 and an isoelectric point of 6.67. Transmembrane structure analysis showed that there was no transmembrane structure, and the secondary structure of AlCMK was predicted to be mainly composed of random coil. Homologous alignment revealed that AlCMK shared high sequence identity with the CMK proteins of Tanacetum cinerariifolium, Osmanthus fragrans, Eucommia ulmoides, Lonicera japonica and Salvia miltiorrhiza. Phylogenetic analysis indicated that AlCMK protein had the higher homology with CMK protein of Compositae. The pET-32a-AlCMK prokaryotic expression vector was constructed and a fusion protein with molecular mass of about 65 kDa was expressed in the E. coli BL21 (DE3). The qRT-PCR was used to analyze the expression pattern of AlCMK gene in different tissues and after MeJA treatment. Meanwhile, the enzyme activity was determined by ELISA kit. The results showed that AlCMK gene was tissue-expressed in different origins and its expression was induced by MeJA, and the results of the enzyme activity assay showed that the AlCMK enzyme activity in different regions was higher in the leaves. The subcellular localization showed that AlCMK was located in the chloroplast. This study provides a reference for further elucidating the biological function of AlCMK gene in terpenoid synthesis pathway in Atractylodes lancea.
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Objective : To study the effect of temperature and light intensity on photosynthetic fluorescence parameters, volatile oil content, and growth of Atractylodes lancea and provide reference for the rational selection of cultivation environment for A. lancea. MethodWe determined the photosynthetic indexes (such as net photosynthetic rate, water use efficiency, and carboxylation rate), light response curve, CO2 response curve, fluorescence parameters, and the content of four volatile oils in A. lancea under two temperature treatments (32 °C and 22 °C) and two light treatments (full light and shade). ResultThe net photosynthetic rate and water use efficiency of A. lancea under high temperature + strong light were significantly higher than those under high temperature + weak light and low temperature + strong light. The ability of A. lancea to use weak light at low temperature was the strongest, while the utilization rate of weak light under strong light significantly reduced. The photosynthetic rate of A. lancea at low temperature was more susceptible to light intensity and CO2 concentration than that at high temperature. The maximum photosynthetic rate and apparent quantum efficiency under weak light were significantly higher than those under strong light. The photoreaction efficiency at high temperature was higher than that at low temperature. The total amount of volatile oil in A. lancea treated with high temperature + weak light was the highest, reaching 4.582%. Compared with high temperature + strong light, high temperature + weak light significantly increased the content of hinesol and β-eudesmol in A. lancea by 91.7% and 35.7%, respectively, and low temperature + strong light significantly increased the content of hinesol by 87.5%. The content of β-eudesmol in low temperature + weak light treatment was significantly lower than that in high temperature + weak light treatment. ConclusionTThe growth of A. lancea was affected by the interaction between temperature and light. The light and temperature conditions required for the accumulation of volatile oil were not consistent with those suitable for the growth and development of A. lancea. A. lancea responded to the changes of light and temperature conditions by regulating the synthesis and accumulation of volatile oil.
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The endophytes of medicinal plants play an important role in promoting the quality formation of the host. Therefore, this paper made a review of endophytes of medicinal plant Atractylodes lancea. According to previous studies, A. lancea boasts endophytes, such as fungi, bacteria, and actinomycetes, among which the beneficial microorganisms help the growth and development of A. lancea. There is a close interaction between the volatile oil of A. lancea and endophytes. Different endophytes vary in regulating the composition and content of the volatile oil of A. lancea, which might contribute to the quality formation of A. lancea. However, the information of the endophytic flora of A. lancea obtained by traditional culture and isolation is not enough to reflect the real situation of the endophytes of A. lancea. Little is known about the endophytes of A. lancea from different chemical types and different habitats, which is not conducive to the study of the ecological relationship between A. lancea and endophytes and limits the development and utilization of the endophytes. Therefore, at the end of this paper, the authors put forward suggestions for future research on endophytes in A. lancea, including:(1)mining the core endophyte resources of A. lancea by combining high-throughput sequencing with traditional culture and isolation;(2)exploring the relationship between the diversity of endophytes and chemical types of A. lancea;(3)strengthening the application of endophytes in A. lancea cultivation, in order to facilitate the cultivation efficiency and quality of A. lancea.
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Atractylodes , Endophytes , Fungi , Oils, Volatile , Plants, MedicinalABSTRACT
In this study, the gene encoding the key enzyme 3-ketoacyl-CoA thiolase(KAT) in the fatty acid β-oxidation pathway of Atractylodes lancea was cloned. Meanwhile, bioinformatics analysis, prokaryotic expression and gene expression analysis were carried out, which laid a foundation for the study of fatty acid β-oxidation mechanism of A. lancea. The full-length sequence of the gene was cloned by RT-PCR with the specific primers designed according to the sequence information of KAT gene in the transcriptomic data of A. lancea and designated as AIKAT(GenBank accession number MW665111). The results showed that the open reading frame(ORF) of AIKAT was 1 323 bp, encoding 440 amino acid. The deduced protein had a theoretical molecular weight of 46 344.36 and an isoelectric point of 8.92. AIKAT was predicted to be a stable alkaline protein without transmembrane segment. The secondary structure of AIKAT was predicted to be mainly composed of α-helix. The tertiary structure of AIKAT protein was predicted by homology modeling method. Homologous alignment revealed that AIKAT shared high sequence identity with the KAT proteins(AaKAT2, CcKAT2, RgKAT and AtKAT, respectively) of Artemisia annua, Cynara cardunculus var. scolymus, Rehmannia glutinosa and Arabidopsis thaliana. The phylogenetic analysis showed that AIKAT clustered with CcKAT2, confirming the homology of 3-ketoacyl-CoA thiolase genes in Compositae. The prokaryotic expression vector pET-32 a-AIKAT was constructed and transformed into Escherichia coli BL21(DE3) for protein expression. The target protein was successfully expressed as a soluble protein of about 64 kDa. A real-time quantitative PCR analysis was performed to profile the AIKAT expression in different tissues of A. lancea. The results demonstrated that the expression level of AIKAT was the highest in rhizome, followed by that in leaves and stems. In this study, the full-length cDNA of AIKAT was cloned and expressed in E. coli BL21(DE3), and qRT-PCR showed the differential expression of this gene in different tissues, which laid a foundation for further research on the molecular mechanism of fatty acid β-oxidation in A. lancea.
Subject(s)
Amino Acid Sequence , Atractylodes/genetics , Cloning, Molecular , Coenzyme A , Escherichia coli/genetics , PhylogenyABSTRACT
Objective:The effects of Atractylodes lancea, A. coreana, A. japonica, A. chinensis and Atractylodis Macrocephalae Rhizoma on spleen-Qi deficiency rats were compared. Method:A model of spleen-Qi deficiency was induced in rats by diet and overwork.The rats are given different suspensions of A.japonica, A.chinesis, A.coreana, A.lancea and Atractylodis Macrocephalae Rhizoma To test the indicators of the digestive system, immune system and antioxidant enzyme system related to spleen deficiency.Compare the similarities and differences between Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma from four different sources. Result:All the drug-administered groups can increase the levels of γ interferon (IFN-γ), gastric secrete element(GAS),serum amylase (AMS) and catalase(CAT) in rats with spleen-deficiency syndrome,in addition to the CAT index, the other indicators of the A. coreana, A.japonica, and A.lancea were significantly increased(P<0.05). The MTL content of the A.chinesis, A.lancea, A.coreana and A.japonica increased and was significant(P<0.05). The SDH content of A.japonica.and A.chinesis increased, and the difference was not significant.The increase of GSH-Px in the A.chinesis is significant(P<0.05). All the drug-administered groups can reduce the content of IgG, TNF-α and MDA in rats with spleen deficiency and deficiency syndrome. Among them, the IgG content of the A.chinesis. and the A.lancea was significantly decreased(P<0.05). The content of TNF-α in A.japonica group was significantly decreased(P<0.05). The content of MDA in the A.chinesis, the A.lancea, the A.coreana,the A.japonica and the Atractylodis Macrocephalae Rhizoma were significantly decreased(P<0.05).The decrease of IL-6 in the A.lancea was significant(P<0.05). Conclusion:Four different sources of Atractylodes Rhizome and A.macrocephala have certain therapeutic effects on spleen-deficiency rats with deficiency syndrome.The therapeutic effect of A.lancea and A.japonica is basically the same,regulating the absorption,secretion and elimination of inflammation in the digestive system of rats with spleen deficiency A.coreana, A.chinesis, and Atractylodis Macrocephalae Rhizoma have certain regulatory effects in the digestive system, digestive tract inflammation, and antioxidant enzymes.
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Objective::To explore the effect of strong light stress on the growth, physiological and biochemical and key enzyme gene expression of the Atractylodes lancea, in order to provide the scientific basis for the standardized cultivation of the A. lancea. Method::The two-year-old A. lancea seedlings were taken as experimental materials. Poplar forest (light transmittance between 18.26%-36.04%) was taken as control group(ck). Different density shading networks were used to simulate different degrees of high light stress (51.10%, 80.73%, 100%) in late July. The growth state of A. lancea was observed. On the 0th, 5th, 10th, 15th, 20th days, the physiological and biochemical indexes of malondialdehyde (MDA) content, cell membrane permeability, proline (Pro) content, antioxidant enzyme activity and chlorophyll content in the leaves of A. lancea were measured. The relative expression levels of 3-hydroxy-3-methylglutarate monoacyl coenzyme A reductase (3-hydroxy-3-methylglutaryl coenzyme A, HMGR) and farnesyl pyrophosphate synthase gene (farnesyl pyrophosphate synthase, FPPS) in leaves of A. lancea under intense light stress were determined by real-time fluorescence quantitative PCR(Real-time PCR). Result::After strong light stress, the color of the leaves of A. lancea changed from dark green to light green and yellowish green, and the burn of leaves became more and more serious. The contents of MDA, conductivity and Pro showed an upward trend with the increase of transmittance. Peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT) tended to increase first and then decrease. The chlorophyll content decreased with the increase of light transmittance. The relative expression of HMGR in leaves of A. lancea decreased with the increase of light transmittance, while FPPS increased first and then decreased. Conclusion::The results showed that A. lanceaa could alleviate the inhibition of strong light stress by increasing the activity of antioxidant enzymes and regulating the content of osmotic pressure under certain strong light stress. Excessively strong intensity light stress leads to disequilibrium of metabolic mechanism of A. lancea, and seriously inhibits the plant growth.
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Objective: To investigate the chemical components from the 80% EtOH extract of Atractylodes lancea, as well as the inhibitory activities of the isolated compounds on LPS-induced NO production of microglia BV2 cells. Methods: The n-BuOH-soluble fraction of the crude extract was successively chromatographed with Diaion HP-20, Sephadex LH-20, and preparative HPLC C18-column. At last, the planar and stereochemical structures of these obtained compounds were established on the basis of extensive spectroscopic data (HRESIMS, NMR, and ECD, etc). Results: Ten glycosides were isolated from the n-BuOH-soluble fraction of the 80% EtOH extract of A. lancea, including (2E,8R)-decene-4,6-diyne-1,8-diol-1-O-β-D- apiofuranosyl-(1→6)-β-D-glucopyranoside (1), (8S)-decane-4,6-diyne-1,8-diol-8-O-β-D-glucopyranoside (2), (2E,8R)-decene-4,6- diyne-1,8-diol-8-O-β-D-glucopyranoside (3), (2E,8S)-decene-4,6-diyne-1,8-diol-8-O-β-D-glucopyranoside (4), (2E,8E)-2,8- decadiene-4,6-diyne-1,10-diol-1-O-β-D-glucopyranoside (5), (7R,8S)-3',9,9'-trihydroxyl-3-methoxyl-1'-propanol-7,8-dihydrobenzo- funanneoligan-4-O-β-D-glucopyranoside (6), (7'R*,8S*,8'S*)-lyoniresinol 9'-O-β-D-glucopyranoside (7), (7S,8R)-4,9,9'-trihydroxy- 3'-methoxy-8-O-4'-neolignan 7-O-β-D-glucopyranoside (8), methyl salicylate 2-O-α-L-xylopyranosyl-(1→6)-β-D-glucopyranoside (9), and phenylmethanol 7-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside (10). Conclusion: Compounds 1 and 2 are named atractyeneyneglycoside A and atractyeneyneglycoside B, while compounds 6, 8-10 are first isolated from the rhizomes of A. lancea. At the concentration of 10 μmol/L, compound 10 exhibited the strongest inhibitory effects on LPS-induced NO production of microglia BV2 cells with the value of 31.18%, while compounds 1 and 2 just showed weaker inhibitory effects with values of 22.01% and 14.09%, respectively.
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Objective:To investigate the dryness effect of Atractylodes lancea and A. chinensis. Method:Sixty normal and healthy SD rats were randomly divided into 6 groups(10 in each group), including normal saline group, soybean oil group, low-dose(46.25 mg·kg-1·d-1) group and high-dose(500 mg·kg-1·d-1) group of A. lancea, low-dose(46.25 mg·kg-1·d-1) group and high-dose(500 mg·kg-1·d-1) group of A. chinensis, the dosing volume was 0.01 mL·g-1, and the drug was administered orally for 21 days. Taking average daily water intake, submandibular gland tissue, urine volume and expression of aquaporin 2(AQP2) in the kidney, and whole blood viscosity as the evaluation indexes, the dryness effect of long-term administration of equal doses of volatile oil from A. lancea and volatile oil from A. chinensis on rats was observed. Result:Compared with the soybean oil group, long-term administration of high doses of volatile oil from A. lancea and volatile oil from A. chinensis could significantly increase average daily water intake, urine volume and whole blood viscosity; decrease the expression of AQP2, and atrophy the acini of submandibular gland, but there was no significant difference between the two groups. Effects of volatile oil from A. lancea and A. chinensis with low dose on dryness of rats were not significant. Conclusion:There is no significant difference between the dryness effect of volatile oil from A. lancea and A. chinensis in the same dose. It is proved that the rationality of A. lancea and A. chinensis are universal in clinical practice, and this study provides experimental basis for rational use of Atractylodis Rhizoma.
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Objective: To explore the effect on the accumulation of medicinal compositions β-eudesmol, atractylon, atractylodin and key enzyme genes 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) and farnesyl pyrophosphate synthase (FPPS) expression in biosynthesis of Atractylodes lancea under copper stress. Methods: Under copper stress, the expression of key enzyme genes HMGR and FPPS in A. lancea was determined by real-time fluorescence quantitative PCR; the content of three medicinal components in A. lancea were determined by HPLC; The correlation analysis was performed with SPSS, and DPS software for grey correlation analysis. Results: When the copper stress concentration was within 100 mg/kg, the expression of FPPS and the content of atractylon in the rhizomes of A. lancea increased slightly. However, when the copper concentration continued to increase, the expression levels of HMGR and FPPS and three medicinal components content of A. lancea showed a different degrees of downward trend. The expression levels of HMGR and FPPS were positively correlated with the content of β-eudesmol, atractylon, and atractylodin (P < 0.05) under copper stress. Grey relational analysis showed that the content of β-eudesmol and atractylon in the rhizomes was significantly correlated with the expression of HMGR and FPPS of A. lancea under copper stress. The expression of FPPS gene had the larger contribution on the composition of β-eudesmol and atractylon. However, the correlation between the content of atractylodin and the expression of these two key enzyme genes was relatively small. Conclusion: This study clarified the change regulation of two key enzyme gene expression and the content of three medicinal compositions, and revealed the relationship between β-eudesmol, atractylon and HMGR and FPPS, the key enzymes in terpene biosynthesis of A. lancea under copper stress. It contributed to the further study of the molecular regulation mechanism of the synthesis of medicinal constituents under copper stress and provided a theoretical basis for improving the quality of A. lancea.
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Objective: To clone the full-length cDNA encoding farnesyl pyrophosphate synthase (FPPS) from Atractylodes lancea and analyze its expression. Methods: The full-length cDNA of FPPS in A. lancea was cloned via homology-based cloning and rapid amplification of cDNA ends approach. Also, the characterization of gene was revealed by bioinformatic analysis. The expression of FPPS was determined by qRT-PCR while the content of sesquiterpenes of rhizome in different growth stages in A. lancea was measured by GC-MS. Meanwhile the correlation between them was analyzed. Results: The full-length cDNA (1320 bp) of FPPS gene was obtained (AlFPPS, GenBank accession number KX443242), with an open reading frame of 1029 bp, encoding 342 amino acids. The deduced AlFPPS protein sequence contained five conserved motifs, two of which is full of Asp (DDXXD). qRT-PCR and GC-MS results showed a significant positive correlation between the content of sesquiterpenes and AlFPPS expression level. Conclusion: It can be primarily deduced that AlFPPS gene should be an important control point in the biosynthetic pathway of sesquiterpenes in A. lancea. This work provides scientific basis for clarifying the biosynthetic pathway of sesquiterpenes and application of biological engineering.
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Objective:To establish a surface enhanced Raman spectroscopy ( SERS) technique for the determination of atropine sulfate. Methods:The Raman peaks of atropine sulfate molecules were classified by theoretical calculations and experimental tests, and the 1002 cm-1 Raman peaks were selected as the characteristic peaks for the quantitative analysis. Results:The detection limit of atropine sulfate in aqueous solution was below 0. 5 μg·ml-1 . The relationship between the intensity of characteristic peaks at 1002 cm-1 and the concentration of aqueous solution was linear within the range of 1-8 μg ml-1 with the linear correlation coefficient r of 0. 9839. The recovery rates of 2, 5 and 7μg ml-1 were measured, which were 97. 1%-109. 8%. The average recovery was 103. 3%, and the RSD was 4. 5% (n=9). At the same time, the stability of the method among the batches was tested, and the relative standard deviation was 5. 7 %. In addition, the atractylodes rhizome samples containing atropine sulfate were detected,and the characteristic peaks still could be detected at 1002 cm-1 . Conclusion:The method is rapid, accurate and nondestructive with easy operation, which can be used for the detection of atropine sulfate.
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The present study was designed to explore the influence of water extracts of Atractylodes lancea rhizomes on the toxicity and anti-inflammatory effects of triptolide (TP). A water extract was prepared from A. lancea rhizomes and co-administered with TP in C57BL/6 mice. The toxicity was assayed by determining serum biochemical parameters and visceral indexes and by liver histopathological analysis. The hepatic CYP3A expression levels were detected using Western blotting and RT-PCR methods. The data showed that the water extract of A. lancea rhizomes reduced triptolide-induced toxicity, probably by inducing the hepatic expression of CYP3A. The anti-inflammatory effects of TP were evaluated in mice using a xylene-induced ear edema test. By comparing ear edema inhibition rates, we found that the water extract could also increase the anti-inflammatory effects of TP. In conclusion, our results suggested that the water extract of A. lancea rhizomes, used in combination with TP, has a potential in reducing TP-induced toxicity and enhancing its anti-inflammatory effects.
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Animals , Male , Mice , Anti-Inflammatory Agents , Pharmacology , Atractylodes , Chemistry , Cytochrome P-450 Enzyme System , Genetics , Diterpenes , Toxicity , Edema , Pathology , Enzyme Induction , Epoxy Compounds , Toxicity , Gene Expression Regulation , Herb-Drug Interactions , Liver , Pathology , Mice, Inbred C57BL , Phenanthrenes , Toxicity , Plant Extracts , Pharmacology , Plants, Medicinal , Chemistry , Rhizome , Chemistry , Water , ChemistryABSTRACT
Objective: To study the mechanism of the content of 5-hydroxymethylfurfural (5-HMF) increasing after Atractylodes lancea in bran-processed. Methods: The fructose and glucose were processed with A. lancea bran-processed conditions and their contents were determined by HPLC. Besides, the content of fructose was measured and the changes were compared before and after processing. Results: Glucose samples did not detect 5-HMF in the processing conditions, but fructose samples detected 5-HMF in the same conditions. In addition, the content of fructose was significantly decreased in the best conditions while the content of 5-HMF increased the most. Conclusion: The content of 5-HMF remarkably increases mainly due to the conversion of fructose.
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This article mainly summarises the results of the chemical compositions and its pharmacological activities of Atractylodes Radix. The chemistry components isolated from Atractylodes Radix are mainly sesquiterpenoids, enediynes, triterpenoids, aromatic glycosides, and etc. Pharmacological results showed that Atractylodes Radix has inhibition of gastric acid secretion, promoting gastrointestinal movement and gastric emptying, hpyerglycemic, antibacterial, anti-inflammatory, cardiovascular protection and nervous system activity, etc. This article hopefully to provide a reference for further research, development and utilization of Atractylodes Radix.
ABSTRACT
To compare the anti-inflammatory activity of the crude Atractylodes lancea (AL) and AL processed products by stir-baking with bran in rat models of gastric ulcer, and preliminarily explore the anti-ulcer mechanisms of AL, the model of gastric ulcer was imitated by local acetic acid injection into gastric mucosa in rats by surgery according to the modified Okabe method. All rats were randomly divided into the following 10 groups: sham-operation group, model group, omeprazole group, Sanjiu Weitai granule group, crude AL low dose group, crude AL middle dose group, crude AL high dose group, processed AL low dose group, processed AL middle dose group, and processed AL high dose group. Rats were administered via intragastric (ig) two times each day, for 10 consecutive days. Blood was collected from the abdominal aorta, serum was separated, and the ulcer tissues were taken. The levels of inflammatory factors interleukin 6, 8 (IL-6, 8), tumor necrosis factor-α (TNF-α), and prostaglandin E2 (PGE2) in serum and gastric tissues were determined by enzyme-linked immunosorbent assay (ELISA), and the mRNA expressions of TNF-α and IL-8 in gastric tissues were detected by quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). The protein expressions of TNF-α and IL-8 in gastric tissues were detected by immunohistochemistry. Compared with sham-operation group, the levels of TNF-α, IL-8, IL-6, PGE2 as well as the mRNA expressions and protein expressions of TNF-α, IL-8 in gastric tissues were significantly higher in model group. The above levels were reduced in different degrees in all treatment groups. Compared with the crude AL, same dose of processed AL was more effective in decreasing the levels of TNF-α, IL-8, IL-6, PGE2 in serum and gastric tissues and down-regulating the mRNA expressions of TNF-α and IL-8 in gastric tissues, with significant difference in middle dose groups and high dose groups. The results showed that AL had potent anti-inflammatory effects in rat models of gastric ulcer induced by acetic acid, and the processed AL had more obvious effect. The anti-ulcer action of AL could be attributed partly to down-regulating the levels of TNF-α, IL-8, IL-6 and PGE2.
ABSTRACT
Based on current progress and field investigations,this review summarized the symptoms,epidemiology and control methods of 11 diseases on Atractylodes lancea, including the most severely harmed root diseases such as root rot and southern blight, as well as the sclerotinia rot that was newly happened. This review aims to demonstrate the progress of studies on diseases of A. lancea, providing guidance for field production. Sclerotonia disease and leaf spot disease are new diseases,suggesting the awareness of this disease on plant quarantine.