CN104313061B - A kind of culture medium for improving Resveratrol in Rhizoma Polygoni Cuspidati content and preparation method thereof - Google Patents
A kind of culture medium for improving Resveratrol in Rhizoma Polygoni Cuspidati content and preparation method thereof Download PDFInfo
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Abstract
The invention discloses it is a kind of improve Resveratrol in Rhizoma Polygoni Cuspidati content culture medium, its component based on the quality of contained substance in every liter of culture medium aqueous solution including：KNO3 1900mg,NH4NO31650 mg, CaCl2H2O 440 mg, MgSO4·7H2O 370 mg, KH2PO4170 mg, FeSO4·7H2O 27.8 mg, Na2EDTA 37.3 mg, MnSO4·4H2O 22.3 mg, ZnSO4·7H2O 8.6 mg, H3BO36.2 mg, KI 0.83 mg, Na2MoO4·2H2O 0.25 mg, CuSO4·5H2O 0.025 mg, CoCl2·6H20.025 mg of O, 100 mg of inositol, 2 mg of glycine, 0.5 mg of puridoxine hydrochloride, 0.5 mg of thiamine hydrochloride, nicotinic acid 0.5 mg, agar 6.5g, sucrose 30g, 5 20 mg of brufen, 1 5mg of acetylsalicylic acid, 15 25 mg of glutamine 30 70 mg, S nitroso N N-acetylpenicillamines.Growth and development of the culture medium to giant knotweed has preferable facilitation effect, can significantly improve the aggregate velocity and content of Resveratrol in Rhizoma Polygoni Cuspidati.
Description
Technical field
The present invention relates to a kind of culture medium for improving Resveratrol in Rhizoma Polygoni Cuspidati content and preparation method thereof.
Background technology
Numerous studies in recent years find that resveratrol can reduce blood viscosity, suppress coagulating platelets, and can promote
Vasodilation, keeps unobstructed blood, and the generation and development of pre- anti-cancer, have antiatherosclerosis and prevention and treatment of coronary heart disease, lacks
Courageous and upright heart disease, the also effect of hyperlipidemia, tool suppress effect and the estrogen-like action of tumour, available for diseases such as treatment breast cancer
Disease, also demonstrates that resveratrol has anti-oxidant and anti-aging function, therefore becomes very promising health medicine.
Content of the resveratrol in plant is generally all very low, and giant knotweed is the highest plant of Resveratrol content, perennial
Resveratrol content reaches as high as 1mg/g in root, and content also can reach 0.7647 mg/g in leaf, therefore be extracted as batch production
The main plant of resveratrol.However, giant knotweed is a kind of Wild Medicinal, the speed of growth is slow, much full by traditional excavation
Foot not demands of the people to resveratrol.
Using plant tissue culture technique, amount reproduction giant knotweed is a very promising method indoors.Plant group
Knit culture to refer to aseptically, by vitro plant organ, tissue, cell and protoplast, cultivate and manually preparing
On culture medium, appropriate condition of culture is given, makes cell Proliferation or it is grown up to complete plant.
A kind of culture medium of suitable giant knotweed growth is provided, is exactly the purpose of the present invention to improve Resveratrol content.
The content of the invention
An object of the present invention is to provide a kind of culture medium for improving Resveratrol in Rhizoma Polygoni Cuspidati content.
For achieving the above object, the technical solution adopted by the present invention is：
Form the component of the culture medium based on the quality of contained substance in every liter of culture medium aqueous solution including：
KNO3 1900mg,NH4NO31650 mg, CaCl2·2H2O 440 mg, MgSO4·7H2O 370 mg, KH2PO4
170 mg, FeSO4·7H2O 55.6 mg, Na2EDTA 74.6 mg, MnSO4·4H2O 22.3 mg, ZnSO4·7H2O 8.6
Mg, H3BO36.2 mg, KI 0.83 mg, Na2MoO4·2H2O0.25 mg, CuSO4·5H2O 0.025 mg, CoCl2·6H2O
0.025 mg, 100 mg of inositol, 2 mg of glycine, 0.5 mg of puridoxine hydrochloride, 0.5 mg of thiamine hydrochloride, nicotinic acid 0.5
Mg, agar 6.5g, sucrose 30g, brufen 5-20 mg, acetylsalicylic acid 1-5 mg, glutamine 30-70 mg, S- are sub-
Nitro-N- N-acetylpenicillamine 15-25 mg.
The second object of the present invention is to provide a kind of preparation method for the culture medium for improving Resveratrol in Rhizoma Polygoni Cuspidati content.
The present invention reaches the purpose of the present invention using the preparation method comprised the steps of：
（1）The preparation of a great number of elements mother liquor
Weigh KNO319000mg, NH4NO316500mg, MgSO4•7H2O 3700mg, KH2PO41700mg, CaCl2•
2H2O 4400mg, after fully being dissolved with distilled water respectively, sequentially pour into the volumetric flask of 1000ml, are sufficiently mixed, most
Constant volume is to 1000ml afterwards；
（2）The preparation of micro- mother liquor
Weigh MnSO4•4H2O 2230mg, ZnSO4•7H2O 860mg, H3BO3620mg, KI 83mg, Na2MoO4•2H2O
25mg, CuSO4•5H2O 2.5mg, CoCl2•6H2O 2.5mg, after fully being dissolved with distilled water respectively, are sequentially poured into
In the volumetric flask of 1000ml, it is sufficiently mixed, last constant volume to 1000ml；
（3）The preparation of mother liquid of iron salt
Weigh Na2EDTA 3730mg, FeSO4•7H2O 2780mg, after fully being dissolved with distilled water respectively, by successively secondary
Sequence is poured into the volumetric flask of 500ml, is sufficiently mixed, last constant volume to 500ml；
（4）The preparation of organic compound mother liquor
Glycine 100mg, puridoxine hydrochloride 25mg, thiamine hydrochloride 25mg, nicotinic acid 25mg, inositol 5000mg are weighed, point
After not dissolved fully with distilled water, sequentially pour into the volumetric flask of 500ml, be sufficiently mixed, last constant volume to 500ml；
（5）The preparation of plant growth regulator mother liquor
Brufen, acetylsalicylic acid and s-nitroso-N-acetylpenicillamine are configured to concentration respectively with distilled water is
1mg•ml-1Brufen mother liquor, acetyl salicylic acid mother liquor and s-nitroso-N-acetylpenicillamine mother liquor；Glutamine is steamed
It is 10mg ml that distilled water, which is configured to concentration,-1Glutamine mother liquor；
（6）The preparation of culture medium
（a）6.5g agar and 30g sucrose are added in the distilled water less than 1000ml, heating makes agar and sucrose complete
Melt；(b) a great number of elements mother liquor 100ml, micro- mother liquor 10ml, mother liquid of iron salt 10ml, organic compound are added and then
Mother liquor 10ml；(c) after high-pressure sterilizing pot sterilizing, when being cooled to 60-70 DEG C, brufen mother liquor 5-20ml, acetylsalicylic acid are added
Mother liquor 1-5ml, glutamine mother liquor 3-7ml, s-nitroso-N-acetylpenicillamine 15-25ml, are settled to distilled water
1000ml, it is 5.8~6.4 to adjust pH value, up to gel culture medium after cooled and solidified.
The micro members such as a great number of elements, copper, iron, manganese such as the required nitrogen of giant knotweed growth, phosphorus, potassium are provided in prescription of the present invention
Element, and the organic compound such as inositol, glycine, abundant nutriment is provided for the growth of giant knotweed.Especially plant growth
The introducing of conditioning agent brufen, acetylsalicylic acid, glutamine and s-nitroso-N-acetylpenicillamine, is improving the life of giant knotweed
Long metabolism, shortening growth cycle and raising resveratrol aggregate velocity etc. have positive meaning；Wherein, brufen (
Ibuprofen) and acetylsalicylic acid (Aspirin) is all artificial synthesized antipyretic-antalgic anti-inflammatory agent, has preferable antipyretic, town
Pain acts on, available for catching a cold, have a fever.Both medical structures and some auxins plant growth regulator have it is common it
Place, is a kind of plant growth regulating substance, is not only involved in the physiological activity such as flowering of plant, seed development, membrane permeability, absorption,
It is also related with disease resistance of plant.Glutamine is extremely important antioxidant-reduced glutathione in synthesising biological body
Precursor substance, is the nitrogen source for forming Amino Acids in Proteins and synthesizing the biological substance containing ammonia, has closely with tissue growth and repairing
Relation, play an important role in vital movement；S-nitroso-N-acetylpenicillamine is nitric oxide production donor, nitric oxide
It is a kind of gas active molecule for being distributed widely in organism, there are different physiological roles, nitric oxide is distribution in organism
One of extremely wide signaling molecule, in addition to mammal, nitric oxide is also made in protozoan, bacterium, yeast and plant
Work for a kind of signaling molecule of generally existing.
The present invention has the following advantages：（1）It is full of nutrition, giant knotweed growth can be made more vigorous,（2）Improve white lamb's-quarters in giant knotweed
The aggregate velocity of reed alcohol, makes Resveratrol in Rhizoma Polygoni Cuspidati content substantially increase, and reaches wild giant knotweed Resveratrol content, and than wild
The growth cycle of raw giant knotweed shortens 10 times or so.
Embodiment
Technical solution of the present invention is further illustrated by the following examples, these embodiments are intended merely to show this hair in detail
Bright technical concept and its exploitativeness, is not limiting the scope of the invention, is made using the technology of the present invention design
Equivalence replacement and accommodation still within protection scope of the present invention.
Embodiment 1
The culture medium of Resveratrol in Rhizoma Polygoni Cuspidati content is improved, its component presses the matter of contained substance in every liter of culture medium aqueous solution
Amount is calculated as：
KNO3 1900mg,NH4NO31650 mg, CaCl2·2H2O 440 mg, MgSO4·7H2O 370 mg, KH2PO4
170 mg, FeSO4·7H2O 27.8 mg, Na2EDTA 37.3 mg, MnSO4·4H2O 22.3 mg, ZnSO4·7H2O 8.6
Mg, H3BO36.2 mg, KI 0.83 mg, Na2MoO4·2H2O 0.25 mg, CuSO4·5H2O 0.025 mg, CoCl2·
6H20.025 mg of O, 100 mg of inositol, 2 mg of glycine, 0.5 mg of puridoxine hydrochloride, 0.5 mg of thiamine hydrochloride, cigarette
Acid 0.5 mg, agar 6.5g, sucrose 30g, 5 mg of brufen, 1 mg of acetylsalicylic acid, glutamine 30 mg, S- are sub-
15 mg of nitro-N- N-acetylpenicillamines.
Embodiment 2
The culture medium of Resveratrol in Rhizoma Polygoni Cuspidati content is improved, its component presses the matter of contained substance in every liter of culture medium aqueous solution
Amount is calculated as：
KNO3 1900mg,NH4NO31650 mg, CaCl2·2H2O 440 mg, MgSO4·7H2O 370 mg, KH2PO4
170 mg, FeSO4·7H2O 27.8 mg, Na2EDTA 37.3 mg, MnSO4·4H2O 22.3 mg, ZnSO4·7H2O 8.6
Mg, H3BO36.2 mg, KI 0.83 mg, Na2MoO4·2H2O 0.25 mg, CuSO4·5H2O 0.025 mg, CoCl2·
6H20.025 mg of O, 100 mg of inositol, 2 mg of glycine, 0.5 mg of puridoxine hydrochloride, 0.5 mg of thiamine hydrochloride, cigarette
Acid 0.5 mg, agar 6.5g, sucrose 30g, brufen 10mg, 2 mg of acetylsalicylic acid, glutamine 50 mg, S- are sub-
20 mg of nitro-N- N-acetylpenicillamines.
Embodiment 3
The culture medium of Resveratrol in Rhizoma Polygoni Cuspidati content is improved, its component presses the matter of contained substance in every liter of culture medium aqueous solution
Amount is calculated as：
KNO3 1900mg,NH4NO31650 mg, CaCl2·2H2O 440 mg, MgSO4·7H2O 370 mg, KH2PO4
170 mg, FeSO4·7H2O 27.8 mg, Na2EDTA 37.3 mg, MnSO4·4H2O 22.3 mg, ZnSO4·7H2O 8.6
Mg, H3BO36.2 mg, KI 0.83 mg, Na2MoO4·2H2O 0.25 mg, CuSO4·5H2O 0.025 mg, CoCl2·
6H20.025 mg of O, 100 mg of inositol, 2 mg of glycine, 0.5 mg of puridoxine hydrochloride, 0.5 mg of thiamine hydrochloride, cigarette
Acid 0.5 mg, agar 6.5g, sucrose 30g, 20 mg of brufen, 5 mg of acetylsalicylic acid, glutamine 70 mg, S- are sub-
25 mg of nitro-N- N-acetylpenicillamines.
Embodiment 4
The preparation method of the culture medium of Resveratrol in Rhizoma Polygoni Cuspidati content is improved, step is as follows：
（1）The preparation of a great number of elements mother liquor
Weigh KNO319000mg, NH4NO316500mg, MgSO4•7H2O 3700mg, KH2PO41700mg, CaCl2•
2H2O 4400mg, after fully being dissolved with distilled water respectively, sequentially pour into the volumetric flask of 1000ml, are sufficiently mixed, most
Constant volume is to 1000ml afterwards；
（2）The preparation of micro- mother liquor
Weigh MnSO4•4H2O 2230mg, ZnSO4•7H2O 860mg, H3BO3620mg, KI 83mg, Na2MoO4•2H2O
25mg, CuSO4•5H2O 2.5mg, CoCl2•6H2O 2.5mg, after fully being dissolved with distilled water respectively, are sequentially poured into
In the volumetric flask of 1000ml, it is sufficiently mixed, last constant volume to 1000ml；
（3）The preparation of mother liquid of iron salt
Weigh Na2EDTA 3730mg, FeSO4•7H2O 2780mg, after fully being dissolved with distilled water respectively, by successively secondary
Sequence is poured into the volumetric flask of 500ml, is sufficiently mixed, last constant volume to 500ml；
（4）The preparation of organic compound mother liquor
Glycine 100mg, puridoxine hydrochloride 25mg, thiamine hydrochloride 25mg, nicotinic acid 25mg, inositol 5000mg are weighed, point
After not dissolved fully with distilled water, sequentially pour into the volumetric flask of 500ml, be sufficiently mixed, last constant volume to 500ml；
（5）The preparation of plant growth regulator mother liquor
Brufen, acetylsalicylic acid and s-nitroso-N-acetylpenicillamine are configured to concentration respectively with distilled water is
1mg•ml-1Brufen mother liquor, acetyl salicylic acid mother liquor and s-nitroso-N-acetylpenicillamine mother liquor；Glutamine is steamed
It is 10mg ml that distilled water, which is configured to concentration,-1Glutamine mother liquor；
（6）The preparation of culture medium
（a）6.5g agar and 30g sucrose are added in the distilled water less than 1000ml, heating makes agar and sucrose complete
Melt；(b) a great number of elements mother liquor 100ml, micro- mother liquor 10ml, mother liquid of iron salt 10ml, organic compound are added and then
Mother liquor 10ml；(c) after high-pressure sterilizing pot sterilizing, when being cooled to 60-70 DEG C, it is female that brufen mother liquor 10ml, acetylsalicylic acid are added
Liquid 2ml, glutamine mother liquor 5ml, s-nitroso-N-acetylpenicillamine 20ml, are settled to 1000ml with distilled water, adjust pH value
For 6, up to gel culture medium after cooled and solidified.
Embodiment 5
The preparation method of the culture medium of Resveratrol in Rhizoma Polygoni Cuspidati content is improved, step is as follows：
（1）The step of preparation of a great number of elements mother liquor, the step is with embodiment 4（1）It is identical；
（2）The step of preparation of micro- mother liquor, the step is with embodiment 4（2）It is identical；
（3）The step of preparation of mother liquid of iron salt, the step is with embodiment 4（3）It is identical；
（4）The step of preparation of organic compound mother liquor, the step is with embodiment 4（4）It is identical；
（5）The step of preparation of plant growth regulator mother liquor, the step is with embodiment 4（5）It is identical；
（6）The preparation of culture medium
（a）6.5g agar and 30g sucrose are added in the distilled water less than 1000ml, heating makes agar and sucrose complete
Melt；(b) a great number of elements mother liquor 100ml, micro- mother liquor 10ml, mother liquid of iron salt 10ml, organic compound are added and then
Mother liquor 10ml；(c) after high-pressure sterilizing pot sterilizing, when being cooled to 60-70 DEG C, it is female that brufen mother liquor 5ml, acetylsalicylic acid are added
Liquid 1ml, glutamine mother liquor 3ml, s-nitroso-N-acetylpenicillamine 15ml, are settled to 1000ml with distilled water, adjust pH value
For 5.8, up to gel culture medium after cooled and solidified.
Embodiment 6
The preparation method of the culture medium of Resveratrol in Rhizoma Polygoni Cuspidati content is improved, step is as follows：
（1）The step of preparation of a great number of elements mother liquor, the step is with embodiment 4（1）It is identical；
（2）The step of preparation of micro- mother liquor, the step is with embodiment 4（2）It is identical；
（3）The step of preparation of mother liquid of iron salt, the step is with embodiment 4（3）It is identical；
（4）The step of preparation of organic compound mother liquor, the step is with embodiment 4（4）It is identical；
（5）The step of preparation of plant growth regulator mother liquor, the step is with embodiment 4（5）It is identical；
（6）The preparation of culture medium
（a）6.5g agar and 30g sucrose are added in the distilled water less than 1000ml, heating makes agar and sucrose complete
Melt agar until boiling；(b) a great number of elements mother liquor 100ml, micro- mother liquor 10ml, mother liquid of iron salt are added and then
10ml, organic compound mother liquor 10ml；(c) after high-pressure sterilizing pot sterilizing, when being cooled to 60-70 DEG C, brufen mother liquor is added
20ml, acetyl salicylic acid mother liquor 5ml, glutamine mother liquor 7ml, s-nitroso-N-acetylpenicillamine 25ml, with distilled water constant volume
To 1000ml, it is 6.4 to adjust pH value, up to gel culture medium after cooled and solidified.
There is good effect below by contrast test to verify the present invention to improving Resveratrol in Rhizoma Polygoni Cuspidati content.
First, culture medium is prepared
1. the preparation of control medium：Prepare without brufen, acetylsalicylic acid, glutamine, S-nitrosoglutathione-N- second
The culture medium of acyl penicillamine.The culture medium is prepared substantially according to the method for embodiment 4, its preparation process includes（1）-（5）Five
Step, wherein step（1）-（4）It is identical with embodiment 4, step（5）Also by（a）, (b) and (c) three small step structure
Into therein（a）The step of (b) with embodiment 4（6）In（a）Identical, difference be step (c) (b),
Step (c) presses operations described below：After high-pressure sterilizing pot sterilizing, 1000ml is settled to distilled water, it is 6 to adjust pH value, loads culture
Bottle cooled and solidified, is denoted as No. 1 culture medium.
2. the preparation containing brufen and acetylsalicylic acid culture medium：Prepare the culture medium containing brufen and acetylsalicylic acid.
The culture medium is prepared substantially according to the method for embodiment 4, also including step（1）-（6）, wherein step（1）-（4）With embodiment 4
The step of（1）-（4）It is identical, step（5）The step of according to embodiment 4（5）The method only prepares brufen and acetyl
Salicylic mother liquor, step（6）In（a）The step of (b) with embodiment 4（6）In（a）It is identical (b), difference
It is in and presses operations described below in step (c), step (c)：After high-pressure sterilizing pot sterilizing, when being cooled to 60-70 DEG C, brufen is added
Mother liquor 10ml and acetyl salicylic acid mother liquor 2ml, 1000ml is settled to distilled water, and it is 6 to adjust pH value, and it is solidifying to load blake bottle cooling
Gu it is denoted as No. 2 culture mediums.
3. the preparation containing acetylsalicylic acid, glutamine and s-nitroso-N-acetylpenicillamine culture medium：Preparation contains second
The culture medium of acyl salicylic acid, glutamine and s-nitroso-N-acetylpenicillamine.Side of the culture medium substantially according to embodiment 4
Prepared by method, also including step（1）-（6）, wherein step（1）-（4）The step of with embodiment 4（1）-（4）It is identical, step
（5）The step of according to embodiment 4（5）It is blue or green that the method only prepares acetylsalicylic acid, glutamine and S-nitrosoglutathione-N- acetyl
The mother liquor of mould amine, step（6）In（a）The step of (b) with embodiment 4（6）In（a）It is identical (b), difference
It is step (c), step (c) presses operations described below：After high-pressure sterilizing pot sterilizing, when being cooled to 60-70 DEG C, acetyl water is added
Poplar acid mother liquor 2ml, glutamine mother liquor 5ml and s-nitroso-N-acetylpenicillamine 20ml, 1000ml is settled to distilled water,
It is 6 to adjust pH value, loads blake bottle cooled and solidified, is denoted as No. 3 culture mediums.
4. the preparation containing brufen, acetylsalicylic acid, glutamine and s-nitroso-N-acetylpenicillamine culture medium：I.e.
Prepare the culture medium containing brufen, acetylsalicylic acid, glutamine and s-nitroso-N-acetylpenicillamine.The culture medium is complete
Prepared according to the method for embodiment 4, simply culture medium solution is fitted into cooled and solidified in blake bottle, is denoted as No. 4 culture mediums.
2nd, the inoculation of giant knotweed tissue-cultured seedling
The giant knotweed aseptic seedling stem sections with bud of about 0.5 centimeter length are taken, its morphology lower end is inserted into above-mentioned four kinds of cultures
In base, 4 sections are inserted into every bottle, and is uniformly distributed, bottle closure will be cultivated with sealed membrane after having inserted.
3rd, cultivate and observe
The blake bottle for being inoculated with aseptic seedling is placed in constant incubator or culturing room and is cultivated about 1 month.Condition of culture is
25 DEG C or so, illumination 2000lx of temperature, when daily light application time is 14 small.After four weeks, 3 are randomly selected from every kind of culture medium
Strain giant knotweed tissue-cultured seedling, observes the upgrowth situation of giant knotweed tissue-cultured seedling, its height is measured with ruler, and counts to its blade, so
After weigh fresh weight, counted data.
After culture 1 month, bud number, the number of sheets, plant height and the fresh weight of measured giant knotweed tissue-cultured seedling are shown in Table 1.
Bud number, the number of sheets, plant height and the fresh weight table of 1 giant knotweed tissue-cultured seedling of table
| Culture medium | Average bud number | Mean number of sheets | Average plant height（cm） | Mean fresh（g） |
| 1 | 10.33333 | 10.00 | 3.33333 | 1.53910 |
| 2 | 12.66667 | 18.00 | 3.60000 | 1.97713 |
| 3 | 16.00000 | 17.00 | 3.23333 | 1.93303 |
| 4 | 18.33333 | 23.00 | 3.73333 | 2.17120 |
4th, conclusion
As can be seen from the above table：Compared with No. 1 culture medium, No. 2 culture mediums（Containing brufen and acetylsalicylic acid）With No. 3
Culture medium（Containing acetylsalicylic acid, glutamine and s-nitroso-N-acetylpenicillamine）Growth to giant knotweed tissue-cultured seedling has rush
Into effect.No. 4 culture mediums（Containing brufen, acetylsalicylic acid, glutamine and s-nitroso-N-acetylpenicillamine）To giant knotweed group
The facilitation of training seedling growth is most obvious, and compared with No. 1 culture medium, bud number, the number of blade, plant height, fresh weight are respectively increased
77%、130%、12%、41%。
Using high performance liquid chromatography detection Resveratrol content, the resveratrol of giant knotweed seedling is averaged in No. 1 culture medium
Content is 0.1496mg/g, and the average content of the resveratrol of giant knotweed seedling is 0.2824 mg/g in No. 2 culture mediums, No. 3 culture mediums
The average content of the resveratrol of middle giant knotweed seedling is 0.3191 mg/g, and being averaged for resveratrol of giant knotweed seedling contains in No. 4 culture mediums
Measure as 0.6413 mg/g.By data comparison as can be seen that through the Resveratrol content in the giant knotweed seedling of the invention cultivated most
Height, the significant difference compared with other culture mediums.
By above-mentioned contrast test, it is not difficult to draw to draw a conclusion：Growth and development of the present invention to giant knotweed has preferable promote
Into effect, the aggregate velocity and content of Resveratrol in Rhizoma Polygoni Cuspidati can be significantly improved.
Claims (2)
1. a kind of culture medium for improving Resveratrol in Rhizoma Polygoni Cuspidati content, it is characterised in that form the component of the culture medium by every liter of training
The quality for supporting contained substance in base aqueous solution is calculated as：
KNO3 1900mg,NH4NO31650 mg, CaCl2·2H2O 440 mg, MgSO4·7H2O 370 mg, KH2PO4 170
Mg, FeSO4·7H2O 55.6 mg, Na2EDTA 74.6 mg, MnSO4·4H2O 22.3 mg, ZnSO4·7H28.6 mg of O,
H3BO36.2 mg, KI 0.83 mg, Na2MoO4·2H2O 0.25 mg, CuSO4·5H2O 0.025 mg, CoCl2·6H2O
0.025 mg, 100 mg of inositol, 2 mg of glycine, 0.5 mg of puridoxine hydrochloride, 0.5 mg of thiamine hydrochloride, nicotinic acid 0.5
Mg, agar 6.5g, sucrose 30g, brufen 5-20 mg, acetylsalicylic acid 1-5 mg, glutamine 30-70 mg, S- are sub-
Nitro-N- N-acetylpenicillamine 15-25 mg.
2. the preparation method of the culture medium of Resveratrol in Rhizoma Polygoni Cuspidati content is improved according to claim 1, it is characterised in that bag
Include following steps：
（1）The preparation of a great number of elements mother liquor
Weigh KNO319000mg, NH4NO316500mg, MgSO4•7H2O 3700mg, KH2PO41700mg, CaCl2•2H2O
4400mg, after fully being dissolved with distilled water respectively, sequentially pours into the volumetric flask of 1000ml, is sufficiently mixed, finally fixed
Hold 1000ml；
（2）The preparation of micro- mother liquor
Weigh MnSO4•4H2O 2230mg, ZnSO4•7H2O 860mg, H3BO3620mg, KI 83mg, Na2MoO4•2H2O
25mg, CuSO4•5H2O 2.5mg, CoCl2•6H2O 2.5mg, after fully being dissolved with distilled water respectively, are sequentially poured into
In the volumetric flask of 1000ml, it is sufficiently mixed, last constant volume to 1000ml；
（3）The preparation of mother liquid of iron salt
Weigh Na2EDTA 3730mg, FeSO4•7H2O 2780mg, after fully being dissolved with distilled water respectively, sequentially fall
In the volumetric flask for entering 500ml, it is sufficiently mixed, last constant volume to 500ml；
（4）The preparation of organic compound mother liquor
Glycine 100mg is weighed, puridoxine hydrochloride 25mg, thiamine hydrochloride 25mg, nicotinic acid 25mg, inositol 5000mg, is used respectively
After distilled water fully dissolves, sequentially pour into the volumetric flask of 500ml, be sufficiently mixed, last constant volume to 500ml；
（5）The preparation of plant growth regulator mother liquor
It is 1mg that brufen, acetylsalicylic acid and s-nitroso-N-acetylpenicillamine are configured to concentration respectively with distilled water
ml-1Brufen mother liquor, acetyl salicylic acid mother liquor and s-nitroso-N-acetylpenicillamine mother liquor；By glutamine distilled water
It is 10mg ml to be configured to concentration-1Glutamine mother liquor；
（6）The preparation of culture medium
（a）6.5g agar and 30g sucrose are added in the distilled water less than 1000ml, heating makes agar and sucrose melt completely
Change；(b) and then to add a great number of elements mother liquor 100ml, micro- mother liquor 10ml, mother liquid of iron salt 10ml, organic compound female
Liquid 10ml；(c) after high-pressure sterilizing pot sterilizing, when being cooled to 60-70 DEG C, it is female that brufen mother liquor 5-20ml, acetylsalicylic acid are added
Liquid 1-5ml, glutamine mother liquor 3-7ml, s-nitroso-N-acetylpenicillamine 15-25ml, 1000ml is settled to distilled water,
It is 5.8~6.4 to adjust pH value, up to gel culture medium after cooled and solidified.
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| CN1292415A (en) * | 2000-08-30 | 2001-04-25 | 尉亚辉 | Production method of cell containing resveratrol |
| CN1759665A (en) * | 2005-09-13 | 2006-04-19 | 河北医科大学 | Method for preparing polygonin through tissue culture and inducement of hairy roots of giant knotweed |
| CN103627736A (en) * | 2013-12-24 | 2014-03-12 | 湖南科源生物制品有限公司 | Method for extracting resveratrol in polygonum cuspidatum fermentation liquor |
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| CN1292415A (en) * | 2000-08-30 | 2001-04-25 | 尉亚辉 | Production method of cell containing resveratrol |
| CN1759665A (en) * | 2005-09-13 | 2006-04-19 | 河北医科大学 | Method for preparing polygonin through tissue culture and inducement of hairy roots of giant knotweed |
| CN103627736A (en) * | 2013-12-24 | 2014-03-12 | 湖南科源生物制品有限公司 | Method for extracting resveratrol in polygonum cuspidatum fermentation liquor |
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| 乙酰水杨酸和布洛芬对甘蓝试管苗生根的影响;夏海武;《植物生理学通讯》;20020620;第38卷(第3期);第305页左栏最后一段至右栏第一段、表1，第306页左栏第3段 * |
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