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Objective: To establish quantitative analysis of multi-components with single marker (QAMS) for determination of 10 phloroglucinol contents in effective fraction of Dryopteris fragrans. Methods: The relative correction factors of nine phloroglucinol (aspidin PB, aspidin AB, flavaspidic acid BB, saroaspidin A, flavaspidic acid PB, disflavapidic acid PB, flavaspidic acid AB, compound VI, and aspidinol B) were determined by HPLC method with the aspidinn BB as the internal standard, which were to calculate the content of each. At the same time, external standard method (ESM) was used to determine the contents of 10 components in effective fraction, and the differences between the two methods were compared to verify the feasibility and accuracy of QAMS method. Results: The relative correction factor (RCF) was good. There was no significant difference between the quantitative results with the two methods in the 12 batches of 10 phloroglucinols. Conclusion: The established QAMS method can be used for quantitative analysis of D. fragrans with aspidinn BB as the internal standard in the absence of reference substances, and provided a reference for the multi-index quality evaluation in effective fraction of D. fragrans.
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Objective: To study the chemical constituents and antifungal activitiy of phloroglucinol derivatives from Dryopteris fragrans. Methods: The phloroglucinol derivatives were isolated by silica gel column chromatography, Sephadex LH-20 column chromatography, and pre-HPLC. The chemical structures were elucidated on the basis of physicochemical and spectral properties. The antifungal activities against Trichophyton rubrum and Microsporum gypseum were tested by CLSI M38-A2 broth microdilution methods. Results: Two phloroglucinol derivatives were isolated and identified as disflavaspidic acid PB (1) and flavaspidic acid PB (2). The minimum inhibitory concentration (MIC) of compounds 1 and 2 were 13.33 and 1.875 μg/mL against T. rubrum respectively, 10 and 1.325 μg/mL against M. gypseum. Conclusion: Compound 1 is a new compound named disflavaspidic acid PB. Compound 2 is isolated from D. fragrans for the first time. Compounds 1 and 2 are a pair of isomers with promising antifungal activity.
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Objective: Expression and analysis of 4CL gene of ferns, which lays the theoretical foundation for the accumulation and regulation of secondary metabolites of ferns. Methods: Cloned 4CL gene (Df4CL) sequence from Dryopteris fragrans was analyzed by CHIPS, CUSP, and Codon W programs online, and then compared with Escherichia coli, yeast, Arabidopsis thaliana, and Nicotiana tabacum L. It is important to identify the codon usage of Df4CL gene and select appropriate expression systems. Results: Df4CL gene had not the bias toward the synonymous codons with ATGC at the third codon position, but had the preference consistency with Physcomitrella patens codon usage. The differences in codon usage frequency between Df4CL gene and A. thaliana were less than those in N. tobacum, E. coli, and yeast. Based on 4CL gene codon usage bias of cluster analysis, it was shown that D. fragrans, P. patens, and A. thaliana were classed into one group. Conclusion: The A. thaliana expression system may be more suitable for the exogenous expression of Df4CL gene of D. fragrans.
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Objective: To study the chemical constituents from the whole plants of Dryopteris fragrans. Methods: The chemical constituents were isolated by various column chromatographic methods. The structures of the compounds were elucidated on the basis of physiochemical properties and spectral analyses. Results: Three compounds named 2-isopropyl-5, 7-dihydroxy-1'-O-β-D-glucopyranosyl chromone (1), (6R, 9R)-3-oxo-α-ionol-9-O-β-D-glucopyranoside (2), and taxicatin (3) were isolated and identified. Conclusion: Compound 1 is a new compound named frachromone B, compounds 2 and 3 are reported for the first time from family Dryopteridaceae.
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Objective: To study the chemical constituents from the whole plants of Dryopteris fragrans. Methods: The chemical constituents were isolated by various column chromatographic methods. The structures of the compounds were elucidated on the basis of physiochemical properties and spectral analysis. Results: Four compounds named 2-ethyl-5, 7-dihydroxy-1'-O-β-Dglucopyranosyl chromone (1), vitexin (2), isoquercitrin (3), and divarin-3-O-β-glucopyranoside (3-hydroxy-5-propylphenyl-O-β-Dglucopyranoside, 4) were isolated and identified. Conclusion: Compound 1 is a new compound named as frachromone A, compound 2 is obtained from this plant for the first time, and compound 4 is reported for the first time from the plants of family Dryopteridaceae.
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Objective To determine the genetic diversity of germplasm resource in Dryopteris fragrans by amplified fragment length polymorphism(AFLP) technique.Methods Forty-six samples from six populations of D.fragrans were analyzed by AFLP DNA markers,and the genetic diversity was evaluated by NTSYS-PC 2.10 and PopGen32.Results The percentage of polymorphic bands(PPB) reached to 76.67%,observed number of alleles(Na) was 1.766 7, effective number of alleles(Ne) was 1.504 6,Nei′s gene diversity index(H) was 0.292 1,Shannon information index(I) was 0.431 6,and genetic differentiation index(Gst) was 0.412 5.Conclusion Genetic diversity is high,gene diversity in populations(Hs) is 0.170 5,and gene diversity among populations(Dst) is 0.119 8.Because of environmental specificity,its resource on the spot should be protected.
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Objective To study the chemical components from Dryopteris fragrans and their biological activities.Methods By means of HPLC and other column chromatography,the chemical components were isolated from D.fragrans and their chemical structures were identified by spectral and chemical tests in accordance with their physicochemical characteristics.Furthermore,the cytotoxicity of the chemical components for Bel-7402,HCT-8,and A-549 cells was measured by MTT method.Results Four compounds,geniposide(Ⅰ),3S,5R,6R,7E,9S-megastigman-7-ene-3,5,6,9-tetrol-3-O-?-D-glucopyranoside(Ⅱ),(6S,9R)-3-oxo-?-ionol-9-O-?-D-glucopyranoside(Ⅲ),and fragranoside a(Ⅳ) were iso-lated and identified.Conclusion Compound Ⅳ is a new compound.And all the other three compounds Ⅰ-Ⅲ are found in plants of Dryopteris Adanson for the first time.The results of pharmacological experiments indicate that compounds Ⅰ and Ⅲ show strong cytotoxicity.