CN106962199A - A kind of beautiful millettia root antibacterial tissue culture medium (TCM) and preparation method thereof - Google Patents
A kind of beautiful millettia root antibacterial tissue culture medium (TCM) and preparation method thereof Download PDFInfo
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- CN106962199A CN106962199A CN201710306804.8A CN201710306804A CN106962199A CN 106962199 A CN106962199 A CN 106962199A CN 201710306804 A CN201710306804 A CN 201710306804A CN 106962199 A CN106962199 A CN 106962199A
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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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Abstract
The invention discloses a kind of beautiful millettia root antibacterial tissue culture medium (TCM) and preparation method thereof, belong to tissue culture medium (TCM) technical field, antibacterial tissue culture medium (TCM) includes following raw material：Agar, glucose, hormone, inorganic salt solution, αnaphthylacetate sodium, silver carboxymethyl chitosan, riboflavin, zeatin, caseinhydrolysate, organic matter, tri-distilled water.The antibacterial tissue culture medium (TCM) being prepared into using raw material of the present invention and method, it is to be prepared according to Nutrient Properties needed for the growth of beautiful millettia root tissue-cultured seedling, enhancing development can be played a part of to beautiful millettia root tissue-cultured seedling, while having antibacterial functions concurrently, can effectively suppress the growth of bacterium in culture medium.
Description
Technical field
The invention belongs to culture medium preparing technical field, and in particular to a kind of beautiful millettia root antibacterial tissue culture medium (TCM).
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 media, strong sprout tissue culture medium (TCM).Tissue culture medium (TCM) plays promotion tissue culture
The purpose that seedling grows, while tissue-cultured seedling nutrient is supplied to, moreover it is possible to lift absorbability of the tissue-cultured seedling to nutrient, indirectly
Ground transplanting survival rate.For different plants, the composition of tissue culture medium (TCM) also should be differentiated.Due to containing in tissue culture medium (TCM)
A large amount of inorganic matters and organic matter, easily cause bacteria breed, are unfavorable for the healthy growth of tissue-cultured seedling.Silver carboxymethyl chitosan is one
The high antibacterial material of biocompatibility is planted, tissue culture medium (TCM) is added to, can effectively suppress the growth of bacterium in culture medium.
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 antibacterial tissue culture medium (TCM), the culture medium is providing foster
While dividing, antibacterial effect can be played.
In order to solve the above technical problems, the present invention uses following technical scheme：
A kind of beautiful millettia root antibacterial tissue culture medium (TCM), including following raw material：40~110g of agar, 15~45g of glucose, hormone
0.3~1.5g, 3~15g of inorganic salt solution, 0.2~1.2g of α-naphthaleneacidsodium, 0.1~0.6g of silver carboxymethyl chitosan, riboflavin
0.2~1.2g, 0.2~1.5g of zeatin, 10~30g of caseinhydrolysate, 55~125g of organic matter, 20~60g of tri-distilled water.
Preferably, described beautiful millettia root antibacterial tissue culture medium (TCM), including following raw material：50~100g of agar, glucose 18
~40g, 0.4~1.2g of hormone, 5~12g of inorganic salt solution, 0.3~1g of α-naphthaleneacidsodium, silver carboxymethyl chitosan 0.2~
0.5g, 0.3~0.9g of riboflavin, 0.3~1.2g of zeatin, 12~26g of caseinhydrolysate, 65~110g of organic matter, tri-distilled water
25~50g.
Preferably, described beautiful millettia root antibacterial tissue culture medium (TCM), including following raw material：60~90g of agar, glucose 20~
35g, 0.5~1g of hormone, 6~10g of inorganic salt solution, 0.4~0.8g of α-naphthaleneacidsodium, 0.3~0.4g of silver carboxymethyl chitosan,
0.4~0.8g of riboflavin, 0.4~1g of zeatin, 14~22g of caseinhydrolysate, 75~95g of organic matter, 28~42g of tri-distilled water.
Preferably, described beautiful millettia root antibacterial tissue culture medium (TCM), the organic matter includes mashed potatoes, apple butter, banana puree
In one kind.
Preferably, described beautiful millettia root antibacterial tissue culture medium (TCM), the hormone includes one in NAA, 6-BA, IBA, ABA
Kind.
It is highly preferred that the inorganic salts in described beautiful millettia root antibacterial tissue culture medium (TCM), described inorganic salt solution include boron
One or more in sour sodium, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, manganese chloride, calcium chloride.
It is highly preferred that the preparation method of described beautiful millettia root antibacterial tissue culture medium (TCM), 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：Glucose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, be subsequently added into hormone solution obtained by S1, inorganic salt solution, α-naphthaleneacidsodium,
Silver carboxymethyl chitosan, riboflavin, zeatin, caseinhydrolysate, organic matter, are sufficiently stirred for；
S4：The glucose solution obtained by S2 is added into the solution made by S3, is sufficiently stirred for, using 10%NaOH and
It is 5~6.6 that 10%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,
Antibacterial tissue culture medium (TCM) is made.
Further, the preparation method of described beautiful millettia root antibacterial tissue culture medium (TCM), used reagent grade is point
Analysis is pure.
Further, the preparation method of described beautiful millettia root antibacterial tissue culture medium (TCM), blake bottle described in step S5 is tank
Head bottle.
Further, sterilising temp is 120 in the preparation method of described beautiful millettia root antibacterial tissue culture medium (TCM), step S6
~135 DEG C, sterilization time is 5~15min.
The invention has the advantages that：
(1) antibacterial tissue culture medium (TCM) of the present invention, its raw material components wide material sources, preparation method is easily operated, and storage
Deposit the phase long；
(2) antibacterial tissue culture medium (TCM) of the present invention, contains the hormone that beautiful millettia root tissue-cultured seedling can be promoted to grow
The silver carboxymethyl chitosan of composition and antibacterial functions, culture medium bacteria breed is suppressed while beautiful millettia root seedling is cultivated.
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 antibacterial tissue culture medium (TCM), including following raw material：Agar 40g, glucose 15g, NAA0.3g, contain
Boratex, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, manganese chloride, inorganic salt solution 3g, the α-naphthaleneacidsodium of calcium chloride
0.2~1.2g, silver carboxymethyl chitosan 0.1g, riboflavin 0.2g, zeatin 0.2g, caseinhydrolysate 10g, mashed potatoes 55g, three
Steam water 20g.
The preparation method of described beautiful millettia root antibacterial tissue culture medium (TCM), comprises the following steps：
S1：After NNA is first dissolved with a small amount of 95% alcohol, add deionized water and be configured to solution；
S2：Glucose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, be subsequently added into NNA solution obtained by S1, inorganic salt solution, α-naphthaleneacidsodium,
Silver carboxymethyl chitosan, riboflavin, zeatin, caseinhydrolysate, mashed potatoes, used reagent grade are that analysis is pure, are filled
Divide stirring；
S4：The glucose solution obtained by S2 is added into the solution made by S3, is sufficiently stirred for, using 10%NaOH and
It is 5.8 that 10%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 antibacterial tissue culture medium (TCM).
Embodiment 2
A kind of beautiful millettia root antibacterial tissue culture medium (TCM), including following raw material：Agar 110g, glucose 45g, 6-BA1.5g, contain
There are inorganic salt solution 15g, α-naphthalene second of Boratex, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, manganese chloride, calcium chloride
Sour sodium 1.2g, silver carboxymethyl chitosan 0.6g, riboflavin 1.2g, zeatin 1.5g, caseinhydrolysate 30g, apple butter 125g, three
Steam water 60g.
The preparation method of described beautiful millettia root antibacterial tissue culture medium (TCM), 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：Glucose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, be subsequently added into 6-BA solution obtained by S1, inorganic salt solution, α-naphthaleneacidsodium,
Silver carboxymethyl chitosan, riboflavin, zeatin, caseinhydrolysate, apple butter, used reagent grade are that analysis is pure, are filled
Divide stirring；
S4：The glucose solution obtained by S2 is added into the solution made by S3, is sufficiently stirred for, using 10%NaOH and
It is 6.6 that 10%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 antibacterial tissue culture medium (TCM).
Embodiment 3
A kind of beautiful millettia root antibacterial tissue culture medium (TCM), including following raw material：Agar 50g, glucose 18g, IBA 0.4g, contain
Boratex, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, manganese chloride, inorganic salt solution 5g, the α-naphthaleneacidsodium of calcium chloride
0.3g, silver carboxymethyl chitosan 0.2g, riboflavin 0.3g, zeatin 0.3g, caseinhydrolysate 12g, banana puree 65g, tri-distilled water
25g。
The preparation method of described beautiful millettia root antibacterial tissue culture medium (TCM), comprises the following steps：
S1：After IBA is first dissolved with a small amount of 95% alcohol, add deionized water and be configured to solution；
S2：Glucose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, be subsequently added into IBA solution obtained by S1, inorganic salt solution, α-naphthaleneacidsodium,
Silver carboxymethyl chitosan, riboflavin, zeatin, caseinhydrolysate, banana puree, used reagent grade are that analysis is pure, are filled
Divide stirring；
S4：The glucose solution obtained by S2 is added into the solution made by S3, is sufficiently stirred for, using 10%NaOH and
It is 5.5 that 10%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 antibacterial tissue culture medium (TCM).
Embodiment 4
A kind of beautiful millettia root antibacterial tissue culture medium (TCM), including following raw material：Agar 100g, glucose 40g, ABA1.2g, contain
Boratex, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, manganese chloride, the inorganic salt solution 12g of calcium chloride, α-naphthylacetic acid
Sodium 1g, silver carboxymethyl chitosan 0.5g, riboflavin 0.9g, zeatin 1.2g, caseinhydrolysate 26g, mashed potatoes 110g, tri-distilled water
50g。
The preparation method of described beautiful millettia root antibacterial tissue culture medium (TCM), 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：Glucose 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, α-naphthaleneacidsodium,
Silver carboxymethyl chitosan, riboflavin, zeatin, caseinhydrolysate, mashed potatoes, used reagent grade are that analysis is pure, are filled
Divide stirring；
S4：The glucose solution obtained by S2 is added into the solution made by S3, is sufficiently stirred for, using 10%NaOH and
It is 5.2 that 10%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 antibacterial tissue culture medium (TCM).
Embodiment 5
A kind of beautiful millettia root antibacterial tissue culture medium (TCM), including following raw material：Agar 70g, glucose 28g, NNA0.7g, contain
Boratex, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, manganese chloride, inorganic salt solution 8g, the α-naphthaleneacidsodium of calcium chloride
0.5g, silver carboxymethyl chitosan 0.3g, riboflavin 0.6g, zeatin 0.6g, caseinhydrolysate 18g, apple butter 82g, tri-distilled water
36g。
The preparation method of described beautiful millettia root antibacterial tissue culture medium (TCM), comprises the following steps：
S1：After NNA is first dissolved with a small amount of 95% alcohol, add deionized water and be configured to solution；
S2：Glucose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, be subsequently added into NNA solution obtained by S1, inorganic salt solution, α-naphthaleneacidsodium,
Silver carboxymethyl chitosan, riboflavin, zeatin, caseinhydrolysate, apple butter, used reagent grade are that analysis is pure, are filled
Divide stirring；
S4：The glucose solution obtained by S2 is added into the solution made by S3, is sufficiently stirred for, using 10%NaOH and
It is 5.8 that 10%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
7min, that is, be made antibacterial tissue culture medium (TCM).
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.After every group sterilizes through mercuric chloride, A~E groups seedling is moved into using formula and method described in the embodiment of the present invention 1~5
In the antibacterial tissue culture medium (TCM) being prepared into, F groups are accessed in commercially available ordinary culture medium, after cultivating 10 days, compare each group culture medium shape
Condition, and it is as shown in the table to record experimental data.
| Group | Bacterium colony pollution rate/% | Tissue-cultured seedling induces differentiation rate/% |
| A | 8 | 675 |
| B | 6 | 733 |
| C | 11 | 751 |
| D | 9 | 697 |
| E | 9 | 720 |
| F | 21 | 543 |
As can be seen here, beautiful millettia root antibacterial tissue culture medium (TCM) of the present invention is compared with commercially available ordinary culture medium, culture medium
Bacterium colony pollution rate it is relatively low, induction differentiation rate is high, can play the effect grown for promoting beautiful millettia root seedling.
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 antibacterial tissue culture medium (TCM), it is characterised in that including following raw material：40~110g of agar, glucose 15
~45g, 0.3~1.5g of hormone, 3~15g of inorganic salt solution, 0.2~1.2g of α-naphthaleneacidsodium, silver carboxymethyl chitosan 0.1~
0.6g, 0.2~1.2g of riboflavin, 0.2~1.5g of zeatin, 10~30g of caseinhydrolysate, 55~125g of organic matter, tri-distilled water
20~60g.
2. beautiful millettia root antibacterial tissue culture medium (TCM) according to claim 1, it is characterised in that including following raw material：Agar 50
~100g, 18~40g of glucose, 0.4~1.2g of hormone, 5~12g of inorganic salt solution, 0.3~1g of α-naphthaleneacidsodium, carboxymethyl
0.2~0.5g of chitosan silver, 0.3~0.9g of riboflavin, 0.3~1.2g of zeatin, 12~26g of caseinhydrolysate, organic matter 65
~110g, 25~50g of tri-distilled water.
3. beautiful millettia root antibacterial tissue culture medium (TCM) according to claim 2, it is characterised in that including following raw material：Agar 60
~90g, 20~35g of glucose, 0.5~1g of hormone, 6~10g of inorganic salt solution, 0.4~0.8g of α-naphthaleneacidsodium, carboxymethyl shell
0.3~0.4g of glycan silver, 0.4~0.8g of riboflavin, 0.4~1g of zeatin, 14~22g of caseinhydrolysate, organic matter 75~
95g, 28~42g of tri-distilled water.
4. the beautiful millettia root antibacterial tissue culture medium (TCM) according to any one of claims 1 to 3, it is characterised in that the organic matter
Including one kind in mashed potatoes, apple butter, banana puree.
5. the beautiful millettia root antibacterial tissue culture medium (TCM) according to any one of claims 1 to 3, it is characterised in that the hormone bag
Include one kind in NAA, 6-BA, IBA, ABA.
6. the beautiful millettia root antibacterial tissue culture medium (TCM) according to any one of claims 1 to 3, it is characterised in that described is inorganic
Inorganic salts in salting liquid are included in Boratex, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, manganese chloride, calcium chloride
It is one or more of.
7. a kind of preparation method of beautiful millettia root antibacterial tissue culture medium (TCM) 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：Glucose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, hormone solution obtained by S1, inorganic salt solution, α-naphthaleneacidsodium, carboxylic first is subsequently added into
Base enclosure glycan silver, riboflavin, zeatin, caseinhydrolysate, organic matter, are sufficiently stirred for；
S4：The glucose 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 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 antibacterial tissue culture medium (TCM).
8. the preparation method of beautiful millettia root antibacterial tissue culture medium (TCM) 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 antibacterial tissue culture medium (TCM) 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 antibacterial tissue culture medium (TCM) 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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| 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 |
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