CN107996374A - A kind of method that tanshinone content in red sage root is improved by salt stress - Google Patents
A kind of method that tanshinone content in red sage root is improved by salt stress Download PDFInfo
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- CN107996374A CN107996374A CN201711303277.1A CN201711303277A CN107996374A CN 107996374 A CN107996374 A CN 107996374A CN 201711303277 A CN201711303277 A CN 201711303277A CN 107996374 A CN107996374 A CN 107996374A
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- tanshinone
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- red sage
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
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Abstract
The present invention provides a kind of method for improving tanshinone content.Specifically include following steps：The Radix Salviae Miltiorrhizae plant of robust growth is chosen, Liquid Culture adapts it to aquatic environment in two months；Then NaCl is added in nutrient solution, when Stress treatment 8 is small after, can improve in red sage root tanshinone content.The method that the present invention improves tanshinone content in red sage root by NaCl Stress treatments, using liquid cultivating method, NaCl is added into nutrient solution, obtains the Radix Salviae Miltiorrhizae plant of tanshinone content raising.The total-tanshinone detected after NaCl Stress treatments in red sage root（Tanshinone I, tanshinone IIA, Cryptotanshinone）Content is significantly improved compared to control group.The average content of tanshinone is more than 1.3 times of control group in the red sage root of NaCl Stress treatments.The present invention can effectively improve tanshinone content in red sage root, and easy to operate, cost is low, can be cultivated for the salt-soda soil of Salvia miltiorrhiza Bge and provide Technical Reference.
Description
Technical field
The invention belongs to Radix Salviae Miltiorrhizae technical field of cultivation, and in particular to a kind of method for improving tanshinone content in red sage root.
Background technology
Radix Salviae Miltiorrhizae（Salvia miltiorrhiza Bge.）For Labiatae（Labiatae）Salvia（Salvia）For many years
This medicinal plant of sward, is used as medicine with root and rhizome, and being that China is a kind of commonly uses large Chinese medicine.Radix Salviae Miltiorrhizae is mainly comprising two major classes activity
Component：One kind is water-soluble phenolic acids material；Another kind of is fat-soluble diterpene quinone, i.e. tanshinone material.It is modern
Medical research shows：Tanshinone has significant curative effect in terms of cardiovascular and cerebrovascular disease is treated.
Salinity is one of the important environmental factor for influencing growth and development of plants, can be reduced in adverse circumstance environment osmotic potential and
Cause plant cell dehydration, osmotic stress is produced, so as to force plant to carry out corresponding osmotic adjustment in the range of its own.
Tripathi etc. thinks that appropriate Salt Strees Condition can promote the secondary metabolism of plant, improves the content of secondary metabolite.
Tanshinone is a kind of important secondary metabolite in Radix Salviae Miltiorrhizae as terpene substances.The biosynthesis of tanshinone by
To the regulation and control and induction of various envirment factors (biotic factor and abiotic component).Recent study finds that sorbierite, infiltration are coerced
Compel YE, Ag+、Co2+And Cd2+There is facilitation Deng the accumulation all to tanshinone.But related salt stress is to tanshinone dynamic product
The tired research influenced has not been reported.Therefore, contacting between salt stress and tanshinone Dynamic Accumulation is explored, can be that research is red
The synthesis mechanism for joining ketone provides theoretical foundation, and Technical Reference is provided for the good Radix Salviae Miltiorrhizae of salt-soda soil high-yield culture, it helps passes
The standardization and targeted plantation of system the traditional Chinese medicine medicinal herbs resource plant.
The content of the invention
It is easy to operate it is an object of the invention to provide a kind of method for improving tanshinone content in red sage root, easy to advise
Modelling produces.
The method of tanshinone content in raising red sage root proposed by the present invention, is that addition NaCl is molten in the nutrient solution of Radix Salviae Miltiorrhizae
Liquid carries out Stress treatment, and tanshinone content has notable rise after 8h.
NaCl solution of the present invention, its concentration are preferably 50-100 mM.
In the present invention, Radix Salviae Miltiorrhizae Liquid Culture is concretely comprised the following steps, the good annual Radix Salviae Miltiorrhizae of growth potential is selected first and plants
Strain, water planting culture 2 months or so, adapts it to Liquid Culture environment, NaCl solution is then added in liquid medium；It is described
Liquid medium, be by 1/2MS nutrient solutions add 30g/L sucrose and.
The key point that the present invention studies is to use 50-100 mM NaCl solution Stress treatment Radix Salviae Miltiorrhizae plant, makes red sage root
In tanshinone content improve.NaCl concentration is less than 50mM, coerces effect unobvious；NaCl concentration is higher than 100mM, influences Radix Salviae Miltiorrhizae
The growth and development of plant.
The method of tanshinone content is possessed compared with prior art in raising red sage root disclosed by the invention actively imitates
Fruit is：
（1）The present invention establishes the method that tanshinone content is improved by NaCl Stress treatments, can effectively improve in red sage root
The content of tanshinone, the content of tanshinone is more than 1.3 times of control in processed red sage root.The method of the present invention need not
Higher early investment, it is easy to operate, Technical Reference can be provided for the good Radix Salviae Miltiorrhizae of salt-soda soil high-yield culture.
（2）The method that the present invention improves tanshinone content in red sage root by NaCl Stress treatments, using Liquid Culture side
Method, NaCl is added into nutrient solution, obtains the Radix Salviae Miltiorrhizae plant of tanshinone content raising.After NaCl Stress treatments in red sage root
The total-tanshinone detected（Tanshinone I, tanshinone IIA, Cryptotanshinone）Content is significantly improved compared to control group.NaCl is coerced
The average content of tanshinone is more than 1.3 times of control group in the red sage root of processing.
（3）This method improves tanshinone content using NaCl Stress treatments, has the characteristics that method simplicity, significant effect,
It can be cultivated for the salt-soda soil of Radix Salviae Miltiorrhizae and Technical Reference is provided.
Embodiment
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention
It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention
Scope, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this
The various changes carried out on the premise of invention spirit and scope to the material component in these embodiments and dosage or change
Belong to protection scope of the present invention.Raw materials used and reagent of the invention is commercially available.
Implementation column 1
The culture of Radix Salviae Miltiorrhizae material
It is healthy and strong to choose seedling, leaf 3-5 pieces/strain, root is sturdy, high 15-20 cm, the good annual Radix Salviae Miltiorrhizae plant of growth potential, liquid
Body culture 2 months or so, replaces a nutrient solution in every 5 days, adapts it to growing environment.The Liquid Culture refers to passing through
The sucrose of 30g/L is added in 1/2MS nutrient solutions and is obtained.Measure Tanshinone I, tanshinone IIA, the content of Cryptotanshinone and use height
Effect liquid phase chromatogram method.
Embodiment 2
Stress treatments of the NaCl to Radix Salviae Miltiorrhizae
The Radix Salviae Miltiorrhizae plant of the robust growth in Liquid Culture environment is selected, salt stress processing is carried out to it, makes its final concentration of 50-
100mM；At the same time with the Radix Salviae Miltiorrhizae of water process as control.Red sage root is collected when 8 is small after processing, Radix Salviae Miltiorrhizae is measured with high performance liquid chromatography
Ketone content, each handles 5 repetitions.The Liquid Culture refers to the sucrose by adding 30g/L in 1/2MS nutrient solutions
And obtain.
Embodiment 3
Tanshinone content in red sage root is measured using HPLC：
（1）Liquid phase chromatogram condition：
The assay of tanshinone utilizes high performance liquid chromatography, 5 μm of Eclipse Plus C18 silica gel reversed-phase columns, 4.6 ×
150 mm, mobile phase are methanol:Water（75:25）, flow velocity 1mL/ min, 280 nm of Detection wavelength, it is Radix Salviae Miltiorrhizae to predominantly detect component
Ketone I, tanshinone IIA, Cryptotanshinone.
（2）The making of standard curve
Tanshinone I, tanshinone IIA, Cryptotanshinone standard items are all purchased from Chinese pharmaceutical biological product and appraise and decide institute.Chromatography first is used respectively
Alcohol compound concentration is the standard sample of 1,10,20,50,100 μ g/mL, and appearance time and peak area are recorded respectively after sample introduction.It is red
Ginseng ketone I, tanshinone IIA, Cryptotanshinone retention time are respectively 7.8,13.8,8.5 min, regression equation be respectively y=
20.099x+54.393, y=20.099x+54.393, y=195.53x -88.845, linear distribution is good, can be with
Quantitative analysis for subsequent sample.
（3）The tanshinone assay of Radix Salviae Miltiorrhizae sample
The Radix Salviae Miltiorrhizae plant in example 2 is collected, accurately weighed 2 g Radix Salviae Miltiorrhizae fresh quench, is placed in mortar, adds 5 mL methanol triturations 5
Min, is transferred in centrifuge tube, then is added into mortar after 5 mL methanol rinse and be transferred to centrifuge tube.1 h of ultrasonic extraction, takes out
Let cool, 14000 rpm centrifuge 15 min, take supernatant, cross 0.45 μm of microporous barrier up to sample to be tested extract.Sample measures
It is next qualitative with retention time using external standard method, quantified with peak area, each sample sets 3 repetitions, is averaged.
（4）Statistical analysis
Standard curve making is carried out using Excel, statistical analysis is carried out using SPSS, as a result with average value ± standard deviation
（Mean±SD）Represent, the equal replication of all samples 3 times.
The tanshinone detected in the red sage root handled in the present invention with NaCl（Tanshinone I, tanshinone IIA, hidden Radix Salviae Miltiorrhizae
Ketone）Content is significantly improved compared to control group.Tanshinone I is 1.44 times of control after processing, and tanshinone IIA content is control
1.30 times, Cryptotanshinone content is 2.15 times of control；
Conclusion：The present embodiment determines tanshinone content in red sage root using HPLC methods.The result shows that the pellet of NaCl Stress treatments
Tanshinone content is significantly improved than control in ginseng root.
Claims (2)
- A kind of 1. method for improving tanshinone content in red sage root, it is characterised in that adding concentration in the nutrient solution of Radix Salviae Miltiorrhizae is 50-100 mM NaCl。
- 2. the method for improving tanshinone content in red sage root described in claim 1, it is characterised in that select annual Radix Salviae Miltiorrhizae to plant Strain, Liquid Culture 2 months, adapts it to liquid environment；Then NaCl is added in nutrient solution and coercing cultivation is carried out to Radix Salviae Miltiorrhizae；Institute The nutrient solution stated, is obtained by addition adding the sucrose of 30g/L in 1/2MS nutrient solutions.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109924090A (en) * | 2019-04-23 | 2019-06-25 | 河南省文医堂药业有限公司 | A kind of Radix Salviae Miltiorrhizae implantation methods improving tanshinone content |
| CN111742828A (en) * | 2020-06-24 | 2020-10-09 | 湖北民族大学 | Method for directionally cultivating longtube ground ivy |
| CN115226519A (en) * | 2022-08-31 | 2022-10-25 | 青岛农业大学 | Method for promoting increase of total flavone content in scutellaria baicalensis |
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| CN102776223A (en) * | 2012-06-05 | 2012-11-14 | 上海师范大学 | Method for improving tanshinone content of hairy roots of salvia miltiorrhiza through quantum dots |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109924090A (en) * | 2019-04-23 | 2019-06-25 | 河南省文医堂药业有限公司 | A kind of Radix Salviae Miltiorrhizae implantation methods improving tanshinone content |
| CN111742828A (en) * | 2020-06-24 | 2020-10-09 | 湖北民族大学 | Method for directionally cultivating longtube ground ivy |
| CN115226519A (en) * | 2022-08-31 | 2022-10-25 | 青岛农业大学 | Method for promoting increase of total flavone content in scutellaria baicalensis |
| CN115226519B (en) * | 2022-08-31 | 2023-11-03 | 青岛农业大学 | Method for promoting increase of total flavone content in radix scutellariae |
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