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Soybean mosaic virus (SMV) is a critical pathogen that reduces the soybean yield and seed quality worldwide, and SMV has a restricted natural host range. In this study, we used sequence-independent amplification (SIA) methods to identify the viruses that may cause the Atractylodes macrocephala Koidz disease. Results revealed that there is SMV in diseased Atractylodes macrocephala leaves, and we named this isolate as SMV-Am. To further characterize the genomic structural and phylogenetic relationships of SMV-Am, genomic dsRNA was extracted, and the genomic sequence was amplified by RT-PCR and RACE. The results of bioinformatics analysis showed that the genomic RNA of SMV-Am is 9587 nucleotides in length, which conforms to the typical characteristics of Potyvirus. According to the sequence alignment of complete nucleotide sequences, SMV-Am showed the highest level of nucleotide homology and amino acid sequences to SMV-Liaoning, 96. 57% and 98. 86%, respectively. In addition, phylogenetic analysis based on the nucleotide sequences of other SMV isolates of SMV-Am revealed that SMV-Am was most closely related to SMV-Liaoning. Then, the SMV-Am protein was further analyzed by I-TASSER and PyMOL software, revealing that the key amino acid mutations led to the structural changes of P1, HC-Pro, P3, 6K2, NIa-pro and NIb proteins, with P1 the most obvious. Finally, recombination has been detected at the position of 6560-8950 nucleotides, the main parent is the isolate SMV-XFQ012 (accession number KP710875. 1), and the secondary parent is isolate SMV-pCB301-SC15 (accession number MH919386. 1). This study is the first report of SMV in Atractylodes macrocephala Koidz, and these results are expected to provide a scientific basis for the prevention and control of SMV infection on Atractylodes macrocephala Koidz.
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Objective: To summarize the prescription rules of traditional Chinese medicine (TCM) for treating alcoholic liver disease (ALD) based on TCM inheritance support system (V2.50). Methods: The literatures about TCM prescriptions for treating ALD were collected from CNKI, Wanfang database, and VIP database. The TCM inheritance platform system was used to analyze the prescription rules of TCM in the treatment of alcoholic liver disease. Results: Statistics showed that the majority of prescriptions were used to treat alcoholic fatty liver, alcoholic hepatitis, and alcoholic cirrhosis. Through "frequency statistics" analysis, 107 prescriptions were found involving 149 flavors of TCM, with a cumulative frequency of 1 195 times. Twenty-three Chinese medicines with a frequency of ≥ 15 times were used, and the cumulative frequency was 737 times (62%). The most frequently used medicines were blood-activating and stasis-removing drugs, water-diffusing and damp-permeating drugs, tonics, heat-clearing drugs, antialcoholic poisons and qi-regulating drugs. The commonly used doses of Salvia miltiorrhiza, Poria cocos, Bupleurum chinense, Glycyrrhiza uralensis, Atractylodes macrocephala, Alismatis Rhizoma, and Curcumae Radix in the top 10 medicines ranked in the frequency of medication accorded with the prescribed doses in the Pharmacopoeia of the People's Republic of China (2015 edition), while Crataegi Fructus, Artemisiae Scopariae Herba, and Puerariae Lobatae Radix exceeded the prescribed doses. In the frequency analysis of drug pairs, the combination of S.miltiorrhiza and B. chinense was the most widely used. According to the association rules of drug combination, the correlation between Curcumae Radix and S. miltiorrhiza was the strongest, that was, the probability of S. miltiorrhiza appearing with the emergence of Curcumae Radix was 88%. From the network display chart, it was indicated that S. miltiorrhiza and P. cocos were the main herbs for treatment. Through unsupervised entropy hierarchical clustering algorithm, 14 core combinations for new clustering were extracted, and seven new prescriptions can be obtained by further clustering. Conclusion: The basic principles of TCM treatment of ALD include promoting blood circulation and removing blood stasis, removing dampness, tonifying, detoxifying alcohol, and promoting qi, and with "protecting spleen and stomach function" as its purpose, which accords with the theoretical basis of traditional Chinese medicine in treating alcoholic liver disease. Core combinations and new prescriptions provide references for clinical drug use and new drug research and development, but new prescriptions must be further evaluated with the combination of traditional Chinese medicine theory and clinical practice.
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To study the quality marker (Q-marker) of Atractylodes macrocephala based on the concept, standard, and research model of Q-marker. The chemical constituents of A. macrocephala were identified by UPLC-Q-TOF-MS/MS. The source and specificity of chemical constituents were confirmed by analyzing biosynthetic pathway and component specificity. The major effective components were clarified through efficacy, drug property, and correlation analysis of chemical constituents. The possible Q-markers of A. macrocephala are estimated based on the results of the study.
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Atractylodes macrocephala, belonging to Compositae family, whose dry rhizome as its medicinal parts, is a famous traditional Chinese medicine, with the function of tonifying spleen, aesthetic improvements and so on. Its main medicinal ingredient is volatile oil, which has obviously effect of antitumor. Recently, the market demand of this herb gradually increased, while its quality has not been guaranteed. This paper concluded the process of biosynthesis and transformation of sesquiterpenoids, an important secondary metabolite in violate oil; And firstly reviewed the factors affecting the accumulation of volatile oil in A. macrocephala in the aspects of inheritance and ecology. It is expected to provide a reference for the further study of the biosynthesis and biotransformation of sesquiterpenoids in A. macrocephala and help to improve the yield and quality of the volatile oil.
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OBJECTIVE: To establish a quantitative analysis of multi-components by single marker(QAMS) for the determination of three atractylenolides in Atractylodes macrocephala Koidz. METHODS: UPLC method was applied to determine atractylenolide Ⅰ, Ⅱ and Ⅲ by external standard method (ESM) first. With this established UPLC method, atractylenolide Ⅲ was used as the internal standard (IS) to determine the correction factors (RCFs) of the two other atractylenolides, and their contents in all the samples were calculated by their RCFs at the same time. By comparing the contents determined by the ESM and QAMS methods, the feasibility and accuracy of QAMS method were verified. RESULTS: Within a certain range(atractylenolideⅠ: 3.7 - 59.3 μg•mL-1, atractylenolide Ⅱ: 3.9 - 63.5 μg•mL-1, atractylenolide Ⅲ: 7.3 - 116.1 μg•mL-1), the RCFs of atractylenolide Ⅰ, Ⅱ to atractylenolide Ⅲ were 0.633 and 1.895, respectively, which had good repeatability in different experimental conditions. There was no significant difference between the QAMS method and ESM method(P > 0.05). CONCLUSION: The QAMS method is feasible and accurate for the determination of the three atractylenolides and is beneficial to the quality control of Atractylodes macrocephala Koidz..
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Objective: To establish an ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous determination of 23 plant growth regulator (PGR) residues in Chinese materia medica (CMM). Methods: Samples were extracted with acetontrile, and then determined by UPLC-MS/MS directly without further clean up. The matrix-matched external standard method was used for quantitative analysis. Results: The calibration curves showed good linearity in each range with correlation coefficients greater than 0.99. The limits of detection (LODs) were 0.01-20.80 ng/mL for the 23 PGRs spiked in Codonopsis pilosula and Angelica dahurica. The recoveries of the 23 PGRs spiked in C. pilosula and A. dahurica at three levels of 0.01, 0.04, and 0.1 mg/kg were in the range of 71.0%-101.4%, the relative standard deviation (RSDs) were 0.8%-15.2%. Commonly used CMM including eight species (63 batches) was analyzed by this method. Conclusion: The method proved to be simple, rapid, sensitive, accurate, and suitable for the simultaneous determination of 23 PGR residues in CMM
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Atractylodes macrocephala Koidz. is a commonly used Chinese materia medica in clinical treatment, and its chemical constituents mainly include volatile oil, lactones, glycosides, atractylodes polysaccharide, and amino acids. In addition to the pharmacological effects of anti-aging, anti-inflammatory, anti-tumor, regulating gastrointestinal function, and liver protection, A. macrocephala also has the function of treating and improving many kinds of diseases in nervous-mental system. In this paper, the role of A. macrocephala in nervous-mental system and its research mechanisms were summarized by literature analysis, in order to provide a reference for further explaining the mechanism and clinical medication of A. macrocephala in nervous-mental system.
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Rhizome atractylodes macrocephalae is the dry roots of Atractylodes macrocephala Koidz. It is one of commonly used Chinese medicine. In the ″Shennong's Herbal″, it was listed as the top grade. It mainly contains volatile oil, atractylenolides, atractylodes polysaccharides, glycosides and amino acids.And it has the medical functions of good for spleen and intestine, diuretic and dehumidifi?cation, hidroschesis, miscarriage prevention and soon.In order to provide references for further devel?opment and utilization, this paper systematic arranged the Chinese medicine atractylodes chemical composition, pharmacological effects, processing technology and effect of processing technology on chemical composition and pharmacological action.
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Objective:To develop an HPLC determination method for ferulic acid and caffeic acid in Atractylodes macrocephala Koidz fromdifferentorigins.Methods:ThecolumnwasAgilent-C18(150mm×4.6mm,5μm),themobilephasewas0.2% acetonitrile-aceto-nitrile(70 ∶30),the UV detection was performed at 320 nm,the flow rate was 1. 0 ml·min-1, and the drift tube temperature was set at 35℃. Results:The calibration curve was linear within the range of 0. 040-0. 500μg for ferulic acid(r=0. 999 8)and 0. 060-0. 750 μg for caffeic acid(r=0.999 8). The average recovery of ferulic acid and caffeic acid was 98.10%(RSD =1.08%)and 98.66%(RSD =0. 61%)(n=6), respectively. Conclusion: The method is simple and accurate. It is suitable for the determination of ferulic acid and caffeic acid in Atractylodes macrocephala Koidz.
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Objective: The aim of present study was to identify Atractylodes lancea and its closely related species (Atractylodes chinesis and Atractylodes macrocephala) using the ITS2 barcode. Methods: The total genomic DNAs were extracted from twenty-nine samples of A. lancea and its closely related species from different habitats. The ITS2 sequences of these samples were amplified and bidirectional sequenced by PCR. The obtained sequences were submitted to the GenBank and the ITS2 sequences of 45 samples belonging to ten species were downloaded from the GenBank. Total 73 ITS2 sequences were aligned and the genetic distances were analyzed using the MEGA 5.1. Identification analyses were performed using the BLAST1 and the nearest distance methods, and were presented intuitively by constructing Neighbor-joining (NJ) tree. Results: The lengths of all ITS2 sequences of A. lancea were 229 bp presented as one haplotype pattern. There was significant divergence between the interspecific and intraspecific genetic distances of the ITS2 sequences. The NJ tree showed that A. lancea could differed obviously from its closely related species, which showed high monophyly. The secondary structures of ITS2 in the helical regions displayed clear differences in stem loop number, size, position, and screw angle among the medicinal plants of Compositae. Conclusion: As a DNA barcode, ITS2 sequences can stably and accurately distinguish A. lancea from its closely related species and also provide a new technique to ensure the clinical safety in utilization of Chinese materia medica.
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Objective: To study the morphological characteristics and genetic diversity of Atractylodes macrocephala from different varieties, in order to protect and select the germplasm resources and optimize the cultivated technology. Methods: The inter-simple sequence repeat (ISSR), morphological characteristics, and physiology were used to analyze the genetic diversity among 11 varieties of A. macrocephala in Dapan mountain, a national natural reserve in Zhejiang Province and the cluster analysis was performed by UPCMA method. Results: The varieties differed greatly in leaf-shape and leaf-color, stem and rhizomatous configuration, production and disease resistance, and no correlation was observed in these differences. However, in all samples, the photosynthesis rate was closely correlated to the biomass production, despite the great differences existing among different varieties. Simultaneously, fifteen screened ISSR primers were served as ISSR-PCR markers, and 129 sites were amplified, which included 107 polymorphic sites (82.95% of the total bands). At molecular level, the genetic differentiations occured among 11 varieties, some differentiations were quite distinctive according to cluster analysis. Conclusion: The germplasm resources of A. macrocephala. are abundant and the genetic diversity basis exists in all kinds of A. macrocephala in Dapan mountain.
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OBJECTIVE:To improve the method of Chinese Pharmacopoeia on the extraction of volatile oil from Atractylodes macrocephala Koidz..METHODS:Volatile oil was extracted from Atractylodes macrocephala Koidz.and determined by the extraction method and determination method stated in Chinese Pharmacopoeia and by the new improved method respectively.And the components of volatile oil obtained in these two methods were respectively separated and identified by GC-MS.RESULTS:The extraction rates of volatile oil by the method of Chinese Pharmacopoeia vs.the new method were(1.28? 0.03) %(n=6) vs.(1.39? 0.03) %(n=6).As compared with the new method,seven components were missed by the method of Chinese Pharmacopoeia.CONCLUSION:The new extraction method of volatile oil can increase both the extraction rate of volatile oil and the kinds of the components in volatile oil,and it is obviously better than the method stated in Chinese Pharmacopoeia.
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Objective To study the function of atractylodes macrocephala koidz volatile oil for transplanted tumor in animals. Methods The animal model of transplanted tumor-ECA was used. The anti-tumor function of atractylodes macrocephala koidz volatile oil was observed. Results Atractylodes macrocephala koidz volatile oil can suppress ECA when given at 100 mg/kg and 50 mg/kg doses into abdominal cavity. When a larger dose at 150 mg/kg was given, the life-span of mice can extended to 197 %. Conclusion Atractylodes macrocephala koidz volatile oil can suppress tumor effectively.
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Objective To research the effect of atractylodes macrocephala koidz on immune function of tumor-bearing mice.Methods With S_ 180 bearing C_ 57 BL/6 mice as model.Effects of atractylodes macrocephala koidz on tumor weight and immunological function of spleen were cells systematically investigated.Results Atractylodes macrocephala koidz could obviously restore immune function of tumor-bearing mice reduced by chemiotherapy,promote proliferation of activated T cell and interleukin-2(IL-2) levels of tumor-bearing mice.Conclusion Atractylodes macrocephala koidz can reverse the immunological inhibition caused by tumor antigen and chemiotherapy.
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Study on the symptoms of the diseases on infected Atractylodes macrocephala Koidz. in Ha Noi, Hung Yen and Lao Cai province. Results: Atractylodes macrocephala Koidz. was infected with harmful disease such as root ulcer caused by Rhizoctonia solani, tuber stink caused by Fusarium solani, brown spot and leaf burn due to Pestalozia sp. The fungus Sclerotium rolfssi isolated from A.macrocephala grew best on PGA medium with the most sclerotia produced at pH=6, was similar with those isolated from tomatoes. Trichoderma viride, a biological product can inhibit the growth of S.rolfsii with 85.49% efficacy and can be applied to control this fungus in the field. Chemical material Ridomil 0.05% can annihilate Sclerotium rolfsii fungus with 100% efficacy after 3 cultured days
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Plants, Medicinal , EpidemiologyABSTRACT
Object To observe the effects of Astragali Injection (Hi) and extracts from Atractylodes macrocephala Koidz. (B1, B4, B9, B13) on the migration of rat small intestinal crypt like cells line (IEC 6). Methods IEC 6 cells were inoculated in 6 well microplates and injured in 72 h, and added with Hi, B1, B4, B9, B13. Then D PBS to negative control in the same amount and EGF to positived control, the cell migration was observed in 24, 48 and 72 h after treatment. Results B4?B9?B13 and EGF significantly promoted the migration of wounded IEC 6 cells (P
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Objective The transgenic Atractylodes macrocephala resistance to Rhizoctonia solani was obtained by gene engineering. Methods On the base of the efficient regeneration system of Baizhu via shoot organogenesis, the rice chitinase gene (RCH10) and the alfalfa ?-1, 3-glucanase gene (AGLU) were tandem-inserted into the transformation vector pB101, which was transformed into A. macrocephala with gene gun. Transformants were confirmed by PCR, GUS assay, and disease resistant. Results Twenty-five independent transformants possessed desired genes were observed by PCR detection, and among them five transformants exhibiting resistance to Rhizoctonia solani. Conclusion The disease resistant variety has been obtained by transformation, which provides a shorten avenue for the direct introduction of novel traits into A. macrocephala through genetic engineering without the need for numerous back-crossing in breeding programs that slow down cultivar improvement, meanwhile it improves disease resistant gene pool.
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Objective To investigate the extraction technique for atractylenolide Ⅰ in Atractylodes macrocephala by supercritical CO_2 fluid extraction and develop a method used for determining the content of atractylenolide Ⅰ in the extract by HPLC. Methods The effects of seven facters, such as the extracting pressure, resolving pressure etc, to the extraction rate of atractylenolide Ⅰ in A. macrocephala by supercritical CO_2 extraction were investigated. RP-HPLC was used to determine the content of atractylenolide Ⅰ in extraction of A. macrocephala. The separation was performed on Hypersil ODS2 column with methanol-water (70∶30) as mobile phase. The flow rate was 1.0 mL/min and the wavelength of UV detector was 220 nm. Results The optimal extracting conditions: taking 10% alcohol as entraiter, the particle size of medicinal substances was 60 screen meshes, extracting pressure 25 MPa, resolving pressure 5 MPa, extracting temperature 40 ℃, resolving temperature 30 ℃, and the extracting time 4 h. Conclusion Supercritical extraction is time-shorter and efficient in extracting atractylenolide Ⅰ from A. macrocephala. It is suitable to both trial and industrialized production. The method established to determine the content of atractylenolide Ⅰ of A. macrocephala by supercritical extraction is simple, sensitive, and reliable.