CN106613955A - Chinese herbal medicine extracting solution used for stevia rebaudiana culture medium and preparation method - Google Patents

Chinese herbal medicine extracting solution used for stevia rebaudiana culture medium and preparation method Download PDF

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CN106613955A
CN106613955A CN201610950090.XA CN201610950090A CN106613955A CN 106613955 A CN106613955 A CN 106613955A CN 201610950090 A CN201610950090 A CN 201610950090A CN 106613955 A CN106613955 A CN 106613955A
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culture medium
parts
herbal medicine
chinese herbal
stevia rebaudiana
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贾德军
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Mingguang City A Total Of Stevia Professional Cooperatives
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Mingguang City A Total Of Stevia Professional Cooperatives
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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
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/40—Liliopsida [monocotyledons]
    • A01N65/48—Zingiberaceae [Ginger family], e.g. ginger or galangal

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Environmental Sciences (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Agronomy & Crop Science (AREA)
  • Plant Pathology (AREA)
  • Dentistry (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Developmental Biology & Embryology (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

The invention belongs to the technical field of stevia rebaudiana culture mediums and provides a Chinese herbal medicine extracting solution used for a stevia rebaudiana culture medium and a preparation method. The Chinese herbal medicine extracting solution used for the stevia rebaudiana culture medium is prepared from the following Chinese herbal medicine components: lignum dalbergiae odoriferae, piper longum, pseudobulbus cremastrae seu pleiones, spora lygodii, Bianjiang and mahonia fortunei; the Chinese herbal medicine extracting solution is used for any one of a stevia rebaudiana callus culture medium, a protocorm culture medium, a subculture culture medium, a passage culture medium and a rooting culture medium. All the Chinese herbal medicines are prepared through techniques such as ethanol digestion, boiling and concentration. Aiming at disadvantages of a bacterium-inhibiting measure in existing stevia rebaudiana tissue culture, the Chinese herbal medicines are added into a basic culture medium and the culture medium is enabled to be located in an immune environment, so that pollution caused by microorganisms including external bacteria and the like is prevented, and furthermore, the efficient and lasting bacterium inhibition effect is realized; meanwhile, the resistance of stevia rebaudiana explants is improved, the investment of manpower and material resources is reduced and the cost is reduced. The method is simple to operate and has a good extracting effect.

Description

A kind of extracts of Chinese herbal medicine and preparation method for STEVIA REBAUDIANA culture medium
Technical field
The invention belongs to STEVIA REBAUDIANA culture medium technical field, in particular it relates to a kind of medium-height grass for STEVIA REBAUDIANA culture medium Medicament extracting solution and preparation method.
Background technology
In recent years STEVIA REBAUDIANA tissue culture technique have developed rapidly, and in tissue culture procedures, be commonly present the microorganisms such as bacterium Pollution problem, causes that STEVIA REBAUDIANA tissue cultures are of poor quality, and efficiency is low, the plantation yield of serious nutrition STEVIA REBAUDIANA, is to perplex this skill The difficult problem of art area research personnel.
The microorganism pollution produced in STEVIA REBAUDIANA tissue culture procedures is essentially from two aspects：One is vegetable material band itself Bacterium；Two be in operator and air institute band microorganism caused by；In addition, pollution is also possible to because culture medium carries disease germs, operation tool band Caused by the cross pollution of bacterium.The microorganism of pollution is divided into bacterium and the big class of fungi two.The characteristics of germ contamination is：Bacterial plaque is in mucus Shape, and after inoculation 2～3d can have found.The link of microorganism is produced in STEVIA REBAUDIANA tissue culture procedures mainly to be had：Pollution Staff has been used without caused by the bacterium discharged when adequately disinfected instrument and breathing, sometimes also because operating personnel use Hand contact material or vessel edge, which increases microorganism and fall into the chance of material or vessel, it should be specifically noted that avoiding intersecting Pollution, such as sterilized inoculating tool be sterilized not enough thoroughly pollute again after the pollution of superclean bench face the material sterilized with Sterilized culture medium etc..Fungal contamination is typically caused by the air pollution of transfer room more.The reason for formation, one is transfer room Air itself without very well sterilization, two when being operation due to when opening bottle cap fungal spore fall into bottle.
Therefore it is intended in the links of tissue cultures under normal circumstances in strict accordance with the requirement of sterile working, carries out nothing Bacterium cultivate, explant after sterilization, in being inoculated in the culture medium for sterilizing, i.e., into the link of sterile culture.Culture Interior need to clean few bacterium, and the method for employing has and fill indoors uviol lamp, sterilizing of periodically turning on light, and also can use potassium permanganate and formaldehyde mixed Close liquid periodically stifling etc..It is above-mentioned to prevent the measure of microorganism pollution in tissue culture procedures from having certain effect, its weak point Be that each antibacterial link isolates, it is impossible to form complete unified antibacterial chain, if during any one link negligence of operation, Microorganism is just taken advantage of a weak point, and causes tissue cultures to fail, and tissue cultures quality and efficiency is reduced, additionally, by going out to culture medium Bacterium, aseptic inoculation is cultivated in gnotobasis, is and is reached the antibacterial purpose of sterilizing by external operation, one in these links The microorganisms such as the then bacterial infection not in place that denier any one link is done, it is therefore desirable to a kind of persistently to play arranging for bacteria resistance function Apply, make STEVIA REBAUDIANA explant be chronically at a gnotobasis, while strengthen the inherent bacteriostasis of STEVIA REBAUDIANA explant, opposing The pollution of external microorganism.
The content of the invention
For defect of the prior art, on the one hand the purpose of the present invention provides a kind of medium-height grass for STEVIA REBAUDIANA culture medium Medicament extracting solution, for the deficiency of antibacterial measure in existing STEVIA REBAUDIANA tissue cultures, by adding in STEVIA REBAUDIANA minimal medium Chinese herbal medicine, makes culture medium be in immune environment, prevents and treats the pollution of the microorganisms such as alien bacteria, presses down so as to reach efficiently and persistently The effect of bacterium, while improving STEVIA REBAUDIANA explant itself resistance, reduces manpower and materials input, reduces cost.On the other hand carry For the preparation method of the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium, methods described is simple to operate, and extraction effect is good.
A kind of extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium is provided according to an aspect of the present invention, it is described for sweet The Chinese herbal medicine contained in the extracts of Chinese herbal medicine of leaf chrysanthemum culture medium includes the component of following parts by weight：10～15 parts of dalbergia wood, Bi Pull out 9～14 parts, 6～10 parts of edible tulip, 7～10 parts of Lygodium japonicum, 4～8 parts of flat ginger, 7～11 parts of Chinese mahonia.
Preferably, the Chinese herbal medicine for containing in the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium includes following weight portion Several components：It is 12～14 parts of dalbergia wood, 11～13 parts of Piper longum, 7～9 parts of edible tulip, 8～9 parts of Lygodium japonicum, 5～7 parts of flat ginger, ten big 8～10 parts of contribution.
Preferably, the Chinese herbal medicine for containing in the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium includes following weight portion Several components：13 parts of dalbergia wood, 12 parts of Piper longum, 8 parts of edible tulip, 8 parts of Lygodium japonicum, 6 parts of flat ginger, 9 parts of Chinese mahonia.
Preferably, the extracts of Chinese herbal medicine is used for the STEVIA REBAUDIANA callus tissue culture base, Protocorm base, subculture Arbitrary culture medium in culture medium, Secondary Culture base or root media.
Preferably, the callus tissue culture base, Protocorm base, subculture medium, Secondary Culture base or training of taking root Arbitrary culture medium in foster base is used to adopt stem or leaf as the STEVIA REBAUDIANA tissue cultures of explant.
According to another aspect of the present invention, there is provided the described extracts of Chinese herbal medicine preparation side for STEVIA REBAUDIANA culture medium Method, methods described comprises the steps：
（1）The dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia are soaked 20～30 minutes respectively with clear water, Pull rear clear water out to rinse 2～3 times, being separately added into ethanol carries out extraction 2～3 hours, after filtration the dalbergia wood, Piper longum, mountain are kind Mushroom, Lygodium japonicum, flat ginger and Chinese mahonia preextraction liquid；
（2）By the step（1）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia preextraction liquid add 2 respectively ～3 times of weight waters, boiling to the ethanol is all evaporated completely, and the dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and ten is obtained Big contribution extract；
（3）By the step（2）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia extract mixing, stirring Uniformly, the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium is obtained final product.
Compared with prior art, the present invention has following beneficial effect：
（1）Chinese herbal medicine of the present invention includes dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia, said herbal medicine To antibacterial its extract is mixed with basal medium with positive effect, can persistently play bacteria resistance function, resisted due to people The microorganism pollution in STEVIA REBAUDIANA incubation is caused for factor or environmental factor, while improve STEVIA REBAUDIANA explant itself exempting from Epidemic disease function, ensures that tissue cultures are smoothed out, and improves tissue cultures quality and efficiency.
（2）Methods described is simple to operate, good using ethanol as extract extraction effect, with reference to boiling concentration, Ke Yibao The activity of barrier medicinal herb componentses.
Specific embodiment
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention Rather than restriction the scope of the present invention.
Embodiment 1
The Chinese herbal medicine for being used to contain in the extracts of Chinese herbal medicine of STEVIA REBAUDIANA culture medium described in the present embodiment includes following parts by weight Component：15 parts of dalbergia wood, 9 parts of Piper longum, 10 parts of edible tulip, 7 parts of Lygodium japonicum, 8 parts of flat ginger, 7 parts of Chinese mahonia.The Chinese herbal medicine is carried Liquid is taken for the STEVIA REBAUDIANA callus tissue culture base；The callus tissue culture base is used for the sweetleaf using stem as explant Chrysanthemum tissue cultures.
It is used to prevent and treat the feed addictive preparation method of sucking pig roundworm disease described in the present embodiment, methods described includes following step Suddenly：
（1）The dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia are soaked 30 minutes respectively with clear water, is pulled out Afterwards clear water rinse 2 times, be separately added into ethanol carry out extraction 3 hours, after filtration the dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, Flat ginger and Chinese mahonia preextraction liquid；
（2）By the step（1）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia preextraction liquid add 2 respectively ～3 times of weight waters, boiling to the ethanol is all evaporated completely, and the dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and ten is obtained Big contribution extract；
（3）By the step（2）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia extract mixing, stirring Uniformly, the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium is obtained final product.
Embodiment 2
The Chinese herbal medicine for being used to contain in the extracts of Chinese herbal medicine of STEVIA REBAUDIANA culture medium described in the present embodiment includes following parts by weight Component：10 parts of dalbergia wood, 14 parts of Piper longum, 6 parts of edible tulip, 10 parts of Lygodium japonicum, 4 parts of flat ginger, 11 parts of Chinese mahonia.The Chinese herbal medicine Extract is used for the STEVIA REBAUDIANA Protocorm base；The Protocorm base is used for the STEVIA REBAUDIANA using leaf as explant Tissue cultures.
It is used to prevent and treat the feed addictive preparation method of sucking pig roundworm disease described in the present embodiment, methods described includes following step Suddenly：
（1）The dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia are soaked 20 minutes respectively with clear water, is pulled out Afterwards clear water rinse 3 times, be separately added into ethanol carry out extraction 2 hours, after filtration the dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, Flat ginger and Chinese mahonia preextraction liquid；
（2）By the step（1）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia preextraction liquid add 3 respectively Times weight water, boiling to the ethanol is all evaporated completely, and the dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and ten is obtained big Contribution extract；
（3）By the step（2）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia extract mixing, stirring Uniformly, the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium is obtained final product.
Embodiment 3
The Chinese herbal medicine for being used to contain in the extracts of Chinese herbal medicine of STEVIA REBAUDIANA culture medium described in the present embodiment includes following parts by weight Component：14 parts of dalbergia wood, 11 parts of Piper longum, 9 parts of edible tulip, 8 parts of Lygodium japonicum, 7 parts of flat ginger, 8 parts of Chinese mahonia.The Chinese herbal medicine is carried Liquid is taken for the STEVIA REBAUDIANA subculture medium；The subculture medium is used for the STEVIA REBAUDIANA tissue training using stem as explant Support.
It is used to prevent and treat the feed addictive preparation method of sucking pig roundworm disease described in the present embodiment, methods described includes following step Suddenly：
（1）The dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia are soaked 30 minutes respectively with clear water, is pulled out Afterwards clear water rinse 2 times, be separately added into ethanol carry out extraction 3 hours, after filtration the dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, Flat ginger and Chinese mahonia preextraction liquid；
（2）By the step（1）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia preextraction liquid add 2 respectively Times weight water, boiling to the ethanol is all evaporated completely, and the dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and ten is obtained big Contribution extract；
（3）By the step（2）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia extract mixing, stirring Uniformly, the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium is obtained final product.
Embodiment 4
The Chinese herbal medicine for being used to contain in the extracts of Chinese herbal medicine of STEVIA REBAUDIANA culture medium described in the present embodiment includes following parts by weight Component：12 parts of dalbergia wood, 13 parts of Piper longum, 7 parts of edible tulip, 9 parts of Lygodium japonicum, 5 parts of flat ginger, 10 parts of Chinese mahonia.The Chinese herbal medicine Extract is used for the STEVIA REBAUDIANA Secondary Culture base；The Secondary Culture base is used for the STEVIA REBAUDIANA using leaf as explant to be organized Culture.
It is used to prevent and treat the feed addictive preparation method of sucking pig roundworm disease described in the present embodiment, methods described includes following step Suddenly：
（1）The dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia are soaked 30 minutes respectively with clear water, is pulled out Afterwards clear water rinse 3 times, be separately added into ethanol carry out extraction 2 hours, after filtration the dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, Flat ginger and Chinese mahonia preextraction liquid；
（2）By the step（1）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia preextraction liquid add 3 respectively Times weight water, boiling to the ethanol is all evaporated completely, and the dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and ten is obtained big Contribution extract；
（3）By the step（2）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia extract mixing, stirring Uniformly, the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium is obtained final product.
Embodiment 5
The Chinese herbal medicine for being used to contain in the extracts of Chinese herbal medicine of STEVIA REBAUDIANA culture medium described in the present embodiment includes following parts by weight Component：13 parts of dalbergia wood, 12 parts of Piper longum, 8 parts of edible tulip, 8 parts of Lygodium japonicum, 6 parts of flat ginger, 9 parts of Chinese mahonia.The Chinese herbal medicine is carried Liquid is taken for the STEVIA REBAUDIANA root media；The root media is used for the STEVIA REBAUDIANA tissue training using stem as explant Support.
It is used to prevent and treat the feed addictive preparation method of sucking pig roundworm disease described in the present embodiment, methods described includes following step Suddenly：
（1）The dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia are soaked 25 minutes respectively with clear water, is pulled out Afterwards clear water rinse 3 times, be separately added into ethanol carry out extraction 2 hours, after filtration the dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, Flat ginger and Chinese mahonia preextraction liquid；
（2）By the step（1）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia preextraction liquid add 3 respectively Times weight water, boiling to the ethanol is all evaporated completely, and the dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and ten is obtained big Contribution extract；
（3）By the step（2）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia extract mixing, stirring Uniformly, the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium is obtained final product.
Experimental example
1st, material process
STEVIA REBAUDIANA stem explant after selection is sterile-processed, is randomly divided into 6 groups, 2～3 per group.
2nd, culture medium is prepared
Basal medium is prepared, the basal medium formulation is as follows：MS+0.12mg/LKT+0.13mg/LGA3+ 1.7mg/ LMgSO4+3.5mg/LNH4NO3+ 13mg/L syrup of organge+130mg/L agar mediums, adjust PH to be 6.2～7.0, prepare identical 6 Group, one of which is as a control group.
Extracts of Chinese herbal medicine is prepared according to ratio described in embodiment 1,2,3,4,5 and be added to the basis culture for preparing In base, 1,2,3,4,5 groups are denoted as.
3rd, experimental technique
Aseptically STEVIA REBAUDIANA stem explant is inoculated into respectively 5 groups of culture mediums for containing extracts of Chinese herbal medicine and control group On, cultivate to explant in identical gnotobasis and under management condition and take root, the bacterial plaque number of 5 groups of generations is counted respectively, tie Fruit is shown in Table 1.
The bacterial plaque of table 1 produces quantity situation list
Title Amount of bacterial plaque
1 group 0
2 groups 0
3 groups 0
4 groups 0
5 groups 1
Control group 5
As can be seen from Table 1, the present invention has efficient bacteriostasis.Simultaneously by experimental result can draw embodiment 1,2,3, Culture medium described in 4, fungistatic effect is more excellent.
The specific embodiment of the present invention is described above.It is to be appreciated that the invention is not limited in above-mentioned Particular implementation, those skilled in the art can within the scope of the claims make various modifications or modification, this not shadow Ring the flesh and blood of the present invention.

Claims (6)

1. a kind of extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium, it is characterised in that：In the culture medium for STEVIA REBAUDIANA The Chinese herbal medicine contained in herbal medicine extract includes the component of following parts by weight：10～15 parts of dalbergia wood, 9～14 parts of Piper longum, mountain are kind 6～10 parts of mushroom, 7～10 parts of Lygodium japonicum, 4～8 parts of flat ginger, 7～11 parts of Chinese mahonia.
2. the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium according to claim 1, it is characterised in that：It is described for sweet The Chinese herbal medicine contained in the extracts of Chinese herbal medicine of leaf chrysanthemum culture medium includes the component of following parts by weight：12～14 parts of dalbergia wood, Bi Pull out 11～13 parts, 7～9 parts of edible tulip, 8～9 parts of Lygodium japonicum, 5～7 parts of flat ginger, 8～10 parts of Chinese mahonia.
3. the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium according to claim 1, it is characterised in that：It is described for sweet The Chinese herbal medicine contained in the extracts of Chinese herbal medicine of leaf chrysanthemum culture medium includes the component of following parts by weight：13 parts of dalbergia wood, Piper longum 12 Part, 8 parts of edible tulip, 8 parts of Lygodium japonicum, 6 parts of flat ginger, 9 parts of Chinese mahonia.
4. the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium according to claim 1, it is characterised in that：The Chinese herbal medicine Extract is used for the STEVIA REBAUDIANA callus tissue culture base, Protocorm base, subculture medium, Secondary Culture base or training of taking root Arbitrary culture medium in foster base.
5. the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium according to claim 4, it is characterised in that：The callus group Knitting the arbitrary culture medium in culture medium, Protocorm base, subculture medium, Secondary Culture base or root media is used to adopt The STEVIA REBAUDIANA tissue cultures of stem or leaf as explant.
6. the extracts of Chinese herbal medicine preparation method for STEVIA REBAUDIANA culture medium as any one of Claims 1 to 5, it is special Levy and be, methods described comprises the steps：
The dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia are soaked 20～30 minutes respectively with clear water, is pulled out Afterwards clear water is rinsed 2～3 times, and being separately added into ethanol carries out extraction 2～3 hours, and the dalbergia wood, Piper longum, edible tulip, sea are obtained after filtration Jinsha, flat ginger and Chinese mahonia preextraction liquid；
（2）By the step（1）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia preextraction liquid add 2 respectively ～3 times of weight waters, boiling to the ethanol is all evaporated completely, and the dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and ten is obtained Big contribution extract；
（3）By the step（2）In dalbergia wood, Piper longum, edible tulip, Lygodium japonicum, flat ginger and Chinese mahonia extract mixing, stirring Uniformly, the extracts of Chinese herbal medicine for STEVIA REBAUDIANA culture medium is obtained final product.
CN201610950090.XA 2016-11-03 2016-11-03 Chinese herbal medicine extracting solution used for stevia rebaudiana culture medium and preparation method Withdrawn CN106613955A (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101946809A (en) * 2010-08-19 2011-01-19 天津市农业生物技术研究中心 Bacteriostat composition applicable to plant tissue culture and using method thereof
CN102308750A (en) * 2011-07-26 2012-01-11 天津滨城龙达集团有限公司 Plant tissue culture bacteriostatic (antiseptic) agent containing botanical fungicide
CN103875732A (en) * 2014-03-11 2014-06-25 四川农业大学 Bacteriostatic agent for open type tissue culture of unibract fritillary bulb, and using method thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101946809A (en) * 2010-08-19 2011-01-19 天津市农业生物技术研究中心 Bacteriostat composition applicable to plant tissue culture and using method thereof
CN102308750A (en) * 2011-07-26 2012-01-11 天津滨城龙达集团有限公司 Plant tissue culture bacteriostatic (antiseptic) agent containing botanical fungicide
CN103875732A (en) * 2014-03-11 2014-06-25 四川农业大学 Bacteriostatic agent for open type tissue culture of unibract fritillary bulb, and using method thereof

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Application publication date: 20170510