High-throughput sequencing-based analysis of the composition and diversity of the endophyte community in roots of Stellera chamaejasme.

Stellera chamaejasme (S. chamaejasme) is an important medicinal plant with heat-clearing, detoxifying, swelling and anti-inflammatory effects. At the same time, it is also one of the iconic plants of natural grassland degradation in northwest China, playing a key role in the invasion process. Plant endophytes live in healthy plant tissues and can synthesize substances needed for plant growth, induce disease resistance in host plants, and enhance plant resistance to environmental stress. Therefore, studying the root endophytes of S. chamaejasme is of great significance for mining beneficial microbial resources and biological prevention and control of S. chamaejasme. This study used Illumina MiSeq high-throughput sequencing technology to analyze the composition and diversity of endophytes in the roots of S. chamaejasme in different alpine grasslands (BGC, NMC and XGYZ) in Tibet. Research results show that the main phylum of endophytic fungi in the roots of S. chamaejasme in different regions is Ascomycota, and the main phyla of endophytic bacteria are Actinobacteria, Proteobacteria and Firmicutes (Bacteroidota). Overall, the endophyte diversity of the NMC samples was significantly higher than that of the other two sample sites. Principal coordinate analysis (PCoA) and permutational multivariate analysis of variance (PERMANOVA) results showed significant differences in the composition of endophytic bacterial and fungal communities among BGC, NMC and XGYZ samples. Co-occurrence network analysis of endophytes showed that there were positive correlations between fungi and some negative correlations between bacteria, and the co-occurrence network of bacteria was more complex than that of fungi. In short, this study provides a vital reference for further exploring and utilizing the endophyte resources of S. chamaejasme and an in-depth understanding of the ecological functions of S. chamaejasme endophytes.

Preparative separation and purification of liensinine, isoliensinine and neferine from seed embryo of Nelumbo nucifera GAERTN using high-speed counter-current chromatography.

A preparative high-speed counter-current chromatography method for separation and purification of liensinine, isoliensinine and neferine from seed embryo of Nelumbo nucifera GAERTN was successfully established by using n-hexane-ethyl acetate-methanol-water (5:8:4:5, v/v, containing 0.5% NH(4)OH) as the two-phase solvent system. From 200 mg of crude extract, 18.4 mg of liensinine, 19.6 mg of isoliensinine and 58.4 mg of neferine were obtained with the purity of 96.8, 95.9, and 98.6%, respectively. The identification of the three alkaloids was performed with (1)H NMR and (13)C NMR.

[Comparison of ferulic acid and paeoniflorin between traditional slice decoction and dispensing granule decoction of Siwu Tang].

OBJECTIVE: To compare the contents of ferulic acid and paeoniflorin between traditional slice decoction and dispensing granule decoction of Siwu Tang. METHODS: A HPLC method detected ferulic acid and paeoniflorin with Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column. The column temperature was setup at 35 degrees C and the flow rate was 1 ml/min. The mobile phase for ferulic acid was a mixed solution of MeOH-H2O-0.05% H3PO4 (40:50:10) and the wavelength was 327 nm, while for paeoniflorin was a mixed solution of CH3CN-H2O-0.05% H3PO4 (16:74:10) and the wavelength was 230 nm. RESULTS: The contents of ferulic acid and paeoniflorin in the dispensing granule decoction were 8.91 mg/dose and 83.57 mg/dose, in the traditional slice decoction were 8. 90 mg/dose and 78. 51 mg/dose, and in the dispensing granule decoction from different factories were from 3.36 to 7.67 mg/dose and from 48.13 to 67.52 mg/dose. CONCLUSION: The HPLC chromatotograms are similar in the two forms of Siwu Tang and the contents of ferulic acid and paeoniflorin are equivalent, but discrepant in the dispensing granule decoction from different factories.

[Separation and purification of alkaloids from radix of Zanthoxylum nitidum by macroporous adsorption resin].

OBJECTIVE: To obtain the optimal conditions for separating the alkaloids from the extract of Radix of Zanthoxylum nitidum by selecting appropriate macroporous adsorption resins. METHOD: Eight types of macroporous adsorption were evaluated in separating efficiency with measuring the adsorption ratio and eluting ratio of Alkaloids as indexes. RESULT: The XDA-5 macroporous adsorption resin had the best separating efficiency. After enrichment and purification with it, the product purity and yield of alkaloids were up to 33.25% and 90.15%, respectively. CONCLUSION: This method is simple, feasible and fit for industry production.

[Separation and purification processes of alkaloids from Lotus plumule with macroporous adsorption resin].

OBJECTIVE: To explore the optimal separation of the alkaloids from Lotus plumule by selecting appropriate macroporous adsorption resins. METHOD: To evaluated the separating efficiency by measuring the adsorption ratio, eluting ratio and the concentration of neferine. RESULT: The LSA -5B macroporous adsorption resin prolided the optimal separating efficiency. The best adsorption capacity was achieved with the following conditions: the concentration of extact liquid was 0. 125 g x mL(-1) (equivalence to raw material), then the resin was washed by water to remove impurity and alkaloids were desorbed by 50% ethanol. CONCLUSION: This method is simple, feasible and suitable for industry production.

[Determination of 3 kinds of components in huanglian jiedu decoction by HPLC under different UV wavelengths].

OBJECTIVE: To determinate 3 kinds of effective components (5 compounds) in Huanglian Jiedu decoction at the same time, namely berberine, palmtine, jatrorrhizine, baicalin and geniposide. METHOD: A HPLC method detected by 3 different UV wavelengths 345 nm for berberine, palmtine and jatrorrhizine, 280 nm for baicalin, 238 nm for geniposide. Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column was used, the mixture of H2O (A) , CH3OH (B) and 0.05% H3PO4 solution (C) as mobile phase in gradient mode. The column temperature was 35 degrees C and the flow rate was 1 mL x min(-1). RESULT: This method was applied to determined 5 compounds in Huanglian Jiedu decoction rapidly and accurately. CONCLUSION: The quality control method is convenient, reasonable and credible for Huanglian Jiedu decoction.