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Separation of Five Flavonoids from Aerial Parts of Salvia Miltiorrhiza Bunge Using HSCCC and Their Antioxidant Activities.

ABSTRACT: The aerial parts of Salvia miltiorrhiza Bunge, as the non-medicinal parts, are always discarded during harvesting, resulting in a huge waste of resources and environmental pressure. Due to the high flavonoid content and their antioxidant activities characteristics, the aerial parts of S. miltiorrhiza can be developed into natural antioxidants and used in foods. A high-speed counter-current chromatography (HSCCC) method, using a two-phase solvent system composed of tert-butyl methyl ether/n-butanol/acetonitrile/water (3:1:1:20, v/v), was the first to successfully isolate five flavonoids from the aerial parts of S. miltiorrhiza in one attempt, and separately categorized as rutin (1), isoquercitrin (2), kaempferol-3-O-?-l-rhamnopyranosyl-(1?6)-?-d-glucopyranoside (3), kaempferol-3-O-?-d-glucopyranoside (4) and apigenin-7-O-?-d-glucopyranoside (5) after identification. The purities of these plant isolates were 97.3%, 99.5%, 92.8%, 98.1% and 98.7%, respectively. All the flavonoids were identified by HR-ESI-MS, 1D and 2D NMR. Compounds 3 and 5 were firstly isolated from the plant of S. miltiorrhiza. Results from antioxidant assays showed that rutin (1) and isoquercitrin (2) had higher antioxidant capacities compared to L-ascorbic acid as the positive control.

SUBMITTER: Yang F

PROVIDER: S-EPMC6804221 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature

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Separation of Five Flavonoids from Aerial Parts of Salvia Miltiorrhiza Bunge Using HSCCC and Their Antioxidant Activities.

Yang Fan F Qi Yingxue Y Liu Wei W Li Jia J Wang Daijie D Fang Lei L Zhang Yongqing Y

Molecules (Basel, Switzerland) 20190923 19

The aerial parts of Salvia miltiorrhiza Bunge, as the non-medicinal parts, are always discarded during harvesting, resulting in a huge waste of resources and environmental pressure. Due to the high flavonoid content and their antioxidant activities characteristics, the aerial parts of S. miltiorrhiza can be developed into natural antioxidants and used in foods. A high-speed counter-current chromatography (HSCCC) method, using a two-phase solvent system composed of tert-butyl ...[more]

PMID: 31547535

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Project description:BackgroundLarge-scale heterosis breeding depends upon stable, inherited male sterility lines. We accidentally discovered a male sterility line (SW-S) in the F1progeny of a Salvia miltiorrhiza Bunge from Shandong, China (purple flowers) crossed with a S. miltiorrhiza f. alba from Sichuan, China (white flowers). We sought to provide insights into the pollen development for male sterility in S. miltiorrhiza.MethodsThe phenotypic and cytological features of the SW-S and fertile control SW-F were observed using scanning electron microscopy and paraffin sections to identify the key stage of male sterility. Transcriptome profiles were recorded for anthers at the tetrad stage of SW-S and SW-F using Illumina RNA-Seq.ResultsThe paraffin sections showed that sterility mainly occurred at the tetrad stage of microspore development, during which the tapetum cells in the anther compartment completely fell off and gradually degraded in the sterile line. There was little-to-no callose deposited around the microspore cells. The tetrad microspore was shriveled and had abnormal morphology. Therefore, anthers at the tetrad stage of SW-S and fertile control SW-F were selected for comparative transcriptome analysis. In total, 266,722,270 clean reads were obtained from SW-S and SW-F, which contained 36,534 genes. There were 2,571 differentially expressed genes (DEGs) in SW-S and SW-F, of which 63.5% were downregulated. Gene Ontology (GO) enrichment analysis indicated that the differentially expressed genes were enriched in 56 functional groups (GO terms); of these, all DEGs involved in microgametogenesis and developmental maturation were downregulated in SW-S. These results were confirmed by quantitative RT-PCR. The two GO terms contained 18 DEGs, among which eight DEGs (namely: GPAT3, RHF1A, phosphatidylinositol, PFAS, MYB96, MYB78, Cals5, and LAT52) were related to gamete development. There were 10 DEGs related to development and maturation, among which three genes were directly related to pollen development (namely: ACT3, RPK2, and DRP1C). Therefore, we believe that these genes are directly or indirectly involved in the pollen abortion of SW-S. Our study provides insight into key genes related to sterility traits in S. miltiorrhiza, and the results can be further exploited in functional and mechanism studies.

Project description:The effect of elicitors on the metabolites in the suspension cells of Salvia miltiorrhiza Bunge was elucidated by comparing and analyzing the metabolites of induced and uninduced suspension cells. The primary metabolites were detected by GC-MS. Twelve types of secondary metabolites, namely, shikimic acid, tanshinol, protocatechuic acid, caffeic acid, p-coumaric acid, rosmarinic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone, cryptotanshinone, tanshinone I, and tanshinone II A were detected by HPLC. Results indicated a total of 90 primary metabolites in the cells. These metabolites consisted of 49 kinds of sugars and their derivatives, 15 organic acids, 9 amino acids and their derivatives, 9 hydrocarbons, and 8 other substances. OPLS-DA results indicated five differential primary metabolites, namely, gluconic acid, mannopyranose, glucose, inositol, and ketoisovalerate, between the callus and suspension cells. SA significantly induced glucose metabolism in the S. miltiorrhiza suspension cells, and the mean contents of glucose, fructose, and mannose were significantly lower in the induced cells than in the control. However, SA, as a plant inducer, could significantly promote the increase in 10 secondary metabolites, except protocatechuic acid and tanshinone I, in the suspension cells at different times or degrees. The effect of NaCl on the S. miltiorrhiza cell mainly depended on the downstream pathway of glucose metabolism. Fructose and glucose were the decomposition products of sucrose, and glucose was processed through monosaccharide metabolism. Induction by NaCl resulted in significantly lower levels of these primary metabolites in the induced cells than in the control group. However, NaCl could significantly promote 10 secondary metabolites, except tanshinol and protocatechuic acid, at different times or degrees. After elicitation by AgNO3, sucrose and proline were higher in the induced cells than in the control group. However, galactose and fructose were lower in the experimental cells than in the control. This phenomenon may have been caused by the induction of plant stress response by AgNO3. AgNO3 could significantly promote shikimic acid, caffeic acid, p-coumaric acid, rosmarinic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone, cryptotanshinone, tanshinone I, and tanshinone IIA. This study will provide a reference for future studies on the effects of primary metabolites on the secondary metabolites and the potential relation of these metabolites to plant stress response in S. miltiorrhiza.

Dataset Information

Separation of Five Flavonoids from Aerial Parts of Salvia Miltiorrhiza Bunge Using HSCCC and Their Antioxidant Activities.

Publications

Separation of Five Flavonoids from Aerial Parts of <i>Salvia Miltiorrhiza</i> Bunge Using HSCCC and Their Antioxidant Activities.

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