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Objective To explore a new identification method for medicinal materials of Dysosma, and analyze the second internal transcribed spacer (ITS2) barcode sequences of Diphylleia sinensis and Dysosma versipellis, Sinopodophyllum hexandrum, Dysosma difformis and Dysosma pleiantha in five kinds of podophyllum. Methods The ITS2 of ribosomal DNA of medicinal materials of podophyllum was amplified and sequenced by bi-directional sequencing of PCR products. Sequence assembly and consensus sequence generation were performed by using CodonCode Aligner. Phylogenetic study was performed using software MEGA 5.1 in accordance with Kimura-2-parameter (K2P) model. Genetic distances were calculated and analyzed and the phylogenetic tree was constructed by using the neighbor-joining (NJ) method. Results There were significant differences among five kinds of Dysosma. Their maximum intraspecific genetic distance (K2P distance) was far lower than their minimum interspecific genetic distance with the other species. In the cluster dendrogram, all species showed monophyletic. Conclusion ITS2 sequence as DNA barcoding technique can be used to identify Chinese herbal materials of Dysosma.
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OBJECTIVE: To examine the diversity and antimicrobial activities of the endophytic fungi of Dysosma versipellis (Hance) M. Cheng. METHODS: The identification of the fungi was carried out by rDNA-ITS sequence analysis. The antimicrobial activities of the isolated endophytic fungi were determined by agar block test. RESULTS: Nineteen strains of various endophytes were obtained from the roots, stems and leaves of D. versipellis, which were molecularly identified to belong to eight catalogues and thirteen genuses. The DV04 had remarkable antimicrobial activities to Staphylococcus aureus, Escherichia coli, and Candida albicans. Morphological and molecular identification demonstrated that DV04 represented Leptodontidium sp.. CONCLUSION: The endophytic fungi in Dysosma versipellis are diverse and rich. Some strains such as DV04 deserve further research as an alternative antimicrobial resource.
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Objective: To study the chemical constitutents from the roots and stems of Dysosma versipellis. Methods: Silica gel column, Sephadex LH-20, and reverse phase HPLC chromatography were used to isolate the chemical constituents, and the structures of the isolated compounds were elucidated by spectral analysis and physicochemical properties. Results: Fourteen compounds were isolated from EtOAc and n-butanol fraction of the 95% ethanol extract in the roots and stems of D. versipellis, including 10 lignans, i.e. 4′,5′- didemethylpodophyllotoxin (1), podophyllotoxin (2), 4′- demethylpodophyllotoxin (3), diphyllin (4), podophyllo-toxone (5), picropodophyll-otoxin-4-O-β-D-glucoside (6), 4′- demethylpodophyllotoxin-4-O-glucoside (7), diphyllin-4-O-β-D-glucoside (8), podophyllotoxin-4-O-β-D-glucoside (9), and dysosmarol (10); three flavonoids, i.e. kaempferol (11), quercetin (12), and kaempferol-3-O-β-D- glucoside (13); and one sterol, i.e. β-sitosterol (14). Conclusion: Compound 1 is a new natural product, and compounds 7 and 8 are isolated from the plants of Dysosma Woodson for the first time.