CN109601382A - Fluid nutrient medium and preparation method thereof suitable for dendrobium candidum tender stem - Google Patents
Fluid nutrient medium and preparation method thereof suitable for dendrobium candidum tender stem Download PDFInfo
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- CN109601382A CN109601382A CN201811620941.XA CN201811620941A CN109601382A CN 109601382 A CN109601382 A CN 109601382A CN 201811620941 A CN201811620941 A CN 201811620941A CN 109601382 A CN109601382 A CN 109601382A
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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
Coconut juice, potato extract and other substances are added the 5-7 times of water measured at room temperature and mixed, obtain A product by the fluid nutrient medium and preparation method thereof that present invention discloses a kind of suitable for dendrobium candidum tender stem；2,4-D and NAA are pre-dissolved using dehydrated alcohol, the solution of 0.5-2mg/ml is formulated as, obtains B product.By A, B product loaded on mixing in tissue culture bottle, sterilize 20-30min under high-temperature steam, and cooling to obtain the final product.The present invention passes through the concentration for adding and controlling coconut juice and potato extract, to promote the arginine content and polyoses content of dendrobium candidum tender stem.
Description
Technical field
The present invention relates to field of agricultural cultivation, specifically, more particularly to a kind of liquid suitable for dendrobium candidum tender stem
Culture medium and preparation method thereof.
Background technique
Dendrobium candidum (Dendrobium officinale) is a kind of many years draft Chinese herbal medicine, and rule belongs to angiosperm
Door (Magnoliophyta), Liliopsida (Liliopsida), Liliales (Asparagales), orchid family (Orchidaceae), stone
Dry measure used in former times category (Dendrobium).It is recorded in Chinese Pharmacopoeia (version in 2015), dendrobium candidum has clearing heat-fire, removing toxic substances, and Li Shui disappears
It is swollen, treat pyreticosis and polydipsia, abscess of throat, edema and little urine, puckery pain of heat gonorrhea and carbuncle, swelling and furuncle and other effects.Equally, modern medicine study
Show that dendrobium candidum has the function of enhancing immune t-cell, can be improved body immunity, removes the thin of human senility and mutation
Born of the same parents, stabilizing blood sugar improve cell metabolic rates, the function of maintaining organism physiology stable.
Dendrobium officinale is generally grown on stone precipice or ancient tree, and the dendrobium candidum habit for being grown on stone precipice is referred to as stone
Dry measure used in former times, pharmaceutical activity are good；The dendrobium candidum habit for being grown on ancient tree is referred to as the wooden dry measure used in former times, drug activity relative mistake.Still regardless of dendrobium nobile
The wooden dry measure used in former times, they grow relatively slowly, and very rare, along with to the immoderate acquisition of dendrobium candidum milli and abuse, wild iron
Skin dendrobium nobile is classified as secondary plant protection kind by country.
Though dendrobium candidum shows extensive artificial growth, since dendrobium candidum own growth is slow and cultivated area is few,
Its annual output is only able to satisfy the demand in a province.In addition, the dendrobium candidum of outdoor plantation needs to occupy a large amount of cultivated area,
The cross pollination between the pollution of pesticide residue and heavy metal, and different dendrobium species is also faced simultaneously and causes hereditary shakiness
The problems such as fixed.In addition, will use a large amount of inorganic fertilizer when dendrobium candidum artificial growth, easily causing environmental pollution and iron
Skin dendrobium nobile quality decline.Using plant tissue culture technique, traditional iron sheet growing dendrobium problem encountered not only can solve,
But also purposeful orientation regulation can be carried out to cultivating system, to improve plant tissue nutritional ingredient or obtain specific cells generation
Thank to product.
Therefore, there are the more patented technology manually cultivated for dendrobium candidum, such as Chinese patent Shen at present
Please CN201510371264.2, CN201510375351.5, CN201710368696.7,201710375637.2,
CN201711119122.2, etc..But these culture mediums belong to earth culture culture both for the growth of dendrobium candidum first
Base, without the fluid nutrient medium specifically for dendrobium candidum tender stem growth；Secondly, organic additive is added to also fewerly
In the culture medium of dendrobium candidum；Moreover, the arginine content and polyoses content of current dendrobium candidum tender stem are also not satisfactory.
Summary of the invention
The purpose of the present invention is overcoming the shortcomings of the prior art, a kind of liquid suitable for dendrobium candidum tender stem is provided
Culture medium and preparation method thereof.
The purpose of the present invention is achieved through the following technical solutions:
A kind of fluid nutrient medium suitable for dendrobium candidum tender stem, formula components composition are as follows:
PH value is 5.6-6.0, remaining is Main elements, microelement, molysite and organic matter.
Preferably, the specific formula of the Main elements, microelement, molysite and organic matter is as follows at being grouped as:
Specifically, being suitable for the fluid nutrient medium of dendrobium candidum tender stem, formula components composition is as follows:
Present invention discloses a kind of preparation methods of fluid nutrient medium suitable for dendrobium candidum tender stem, including walk as follows
Suddenly,
S1, by coconut juice, potato extract, the Main elements including potassium, nitrogen, phosphorus, magnesium, including iodine, boron, molybdenum
The 5-7 times of water measured is added at room temperature and mixes for microelement, molysite and organic matter, obtains A product；
2,4-D and NAA are pre-dissolved using dehydrated alcohol, are formulated as the solution of 0.5-2mg/ml, obtain B product by S2；
S3, by A, B product loaded on mixing in tissue culture bottle, sterilize 20-30min under high-temperature steam, and cooling to obtain the final product.
Preferably, the sucrose is carbon source；Potassium nitrate is nitrogen source, potassium resource；Ammonium nitrate is nitrogen source；Magnesium sulfate is magnesium source；Iodine
Change potassium is propiodal；Boric acid is boron source, and sodium molybdate is molybdenum source, and potassium dihydrogen phosphate is phosphorus source.
The beneficial effects are mainly reflected as follows: by adding and controlling potato extract, coconut juice, to improve iron sheet stone
The arginine content and polyoses content of dry measure used in former times tender stem.In addition, and being insoluble in water since 2,4-D and NAA belong to micro, needing
It is diluted with water after being pre-dissolved in alcohol, therefore fluid nutrient medium of the invention uses mutually applicable specific preparation method.
Specific embodiment
Below with reference to embodiment, the present invention is further illustrated, but is not intended as the foundation limited the present invention.
It is different from the existing earth culture culture medium about dendrobium candidum, the liquid suitable for dendrobium candidum tender stem of the invention
The formula components composition of culture medium is as follows:
In the present invention, the effect of sucrose is carbon source；Potassium nitrate is nitrogen source, potassium resource；Ammonium nitrate is nitrogen source；The effect of magnesium sulfate
It is magnesium source, magnesium is the important component of chlorophyll；Potassium iodide is as propiodal；Boric acid plays promotion, adjustment effect as boron source；
Catalyst of the manganese sulfate as synthesis chlorophyll；Zinc sulfate has the function of the synthesis for promoting photosynthesis, participating in heteroauxin；
Sodium molybdate provides molybdenum source, and molybdenum is the ingredient of nitrate reductase, participates in the reduction of nitrate and nitrogen metabolism in plant；Ethylenediamine
Tetraacethyl disodium is the good compounding agent that the present invention selects；Ferrous sulfate supplements outside the intracorporal ferro element of plant as source of iron,
It can also promote its absorption to nitrogen and P elements；Inositol alcohol works in the mutually converting of carbohydrate, and is the building material of cell wall
Material；Glycine has unique effect to the photosynthesis of crop, is conducive to plant growth, increases the content of crop sugar, native metal
Chelating agent；Thiamine hydrochloride is that organism aerobic respiration must substance；Niacin has good influence to the formation of embryoid and bud；Phosphorus
Acid dihydride potassium is as phosphorus source；2,4-D and NAA are as growth regulator.Inventor it has been investigated that: Main elements, microelement,
The specific formula specific gravity of molysite and organic matter is also extremely important for the performance of fluid nutrient medium of the invention.
It should be noted that potato extract and coconut juice contain certain plant hormone, be conducive to the growth of tissue culture plant,
And containing certain peroxidase and superoxide radical dismutase and vitamin C, oxygen excessive in air can be prevented,
Negative oxygen ion is generated to damage tissue culture plant.Coconut juice also can promote tissue cultures activity；Moreover, starch in potato extract
Rich content can also be used as the carbon source supplement of culture.
Present invention further teaches a kind of preparation methods of fluid nutrient medium suitable for dendrobium candidum tender stem, including walk as follows
It is rapid:
S1, by coconut juice, potato extract, the Main elements including potassium, nitrogen, phosphorus, magnesium, including iodine, boron, molybdenum
The 5-7 times of water measured is added at room temperature and mixes for microelement, molysite and organic matter, obtains A product；
2,4-D and NAA are pre-dissolved using dehydrated alcohol, are formulated as the solution of 0.5-2mg/ml, obtain B product by S2；
S3, by A, B product loaded on mixing in tissue culture bottle, sterilize 20-30min under high-temperature steam, and cooling to obtain the final product.
Specific formula embodiment is as follows:
Embodiment 1
Formula: sucrose 20g/L, potassium nitrate 50mg/L, ammonium nitrate 80.04mg/L, magnesium sulfate 246.47mg/L, calcium chloride
147.02mg/L, potassium iodide 166.01mg/L, boric acid 61.83mg/L, manganese sulfate 223.01mg/L, zinc sulfate 287.54mg/L,
Sodium molybdate 241.95mg/L, copper sulphate 249.68mg/L, cobalt chloride 237.93mg/L, disodium ethylene diamine tetraacetate 372.25mg/
L, ferrous sulfate 278.03mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride
0.5mg/L, niacin 0.5mg/L, potassium dihydrogen phosphate 75mg/L, coconut juice 10%, potato extract 10%, 2,4-D 2mg/L, NAA
0.5mg/L。
Embodiment 2
Formula: sucrose 20g/L, potassium nitrate 50mg/L, ammonium nitrate 80.04mg/L, magnesium sulfate 246.47mg/L, calcium chloride
147.02mg/L, potassium iodide 166.01mg/L, boric acid 61.83mg/L, manganese sulfate 223.01mg/L, zinc sulfate 287.54mg/L,
Sodium molybdate 241.95mg/L, copper sulphate 249.68mg/L, cobalt chloride 237.93mg/L, disodium ethylene diamine tetraacetate 372.25mg/
L, ferrous sulfate 278.03mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride
0.5mg/L, niacin 0.5mg/L, potassium dihydrogen phosphate 75mg/L, coconut juice 20%, potato extract 10%, 2,4-D 2mg/L, NAA
0.5mg/L。
Embodiment 3
Formula: sucrose 10g/L, potassium nitrate 50mg/L, ammonium nitrate 80.04mg/L, magnesium sulfate 246.47mg/L, calcium chloride
147.02mg/L, potassium iodide 166.01mg/L, boric acid 61.83mg/L, manganese sulfate 223.01mg/L, zinc sulfate 287.54mg/L,
Sodium molybdate 241.95mg/L, copper sulphate 249.68mg/L, cobalt chloride 237.93mg/L, disodium ethylene diamine tetraacetate 372.25mg/
L, ferrous sulfate 278.03mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride
0.5mg/L, niacin 0.5mg/L, potassium dihydrogen phosphate 75mg/L, coconut juice 20%, potato extract 15%, 2,4-D 2mg/L, NAA
0.5mg/L。
Embodiment 4
Formula: sucrose 10g/L, potassium nitrate 50mg/L, ammonium nitrate 80.04mg/L, magnesium sulfate 246.47mg/L, calcium chloride
147.02mg/L, potassium iodide 166.01mg/L, boric acid 61.83mg/L, manganese sulfate 223.01mg/L, zinc sulfate 287.54mg/L,
Sodium molybdate 241.95mg/L, copper sulphate 249.68mg/L, cobalt chloride 237.93mg/L, disodium ethylene diamine tetraacetate 372.25mg/
L, ferrous sulfate 278.03mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride
0.5mg/L, niacin 0.5mg/L, potassium dihydrogen phosphate 75mg/L, coconut juice 10%, potato extract 20%, 2,4-D 2mg/L, NAA
0.5mg/L。
Embodiment 5
Formula: sucrose 30g/L, potassium nitrate 50mg/L, ammonium nitrate 80.04mg/L, magnesium sulfate 246.47mg/L, calcium chloride
147.02mg/L, potassium iodide 166.01mg/L, boric acid 61.83mg/L, manganese sulfate 223.01mg/L, zinc sulfate 287.54mg/L,
Sodium molybdate 241.95mg/L, copper sulphate 249.68mg/L, cobalt chloride 237.93mg/L, disodium ethylene diamine tetraacetate 372.25mg/
L, ferrous sulfate 278.03mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride
0.5mg/L, niacin 0.5mg/L, potassium dihydrogen phosphate 75mg/L, coconut juice 20%, potato extract 20%, 2,4-D 2mg/L, NAA
0.5mg/L。
Embodiment 6
Formula: sucrose 30g/L, potassium nitrate 50mg/L, ammonium nitrate 80.04mg/L, magnesium sulfate 246.47mg/L, calcium chloride
147.02mg/L, potassium iodide 166.01mg/L, boric acid 61.83mg/L, manganese sulfate 223.01mg/L, zinc sulfate 287.54mg/L,
Sodium molybdate 241.95mg/L, copper sulphate 249.68mg/L, cobalt chloride 237.93mg/L, disodium ethylene diamine tetraacetate 372.25mg/
L, ferrous sulfate 278.03mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride
0.5mg/L, niacin 0.5mg/L, potassium dihydrogen phosphate 75mg/L, coconut juice 15%, potato extract 15%, 2,4-D 2mg/L, NAA
0.5mg/L。
Comparative example 1
Formula: sucrose 20g/L, potassium nitrate 50mg/L, ammonium nitrate 80.04mg/L, magnesium sulfate 246.47mg/L, calcium chloride
147.02mg/L, potassium iodide 166.01mg/L, boric acid 61.83mg/L, manganese sulfate 223.01mg/L, zinc sulfate 287.54mg/L,
Sodium molybdate 241.95mg/L, copper sulphate 249.68mg/L, cobalt chloride 237.93mg/L, disodium ethylene diamine tetraacetate 372.25mg/
L, ferrous sulfate 278.03mg/L, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride
0.5mg/L, niacin 0.5mg/L, potassium dihydrogen phosphate 75mg/L, 2,4-D 2mg/L, NAA 0.5mg/L.
The above culture medium is respectively inoculated with 10 bottles by preparation method of the present invention, is (22 ± 2) DEG C, natural lighting in cultivation temperature,
Light application time is cultivated under the conditions of 14 hour/day.It is tender to be chosen at the good same batch dendrobium candidum of upgrowth situation in above-mentioned culture medium
Stem counts the arginine and polyoses content of dendrobium candidum tender stem after culture 45 days.
| Culture medium | Arginine content | Polyoses content |
| Embodiment 1 | 2.0% | 19.3% |
| Embodiment 2 | 1.7% | 17.6% |
| Embodiment 3 | 1.6% | 18.0% |
| Embodiment 4 | 1.5% | 18.3% |
| Embodiment 5 | 1.7% | 18.8% |
| Embodiment 6 | 1.4% | 17.8% |
| Comparative example 1 | 0.5% | 10.7% |
It can be seen that dendrobium candidum tender stem growth conditions in culture medium of the present invention are good, arginine in above-described embodiment
Content is greatly improved with polyoses content.It should be appreciated that although this specification is described in terms of embodiments, but it is not every
A embodiment only contains an independent technical solution, and this description of the specification is merely for the sake of clarity, ability
Field technique personnel should consider the specification as a whole, and the technical solution in each embodiment may also be suitably combined, shape
The other embodiments that skilled artisan is understood that.
The series of detailed descriptions listed above only for feasible embodiment of the invention specifically
Protection scope bright, that they are not intended to limit the invention, it is all without departing from equivalent implementations made by technical spirit of the present invention
Or change should all be included in the protection scope of the present invention.
Claims (5)
1. a kind of fluid nutrient medium suitable for dendrobium candidum tender stem, it is characterised in that: its formula components composition is as follows:
Sucrose 10g/L-30g/L
Coconut juice 10-20%
Potato extract 10-20%
2,4-D 1mg/L-3mg/L
NAA 0.3mg/L-1mg/L
PH value is 5.6-6.0, remaining is Main elements, microelement, molysite and organic matter.
2. the fluid nutrient medium according to claim 1 suitable for dendrobium candidum tender stem, which is characterized in that the main amount member
Element, microelement, the specific formula of molysite and organic matter are as follows at being grouped as:
Potassium nitrate 50-55mg/L
Ammonium nitrate 80-85mg/L
Magnesium sulfate 245-250mg/L
Calcium chloride 145-150mg/L
Potassium iodide 165-170mg/L
Boric acid 60-65mg/L
Manganese sulfate 220-225mg/L
Zinc sulfate 285-290mg/L
Sodium molybdate 240-245mg/L
Copper sulphate 245-250mg/L
Cobalt chloride 235-240mg/L
Disodium ethylene diamine tetraacetate 370-375mg/L
Ferrous sulfate 275-280mg/L
Inositol 100-105mg/L
Glycine 1-5mg/L
Thiamine hydrochloride 0.05-0.2mg/L
Puridoxine hydrochloride 0.1-1mg/L
Niacin 0.1-1mg/L
Potassium dihydrogen phosphate 70-80mg/L.
3. the fluid nutrient medium according to claim 1 suitable for dendrobium candidum tender stem, which is characterized in that its formula components
It forms as follows:
Sucrose 10g/L-30g/L
Potassium nitrate 50mg/L
Ammonium nitrate 80.04mg/L
Magnesium sulfate 246.47mg/L
Calcium chloride 147.02mg/L
Potassium iodide 166.01mg/L
Boric acid 61.83mg/L
Manganese sulfate 223.01mg/L
Zinc sulfate 287.54mg/L
Sodium molybdate 241.95mg/L
Copper sulphate 249.68mg/L
Cobalt chloride 237.93mg/L
Disodium ethylene diamine tetraacetate 372.25mg/L
Ferrous sulfate 278.03mg/L
Inositol 100mg/L
Glycine 2mg/L
Thiamine hydrochloride 0.1mg/L
Puridoxine hydrochloride 0.5mg/L
Niacin 0.5mg/L
Potassium dihydrogen phosphate 75mg/L
Coconut juice 10-20%
Potato extract 10-20%
2,4-D 2mg/L
NAA 0.5mg/L
PH value is 5.6-6.0.
4. a kind of preparation method of the fluid nutrient medium suitable for dendrobium candidum tender stem as described in claims 1 to 3 is any,
It is characterized in that: including the following steps,
S1, by coconut juice, potato extract, the Main elements including potassium, nitrogen, phosphorus, magnesium, micro including iodine, boron, molybdenum
The 5-7 times of water measured is added at room temperature and mixes for element, molysite and organic matter, obtains A product；
2,4-D and NAA are pre-dissolved using dehydrated alcohol, are formulated as the solution of 0.5-2mg/ml, obtain B product by S2；
S3, by A, B product loaded on mixing in tissue culture bottle, sterilize 20-30min under high-temperature steam, and cooling to obtain the final product.
5. the preparation method according to claim 4, it is characterised in that: the sucrose is carbon source；Potassium nitrate is nitrogen source, potassium
Source；Ammonium nitrate is nitrogen source；Magnesium sulfate is magnesium source；Potassium iodide is propiodal；Boric acid is boron source, and sodium molybdate is molybdenum source, potassium dihydrogen phosphate
For phosphorus source.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811620941.XA CN109601382A (en) | 2018-12-28 | 2018-12-28 | Fluid nutrient medium and preparation method thereof suitable for dendrobium candidum tender stem |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811620941.XA CN109601382A (en) | 2018-12-28 | 2018-12-28 | Fluid nutrient medium and preparation method thereof suitable for dendrobium candidum tender stem |
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Country Status (1)
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103583358A (en) * | 2013-10-24 | 2014-02-19 | 湖南农业大学 | Method for in vitro culturing of regenerated plant of dendrobium officinale |
| CN104396722A (en) * | 2014-07-18 | 2015-03-11 | 皖西学院 | Method for cultivating triploid dendrobium huoshanense by crossbreeding method of tetraploid plants and diploid plants |
-
2018
- 2018-12-28 CN CN201811620941.XA patent/CN109601382A/en not_active Withdrawn
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103583358A (en) * | 2013-10-24 | 2014-02-19 | 湖南农业大学 | Method for in vitro culturing of regenerated plant of dendrobium officinale |
| CN104396722A (en) * | 2014-07-18 | 2015-03-11 | 皖西学院 | Method for cultivating triploid dendrobium huoshanense by crossbreeding method of tetraploid plants and diploid plants |
Non-Patent Citations (1)
| Title |
|---|
| 蒋林等: "添加剂对铁皮石斛组织培养和快速繁殖的影响", 《中药材》 * |
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