CN107135944A - A kind of beautiful millettia root root tissues culture medium and preparation method thereof - Google Patents
A kind of beautiful millettia root root tissues culture medium and preparation method thereof Download PDFInfo
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- CN107135944A CN107135944A CN201710306844.2A CN201710306844A CN107135944A CN 107135944 A CN107135944 A CN 107135944A CN 201710306844 A CN201710306844 A CN 201710306844A CN 107135944 A CN107135944 A CN 107135944A
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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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
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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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a kind of beautiful millettia root root tissues culture medium and preparation method thereof, belong to tissue culture medium (TCM) technical field, root tissues culture medium includes following raw material：Agar, sucrose, hormone, inorganic salt solution, nicotinic acid, activated carbon, cysteine, streptomysin, lactoalbumin hydrolysate, organic matter, tri-distilled water.The root tissues culture medium being prepared into using raw material of the present invention and method is prepared according to Nutrient Properties needed for the growth of beautiful millettia root tissue-cultured seedling, tissue-cultured seedling root induction rate to beautiful millettia root is high, and transplanting survival rate is high, and good basis is provided for the tissue cultures of beautiful millettia root.
Description
Technical field
The invention belongs to culture medium preparing technical field, and in particular to a kind of beautiful millettia root root tissues culture medium.
Background technology
Beautiful millettia root, alias pig's feet large bamboo hat with a conical crown and broad brim, Jin Zhonggen, beautiful millettia root, hangs upside down Jin Zhong, energetically potato, is that pulse family pulse family Millettia is planted
Thing, its main component is protein, starchiness and alkaloid etc..Beautiful millettia root is used as medicine with root, and whole year can excavate, and is traditional medicine food
Homologous plant.Beautiful millettia root is mild-natured, sweet, there is the active effect of kidney tonifying moistening lung, strengthening tendons, is usually used in treatment lumbar muscle strain, rheumatic
The illnesss such as arthritis, pulmonary tuberculosis, chronic bronchitis, chronic hepatitis.Beautiful millettia root is also often among the people for Guangdong in addition to the decoction that is used as medicine
Enter soup and do dietotherapy, herbal cuisine to be used.Beautiful millettia root active ingredient has maackiain, onocerin, chalcone compounds, isoflavones
Class, furfural, beautiful millettia root polysaccharide etc., these compositions have anti-inflammatory sterilization, immunoregulation effect, and by certain anti-oxidant and removing certainly
Acted on by base.Beautiful millettia root is after excavation, and cleaned, drying can directly be used as medicine, it can also be used to edible, can be more made through deep processing
The deep processed products such as beautiful millettia root health liquor, beautiful millettia root jasmine tea.Beautiful millettia root is pharmacy corporation processing Chinese patent drug, the conventional original of health products
Material, in recent years, as China's traditional Chinese medicine flourishes, people's living standard is gradually stepped up, and health care consciousness constantly strengthens, ox
The market demand energetically is also constantly expanding.
At present, the propagation method of the beautiful millettia root several modes such as mainly there are seed growing, cutting propagation, tissue cultures to breed.
Seed growing and cuttage culture require higher in itself to beautiful millettia root seedling, and cultivation period is longer, are unfavorable for beautiful millettia root plantation and push away
Extensively；Therefore, excellent beautiful millettia root seedling is selected, is bred using the pattern of tissue cultures, is that a kind of efficiently breed cultivates ox
Method energetically, can greatly shorten the cultivation time of beautiful millettia root, and improve the survival rate of beautiful millettia root seedling, breed gained plant and be good for
Health, is conducive to improving the tuber formation and yield of beautiful millettia root.
It is different according to the required nutrient of plant growth and development process in plant tissue culture course, three kinds can be divided into
Culture medium, respectively inductive differentiation medium, root tissues culture medium, strong seedling culture base.Wherein, root tissues culture medium is played
The purpose that plant is taken root, directly determines the root system development degree of plant, so as to lift absorption of the tissue-cultured seedling to nutrient
Ability and transplanting survival rate.For different plants, the composition of root tissues culture medium also should be differentiated.
The disclosure of background above technology contents is only used for inventive concept and the technical scheme that auxiliary understands the present invention, and it is not
The prior art of present patent application is necessarily belonged to, the applying date of the above in present patent application is being shown without tangible proof
In the case of disclosed, above-mentioned background technology should not be taken to evaluate the novelty and creativeness of the application.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of beautiful millettia root root tissues culture medium, to realize to beautiful millettia root group
Train the efficient root induction of seedling.
In order to solve the above technical problems, the present invention uses following technical scheme：
A kind of beautiful millettia root root tissues culture medium, including following raw material：35~105g of agar, 12~48g of sucrose, hormone
0.3~1.2g, 3~14g of inorganic salt solution, 0.1~1.2g of nicotinic acid, 8~25g of activated carbon, 0.2~1.2g of cysteine, strepto-
0.1~1.5g of element, 7~18g of lactoalbumin hydrolysate, 40~120g of organic matter, 15~60g of tri-distilled water.
Preferably, described beautiful millettia root root tissues culture medium, including following raw material：40~90g of agar, sucrose 18~
42g, 0.5~1g of hormone, 5~12g of inorganic salt solution, 0.2~1g of nicotinic acid, 10~22g of activated carbon, 0.3~1g of cysteine, chain
0.2~1.2g of mycin, 8~15g of lactoalbumin hydrolysate, 50~110g of organic matter, 20~50g of tri-distilled water.
Preferably, described beautiful millettia root root tissues culture medium, including following raw material：45~80g of agar, sucrose 20~
36g, 0.6~0.8g of hormone, 6~10g of inorganic salt solution, 0.3~0.9g of nicotinic acid, 12~20g of activated carbon, cysteine 0.5~
0.8g, 0.3~0.9g of streptomysin, 10~14g of lactoalbumin hydrolysate, 55~95g of organic matter, 25~45g of tri-distilled water.
Preferably, described beautiful millettia root root tissues culture medium, the organic matter is included in milk, apple butter, banana puree
One kind.
It is highly preferred that described beautiful millettia root root tissues culture medium, the hormone is included in 6-BA, KT, CPPU, ABA
It is a kind of.
It is highly preferred that inorganic salts in described beautiful millettia root root tissues culture medium, its inorganic salt solution include Boratex,
One or more in zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, calcium chloride.
Further, the preparation method of described beautiful millettia root root tissues culture medium, comprises the following steps：
S1：After hormone is first dissolved with a small amount of 95% alcohol, add deionized water and be configured to solution；
S2：Sucrose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, hormone solution obtained by S1, inorganic salt solution, nicotinic acid, activity is subsequently added into
Charcoal, cysteine, streptomysin, lactoalbumin hydrolysate, organic matter, are sufficiently stirred for；
S4：The sucrose solution obtained by S2 is added into the solution made by S3, is sufficiently stirred for, uses 10%NaOH and 10%
It is 5~6.6 that HCl solution, which is adjusted to system pH,；
S5：Agar is added into the solution made by step S4, heating stirring is sub-packed in blake bottle after solution seethes with excitement
In；
S6：Induced tissue blake bottle made by step S5 was placed in high-pressure steam sterilizing pan after a period of time that sterilizes,
Root tissues culture medium is made.
Further, the preparation method of described beautiful millettia root root tissues culture medium, used reagent grade is point
Analysis is pure.
Further, the preparation method of described beautiful millettia root root tissues culture medium, blake bottle described in step S5 is tank
Head bottle.
Further, in the preparation method of described beautiful millettia root root tissues culture medium, step S6 sterilising temp be 120~
135 DEG C, sterilization time is 5~15min.
The invention has the advantages that：
(1) root tissues culture medium of the present invention, extensively, preparation method is easily operated, and Storage period for its ingredient origin
It is long；
(2) root tissues culture medium of the present invention, containing to promote the obvious boron of plant establishment, abscisic acid etc. into
Point；
(3) root tissues culture medium of the present invention can effectively facilitate the beautiful millettia root tissue-cultured seedling young and take root, lifting life
Root rate and transplanting survival rate.
Embodiment
The present invention is described in further detail with reference to embodiment.It is emphasized that the description below is only
Only it is exemplary, the scope being not intended to be limiting of the invention and its application.
Embodiment 1：
A kind of beautiful millettia root root tissues culture medium, including following raw material：Agar 35g, sucrose 12g, 6-BA0.3g, contain boron
Sour sodium, zinc chloride, potassium dihydrogen phosphate, magnesium chloride, the inorganic salt solution 3g of calcium chloride, nicotinic acid 0.1g, activated carbon 8g, cysteine
0.2g, streptomysin 0.1g, lactoalbumin hydrolysate 7g, milk 40g, tri-distilled water 15g.
The preparation method of described beautiful millettia root root tissues culture medium, comprises the following steps：
S1：After 6-BA is first dissolved with a small amount of 95% alcohol, add deionized water and be configured to solution；
S2：Sucrose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, 6-BA solution obtained by S1, inorganic salt solution, nicotinic acid, activity is subsequently added into
Charcoal, cysteine, streptomysin, lactoalbumin hydrolysate, milk, each reagent are that analysis is pure, are then sufficiently stirred for；
S4：The sucrose solution obtained by S2 is added into the solution made by S3, is sufficiently stirred for, uses 10%NaOH and 10%
It is 5.5 that HCl solution, which is adjusted to system pH,；
S5：Agar is added into the solution made by step S4, heating stirring is sub-packed in can culture after solution seethes with excitement
In bottle；
S6：Induced tissue blake bottle made by step S5 is placed in high-pressure steam sterilizing pan and sterilized at 120 DEG C
15min, that is, be made root tissues culture medium.
Embodiment 2
A kind of beautiful millettia root root tissues culture medium, including following raw material：Agar 105g, sucrose 48g, KT 1.2g, contain boron
Sour sodium, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, the inorganic salt solution 14g of calcium chloride, nicotinic acid 1.2g, activated carbon 25g, half Guang ammonia
Sour 1.2g, streptomysin 1.5g, lactoalbumin hydrolysate 18g, apple butter 120g, tri-distilled water 60g.
The preparation method of described beautiful millettia root root tissues culture medium, comprises the following steps：
S1：After KT is first dissolved with a small amount of 95% alcohol, add deionized water and be configured to solution；
S2：Sucrose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, be subsequently added into KT solution obtained by S1, inorganic salt solution, nicotinic acid, activated carbon,
Cysteine, streptomysin, lactoalbumin hydrolysate, apple butter, each reagent are that analysis is pure, are then sufficiently stirred for；
S4：The sucrose solution obtained by S2 is added into the solution made by S3, is sufficiently stirred for, uses 10%NaOH and 10%
It is 5.8 that HCl solution, which is adjusted to system pH,；
S5：Agar is added into the solution made by step S4, heating stirring is sub-packed in can culture after solution seethes with excitement
In bottle；
S6：Induced tissue blake bottle made by step S5 is placed in high-pressure steam sterilizing pan and sterilized at 135 DEG C
5min, that is, be made root tissues culture medium.
Embodiment 3
A kind of beautiful millettia root root tissues culture medium, including following raw material：Agar 40g, sucrose 18g, CPPU 0.5g, contain
Boratex, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, the inorganic salt solution 5g of calcium chloride, nicotinic acid 0.2g, activated carbon 10g, half Guang
Propylhomoserin 0.3g, streptomysin 0.2g, lactoalbumin hydrolysate 8g, banana puree 50g, tri-distilled water 20g.
The preparation method of described beautiful millettia root root tissues culture medium, comprises the following steps：
S1：After CPPU is first dissolved with a small amount of 95% alcohol, add deionized water and be configured to solution；
S2：Sucrose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, CPPU solution obtained by S1, inorganic salt solution, nicotinic acid, activity is subsequently added into
Charcoal, cysteine, streptomysin, lactoalbumin hydrolysate, banana puree, each reagent are that analysis is pure, are then sufficiently stirred for；
S4：The sucrose solution obtained by S2 is added into the solution made by S3, is sufficiently stirred for, uses 10%NaOH and 10%
It is 6.5 that HCl solution, which is adjusted to system pH,；
S5：Agar is added into the solution made by step S4, heating stirring is sub-packed in can culture after solution seethes with excitement
In bottle；
S6：Induced tissue blake bottle made by step S5 is placed in high-pressure steam sterilizing pan and sterilized at 125 DEG C
12min, that is, be made root tissues culture medium.
Embodiment 4
A kind of beautiful millettia root root tissues culture medium, including following raw material：Agar 90g, sucrose 42g, ABA 1g, contain boric acid
Sodium, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, the inorganic salt solution 12g of calcium chloride, nicotinic acid 1g, activated carbon 22g, half
Cystine 1g, streptomysin 1.2g, lactoalbumin hydrolysate 15g, milk 110g, tri-distilled water 50g.
The preparation method of described beautiful millettia root root tissues culture medium, comprises the following steps：
S1：After ABA is first dissolved with a small amount of 95% alcohol, add deionized water and be configured to solution；
S2：Sucrose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, be subsequently added into ABA solution obtained by S1, inorganic salt solution, nicotinic acid, activated carbon,
Cysteine, streptomysin, lactoalbumin hydrolysate, milk, each reagent are that analysis is pure, are then sufficiently stirred for；
S4：The sucrose solution obtained by S2 is added into the solution made by S3, is sufficiently stirred for, uses 10%NaOH and 10%
It is 5.8 that HCl solution, which is adjusted to system pH,；
S5：Agar is added into the solution made by step S4, heating stirring is sub-packed in can culture after solution seethes with excitement
In bottle；
S6：Induced tissue blake bottle made by step S5 is placed in high-pressure steam sterilizing pan and sterilized at 135 DEG C
6min, that is, be made root tissues culture medium.
Embodiment 5
A kind of beautiful millettia root root tissues culture medium, including following raw material：Agar 55g, sucrose 28g, ABA0.7g, inorganic salts
Solution 8g, nicotinic acid 0.6g, activated carbon 16g, cysteine 0.7g, streptomysin 0.5g, lactoalbumin hydrolysate 12g, apple butter 80g, three
Steam water 30g.
It is highly preferred that inorganic salts in described beautiful millettia root root tissues culture medium, its inorganic salt solution include Boratex,
One or more in zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, calcium chloride.
Further, the preparation method of described beautiful millettia root root tissues culture medium, comprises the following steps：
S1：After ABA is first dissolved with a small amount of 95% alcohol, add deionized water and be configured to solution；
S2：Sucrose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, be subsequently added into ABA solution obtained by S1, inorganic salt solution, nicotinic acid, activated carbon,
Cysteine, streptomysin, lactoalbumin hydrolysate, apple butter, each reagent are that analysis is pure, are then sufficiently stirred for；
S4：The sucrose solution obtained by S2 is added into the solution made by S3, is sufficiently stirred for, uses 10%NaOH and 10%
It is 6.2 that HCl solution, which is adjusted to system pH,；
S5：Agar is added into the solution made by step S4, heating stirring is sub-packed in can culture after solution seethes with excitement
In bottle；
S6：Induced tissue blake bottle made by step S5 is placed in high-pressure steam sterilizing pan and sterilized at 130 DEG C
8min, that is, be made root tissues culture medium.
In order to be described in more detail beneficial effects of the present invention, specific result of the test also provided below.
Beautiful millettia root fresh seeds 180 parts of explants as tissue cultures of section are chosen, A, B, C, D, E, F six is randomly divided into
Group, every group 30 parts.Every group sterilized through mercuric chloride after be respectively connected to the inducing culture of same composition, after cultivating 10 days, A~E groups kind
Seedling is moved into the root tissues culture medium being prepared into using formula and method described in the embodiment of the present invention 1~5, and the access of F groups is commercially available
In ordinary student Root tissue culture base, continue after cultivating 7 days, compare the rooting rate of each group.Move into planting site plantation afterwards to cultivate, record
Shoot survival percent is planted, experimental data is as shown in the table.
As can be seen here, beautiful millettia root root tissues culture medium of the present invention is compared with commercially available ordinary culture medium, Neng Gouti
The rooting rate of beautiful millettia root tissue-cultured seedling is risen, and transplanting survival rate is higher.
The specific implementation of the invention is not to be limited to these illustrations for above content, is led for technology belonging to the present invention
For the those of ordinary skill in domain, without departing from the inventive concept of the premise, some simple deduction or replace can also be made,
The scope of patent protection that the present invention is determined by the claims submitted should be all considered as belonging to.
Claims (10)
1. a kind of beautiful millettia root root tissues culture medium, it is characterised in that including following raw material：35~105g of agar, sucrose 12~
48g, 0.3~1.2g of hormone, 3~14g of inorganic salt solution, 0.1~1.2g of nicotinic acid, 8~25g of activated carbon, cysteine 0.2~
1.2g, 0.1~1.5g of streptomysin, 7~18g of lactoalbumin hydrolysate, 40~120g of organic matter, 15~60g of tri-distilled water.
2. beautiful millettia root root tissues culture medium according to claim 1, it is characterised in that including following raw material：Agar 40
~90g, 18~42g of sucrose, 0.5~1g of hormone, 5~12g of inorganic salt solution, 0.2~1g of nicotinic acid, 10~22g of activated carbon, half Guang
0.3~1g of propylhomoserin, 0.2~1.2g of streptomysin, 8~15g of lactoalbumin hydrolysate, 50~110g of organic matter, 20~50g of tri-distilled water.
3. beautiful millettia root root tissues culture medium according to claim 2, it is characterised in that including following raw material：Agar 45
~80g, 20~36g of sucrose, 0.6~0.8g of hormone, 6~10g of inorganic salt solution, 0.3~0.9g of nicotinic acid, 12~20g of activated carbon,
0.5~0.8g of cysteine, 0.3~0.9g of streptomysin, 10~14g of lactoalbumin hydrolysate, 55~95g of organic matter, tri-distilled water 25~
45g。
4. the beautiful millettia root root tissues culture medium according to any one of claims 1 to 3, it is characterised in that the organic matter
Including one kind in milk, apple butter, banana puree.
5. the beautiful millettia root root tissues culture medium according to any one of claims 1 to 3, it is characterised in that the hormone bag
Include one kind in 6-BA, KT, CPPU, ABA.
6. the beautiful millettia root root tissues culture medium according to any one of claims 1 to 3, it is characterised in that described is inorganic
Inorganic salts in salting liquid include one kind or several in Boratex, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, calcium chloride
Kind.
7. a kind of preparation method of beautiful millettia root root tissues culture medium according to any one of Claims 1 to 5, its feature exists
In comprising the following steps：
S1：After hormone is first dissolved with a small amount of 95% alcohol, add deionized water and be configured to solution；
S2：Sucrose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, hormone solution obtained by S1, inorganic salt solution, nicotinic acid, activated carbon, half is subsequently added into
Cystine, streptomysin, lactoalbumin hydrolysate, organic matter, are sufficiently stirred for；
S4：The sucrose solution obtained by S2 is added into the solution made by S3, is sufficiently stirred for, uses 10%NaOH and 10%HCl
It is 5~6.6 that solution, which is adjusted to system pH,；
S5：Agar is added into the solution made by step S4, heating stirring is sub-packed in blake bottle after after solution boiling；
S6：Induced tissue blake bottle made by step S5 was placed in high-pressure steam sterilizing pan after a period of time that sterilizes, that is, made
Into root tissues culture medium.
8. the preparation method of beautiful millettia root root tissues culture medium according to claim 7, it is characterised in that used examination
Agent rank is that analysis is pure.
9. the preparation method of beautiful millettia root root tissues culture medium according to claim 7, it is characterised in that institute in step S5
Blake bottle is stated for can.
10. the preparation method of beautiful millettia root root tissues culture medium according to claim 7, it is characterised in that in step S6
Sterilising temp is 120~135 DEG C, and sterilization time is 5~15min.
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| WO2021109714A1 (en) * | 2019-12-06 | 2021-06-10 | 华南理工大学 | Millettia speciosa polysaccharides, preparation method therefor, and application thereof in antibacterial aspects |
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| CN113416689A (en) * | 2021-08-04 | 2021-09-21 | 海南鑫开源医药科技有限公司 | Method for producing Korean sophoricoside by using plant cell fermentation technology |
| CN113416689B (en) * | 2021-08-04 | 2023-01-10 | 海南鑫开源医药科技有限公司 | Method for producing Korean sophoricoside by using plant cell fermentation technology |
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