CN109287482A - A kind of Yellow Fructus Gardeniae tissue-culturing rapid propagation special culture media - Google Patents
A kind of Yellow Fructus Gardeniae tissue-culturing rapid propagation special culture media Download PDFInfo
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- CN109287482A CN109287482A CN201811279135.0A CN201811279135A CN109287482A CN 109287482 A CN109287482 A CN 109287482A CN 201811279135 A CN201811279135 A CN 201811279135A CN 109287482 A CN109287482 A CN 109287482A
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- Prior art keywords
- fructus gardeniae
- yellow fructus
- promotor
- tissue
- rapid propagation
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Classifications
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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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/22—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing plant material
- A01G24/25—Dry fruit hulls or husks, e.g. chaff or coir
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/30—Growth substrates; Culture media; Apparatus or methods therefor based on or containing synthetic organic compounds
Abstract
The present invention provides a kind of Yellow Fructus Gardeniae tissue-culturing rapid propagation special culture medias, and the rooting rate and transplanting survival rate of Yellow Fructus Gardeniae can be improved.The promotor compounded by the modification pine nut shell and 1-DNJ of special ratios is added in the present invention in conventional root media, and the rooting rate of Yellow Fructus Gardeniae can be improved, especially can be shortened the time taken root.And it can achieve 100% at rear rate after transplanting.In addition, it has been found that promotor of the present invention is added in MS culture medium, additive amount 1.0mg/L, for cultivating Yellow Fructus Gardeniae callus, the content of Gardenoside in callus can be greatly improved, open new purposes for promotor of the present invention.
Description
Technical field
The present invention relates to a kind of Yellow Fructus Gardeniae tissue-culturing rapid propagation special culture medias, belong to Chinese medicine numerous field fastly
Background technique
Yellow Fructus Gardeniae alias cape jasmine, fructus gardeniae, are used as medicine with fruit and root, there is purging fire for removing toxin, clearing heat and promoting diuresis, the effect of cool blood scattered silt.
Be distributed in Zhejiang, Jiangxi, Fujian, Hubei, Sichuan, Guizhou etc. province, Yichuan area be mainly distributed on camphor tree, Fengcheng, the ten thousand years,
The ground such as Fengxin.
Yellow Fructus Gardeniae tradition mode of reproduction is plant division or cuttage, and this method breeding coefficient is low, speed is slow, breeding cycle is long, pole
The earth hinders the cultivation and popularization of cape jasmine.Its breeding coefficient can be improved using the method for tissue cultures, shorten incubation time.But
Existing culture medium still has several drawbacks in terms of improving transplanting survival rate and active constituent content, is further improved.
Summary of the invention
The purpose of the present invention is to provide a kind of Yellow Fructus Gardeniae tissue-culturing rapid propagation special culture medias, and the rooting rate of Yellow Fructus Gardeniae can be improved
And transplanting survival rate.
The technical solution of the present invention is as follows:
A kind of Yellow Fructus Gardeniae tissue-culturing rapid propagation special culture media is then to add the rush of 0.5mg/L using 1/2MS as minimal medium
Into agent, the agar of the sucrose of 30g/L and 7g/L, pH value is 5.8~6.0.
The promotor is to be mixed to get by modified pine nut shell and 1-DNJ with mass ratio 5:1.
The modified pine nut shell the preparation method comprises the following steps: pine nut shell is crushed to 40 mesh, 12h, mistake are impregnated in modified solution
Filter, filter residue are washed to washing lotion repeatedly with distilled water and are in neutrality, and moisture content is then dried at 100 DEG C 8% hereinafter, to obtain the final product.
The modified solvent is the aqueous solution of triethylene diamine and choline chloride, and wherein the mass fraction of triethylene diamine is
6.5%, the mass fraction of choline chloride is 2.5%.
Waste during the red SEMEN PINI KORAIENSIS processing of the pine nut shell contains lignin, stilbene class, volatilization in pine nut shell
The various bioactivators such as oil, vitamins, Palmatine, minerals, polysaccharide, protein, fat and flavonoids.The present invention adopts
It is raw material with pine nut shell, turns waste into wealth, economizes on resources, has also achieved the purpose that protect environment.
Modification pine nut shell and 1-DNJ by special ratios is added in the present invention in conventional root media
Obtained promotor is compounded, the rooting rate of Yellow Fructus Gardeniae can be improved, especially can be shortened the time taken root.And transplant after at rear
Rate can achieve 100%.
In addition, it has been found that promotor of the present invention is added in MS culture medium, additive amount 1.0mg/L, uses
In culture Yellow Fructus Gardeniae callus, the content of Gardenoside in callus can be greatly improved, is opened newly for promotor of the present invention
Purposes.
The present invention has the advantages that adding promotor of the present invention in conventional 1/2MS culture medium, yellow Cape jasmine can be improved
The situation of taking root of son shortens rootage duration, the survival rate after improving transplanting.And the promotor is added in regular MS media,
Determination of Gardenoside in cape jasmine callus can be improved.
Specific embodiment
Embodiment 1: it is minimal medium that a kind of Yellow Fructus Gardeniae tissue-culturing rapid propagation special culture media, which is 1/2MS, then adds 0.5mg/
The agar of the promotor of L, the sucrose of 30g/L and 7g/L, pH value are 5.8~6.0.
The promotor is to be mixed to get by modified pine nut shell and 1-DNJ with mass ratio 5:1.
The modified pine nut shell the preparation method comprises the following steps: pine nut shell is crushed to 40 mesh, 12h, mistake are impregnated in modified solution
Filter, filter residue are washed to washing lotion repeatedly with distilled water and are in neutrality, and moisture content is then dried at 100 DEG C 8% hereinafter, to obtain the final product.
The modified solvent is the aqueous solution of triethylene diamine and choline chloride, and wherein the mass fraction of triethylene diamine is
6.5%, the mass fraction of choline chloride is 2.5%.
Embodiment 2: it is minimal medium that a kind of Yellow Fructus Gardeniae tissue-culturing rapid propagation special culture media, which is 1/2MS, then adds 0.5mg/
The agar of the promotor of L, the sucrose of 30g/L and 7g/L, pH value are 5.8~6.0.
The promotor is 1-DNJ.
Embodiment 3: it is minimal medium that a kind of Yellow Fructus Gardeniae tissue-culturing rapid propagation special culture media, which is 1/2MS, then adds 0.5mg/
The agar of the promotor of L, the sucrose of 30g/L and 7g/L, pH value are 5.8~6.0.
The promotor is modified pine nut shell, preparation method are as follows: pine nut shell is crushed to 40 mesh, is soaked in modified solution
Steep 12h, filtering, filter residue washs to washing lotion repeatedly with distilled water and is in neutrality, moisture content is then dried at 100 DEG C 8% hereinafter,
To obtain the final product.
The modified solvent is the aqueous solution of triethylene diamine and choline chloride, and wherein the mass fraction of triethylene diamine is
6.5%, the mass fraction of choline chloride is 2.5%.
Embodiment 4: it is minimal medium that a kind of Yellow Fructus Gardeniae tissue-culturing rapid propagation special culture media, which is 1/2MS, then adds 0.5mg/
The agar of the promotor of L, the sucrose of 30g/L and 7g/L, pH value are 5.8~6.0.
The promotor is to be mixed to get by being crushed to the pine nut shell after 40 mesh and 1-DNJ with mass ratio 5:1
's.
Embodiment 5: difference from example 1 is that:
The modified solvent is triethylene diamine aqueous solution, and wherein the mass fraction of triethylene diamine is 6.5%.
Remaining is the same as embodiment 1.
Embodiment 6: difference from example 1 is that:
The modified solvent is the aqueous solution of choline chloride, and wherein the mass fraction of choline chloride is 2.5%.
Remaining is the same as embodiment 1.
Embodiment 7: a kind of Yellow Fructus Gardeniae callus culture medium using MS as minimal medium, then adds the rush of 1.0mg/L
Into agent.The formula of the promotor and the preparation method is the same as that of Example 1.
Embodiment 8: a kind of Yellow Fructus Gardeniae callus culture medium using MS as minimal medium, then adds the rush of 1.0mg/L
Into agent.The promotor is the same as embodiment 2.
Embodiment 9: a kind of Yellow Fructus Gardeniae callus culture medium using MS as minimal medium, then adds the rush of 1.0mg/L
Into agent.The formula and preparation method of the promotor are the same as embodiment 3.
Embodiment 10: a kind of Yellow Fructus Gardeniae callus culture medium using MS as minimal medium, then adds 1.0mg/L's
Promotor.The formula and preparation method of the promotor are the same as embodiment 4.
Embodiment 11: a kind of Yellow Fructus Gardeniae callus culture medium using MS as minimal medium, then adds 1.0mg/L's
Promotor.The formula and preparation method of the promotor are the same as embodiment 5.
Embodiment 12: a kind of Yellow Fructus Gardeniae callus culture medium using MS as minimal medium, then adds 1.0mg/L's
Promotor.The formula and preparation method of the promotor are the same as embodiment 6.
Experiment 1: by 3-4cm long, the Yellow Fructus Gardeniae Multiple Buds with 3-5 piece leaf are cut into single plant and are seeded in embodiment 1- respectively
Root induction is carried out on 6 culture medium.Separately set a blank control group and a hormone control group, the culture medium of blank control group
Are as follows: the agar of the sucrose+7g/L of 1/2MS minimal medium+30g/L, the culture medium of hormone control group are as follows: 1/2MS is cultivated substantially
The fine jade of the sucrose+7g/L of base+0.2mg/L NAA (optium concentration that the promotion Yellow Fructus Gardeniae Multiple Buds filtered out are taken root)+30g/L
Rouge.Cultivation temperature is (25+1) DEG C, relative humidity 70%, intensity of illumination 2400lx, daily illumination 12h.Each group after statistics 20d
It takes root situation, then by each group cape jasmine transplantation of seedlings to matrix is fertile soil: land: sandy soil=3:2:1 nutritive cube (nutritive cube bottom
Diameter 8cm, high 10cm) in, it sprinkles profoundly water, is placed in greenhouse after transplanting.Greenhouse temperature is 15~30 DEG C, relative humidity 11-
65%, 50% shading, plant investigated survival rate after 2 weeks.
The influence that 1 different culture medium of table takes root to Yellow Fructus Gardeniae
| Grouping | 20d rooting rate (%) | Most short rootage duration (d) |
| Blank control group | 34.3 | 11 |
| Hormone control group | 100 | 7 |
| Embodiment 1 | 100 | 5 |
| Embodiment 2 | 36.9 | 12 |
| Embodiment 3 | 34.5 | 13 |
| Embodiment 4 | 41.2 | 11 |
| Embodiment 5 | 40.8 | 10 |
| Embodiment 6 | 37.7 | 12 |
Influence of 2 different culture medium of table to Yellow Fructus Gardeniae transplanting survival rate
| Grouping | Transplanting survival rate (%) |
| Blank control group | 62 |
| Hormone control group | 95 |
| Embodiment 1 | 100 |
| Embodiment 2 | 62 |
| Embodiment 3 | 65 |
| Embodiment 4 | 64 |
| Embodiment 5 | 60 |
| Embodiment 6 | 64 |
Experiment 2: it is transferred to after taking Yellow Fructus Gardeniae seed water rinsed clean with 75% (volume fraction, similarly hereinafter) ethanol disinfection 1min
10min is sterilized in 2% liquor natrii hypochloritis, aseptic water washing 5 times, is seeded in MS culture medium (MS+2.0mg/L 6BA+ respectively
+ 0.6% agar of 0.50mg/L NAA+2% sucrose) on sprout, inducing temperature (25 ± 1) DEG C, light application time 16h/d, light intensity l
200Lux obtained aseptic seedling after 2 weeks.Cape jasmine young tender leaf is taken, young leaflet tablet is cut into the fritter of 1cm × 1cm with scissors, is connect respectively
Kind carries out callus tissue culture in the culture medium of embodiment 7-12.A blank control group and a hormone control group separately are set,
The culture medium of blank control group are as follows: MS minimal medium, the culture medium of hormone control group are as follows: MS minimal medium+1.0mg/L6-
BA.It is 25 DEG C in temperature, illumination is 3 000Lx, and culture 30d can be obtained callus under conditions of irradiating 14h daily, is measured
The content of Gardenoside, measuring method are as follows in callus:
(1) Determination of Gardenoside measurement chromatographic condition and system suitability in Yellow Fructus Gardeniae callus: with octadecyl
Silane group silica gel is filler；Own nitrile-water (volume ratio 15:85) is mobile phase；Detection wavelength 238nm；Number of theoretical plate is with Cape jasmine
Sub- glycosides peak, which calculates, should be not less than 3 000.
(2) preparation of reference substance solution: taking Gardenoside reference substance appropriate, accurately weighed, adds methanol that every lmL is made and contains
The solution of 0.1mg to get.
(3) preparation of cape jasmine callus extracting solution: weighing 25mg callus dry powder, accurately weighed, sets 25mL measuring bottle
In, add methanol to dissolve and be diluted to scale, shakes up；Precision draws 2mL, sets in 10mL measuring bottle, adds methanol dilution to scale, shake
It is even to get.
(4) measuring method: it is accurate respectively to draw reference substance solution and each 10 μ L of test solution, liquid chromatograph is injected,
Measurement to get.
Influence of 3 different culture medium of table to Determination of Gardenoside in Yellow Fructus Gardeniae callus
| Grouping | Determination of Gardenoside (mg/g) |
| Blank control group | 17.23 |
| Hormone control group | 21.18 |
| Embodiment 1 | 95.66 |
| Embodiment 2 | 18.41 |
| Embodiment 3 | 17.35 |
| Embodiment 4 | 18.29 |
| Embodiment 5 | 17.80 |
| Embodiment 6 | 17.58 |
Claims (4)
1. a kind of Yellow Fructus Gardeniae tissue-culturing rapid propagation special culture media, it is characterised in that: then added using 1/2MS as minimal medium
The agar of the promotor of 0.5mg/L, the sucrose of 30g/L and 7g/L, pH value are 5.8~6.0.
2. a kind of Yellow Fructus Gardeniae tissue-culturing rapid propagation special culture media as described in claim 1, it is characterised in that: the promotor be by
What modified pine nut shell and 1-DNJ were mixed to get with mass ratio 5:1；
The modified pine nut shell the preparation method comprises the following steps: pine nut shell is crushed to 40 mesh, 12h is impregnated in modified solution, is filtered, filter
Slag is washed to washing lotion repeatedly with distilled water and is in neutrality, and moisture content is then dried at 100 DEG C 8% hereinafter, to obtain the final product.
3. a kind of Yellow Fructus Gardeniae tissue-culturing rapid propagation special culture media as claimed in claim 2, it is characterised in that: the modified solvent is
The aqueous solution of triethylene diamine and choline chloride, wherein the mass fraction of triethylene diamine is 6.5%, the quality point of choline chloride
Number is 2.5%.
4. application of the promotor in improving Yellow Fructus Gardeniae callus in Determination of Gardenoside in claim 2, it is characterised in that:
Promotor of the present invention is added in MS culture medium, additive amount 1.0mg/L, for cultivating Yellow Fructus Gardeniae callus.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811279135.0A CN109287482B (en) | 2018-10-30 | 2018-10-30 | Special culture medium for tissue culture and rapid propagation of gardenia jasminoides |
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| CN201811279135.0A CN109287482B (en) | 2018-10-30 | 2018-10-30 | Special culture medium for tissue culture and rapid propagation of gardenia jasminoides |
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| CN109287482B CN109287482B (en) | 2022-03-01 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114747486A (en) * | 2022-04-08 | 2022-07-15 | 湖南省林业科学院 | Somatic embryogenesis and plant regeneration method for gardenia jasminoides ellis |
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