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Objective: To study the genetic diversity and genetic structure of Paris genus in Shaanxi Province. Methods: Start Codon Targeted Polymorphism (SCoT) molecular markers were performed on 48 samples from six taxas of Paris genus in Shaanxi. Genetic distance was calculated by POPGENE 3.2 software, and cluster analysis was generated by NTSYS 2.10 software based on unweighted mean distance method (UPGMA) method. Results: A total of 152 bands were produced by 12 primers, among which 135 bands were polymorphic bands, and the percentage of polymorphic bands was 88.82%. The average value of Nei’s genetic diversity index (H) was 0.267 4, Shannon’s information index (I) was 0.404 1, genetic differentiation coefficient (Gst) was 0.517 9, and the gene flow (Nm) was 0.465 4. Six taxas were ranked by genetic diversity: Paris polyphylla > Paris polyphylla var. latifolia > Paris fargesii var. petiolata > Paris polyphylla var. stenophylla > Paris polyphylla var. yunnanensis > Paris verticillata. Cluster analysis distinguished others from Paris verticillata; When the genetic distance was certain, six taxas of the genus were completely separated. Conclusion: SCoT molecular markers can obtain polymorphic and clear band amplification map, which indicates that the technology can be used for molecular identification and genetic relationship of major taxas in Paris genus in Shaanxi Province, providing a scientific basis for screening alternative resource types close to the Pharmacopoeia collection and guiding the rational use of local species.
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Paridis Rhizoma is a traditional Chinese medicine, mainly distributed in Yunnan, Sichuan, Guizhou and other areas of China. Because of its analgesic, hemostatic, anti-tumor, anti-oxidant, immune regulation and other functions, it has been widely used in medicine and health products. Therefore, the quality evaluation of Paridis Rhizoma has become an urgent restriction on the development and utilization, which becomes one of the problems to be solved. In this study, the chemical constituents and pharmacological activities of Paris plants were systematically sorted out, and the quality markers of Paris plants were predicted and analyzed based on the specificity of chemical constituents, the correlation between chemical constituents and pharmacodynamics, the traditional medicinal properties and the measurability of chemical constituents, which provides scientific basis for the quality evaluation of the genus Paris.
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Objective: To explore genetic diversity and phylogenic relationship among different types of Paris species and provide an effective method for the rapid identification of germplasm resources. Methods: CDDP marker was used to evaluate the genetic diversity and phylogenic relationship among 13 types of Paris species. And the coding of Paris genus plants was carried out based on method of CODE128 barcode. Results: Our results indicated that 73 polymorphic bands were amplified by 11 primers among 80 bands, and the ratio of polymorphic band was 91.25%. The observed number of alleles (Na), effective number of alleles (Ne), Nei’s gene diversity (H), and Shannon’s information index (I) was 1.912 5, 1.589 6, 0.342 3, and 0.507 0, respectively. The results of UPGMA analysis showed that there were great differences among 13 types of Paris species on genetic diversity. Based on CDDP markers, 13 barcode molecular identity cards were constructed for Paris species based on method of CODE128 barcode. Conclusion: There was abundant polymorphism among Paris species, and CDDP markers were effective to analyze the genetic diversity of Paris species, and the established barcode molecular identity for Paris species was sensitive and fast, which can be used for scientific research and industrial production of Paris genus.
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Objective To investigate the medicinal plant group and resource of Sect. Euthya in Paris L. in Shaanxi Province. Methods Through literature analysis and interview survey, combined with line transect method, the medicinal plant group, the natural distribution and the status of medicinal plant group and resources about Sect. Euthya in the specific county territory in Shaanxi Province were investigated.Results According to related records about the medicinal plants of Sect. Euthya in Paris L. included P. polyphylla, var. stenophylla, var. apetala, and Paris fargesii var. petiolatain in Shaanxi Province. Based on field investigation, it was found that, the medicinal plants of Sect.Euthya in Paris L.also included five variations of P.polyphylla var.latifolia,var.apperdiculata,var. thibetica, var. chinensis, and var. yunnanensis, which were new distribution records. No var. apetala was found under field investigation. Most of the rhizomes of the Sect. Euthya plants were used as Chinese materia medica Paridis Rhizoma, with wide distribution and good growth condition. The natural resources of these plants are endangered. Conclusion In this study, two species and six variations in the Sect. Euthya are identified as new distribution records. Consequently, the medicinal plant distribution record of Paris L. in Shaanxi Province is complete. The natural resources are investigated, which have laid the foundation for further research, development and protection.
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The study was established an HPLC method for simultaneous determination of 8 steroidal saponins (polyphyllins Ⅶ, H, Ⅵ, Ⅱ, Ⅰ, and Ⅴ, dioscin, and gracillin) in Paridis Rhizoma, and made an evaluation by determining steroidal saponins in 15 kinds of genus Paris. The analysis was performed on a Waters Acquity H-ClassTM UPLC ultrafine liquid chromatography system coupled with a PDA detector. The chromatographic separation was achieved through a CAPCELL PAK ADME (4.6 mm× 250 mm, 5 μm) column and the optimal mobile phase consisted of acetonitrile and water. The column was maintained at 21 ℃, and the flow rate was 0.8 mL•min ⁻¹. The UV detection wavelength was 203 nm. The results showed that ① the detected components can be well separated and all with good correlation coefficients. The standard calibration curves were linearly good (R2>0.999 9). The linearity was obtained over 0.041 70-3.812 00 μg. The average recoveries ranged from 95.91% to 103.8%. ② there are significant differences in the content of steroidal saponins from different species. The steroidal saponins are low content or almost none in P. mairei, P. polyphylla var. stenophylla, and P. delavayi have low content or almost did not contain, so these species are not suitable for medicinal use. The contents of steroidal saponins in P. polyphylla var. chinensis are varied from different places. There were high content of steroidal saponins in P. polyphylla var. yunnanensis, P. forrestii, P. daliensis, and P. axialis, even up to 5.0%, which indicated that they had the potential pharmic value of development.
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Objective: DNA barcoding technology, a molecular identification method, is applied to Paris polyphylla collected from Wudang Mountain area in order to solve the differential problem caused by small morphology difference, make authentic origins canonical, and guarantee the quality and clinical curative effect of this medical material in our area. Methods: In this study, the second internal transcribed spacers (ITS2) of ribosomal DNA were amplified and sequenced. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner. To perform a phylogenetic study, the genetic distances were computed using molecular evolutionary genetics analysis 6.0 (MEGA 6.0) in accordance with the Kimura-2-parameter (K2P) model. The phylogenetic tree was constructed using the neighbor-joining (NJ) and Kimura-2-parameter methods. The ITS2 secondary structures were predicted using ITS2 database and website which was found by Keller. Results: Identified by BLAST, P. polyphylla collected in Wudang mountain area was proved to be precise. The results showed that the ITS2 sequence length was 232 bp. It was indicated that their K2P distance were 0-0.073 6 and the average was 0.027 2. Compared with the gene sequence of P. polyphylla, there were 16 variation loci in P. fargesii and only one variation locus in P. polyphylla var. chinensis. As seen from the NJ tree, P. polyphylla in this area was mainly divided into two categories, P. polyphylla and P. fargesii. Obviously, P. polyphylla var. chinensis was a variation of P. polyphylla. Meanwhile, it was not tenable that P. polyphylla var. stenophylla and P. polyphylla var. latifolia were the variations of P. polyphylla because they belonged to a branch. To compare the ITS2 secondary structure of P. polyphylla, we noticed the differences in the amount, size, and position of loop of helix zone. Conclusion: DNA barcoding technology opens up a new way for the identification of P. polyphylla. Regardless of what methods were used, such as similarity search, nearest distance, and NJ tree method, the analysis results have fully demonstrated that ITS2 sequences could be used to correctly identify the different varieties of P. polyphylla. It is identical in 232 base sequence of ITS2 to P. polyphylla var. stenophylla, P. polyphylla var. latifolia, and P. polyphylla. The results computed by Mega 6.0 also show that they belong to a branch. It is more appropriate to say P. polyphylla var. stenophylla and P. polyphylla var. latifolia are variants of P. polyphylla. Hence, as a DNA barcode sequence, ITS2 can be applied to intraspecific discrimination of P. polyphylla. Furthermore, the application of ITS2 in the identification of traditional Chinese medicine has an important prospect. And it can be used to correct identification of germplasm resources in GAP procedures.
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OBJECTIVE: To investigate the differences of the accumulation of the main secondary metabolites in the leaves of 11 kinds of Paris L. medicinal plants. METHODS: The contents of the main secondary metabolites, polyphyllin I, II, VI, and VII in the leaves of 11 kinds of Paris L. medicinal plants were determined by UPLC, and the UPLC fingerprints were established. The accumulation of the main secondary metabolites was evaluated by one-way ANOVA and chromatographic analysis. RESULTS: There was significant difference(P<0.01)in the contents of polyphyllin I, II, VI, and VII in the leaves of 11 kinds of Paris L. medicinal plants, and polyphyllin I, II, VI, VII were simutaneously detected only in var. yunnanensis-1, P. axialis, P. thibetica, P. forrestii and var. yunnanensis; there was significant difference in the UPLC chromatograms of 11 kinds of Paris L. medicinal plants, but the similarities among var. yunnanensis-1, P. axialis, P. thibetica, P. forrestii and var. yunnanensis all reached 0.902 with 16 common peaks, indicating smaller difference in their main secondary metabolites. CONCLUSION: There is significant difference in the abilities of the 11 kinds of Paris L.medicinal plants to accumulate polyphyllin I, II, VI, and VII, which may be the main reason that there are significant differences in the contents and classes of the major secondary metabolites of Paris L. roots.
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Objective: In order to provide a theory basis for rational development and utilization of medicinal plants from Paris L., this study compared nucleosides (cytidine, uridine, inosine, guanosine, thymidine, adenosine and 2'-desoxyadenossine) and bases (including uracil, guanine, and adenine) contents in the different plants of Paris L.which collected from different places. Methods: RP-HPLC was applied in this study, Venusil MP C18(2) chromatographic column (250 mm × 4.6 mm, 5 μm) was used, the mobile phase was a methanol and water gradient, the flow rate was1.0 mL/min, the temperature of column was 35℃ and the UV detector was set at 260 nm. Sixteen batches of samples in Paris L. were measured by above method, and cluster analysis was used to analyze the results. Results: Ten nucleosides and bases from the plants of Paris L. were measured, a good linear relationship was showed (r2 > 0.999 1), and the average recovery was 96.04%-104.34%, RSD ≤ 2.94%. The samples in Paris L. from different places were divided into three categories by cluster analysis. All of the ten nucleosides and bases were detected in the 16 batches of samples; However, the mass fraction and component ratio of the ten substances were significantly different from each variety. The average of total nucleosides and bases content was ranged from 735.390 1 to 1 612.807 3 μg/g, and the order from high to low was: P. bashanensis > P. polyphylla var. pseudothibetica > P. delavayi var. petiolata > P. axialis > P. polyphylla var. stenophylla > P. polyphylla var. chinensis > P. polyphylla var. yunnanensis > P. marmorata > P. polyphylla var. minor > P. mairei. The mass fraction and component ratio of the ten substances also had a significant difference between the samples collected from different regions which were the same variety. Conclusion: This method is simple, accurate and reliable and has good reproducibility. It is appropriate for measuring the contents of the ten nucleosides and bases. Simultaneously, this study provids a scientific basis for an objective and complete knowledge of the material basis for the efficacy, and for an enrichment and development of evaluation indicators system of in the plants Paris L.
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The genus Paris L. (Family Liliaceae) concerns 24 species all over the world, and 19 species are native to China especially in the southwest of China. Steroid saponins, flavonoid, triterpenes, and fatty acids are the major bioactive components in the plants of Paris L. Moden pharmacological researches demonstrate that the plants in this genus have many biological activities, such as antitumor, antibacterial, hemostatic, anthelmintic effect, etc. In this paper, the systematic classification, chemical constituents, and pharmacological effects of plants in Paris L. have been summarized. It may provide the reference for the further studies of this genus.