CN106212284A - A kind of culture medium for improving flavone compound and polysaccharide - Google Patents
A kind of culture medium for improving flavone compound and polysaccharide Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
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- A01H4/001—Culture apparatus for tissue culture
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Abstract
The invention discloses a kind of culture medium for improving flavone compound and polysaccharide, including following raw material: Cantaloupe juice, Fructus Melo juice, prune juice, Sucus Vitis viniferae, tea polyphenols, potassium nitrate, iron phosphate, ammonium nitrate, calcium nitrate, magnesium nitrate, potassium iodate, sodium borate, potassium molybdate, zinc chloride, manganous sulphate, copper chloride, ferric nitrate, vitamin, aminoacid, indolebutyric acid, gibberellins, 6 benzyl gland fat purines, iso-amylene amidopurin, naphthalene acetic acid, agar, kinetins, cosolvent, dispersant, compatilizer, stabilizer, antibacterial, water.The culture medium for improving flavone compound and polysaccharide of the present invention, collocation is reasonable, comprehensive nutrition, it is possible to quickly promotees the raw stem of root, improves transplanting survival rate, also can promote the accumulation of flavone compound simultaneously, increase economic efficiency；The culture medium preparation method for improving flavone compound and polysaccharide of the present invention, simple to operate, it is easy to accomplish industrialization, standardization and standardized production.
Description
[technical field]
The invention belongs to Herba Scutellariae Barbatae planting technology field, be specifically related to a kind of cultivation for improving flavone compound and polysaccharide
Base.
[background technology]
Herba Scutellariae Barbatae (Scutellaria barbate D.Don) is Labiatae Scutellaria plant, another name head leaf Korea Spro careless, narrow letter
Grass, go to mountain whip, toothbrush grass, spoon grass, Orchid etc., genus Labiatae perennial Wild Medicinal herbaceous plant, all herbal medicine.Its property
Acrid in the mouth, hardship, cold, return lung, liver, kidney channel, there is heat-clearing and toxic substances removing, dissipating blood stasis hemostasis, effect of inducing diuresis to remove edema.Containing multiple in Herba Scutellariae Barbatae
Compound, flavones ingredient is the active component that it is main, has antitumor, antioxidation, the multiple pharmacologically active such as antibacterial.
Herba Scutellariae Barbatae is choice drug for cancer, and the consumption in terms of medical science increases year by year, is that State Administration of Traditional Chinese Medicine is near
One of 63 kinds of Chinese crude drugs in short supply of year keypoint recommendation development.Wild resource constantly reduces frequency and faces extinction, and family plants Herba Scutellariae Barbatae and is bullied
The impact of the many factors such as time and geographical conditions, yield is the most unstable, often has the situation that supply falls short of demand to occur.In view of Herba Scutellariae Barbatae
Good efficacy, for preferably development and utilization Scutellaria barbata resource, can be by tissue culture technique tissue culture half
Branch lotus, can provide the Herba Scutellariae Barbatae seedling of a large amount of high-quality in a short time, lowers production cost, improves seedling quality, thus improves kind
Shoot survival percent and root yield.
Patent Application Publication " a kind of Herba Scutellariae Barbatae group cultivates seedling industrialized production method (publication number: CN103518626A) " is public
Open a kind of with MS as minimal medium, added the culture medium of each Plant Hormone, reached quickly tissue culture Herba Scutellariae Barbatae tissue cultured seedling
Effect, and so this patent application is not directed to promote extract from Scutellaria barbatae D and the technology of polysaccharide accumulation.
[summary of the invention]
The present invention provides a kind of culture medium for improving flavone compound and polysaccharide, to solve how to improve the master of Herba Scutellariae Barbatae
Want the content of effective ingredient flavone compound and polysaccharide, the problem such as increase economic efficiency.Being used for of the present invention improves flavonoid
Compound and the culture medium of polysaccharide, collocation is reasonable, comprehensive nutrition, it is possible to quickly promote the raw stem of root, improves transplanting survival rate, the most also
The accumulation of flavone compound can be promoted, increase economic efficiency.
For solve above technical problem, the present invention by the following technical solutions:
A kind of culture medium for improving flavone compound and polysaccharide, in units of weight portion, including following raw material: hami melon
Juice 210-292 part, Fructus Melo juice 158-242 part, prune juice 128-200 part, Sucus Vitis viniferae 105-180 part, tea polyphenols 50-80 part, nitre
Acid potassium 5-8 part, iron phosphate 4-6 part, ammonium nitrate 4-6 part, calcium nitrate 0.8-1 part, magnesium nitrate 0.6-0.8 part, potassium iodate 0.3-0.5
Part, sodium borate 0.3-0.5 part, potassium molybdate 0.3-0.5 part, zinc chloride 0.2-0.4 part, manganous sulphate 0.2-0.4 part, copper chloride
0.2-0.3 part, ferric nitrate 0.2-0.3 part, vitamin 0.1-0.3 part, aminoacid 0.1-0.3 part, indolebutyric acid 0.06-0.1 part,
Gibberellins 0.06-0.1 part, 6-benzyl gland fat purine 0.06-0.1 part, iso-amylene amidopurin 0.08-0.12 part, naphthalene acetic acid 0.04-
0.08 part, agar 0.2-0.4 part, kinetins 0.06-0.09 part, cosolvent 1-3 part, dispersant 0.8-2.5 part, compatilizer 1.2-
2 parts, stabilizer 1-2 part, antibacterial 1.5-3 part, water 8000-12000 part；
Described dispersant is in units of weight portion, including following raw material: potassium carboxymethylcellulose 4-6 part, modified starch 3-6 part,
Polyacrylic acid potassium 2-4 part, potassium alginate 1-3 part；
The Preparation Method of the described culture medium for improving flavone compound and polysaccharide, comprises the following steps:
S1: hami melon, Fructus Melo, prune juice, Fructus Vitis viniferae mixing pulper are blended, gained squeezed the juice 2000-6000 mesh sieve, system
Obtain mixed juice；
The preparation of S2: tea polyphenols, comprises the following steps:
S21: cross 50-90 mesh sieve after being pulverized by green tea, be 1 in mass ratio with the ethanol water that mass fraction is 36%-62%:
13-19 mixes, and is 30-36 DEG C in temperature, and ultrasonic power is under 180-260W, extracts 2-3 time, each 7-15min, filters,
Merging filtrate, concentrates, and prepares tea polyphenols crude extract；
S22: the tea polyphenols crude extract that step S21 prepares is joined in KCl solution, pH regulator to 6.2-6.7, turn centrifugal
Speed is removing impurity under 3000-6000r/min, uses 0.05-0.1 μm filter membrane molten to after centrifugal at temperature is 30-37 DEG C
Liquid filters, and prepares filtrate；
S23: filtrate ether extraction step S22 prepared, freeze concentration at temperature≤-12 DEG C, is 78-88 in temperature
Dried prepared tea polyphenols at DEG C；
S3: by step S1 prepare mixed juice, step S2 prepare tea polyphenols, potassium nitrate, iron phosphate, ammonium nitrate, calcium nitrate, nitre
Acid magnesium, potassium iodate, sodium borate, potassium molybdate, zinc chloride, manganous sulphate, copper chloride, ferric nitrate, vitamin, aminoacid, indole fourth
Acid, gibberellins, 6-benzyl gland fat purine, iso-amylene amidopurin, naphthalene acetic acid, agar, kinetins, cosolvent, dispersant, compatible
Agent, stabilizer, antibacterial, water mix in proportion, and prepare mixed liquor；
S4: mixed liquor pH value step S3 prepared regulates to 5-5.6, then uses pulse vacuum pressure steam sterilizer in temperature
Degree is for 134-136 DEG C, and pressure is under 210-230KPa, sterilizing 1.5-2min, prepares for improving flavone compound and polysaccharide
Culture medium.
Further, described vitamin is composed of the following components: raw element A0.06-0.1 part, vitamin B10.06-0.1
Part, vitamin B20.04-0.05 part, vitamin D 0.04-0.05 part.
Further, described aminoacid is composed of the following components: glycine 0.08-0.1 part, cystine 0.06-0.08 part,
Proline 0.04-0.07 part, serine 0.02-0.05 part.
Further, described cosolvent is polyvinylpyrrolidone.
Further, described compatilizer is Dupont EEA2112AC.
Further, described stabilizer is xanthan gum.
Further, described antibacterial is methyl-furfuryl fumarate.
Present invention effect of fractions in the culture medium improving flavone compound and polysaccharide is as follows:
The accumulation being added with beneficially polyoses content of fruit juice.
Tea polyphenols, as a kind of antioxidant, can make cell poor growth, it is also possible to promotes accumulation of flavonoids.
The nutritive effect of potassium element can improve photosynthetic intensity, promotes starch and the formation of sugar in crop body, increases
Pretend resistance and the resistance against diseases of thing, moreover it is possible to improve crop and nitrogen is absorbed.
Phosphorus is to form the indispensable element such as Nuclear extract, lecithin.P elements can accelerate cell division, promotes root
System and overground part accelerate growth, promote bud differentiation, ahead of time maturation, improve fruit quality.
Nitrogen is the essential element that protein is constituted, and protein is the base substance in cellular plasm composition.Nitrogen element energy
Promoting protein and chlorophyllous formation, make dark green leaf color, leaf area increases, and promotes that the assimilation of carbon, beneficially yield increase, product
Matter is improved.
Lacking the merismatic growth of calcium plant slack-off, calcium deficiency and disease are just at growing point and spire, and time serious, leaf deforms
And chlorosis, at the edge of leaf, necrotic spot occurs.
Magnesium is chlorophyllous constituent.If lacking magnesium, blade loses green, it is impossible to manufacture nutrient substance.
Iodine is the indispensable element of plant growing, is described as biological element, wisdom element.
Boron compares in plant and concentrates in stem apex, the tip of a root, blade and floral organ, and the content in dicotyledon is usually
Higher than monocotyledon.Boron can improve the nitrogenase activity of legume root nodule bacteria, increases amount of nitrogen fixation, and during boron deficiency, root stays not to be sent out
Reach, affect amount of nitrogen fixation.Boron can also strengthen crop anti-adversity.Molybdenum is nitrogen-fixing microorganism, particularly with the root nodule of legume symbiosis
Bacterium is fixed during air nitrogen necessary.Leaf photosynthesis intensity can be promoted again simultaneously.When lacking molybdenum, plant is short and small, between blade arteries and veins
Chlorosis, leaf margin is dried-up, inward curl, becomes wilting state.
Zinc can promote that crop carries out photosynthesis, and it is the constituent of multiple enzyme, and during zinc deficiency, stagnating occurs in crop growth.
Containing manganese in plant chloroplast, manganese can promote germination and seedling growth in period, affects the photosynthesis of crop, exhales during manganese deficiency
The accumulation in vivo of suction effect, nitrate nitrogen.Show as chlorosis yellow between blade arteries and veins, have brown speckle.Copper participates in the photosynthetic of plant
Effect, Repiration, nitrogen metabolism, the resistance of enhancing plant.When lacking copper, plant strain growth is thin and weak, and young leaves turns to be yellow, wilting withered, blade tip
Curling is turned white, and has necrotic spot.Ferrum is that chlorophyll formation is indispensable, is difficult to transfer in plant body, so blade " chlorosis
Disease " it is the performance of plant iron deficiency, and first this chlorosis shows on young leaflet tablet.Plant is taken root effectively by vitamin
Should, it is possible to promote plant establishment.
Aminoacid is nutrient substance necessary to plant growing, due to the characteristic of aminoacid itself, special to plant growing
Being that photosynthesis has unique facilitation, it can increase chlorophyll content of plant, improves the activity of enzyme, promotes titanium dioxide
The infiltration of carbon, makes photosynthesis more vigorous, and to improving crop quality, the content increasing Vc and sugar suffers from important function.
The method have the advantages that
(1) tea polyphenols can promote the accumulation of extract from Scutellaria barbatae D；Tea polyphenols, as antioxidant, can suppress plant thin simultaneously
The growth of born of the same parents, but the present invention is under other nutrient media components synergism, the suppression Herba Scutellariae Barbatae effect of tea polyphenols is less；
(2) the transplanting survival being conducive to improving Herba Scutellariae Barbatae Seedling for improving the culture medium of flavone compound and polysaccharide of the present invention
Rate, reaches 99%-100%；
(3) culture medium for improving flavone compound and polysaccharide of the present invention, collocation is reasonable, comprehensive nutrition, it is possible to quickly
Promote the raw stem of root, improve and transplant survival rate, also can promote the accumulation of flavone compound and polysaccharide simultaneously, increase economic efficiency；
(4) the culture medium preparation method for improving flavone compound and polysaccharide of the present invention is used, it is possible to when shortening sterilization
Between, improve sterilization effect reliability and industrialization production efficiency, reduce production cost；
(5) the culture medium preparation method for improving flavone compound and polysaccharide of the present invention, simple to operate, it is easy to accomplish to produce
Industry, standardization and standardized production.
[detailed description of the invention]
For ease of being more fully understood that the present invention, being illustrated by following example, these embodiments belong to the protection of the present invention
Scope, but it is not intended to protection scope of the present invention.
In an embodiment, the described culture medium for improving flavone compound and polysaccharide, in units of weight portion, including
Following raw material: Cantaloupe juice 210-292 part, Fructus Melo juice 158-242 part, prune juice 128-200 part, Sucus Vitis viniferae 105-180 part, tea
Polyphenol 50-80 part, potassium nitrate 5-8 part, iron phosphate 4-6 part, ammonium nitrate 4-6 part, calcium nitrate 0.8-1 part, magnesium nitrate 0.6-0.8
Part, potassium iodate 0.3-0.5 part, sodium borate 0.3-0.5 part, potassium molybdate 0.3-0.5 part, zinc chloride 0.2-0.4 part, manganous sulphate
0.2-0.4 part, copper chloride 0.2-0.3 part, ferric nitrate 0.2-0.3 part, vitamin 0.1-0.3 part, aminoacid 0.1-0.3 part, Yin
Diindyl butanoic acid 0.06-0.1 part, gibberellins 0.06-0.1 part, 6-benzyl gland fat purine 0.06-0.1 part, iso-amylene amidopurin 0.08-
0.12 part, naphthalene acetic acid 0.04-0.08 part, agar 0.2-0.4 part, kinetins 0.06-0.09 part, cosolvent 1-3 part, dispersant
0.8-2.5 part, compatilizer 1.2-2 part, stabilizer 1-2 part, antibacterial 1.5-3 part, water 8000-12000 part；
Described vitamin is composed of the following components: raw element A0.06-0.1 part, vitamin B10.06-0.1 part, vitamin
B20.04-0.05 part, vitamin D 0.04-0.05 part；
Described aminoacid is composed of the following components: glycine 0.08-0.1 part, cystine 0.06-0.08 part, proline 0.04-
0.07 part, serine 0.02-0.05 part；
Described cosolvent is polyvinylpyrrolidone；
Described dispersant is in units of weight portion, including following raw material: potassium carboxymethylcellulose 4-6 part, modified starch 3-6 part,
Polyacrylic acid potassium 2-4 part, potassium alginate 1-3 part；
Described compatilizer is Dupont EEA2112AC；
Described stabilizer is xanthan gum；
Described antibacterial is methyl-furfuryl fumarate；
The Preparation Method of the described culture medium for improving flavone compound and polysaccharide, comprises the following steps:
S1: hami melon, Fructus Melo, prune juice, Fructus Vitis viniferae mixing pulper are blended, gained squeezed the juice 2000-6000 mesh sieve, system
Obtain mixed juice；
The preparation of S2: tea polyphenols, comprises the following steps:
S21: cross 50-90 mesh sieve after being pulverized by green tea, be 1 in mass ratio with the ethanol water that mass fraction is 36%-62%:
13-19 mixes, and is 30-36 DEG C in temperature, and ultrasonic power is under 180-260W, extracts 2-3 time, each 7-15min, filters,
Merging filtrate, concentrates, and prepares tea polyphenols crude extract；
S22: the tea polyphenols crude extract that step S21 prepares is joined in KCl solution, pH regulator to 6.2-6.7, turn centrifugal
Speed is removing impurity under 3000-6000r/min, uses 0.05-0.1 μm filter membrane molten to after centrifugal at temperature is 30-37 DEG C
Liquid filters, and prepares filtrate；
S23: filtrate ether extraction step S22 prepared, freeze concentration at temperature≤-12 DEG C, is 78-88 in temperature
Dried prepared tea polyphenols at DEG C；
S3: by step S1 prepare mixed juice, step S2 prepare tea polyphenols, potassium nitrate, iron phosphate, ammonium nitrate, calcium nitrate, nitre
Acid magnesium, potassium iodate, sodium borate, potassium molybdate, zinc chloride, manganous sulphate, copper chloride, ferric nitrate, vitamin, aminoacid, indole fourth
Acid, gibberellins, 6-benzyl gland fat purine, iso-amylene amidopurin, naphthalene acetic acid, agar, kinetins, cosolvent, dispersant, compatible
Agent, stabilizer, antibacterial, water mix in proportion, and prepare mixed liquor；
S4: mixed liquor pH value step S3 prepared regulates to 5-5.6, then uses pulse vacuum pressure steam sterilizer in temperature
Degree is for 134-136 DEG C, and pressure is under 210-230KPa, sterilizing 1.5-2min, prepares for improving flavone compound and polysaccharide
Culture medium.
It is illustrated below by more specifically embodiment.
Embodiment 1
A kind of culture medium for improving flavone compound and polysaccharide, in units of weight portion, including following raw material: hami melon
250 parts of juice, 200 parts of Fructus Melo juice, prune juice 160 parts, Sucus Vitis viniferae 140 parts, tea polyphenols 65 parts, potassium nitrate 7 parts, iron phosphate 5 parts, nitre
Acid ammonium 5 parts, calcium nitrate 0.9 part, magnesium nitrate 0.7 part, potassium iodate 0.4 part, sodium borate 0.4 part, potassium molybdate 0.4 part, zinc chloride 0.3
Part, manganous sulphate 03 part, copper chloride 0.2 part, ferric nitrate 0.2 part, vitamin 0.2 part, 0.2 part of aminoacid, indolebutyric acid 0.08
Part, gibberellins 0.08 part, 6-benzyl gland fat purine 0.08 part, iso-amylene amidopurin 0.1 part, naphthalene acetic acid 0.06 part, agar 0.3
Part, kinetins 0.08 part, cosolvent 2 parts, dispersant 1.6 parts, compatilizer 1.6 parts, stabilizer 1.5 parts, antibacterial 2.2 parts, water
10000 parts；
Described vitamin is composed of the following components: raw element A0.08 part, vitamin B10.08 part, vitamin B20.04 part, dimension life
Element D0.04 part；
Described aminoacid is composed of the following components: glycine 0.09 part, cystine 0.07 part, proline 0.06 part, serine
0.04 part；
Described cosolvent is polyvinylpyrrolidone；
Described dispersant is in units of weight portion, including following raw material: potassium carboxymethylcellulose 5 parts, modified starch 5 parts, poly-third
3 parts of olefin(e) acid potassium, potassium alginate 2 parts；
Described compatilizer is Dupont EEA2112AC；
Described stabilizer is xanthan gum；
Described antibacterial is methyl-furfuryl fumarate；
The Preparation Method of the described culture medium for improving flavone compound and polysaccharide, comprises the following steps:
S1: hami melon, Fructus Melo, prune juice, Fructus Vitis viniferae mixing pulper are blended, gained was squeezed the juice 4000 mesh sieve, prepared mixed
Close juice；
The preparation of S2: tea polyphenols, comprises the following steps:
S21: cross 80 mesh sieves after being pulverized by green tea, mix for 1:16 in mass ratio with the ethanol water that mass fraction is 50%,
Temperature is 34 DEG C, and ultrasonic power is under 220W, extracts 2 times, each 11min, filters, merging filtrate, concentrates, and prepares tea polyphenols
Crude extract；
S22: the tea polyphenols crude extract that step S21 prepares is joined in KCl solution, pH regulator to 6.5, at centrifugal rotational speed be
Remove impurity under 5000r/min, use 0.08 μm filter membrane at temperature is 34 DEG C, the solution after being centrifuged to be filtered, prepared
Filtrate；
S23: filtrate ether extraction step S22 prepared, freeze concentration at temperature is-12 DEG C, is 84 DEG C in temperature
Lower dried prepared tea polyphenols；
S3: by step S1 prepare mixed juice, step S2 prepare tea polyphenols, potassium nitrate, iron phosphate, ammonium nitrate, calcium nitrate, nitre
Acid magnesium, potassium iodate, sodium borate, potassium molybdate, zinc chloride, manganous sulphate, copper chloride, ferric nitrate, vitamin, aminoacid, indole fourth
Acid, gibberellins, 6-benzyl gland fat purine, iso-amylene amidopurin, naphthalene acetic acid, agar, kinetins, cosolvent, dispersant, compatible
Agent, stabilizer, antibacterial, water mix in proportion, and prepare mixed liquor；
S4: mixed liquor pH value step S3 prepared regulates to 5.3, then uses pulse vacuum pressure steam sterilizer in temperature
Being 135 DEG C, pressure is under 220KPa, sterilizing 1.8min, prepares the culture medium for improving flavone compound and polysaccharide.
Embodiment 2
A kind of culture medium for improving flavone compound and polysaccharide, in units of weight portion, including following raw material: hami melon
210 parts of juice, 158 parts of Fructus Melo juice, prune juice 128 parts, Sucus Vitis viniferae 105 parts, tea polyphenols 50 parts, potassium nitrate 5 parts, iron phosphate 4 parts, nitre
Acid ammonium 4 parts, calcium nitrate 0.8 part, magnesium nitrate 0.6 part, potassium iodate 0.3 part, sodium borate 0.3 part, potassium molybdate 0.3 part, zinc chloride 0.2
Part, manganous sulphate 0.2 part, copper chloride 0.2 part, ferric nitrate 0.2 part, vitamin 0.1 part, 0.1 part of aminoacid, indolebutyric acid 0.06
Part, gibberellins 0.06 part, 6-benzyl gland fat purine 0.06 part, iso-amylene amidopurin 0.08 part, naphthalene acetic acid 0.04 part, agar 0.2
Part, kinetins 0.06 part, cosolvent 1 part, dispersant 0.8 part, compatilizer 1.2 parts, stabilizer 1 part, antibacterial 1.5 parts, water
8000 parts；
Described vitamin is composed of the following components: raw element A0.06 part, vitamin B10.06 part, vitamin B20.04 part, dimension life
Element D0.04 part；
Described aminoacid is composed of the following components: glycine 0.08 part, cystine 0.06 part, proline 0.04 part, serine
0.02 part；
Described cosolvent is polyvinylpyrrolidone；
Described dispersant is in units of weight portion, including following raw material: potassium carboxymethylcellulose 4 parts, modified starch 3 parts, poly-third
2 parts of olefin(e) acid potassium, potassium alginate 1 part；
Described compatilizer is Dupont EEA2112AC；
Described stabilizer is xanthan gum；
Described antibacterial is methyl-furfuryl fumarate；
The Preparation Method of the described culture medium for improving flavone compound and polysaccharide, comprises the following steps:
S1: hami melon, Fructus Melo, prune juice, Fructus Vitis viniferae mixing pulper are blended, gained was squeezed the juice 2000 mesh sieve, prepared mixed
Close juice；
The preparation of S2: tea polyphenols, comprises the following steps:
S21: cross 50 mesh sieves after being pulverized by green tea, mix for 1:13 in mass ratio with the ethanol water that mass fraction is 36%,
Temperature is 30 DEG C, and ultrasonic power is under 180W, extracts 2 times, each 15min, filters, merging filtrate, concentrates, and prepares tea polyphenols
Crude extract；
S22: the tea polyphenols crude extract that step S21 prepares is joined in KCl solution, pH regulator to 6.2, at centrifugal rotational speed be
Remove impurity under 3000r/min, use 0.05 μm filter membrane at temperature is 30 DEG C, the solution after being centrifuged to be filtered, prepared
Filtrate；
S23: filtrate ether extraction step S22 prepared, freeze concentration at temperature is-18 DEG C, is 78 DEG C in temperature
Lower dried prepared tea polyphenols；
S3: by step S1 prepare mixed juice, step S2 prepare tea polyphenols, potassium nitrate, iron phosphate, ammonium nitrate, calcium nitrate, nitre
Acid magnesium, potassium iodate, sodium borate, potassium molybdate, zinc chloride, manganous sulphate, copper chloride, ferric nitrate, vitamin, aminoacid, indole fourth
Acid, gibberellins, 6-benzyl gland fat purine, iso-amylene amidopurin, naphthalene acetic acid, agar, kinetins, cosolvent, dispersant, compatible
Agent, stabilizer, antibacterial, water mix in proportion, and prepare mixed liquor；
S4: mixed liquor pH value step S3 prepared regulates to 5, then employing pulse vacuum pressure steam sterilizer in temperature is
134 DEG C, pressure is under 210KPa, sterilizing 2min, prepares the culture medium for improving flavone compound and polysaccharide.
Embodiment 3
A kind of culture medium for improving flavone compound and polysaccharide, in units of weight portion, including following raw material: hami melon
292 parts of juice, 242 parts of Fructus Melo juice, prune juice 200 parts, Sucus Vitis viniferae 180 parts, tea polyphenols 80 parts, potassium nitrate 8 parts, iron phosphate 6 parts, nitre
Acid ammonium 6 parts, calcium nitrate 1 part, magnesium nitrate 0.8 part, potassium iodate 0.5 part, sodium borate 0.5 part, potassium molybdate 0.5 part, zinc chloride 0.4
Part, manganous sulphate 0.4 part, copper chloride 0.3 part, ferric nitrate 0.3 part, vitamin 0.3 part, 0.3 part of aminoacid, indolebutyric acid 0.1
Part, gibberellins 0.1 part, 6-benzyl gland fat purine 0.1 part, iso-amylene amidopurin 0.12 part, naphthalene acetic acid 0.08 part, 0.4 part of agar,
Kinetins 0.09 part, cosolvent 3 parts, dispersant 2.5 parts, compatilizer 2 parts, stabilizer 2 parts, antibacterial 3 parts, 12000 parts of water；
Described vitamin is composed of the following components: raw element A0.1 part, vitamin B10.1 part, vitamin B20.05 part, vitamin
D0.05 part；
Described aminoacid is composed of the following components: glycine 0.1 part, cystine 0.08 part, proline 0.07 part, serine 0.05
Part；
Described cosolvent is polyvinylpyrrolidone；
Described dispersant is in units of weight portion, including following raw material: potassium carboxymethylcellulose 6 parts, modified starch 6 parts, poly-third
4 parts of olefin(e) acid potassium, potassium alginate 3 parts；
Described compatilizer is Dupont EEA2112AC；
Described stabilizer is xanthan gum；
Described antibacterial is methyl-furfuryl fumarate；
The Preparation Method of the described culture medium for improving flavone compound and polysaccharide, comprises the following steps:
S1: hami melon, Fructus Melo, prune juice, Fructus Vitis viniferae mixing pulper are blended, gained was squeezed the juice 6000 mesh sieve, prepared mixed
Close juice；
The preparation of S2: tea polyphenols, comprises the following steps:
S21: cross 90 mesh sieves after being pulverized by green tea, mix for 1:19 in mass ratio with the ethanol water that mass fraction is 62%,
Temperature is 36 DEG C, and ultrasonic power is under 260W, extracts 3 times, each 7min, filters, merging filtrate, concentrates, and prepares tea polyphenols
Crude extract；
S22: the tea polyphenols crude extract that step S21 prepares is joined in KCl solution, pH regulator to 6.7, at centrifugal rotational speed be
Remove impurity under 6000r/min, use 0.1 μm filter membrane at temperature is 37 DEG C, the solution after being centrifuged to be filtered, prepared
Filtrate；
S23: filtrate ether extraction step S22 prepared, freeze concentration at temperature is-20 DEG C, is 88 DEG C in temperature
Lower dried prepared tea polyphenols；
S3: by step S1 prepare mixed juice, step S2 prepare tea polyphenols, potassium nitrate, iron phosphate, ammonium nitrate, calcium nitrate, nitre
Acid magnesium, potassium iodate, sodium borate, potassium molybdate, zinc chloride, manganous sulphate, copper chloride, ferric nitrate, vitamin, aminoacid, indole fourth
Acid, gibberellins, 6-benzyl gland fat purine, iso-amylene amidopurin, naphthalene acetic acid, agar, kinetins, cosolvent, dispersant, compatible
Agent, stabilizer, antibacterial, water mix in proportion, and prepare mixed liquor；
S4: mixed liquor pH value step S3 prepared regulates to 5.6, then uses pulse vacuum pressure steam sterilizer in temperature
Being 136 DEG C, pressure is under 230KPa, sterilizing 1.5min, prepares the culture medium for improving flavone compound and polysaccharide.
For the checking present invention for improving the effect of the culture medium of flavone compound and polysaccharide, tested by following experiment
Card.
The often Herba Scutellariae Barbatae stem with bud quantity 100 of the 2-3 cm that group selection specification is basically identical, matched group 1 is with hami melon
Juice, Fructus Melo juice, prune juice, Sucus Vitis viniferae, potassium nitrate, iron phosphate, ammonium nitrate, calcium nitrate, magnesium nitrate, potassium iodate, sodium borate, molybdic acid
Potassium, zinc chloride, manganous sulphate, copper chloride, ferric nitrate, vitamin, aminoacid, indolebutyric acid, gibberellins, 6-benzyl gland fat purine,
Iso-amylene amidopurin, naphthalene acetic acid, agar, kinetins, cosolvent, dispersant, compatilizer, stabilizer, antibacterial, water composition
Culture medium, matched group 2 is individually with tea polyphenols as culture medium, and other use the culture medium of embodiment 1-3.Measure after cultivating 110 days
Herba Scutellariae Barbatae Seedling flavone compound average yield and length, the results are shown in Table 1.
Table 1 cultivates basis set training Herba Scutellariae Barbatae Seedling flavone compound average yield and length situation
| Embodiment | Experimental group number | The average yield (%) of flavone compound | Herba Scutellariae Barbatae Seedling length (cm) |
| 1 | 100 | 0.96 | 5.85 |
| 2 | 100 | 0.94 | 5.83 |
| 3 | 100 | 0.99 | 5.87 |
| Matched group 1 | 100 | 0.78 | 5.96 |
| Matched group 2 | 100 | / | Almost unchanged |
As shown in Table 1, the Herba Scutellariae Barbatae Seedling flavone compound average yield of embodiment 1-3 is than the Herba Scutellariae Barbatae Seedling flavone of matched group 1
Compounds average yield height 20.51%-26.92%；The Herba Scutellariae Barbatae Seedling length of embodiment 1-3 is longer than the Herba Scutellariae Barbatae Seedling of matched group 1
Spending short 1.51%-2.18%, Herba Scutellariae Barbatae grows matched group 2 hardly, illustrates that the existence of tea polyphenols can be greatly improved Herba Scutellariae Barbatae Seedling
Flavone compound average yield, but tea polyphenols is as antioxidant simultaneously, also suppresses the growth of plant cell, but trains at other
Supporting basis set point 5 and join under effect, the inhibitory action of tea polyphenols is less.
The often Herba Scutellariae Barbatae stem with bud quantity 100 of the 2-3cm that group selection specification is basically identical, matched group is with tea polyphenols, nitre
Acid potassium, iron phosphate, ammonium nitrate, calcium nitrate, magnesium nitrate, potassium iodate, sodium borate, potassium molybdate, zinc chloride, manganous sulphate, copper chloride,
Ferric nitrate, vitamin, aminoacid, indolebutyric acid, gibberellins, 6-benzyl gland fat purine, iso-amylene amidopurin, naphthalene acetic acid, agar,
The culture medium of kinetins, cosolvent, dispersant, compatilizer, stabilizer, antibacterial, water composition, other use embodiment 1-3
Culture medium.Measure polysaccharide average yield and the length of tissue cultured seedling after cultivating 110 days, the results are shown in Table 2.
Table 2 cultivates basis set training Herba Scutellariae Barbatae Seedling polysaccharide average yield and length situation
| Embodiment | Experimental group seedlings cultivating number | Polysaccharide average yield (%) | Herba Scutellariae Barbatae Seedling length (cm) |
| 1 | 100 | 13.22 | 5.85 |
| 2 | 100 | 12.85 | 5.83 |
| 3 | 100 | 13.34 | 5.87 |
| Matched group | 100 | 10.23 | 5.81 |
As shown in Table 2, the Herba Scutellariae Barbatae Seedling polysaccharide average yield of embodiment 1-3 is higher than the Herba Scutellariae Barbatae Seedling polysaccharide average yield of matched group
25.61%-30.4%；The Herba Scutellariae Barbatae Seedling length of embodiment 1-3 is than the Herba Scutellariae Barbatae short 0.34%-1.03% of Seedling length of matched group, explanation
By culture medium without fruice, or fruice is joined under effect, to Herba Scutellariae Barbatae height of seedling degree accretion with other nutrient media componentses 5
Not quite.
Table 3 is that embodiment 1-3 cultivates basis set training Herba Scutellariae Barbatae transplantation of seedlings survival condition.
Basis set training Herba Scutellariae Barbatae transplantation of seedlings survival condition cultivated by table 3
| Embodiment | Experimental group seedlings cultivating number | Tissue cultured seedling transplants survival number | Transplant survival rate (%) |
| 1 | 100 | 98 | 100 |
| 2 | 100 | 98 | 99 |
| 3 | 100 | 99 | 100 |
By table 3 it is recognised that with the basis set training Herba Scutellariae Barbatae Seedling cultivated for improving flavone compound and polysaccharide of the present invention
Average survival rate of transplanting is 99%-100%, illustrates that the present invention is conducive to for the culture medium improving flavone compound and polysaccharide
Improve the transplanting survival rate of Herba Scutellariae Barbatae Seedling.
Above content it cannot be assumed that the present invention be embodied as be confined to these explanation, technology belonging to the present invention is led
For the those of ordinary skill in territory, without departing from the inventive concept of the premise, it is also possible to make some simple deduction or replace,
All should be considered as belonging to the scope of patent protection that the present invention is determined by the claims submitted to.
Claims (7)
1. one kind for improving the culture medium of flavone compound and polysaccharide, it is characterised in that in units of weight portion, including with
Lower raw material: Cantaloupe juice 210-292 part, Fructus Melo juice 158-242 part, prune juice 128-200 part, Sucus Vitis viniferae 105-180 part, tea are many
Phenol 50-80 part, potassium nitrate 5-8 part, iron phosphate 4-6 part, ammonium nitrate 4-6 part, calcium nitrate 0.8-1 part, magnesium nitrate 0.6-0.8 part,
Potassium iodate 0.3-0.5 part, sodium borate 0.3-0.5 part, potassium molybdate 0.3-0.5 part, zinc chloride 0.2-0.4 part, manganous sulphate 0.2-
0.4 part, copper chloride 0.2-0.3 part, ferric nitrate 0.2-0.3 part, vitamin 0.1-0.3 part, aminoacid 0.1-0.3 part, indole fourth
Acid 0.06-0.1 part, gibberellins 0.06-0.1 part, 6-benzyl gland fat purine 0.06-0.1 part, iso-amylene amidopurin 0.08-0.12
Part, naphthalene acetic acid 0.04-0.08 part, agar 0.2-0.4 part, kinetins 0.06-0.09 part, cosolvent 1-3 part, dispersant 0.8-
2.5 part, compatilizer 1.2-2 part, stabilizer 1-2 part, antibacterial 1.5-3 part, water 8000-12000 part；
Described dispersant is in units of weight portion, including following raw material: potassium carboxymethylcellulose 4-6 part, modified starch 3-6 part,
Polyacrylic acid potassium 2-4 part, potassium alginate 1-3 part；
The Preparation Method of the described culture medium for improving flavone compound and polysaccharide, comprises the following steps:
S1: hami melon, Fructus Melo, prune juice, Fructus Vitis viniferae mixing pulper are blended, gained squeezed the juice 2000-6000 mesh sieve, system
Obtain mixed juice；
The preparation of S2: tea polyphenols, comprises the following steps:
S21: cross 50-90 mesh sieve after being pulverized by green tea, be 1 in mass ratio with the ethanol water that mass fraction is 36%-62%:
13-19 mixes, and is 30-36 DEG C in temperature, and ultrasonic power is under 180-260W, extracts 2-3 time, each 7-15min, filters,
Merging filtrate, concentrates, and prepares tea polyphenols crude extract；
S22: the tea polyphenols crude extract that step S21 prepares is joined in KCl solution, pH regulator to 6.2-6.7, turn centrifugal
Speed is removing impurity under 3000-6000r/min, uses 0.05-0.1 μm filter membrane molten to after centrifugal at temperature is 30-37 DEG C
Liquid filters, and prepares filtrate；
S23: filtrate ether extraction step S22 prepared, freeze concentration at temperature≤-12 DEG C, is 78-88 in temperature
Dried prepared tea polyphenols at DEG C；
S3: by step S1 prepare mixed juice, step S2 prepare tea polyphenols, potassium nitrate, iron phosphate, ammonium nitrate, calcium nitrate, nitre
Acid magnesium, potassium iodate, sodium borate, potassium molybdate, zinc chloride, manganous sulphate, copper chloride, ferric nitrate, vitamin, aminoacid, indole fourth
Acid, gibberellins, 6-benzyl gland fat purine, iso-amylene amidopurin, naphthalene acetic acid, agar, kinetins, cosolvent, dispersant, compatible
Agent, stabilizer, antibacterial, water mix in proportion, and prepare mixed liquor；
S4: mixed liquor pH value step S3 prepared regulates to 5-5.6, then uses pulse vacuum pressure steam sterilizer in temperature
Degree is for 134-136 DEG C, and pressure is under 210-230KPa, sterilizing 1.5-2min, prepares for improving flavone compound and polysaccharide
Culture medium.
The most according to claim 1 for improving the culture medium of flavone compound and polysaccharide, it is characterised in that described dimension is raw
Element is composed of the following components: raw element A0.06-0.1 part, vitamin B10.06-0.1 part, vitamin B20.04-0.05 part, dimension life
Element D0.04-0.05 part.
The most according to claim 1 for improving the culture medium of flavone compound and polysaccharide, it is characterised in that described amino
Acid is composed of the following components: glycine 0.08-0.1 part, cystine 0.06-0.08 part, proline 0.04-0.07 part, serine
0.02-0.05 part.
The most according to claim 1 for improving the culture medium of flavone compound and polysaccharide, it is characterised in that described hydrotropy
Agent is polyvinylpyrrolidone.
The most according to claim 1 for improving the culture medium of flavone compound and polysaccharide, it is characterised in that described compatible
Agent is Dupont EEA2112AC.
The most according to claim 1 for improving the culture medium of flavone compound and polysaccharide, it is characterised in that described stable
Agent is xanthan gum.
The most according to claim 1 for improving the culture medium of flavone compound and polysaccharide, it is characterised in that described antibacterial
Agent is methyl-furfuryl fumarate.
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| CN106818490A (en) * | 2017-04-08 | 2017-06-13 | 广西科技大学鹿山学院 | A kind of culture medium for improving subprostrate sophora tissue-cultured seedling flavone compound and polysaccharide |
| CN106922535A (en) * | 2017-04-08 | 2017-07-07 | 广西科技大学鹿山学院 | A kind of culture medium for improving Sculellaria barbata tissue-cultured seedling flavone compound and polysaccharide |
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| CN104335903B (en) * | 2014-11-21 | 2017-04-05 | 广西中医药大学 | It is a kind of to promote Pseudobulbus Bletillae (Rhizoma Bletillae) rapid propagation method |
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| CN106818490A (en) * | 2017-04-08 | 2017-06-13 | 广西科技大学鹿山学院 | A kind of culture medium for improving subprostrate sophora tissue-cultured seedling flavone compound and polysaccharide |
| CN106922535A (en) * | 2017-04-08 | 2017-07-07 | 广西科技大学鹿山学院 | A kind of culture medium for improving Sculellaria barbata tissue-cultured seedling flavone compound and polysaccharide |
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