CN106962200A - A kind of beautiful millettia root strong sprout tissue culture medium (TCM) and preparation method thereof - Google Patents
A kind of beautiful millettia root strong sprout tissue culture medium (TCM) and preparation method thereof Download PDFInfo
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- CN106962200A CN106962200A CN201710306843.8A CN201710306843A CN106962200A CN 106962200 A CN106962200 A CN 106962200A CN 201710306843 A CN201710306843 A CN 201710306843A CN 106962200 A CN106962200 A CN 106962200A
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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 strong sprout tissue culture medium (TCM) and preparation method thereof, belong to tissue culture medium (TCM) technical field, strong sprout tissue culture medium (TCM) includes following raw material：Agar, glucose, hormone, inorganic salt solution, αnaphthylacetate sodium, activated carbon, riboflavin, zeatin, caseinhydrolysate, organic matter, tri-distilled water.The strong sprout tissue culture medium (TCM) being prepared into using raw material of the present invention and method, is to be prepared according to Nutrient Properties needed for the growth of beautiful millettia root tissue-cultured seedling, the tissue-cultured seedling strong sprout effect to beautiful millettia root is good, and survival rate is high after transplanting, may advantageously facilitate efficiently breeding for 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 strong sprout 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).Wherein, strong sprout tissue culture medium (TCM) is played
The purpose for promoting tissue-cultured seedling to grow, so as to strengthen the health of tissue-cultured seedling, lifts absorbability of the tissue-cultured seedling to nutrient,
Transplanting survival rate indirectly.For different plants, the composition of strong sprout tissue culture medium (TCM) 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 strong sprout tissue culture medium (TCM), to realize beautiful millettia root tissue culture
The strong seedling cultivation of seedling.
In order to solve the above technical problems, the present invention uses following technical scheme：
A kind of beautiful millettia root strong sprout tissue culture medium (TCM), including following raw material：28~90g of agar, 7~22g of glucose, hormone
0.2~1g, 2~10g of inorganic salt solution, 0.2~0.8g of α-naphthaleneacidsodium, 9~25g of activated carbon, 0.1~0.7g of riboflavin, jade
0.1~0.8g of rice element, 10~30g of caseinhydrolysate, 55~125g of organic matter, 20~60g of tri-distilled water.
Preferably, described beautiful millettia root strong sprout tissue culture medium (TCM), including following raw material：30~85g of agar, glucose 10~
18g, 0.3~0.8g of hormone, 3~9g of inorganic salt solution, 0.3~0.7g of α-naphthaleneacidsodium, 10~22g of activated carbon, riboflavin 0.3
~0.6g, 0.2~0.6g of zeatin, 12~26g of caseinhydrolysate, 65~110g of organic matter, 25~50g of tri-distilled water.
Preferably, described beautiful millettia root strong sprout tissue culture medium (TCM), including following raw material：35~70g of agar, glucose 12~
16g, 0.4~0.6g of hormone, 4~7g of inorganic salt solution, 0.4~0.6g of α-naphthaleneacidsodium, 12~20g of activated carbon, riboflavin 0.4
~0.5g, 0.3~0.5g of zeatin, 14~22g of caseinhydrolysate, 75~95g of organic matter, 28~42g of tri-distilled water.
Preferably, described beautiful millettia root strong sprout tissue culture medium (TCM), the organic matter includes mashed potatoes, apple butter, banana puree
In one kind.
Preferably, described beautiful millettia root strong sprout tissue culture medium (TCM), the hormone includes one in GA, TDZ, CPPU, ABA
Kind.
It is highly preferred that the inorganic salts in described beautiful millettia root strong sprout 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 strong sprout 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,
Activated carbon, 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,
Strong sprout tissue culture medium (TCM) is made.
Further, the preparation method of described beautiful millettia root strong sprout tissue culture medium (TCM), used reagent grade is point
Analysis is pure.
Further, the preparation method of described beautiful millettia root strong sprout 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 strong sprout tissue culture medium (TCM), step S6
~135 DEG C, sterilization time is 5~15min.
The invention has the advantages that：
(1) strong sprout tissue culture medium (TCM) of the present invention, its raw material rents a point wide material sources, and preparation method is easily operated, and storage
Deposit the phase long；
(2) strong sprout tissue culture medium (TCM) of the present invention, promotees containing manganese, α-naphthylacetic acid for promoting beautiful millettia root seedling development etc.
Enter composition, and contained inorganic salt concentration is low, can effectively facilitate the growth of tissue-cultured seedling；
(3) strong sprout tissue culture medium (TCM) of the present invention can effectively facilitate that beautiful millettia root tissue-cultured seedling straw development is sturdy, bud
Growth.
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 strong sprout tissue culture medium (TCM), including following raw material：Agar 28g, glucose 7g, GA0.2g, contain boron
Sour sodium, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, manganese chloride, inorganic salt solution 2g, the α-naphthaleneacidsodium of calcium chloride
0.2g, activated carbon 9g, riboflavin 0.1g, zeatin 0.1g, caseinhydrolysate 10g, mashed potatoes 55g, tri-distilled water 20g.
The preparation method of described beautiful millettia root strong sprout tissue culture medium (TCM), comprises the following steps：
S1：After GA 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, GA solution obtained by S1, inorganic salt solution, α-naphthaleneacidsodium, work is subsequently added into
Property charcoal, riboflavin, zeatin, caseinhydrolysate, mashed potatoes, each reagent be analysis it is pure, be 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 5.5 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 120 DEG C
15min, that is, be made strong sprout tissue culture medium (TCM).
Further, sterilising temp is 120 in the preparation method of described beautiful millettia root strong sprout tissue culture medium (TCM), step S6
~135 DEG C, sterilization time is 5~15min.
Embodiment 2
A kind of beautiful millettia root strong sprout tissue culture medium (TCM), including following raw material：Agar 90g, glucose 22g, TDZ 1g, contain boron
Sour sodium, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, manganese chloride, inorganic salt solution 10g, the α-naphthaleneacidsodium of calcium chloride
0.8g, activated carbon 25g, riboflavin 0.7g, zeatin 0.8g, caseinhydrolysate 30g, apple butter 125g, tri-distilled water 60g.
The preparation method of described beautiful millettia root strong sprout tissue culture medium (TCM), comprises the following steps：
S1：After TDZ 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 TDZ solution obtained by S1, inorganic salt solution, α-naphthaleneacidsodium,
Activated carbon, riboflavin, zeatin, caseinhydrolysate, apple butter, each reagent are that analysis is pure, 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 6.5 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 135 DEG C
5min, that is, be made strong sprout tissue culture medium (TCM).
Embodiment 3
A kind of beautiful millettia root strong sprout tissue culture medium (TCM), including following raw material：Agar 30g, glucose 10g, CPPU0.3g, contain
Boratex, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, manganese chloride, inorganic salt solution 3g, the α-naphthaleneacidsodium of calcium chloride
0.3g, activated carbon 10g, riboflavin 0.3g, zeatin 0.2g, caseinhydrolysate 12g, banana puree 65g, tri-distilled water 25g.
The preparation method of described beautiful millettia root strong sprout tissue culture medium (TCM), 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：Glucose is configured to 10% solution using deionized water；
S3：Tri-distilled water is added into beaker, be subsequently added into CPPU solution obtained by S1, inorganic salt solution, α-naphthaleneacidsodium,
Activated carbon, riboflavin, zeatin, caseinhydrolysate, banana puree, each reagent are that analysis is pure, 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 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 strong sprout tissue culture medium (TCM).
Embodiment 4
A kind of beautiful millettia root strong sprout tissue culture medium (TCM), including following raw material：Agar 85g, glucose 18g, ABA0.8g, contain
Boratex, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, manganese chloride, inorganic salt solution 9g, the α-naphthaleneacidsodium of calcium chloride
0.7g, activated carbon 22g, riboflavin 0.6g, zeatin 0.6g, caseinhydrolysate 26g, mashed potatoes 110g, tri-distilled water 50g.
The preparation method of described beautiful millettia root strong sprout 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,
Activated carbon, riboflavin, zeatin, caseinhydrolysate, mashed potatoes, each reagent are that analysis is pure, 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 5.5 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 135 DEG C
5min, that is, be made strong sprout tissue culture medium (TCM).
Embodiment 5
A kind of beautiful millettia root strong sprout tissue culture medium (TCM), including following raw material：Agar 55g, glucose 15g, GA0.5g, contain boron
Sour sodium, zinc chloride, ammonium chloride, potassium dihydrogen phosphate, magnesium chloride, manganese chloride, inorganic salt solution 6g, the α-naphthaleneacidsodium of calcium chloride
0.5g, activated carbon 15g, riboflavin 0.5g, zeatin 0.4g, caseinhydrolysate 18g, apple butter 85g, tri-distilled water 36g.
The preparation method of described beautiful millettia root strong sprout tissue culture medium (TCM), comprises the following steps：
S1：After GA 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, GA solution obtained by S1, inorganic salt solution, α-naphthaleneacidsodium, work is subsequently added into
Property charcoal, riboflavin, zeatin, caseinhydrolysate, apple butter, each reagent be analysis it is pure, be 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 6.2 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 128 DEG C
12min, that is, be made strong sprout 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.Every group sterilized through mercuric chloride after be respectively connected to the inducing culture of same composition, move into identical life after cultivating 10 days
Cultivate, continue after cultivating 10 days in root culture medium, A~E groups seedling is moved into using formula described in the embodiment of the present invention 1~5 and side
In the strong sprout tissue culture medium (TCM) that method is prepared into, F groups are accessed in commercially available common strong sprout tissue culture medium (TCM), are continued after cultivating 10 days, than
Compared with the upgrowth situation of each group tissue-cultured seedling, experimental data is as shown in the table.
| Group | Stalk average diameter/mm | Average every plant of leaf bud number/ |
| A | 2.8 | 8.2 |
| B | 3.1 | 8.6 |
| C | 2.7 | 7.7 |
| D | 3.0 | 9.3 |
| E | 2.8 | 8.4 |
| F | 2.1 | 6.7 |
As can be seen here, beautiful millettia root strong sprout tissue culture medium (TCM) of the present invention is compared with commercially available ordinary culture medium, Neng Gouti
The sturdy degree of stalk and the average leaf bud number of increase of beautiful millettia root tissue-cultured seedling are risen, the growth hair of beautiful millettia root seedling can be effectively facilitated
Educate.
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 strong sprout tissue culture medium (TCM), it is characterised in that including following raw material：28~90g of agar, glucose 7~
22g, 0.2~1g of hormone, 2~10g of inorganic salt solution, 0.2~0.8g of α-naphthaleneacidsodium, 9~25g of activated carbon, riboflavin 0.1~
0.7g, 0.1~0.8g of zeatin, 10~30g of caseinhydrolysate, 55~125g of organic matter, 20~60g of tri-distilled water.
2. beautiful millettia root strong sprout tissue culture medium (TCM) according to claim 1, it is characterised in that including following raw material：Agar 30
~85g, 10~18g of glucose, 0.3~0.8g of hormone, 3~9g of inorganic salt solution, 0.3~0.7g of α-naphthaleneacidsodium, activated carbon
10~22g, 0.3~0.6g of riboflavin, 0.2~0.6g of zeatin, 12~26g of caseinhydrolysate, 65~110g of organic matter, three steam
25~50g of water.
3. beautiful millettia root strong sprout tissue culture medium (TCM) according to claim 2, it is characterised in that including following raw material：Agar 35
~70g, 12~16g of glucose, 0.4~0.6g of hormone, 4~7g of inorganic salt solution, 0.4~0.6g of α-naphthaleneacidsodium, activated carbon
12~20g, 0.4~0.5g of riboflavin, 0.3~0.5g of zeatin, 14~22g of caseinhydrolysate, 75~95g of organic matter, three steam
28~42g of water.
4. the beautiful millettia root strong sprout 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 strong sprout 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 GA, TDZ, CPPU, ABA.
6. the beautiful millettia root strong sprout 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 strong sprout 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, activity is subsequently added into
Charcoal, 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 strong sprout tissue culture medium (TCM).
8. the preparation method of beautiful millettia root strong sprout 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 strong sprout 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 strong sprout 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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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102893870A (en) * | 2012-10-09 | 2013-01-30 | 冯志祥 | Rapid propagation and seedling raising method for beautiful millettia root seedling tissue culture |
| CN104737905A (en) * | 2015-02-14 | 2015-07-01 | 涂湘炎 | Millettia specisoa tissue culture rooting method |
-
2017
- 2017-05-04 CN CN201710306843.8A patent/CN106962200A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102893870A (en) * | 2012-10-09 | 2013-01-30 | 冯志祥 | Rapid propagation and seedling raising method for beautiful millettia root seedling tissue culture |
| CN104737905A (en) * | 2015-02-14 | 2015-07-01 | 涂湘炎 | Millettia specisoa tissue culture rooting method |
Non-Patent Citations (4)
| Title |
|---|
| 张传海等: "《植物生物技术》", 30 September 2015, 合肥工业大学出版社 * |
| 时群等: "牛大力组织培养瓶内复壮瓶外生根快繁技术", 《林业实用技术》 * |
| 胡颂平等: "《植物细胞组织培养技术》", 31 August 2014, 中国农业大学出版社 * |
| 韦莹等: "牛大力离体培养与植株再生条件的优化", 《湖北农业科学》 * |
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Application publication date: 20170721 |