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In order to develop genomic-SSR markers for species of Saxifraga genus, a mixed plant genomic DNA sample was sequenced based on high-throughput Illumina MiSeq platform. According to genomic sequencing data, SSR loci were identified with MISA software, and then primers were designed with Primer 3 software. A total of 120 pairs of primers were randomly synthesized and amplified in genomic DNA of a few plant samples. Those primers who have yielded polymorphic bands and were considered easy to amplify were identified. After that, transferability of these primers was evaluated, and phylogenetic relationship of 25 species of Saxifraga genus was analyzed with UPGMA (unweighted pair group method analysis). In our results, 587 256 sequences containing SSRs were identified from a total of 1 881 979 combined read pairs obtained in genomic sequencing. Primers were designated to amplify SSRs containing two to six nucleotide repeat units, screened in a small portion of species. Finally, 17 pairs of primers which have produced abundant of polymorphic bands with little problem were amplified in 25 species of Saxifraga genus. A total of 2 687 polymorphic bands were obtained, the average polymorphic rate was 158 bands per pairs of primers. The transferability rate was ranging from 88.0% to 100% across 25 species of Saxifraga. In phylogenetic analysis, the clustering of 25 species based on 17 pairs of SSR primers was different from morphological classification. Our analysis has provided molecular data for genetic relationship of Saxifraga genus, and the transferable and polymorphic SSRs have provided information for genetic diversity research.
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Objective To establish a method to research the fingerprint and determine the ingredients of Saxifraga stolonifera by HPLC, which aimed at providing reference for the quality control of Saxifraga stolonifera. Methods Agilent Eclipse XDB-C18 (150 mm × 4.6 mm, 5 μm) column was used as the stationary phase, and the mobile phase consisted of acetonitrile (A)-0.1% phosphoric acid with gradient elution at the flow rate of 1.0 mL/min. The detection wavelength was 254 nm, and the column temperature was maintained at 35 ℃. The result would be analyzed by SOP of Similarity evaluation system for chromatographic fingerprint of TCM. Results The results obtained by the method of fingerprint analysis are good. In the fingerprints, 11 peaks were selected as the common peaks to evaluate the similarities among 21 batches of S. stolonifera collected from different regions, and five contents of them were indentified as protocatechuic acid, gallic acid, bergenin, quercetin-5-O-β-D-glucoside, and quercitrin. The similarities between standard herb and each determined herb showed a lot of differences from others. The determination method showed that there weregood linear relationships of five figured contents in the range of 0.052 8-0.844 8, 0.020 96-0.335 36, 0.241 6-3.865 6, 0.130 8-2.092 8, and 0.023 68-0.378 88 μg. Moreover, the recoveries rates of five figured contents are 96.64%, 100.72%, 96.62%, 103.71%, 96.75%,and all RSDs were lower than 2%. The contents of 5 components in the gathered herbs were determined by external standard method in the range of 0.07-0.40, 0.19-4.36, 1.42-5.98, 0.42-6.86, and 0.11-1.51 mg/g. Conclusion The method established in this study is simple, reliable and durable, which can provide a scientific basis for the quality control of S. stolonifera.
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Introduction:Saxifraga spinulosa Adams (S.spinulosa) belongs to the Saxifragaceae family and it is used for treating conjunctivitis, gynecopathy, larynx, pleural cavity in traditional Tibetan and Russian medicine. Study on antioxidant activity on this plant have also been carried out previously. However, no work has been reported on its anti-acetylcholinesterasic and anti-quorum sensing activity.Goal: The purpose of this research was to perform phytochemical, anti-acetylcholinesterase and anti-quorum sensing activities on the extracts of aerial parts of Saxifraga spinulosa Adams.Methods and materials: Anti-acetylcholinesterase activity was evaluated by colorimetric method. Extracts of aerial parts of S.spinulosa were tested for anti-quorum sensing activity by the Chromobacterium violaceum assay using the standard disc-diffusion method. Extracts were chromatographed with thin layer chromatography method. Phytochemical screening of crude extracts S. spinulosa Adams was carried out to detect saponins (Kokate, 1999), ferric chloride and lead acetate test was for tannin and phenolic contents and the detection of gum (Whistler, 1993) and Fehling test of carbohydrates respectively. Results: Plant extraction and fractionation was made earlier. Acetylcholinesterase activity screening was performed on its 4 extracts, prepared from aerial parts of Saxifraga spinulosa Adams. Non-polar chloroform fraction showed more active with 45.7% compared to the aceton extract (36%), water residue (35,5%) and n-butanol fraction (37,5%). Anti-quorum sensing activity was observed with 7 extracts of S. spinulosa aerial part. From them, fractions A (21 mm), B (20 mm), C (20 mm) showed significant activity and others were moderate. No antibacterial activity (for Chromobacterium violaceum) was observed. Thin layer chromatography (TLC) fingerprinting indicated the presence of the phenolic components and flavonoids. Phytochemical screening revealed that phenolic contents, tannins, saponins, carbohydrates, gums were present in the whole plant. Conclusion: This study shows that, S. spinulosa has anti-acetylcholinesterase and anti-quorum sensing activity and we determined that this plant mainly contains polyphenolics compounds. Key words: anti-quorum sensing, anti-acethylcholinesterase, Saxifraga spinulosa Adams
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Introduction: Oxidative stress occurs when the production of harmful molecules called free radicals are beyond the protective capability of the antioxidant defenses. Free radicals are known to cause damage to lipids, proteins, enzymes and nucleic acids, they are leading to cell or tissue injury. Therefore, there is an increasing interest in searching antioxidants from natural origin to prevent human body from oxidative stress produced by free radicals. In this research, 52 extractions of Mongolian medicinal plants were tested for their DPPH radical scavenging activity. Out of them, the aerial parts of Saxifraga spinulosa showed significant activity. Phytochemical and biological studies on this plant haven’t been determined previously. Methods: Antioxidant activity was evaluated by DPPH (2,2- diphenyl-1-picrylhydrazyl) radical scavenging assay. The antioxidant activity of these extracts was compared with standard solution of rutin. The total phenolic contents, flavonoid, and flavonols were determined by Folin-Ciocalteu, aluminum chloride, and Yermakov methods, respectively. Results: The acetone extract of the aerial parts of Saxifraga spinulosa exhibited excellent scavenging activity with IC50 (μg/mL) 14.98±0.16 against DPPH radical. Moreover the n-butanol fraction showed greater DPPH scavenging activity with IC50 (μg/mL) 28.69±0.09 compared to the non-polar chloroform fraction. So far the active n-butanol fraction passed through an octadecylsilicagel (3 x 12.5 см) and eluted with H20 (fraction A, 1.9 g), MeOH 20% (fraction B, 0.45 g), MeOH 40% (fraction C, 1.56 g), MeOH 80% (fraction D, 0.268 g), and MeOH (fraction E, 0.0012 g). From them, fractions A, B, C, and D showed significant activity, while fraction E had moderate activity. The total phenolic contents of acetone extract (121.42±14.33), n-butanol (113.42±16.29), and water residue (68.09±20.44), were expressed as Gallic acid equivalents. The flavonoid contents were found to be acetone extract (37.27±3.05), n-butanol (50.6±6.43), and water residue (5.93±2.0) in methanol using rutin equivalents. Conclusions: Antioxidant activity of Saxifraga spinulosa extracts were shown to be related to the phenolic components. Detailed investigation is needed to isolate antioxidant active compounds from this plant and determine their chemical structures as well as to make it accessible to modern clinical practice. Key words: Saxifraga spinulosa, DPPH radical scavenging activity, total phenolic content
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Objective: To optimize the extracting technology for active fractions from Saxifraga stolonifera with anti-prostate cancer activity. Methods: Using the extraction rates of bergenin, quercetin-5-O-β-D-glucopyranoside, quercetin-3-O-L-rhamnopyranoside, quercetin, and flavonoids as indexes, the single factor experiment combined with Box-Behnken design was used to optimize the extracting technology. Results: The optimal extraction conditions were as follows: ethanol volume fraction 60.04%, liquid-solid ratio of 1:13.79 for 89.98 min, and at the temperature of 80°C. Under these conditions, the extraction rates of bergenin, quercetin-5-O-β-D-glucopyranoside, quercetin-3-O-L-rhamnopyranoside, quercetin, and flavonoids were 97.15%, 98.13%, 98.37%, 97.82%, and 96.33%, respectively. Conclusion: The response surface analysis could optimize the extracting technology of active fraction from S. stolonifera with the anti-prostate cancer activity, which provides the basis for the comprehensive utilization and industrialization of S. stolonifera.