CN107736245A - A kind of mountain orchid Fast Asexual Propagation Technique - Google Patents
A kind of mountain orchid Fast Asexual Propagation Technique Download PDFInfo
- Publication number
- CN107736245A CN107736245A CN201711006958.1A CN201711006958A CN107736245A CN 107736245 A CN107736245 A CN 107736245A CN 201711006958 A CN201711006958 A CN 201711006958A CN 107736245 A CN107736245 A CN 107736245A
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- mountain
- orchid
- mountain orchid
- pseudobulb
- illumination
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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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
-
- 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
- A01H4/008—Methods for regeneration to complete plants
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- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
A kind of blue Fast Asexual Propagation Technique in mountain, the breeding blue for carrying out mountain, overcome mountain is blue to breed the problem of difficult, planted for mountain orchid species and substantial amounts of seedling is provided.Operating procedure is：（1）Mountain orchid pseudobulb is taken fully to be cleaned under flowing water.Then aseptically, pseudobulb is carried out disinfection, sterile to wash 5~6 times, aseptic filter paper suck dry moisture.It is cut into small pieces, is inoculated on MS culture mediums.（2）Special callus inducing medium is made after each composition is added by formula.（3）Every bottle is inoculated with 3~5 pieces.（4）（25 soil 1）Cultivated in DEG C constant incubator, intensity of illumination 2000lx, illumination 12h/d.30~60d of incubation time.（5）The callus of vigorous growth grows 30~60d on Clonal regeneration culture medium and can induce and takes root and come into leaves.Obtain product of the present invention --- mountain orchid regeneration plant.The beneficial effect of the inventive method is：In laboratory, lead to too small amount of mountain orchid pseudobulb and be achieved with substantial amounts of vegetative propagation test tube seedling, inductivity is up to more than 90%.
Description
Technical field
The invention belongs to plant breeding field, and in particular to mountain orchid plant in vitro asexual quick breeding content.
Background technology
Mountain is blueOreorchas patens (Lindl.) Lindl. is the perennial raw herbaceous plant of the moon of orchid family mountain Cymbidium, extensively
It is general to be distributed in the ground such as China northeast, northwest, southwest, Central China, it is used as medicine with drying pseudobulb, property of medicine category is cold, acrid flavour, sweet, slightly poisonous,
With the removing toxic substances row stasis of blood, desinsection disappears carbuncle, enriching yin clearing lung-heat, preventing phlegm from forming and stopping coughing and other effects, according to《Dictionary of medicinal plant》Record, mountain Lan Yizuo mountains are kind
Mushroom is used as medicine.Edible tulip is a kind of conventional Chinese medicine, according to《Pharmacopoeia of People's Republic of China》Record：Edible tulip original plant has 3 kinds,
Respectively orchid Cremastra appendiculata [Cremastra appendiculata（D.Don）Makino], pleione bulbocodioides [Pleione bulbocodioides（Franch.）Rolfe] or Pleione yunnanensis（P. yunnanensisRolfe）, it is false with its drying
Bulb is used as medicine, and above-mentioned 3 kinds of plants are in the equal distribution-free in Jilin Province.At present, edible tulip is mainly derived from wild product, because its former plant
（It is blue including mountain）It is the plant that orchid family does not belong to together, the common problem bred all is not easy with orchid, as mountain is kind in recent years
The active anticancer of mushroom is found, and its demand is continuously increased, because the edible tulip in original producton location has been difficult to meet supply, from 2008
Year rise a people start largely from Changbaishan area excavation mountain orchid be used as medicine as edible tulip, cause the mountain orchid wild resource reserves of Changbaishan area
Drastically decline, turn into more rare plant, therefore, to protect the germ plasm resource of this plant, ensure the blue Sustainable Exploitation in mountain
Utilize, it is extremely urgent to carry out its Study on Fast Asexual Propagation Technique.Because the time of this plant exploitation is short, the research of each side
Report is few, especially its reproduction technique, including there has been no people's issue for Fast Asexual Propagation Technique.
The content of the invention
It is an object of the invention to provide the Fast Asexual Propagation Technique that a kind of mountain is blue, the breeding blue for carrying out mountain, overcomes mountain blue
The problem of difficult is bred, is planted for mountain orchid species and substantial amounts of seedling is provided.
The blue Fast Asexual Propagation Technique in the mountain of the present invention comprises the following steps：
1st, the optimal Fiber differentiation based formulas of callus：MS +2.4-D 0.5mg/L+6-BA 1.0mg/L.
2nd, take mountain orchid pseudobulb to be rinsed under flowing water and gently outwash soil, flowing water rinse 40min with hairbrush.Then super
On net workbench, pseudobulb exocuticle is gently peeled off with the scalpel after sterilizing, with 70% ethanol disinfection 0.5min, 0.1%
HgCl2Solution disinfection 10min, sterile to wash 5~6 times, aseptic filter paper suck dry moisture.The fritter of 0.5cm or so square is cut into, is connect
Kind on MS culture mediums, culture medium contains 3.0% sucrose, 0.8% agar, pH6.0, and hormone prescription is shown in step 1.
3rd, every bottle is inoculated with 3~5 pieces.
4th, exist（25 soil 1）Cultivated in DEG C constant incubator, intensity of illumination 2000lx, illumination 12h/d.Incubation time 30~
60d。
5th, the callus of vigorous growth grows 30~60d on Clonal regeneration culture medium and can induce and takes root and come into leaves.
6th, optimal Clonal regeneration culture medium prescription：MS+NAA1.0mg/L+6-BA 1.0mg /L.
The beneficial effect of the inventive method is：In laboratory, lead to too small amount of mountain orchid pseudobulb and be achieved with largely
Vegetative propagation test tube seedling, inductivity is up to more than 90%.
Embodiment
Embodiment 1
1st, take mountain orchid pseudobulb to be rinsed under flowing water and gently outwash soil, flowing water rinse 40min with hairbrush.Then in ultra-clean work
Make on platform, gently peeled off pseudobulb exocuticle with the scalpel after sterilizing, with 70% ethanol disinfection 0.5min, 0.1%HgCl2It is molten
Liquid disinfectant 10min, sterile to wash 5~6 times, aseptic filter paper suck dry moisture.The fritter of 0.5cm or so square is cut into, is inoculated in MS
On culture medium, culture medium contains 3.0% sucrose, 0.8% agar, pH6.0.
2nd, the optimal Fiber differentiation based formulas of callus：MS +2.4-D 0.5mg/L+6-BA 1.0mg/L.
3rd, every bottle is inoculated with 3~5 pieces.
4th, exist（25 soil 1）Cultivated in DEG C constant incubator, intensity of illumination 2000lx, illumination 12h/d.Incubation time 30~
60d。
5th, the callus of vigorous growth grows 30~60d on Clonal regeneration culture medium and can induce and takes root and come into leaves.
Product of the present invention --- mountain orchid regeneration plant.
6th, optimal Clonal regeneration culture medium prescription：MS+NAA1.0mg/L+6-BA 1.0mg /L.
Embodiment 2
1st, mountain orchid blade is taken to rinse 10min under flowing water, then on superclean bench, with 70% ethanol disinfection 0.5min, 0.1%
HgCl2Solution disinfection 5min, sterile to wash 3~4 times, aseptic filter paper suck dry moisture.The small pieces of 0.5cm or so square are cut into, are connect
For kind on MS culture mediums, culture medium contains 3.0% sucrose, 0.8% agar, pH6.0.
2nd, the optimal Fiber differentiation based formulas of callus：MS +2.4-D 0.5mg/L+6-BA 1.0mg/L.
3rd, every bottle is inoculated with 3~5.
4th, exist（25 soil 1）Cultivated in DEG C constant incubator, intensity of illumination 2000lx, illumination 12h/d.Incubation time 30~
60d。
5th, the callus of vigorous growth grows 30~60d on Clonal regeneration culture medium and can induce and takes root and come into leaves.
Product of the present invention --- mountain orchid regeneration plant.
6th, optimal Clonal regeneration culture medium prescription：MS+NAA1.0mg/L+6-BA 1.0mg /L.
Embodiment 3
1st, mountain orchid peduncle is taken, 10min is rinsed under flowing water after removing whole little Hua, then on superclean bench, with 70% second
Alcohol sterilizes 0.5min, 0.1%HgCl2Solution disinfection 5min, sterile to wash 3~4 times, aseptic filter paper suck dry moisture.It is cut into 0.5cm
Long segment, it is inoculated on MS culture mediums, culture medium contains 3.0% sucrose, 0.8% agar, pH6.0.
2nd, the optimal Fiber differentiation based formulas of callus：MS +2.4-D 0.5mg/L+6-BA 1.0mg/L.
3rd, every bottle is inoculated with 3~5 sections.
4th, exist（25 soil 1）Cultivated in DEG C constant incubator, intensity of illumination 2000lx, illumination 12h/d.Incubation time 30~
60d。
5th, the callus of vigorous growth grows 30~60d on Clonal regeneration culture medium and can induce and takes root and come into leaves.
Product of the present invention --- mountain orchid regeneration plant.
6th, optimal Clonal regeneration culture medium prescription：MS+NAA1.0mg/L+6-BA 1.0mg /L.
Claims (8)
- A kind of 1. mountain orchid Fast Asexual Propagation Technique, it is characterised in that：It is big that explant can be made with mountain orchid pseudobulb or blade or peduncle Amount production mountain orchid test tube seedling.
- 2. technology according to claim 1, it is characterised in that：Explant used in being rinsed under flowing water, and use hairbrush Gently outwash soil, flowing water rinse a period of time.
- 3. material according to claim 2, it is characterised in that：Explant after cleaning is on superclean bench, with 70% ethanol Sterilize 0.5min, 0.1%HgCl25~10min of solution disinfection, sterile to wash 3~6 times, aseptic filter paper suck dry moisture.
- 4. material according to claim 3, it is characterised in that：It is cut into the fritter or small pieces or small of 0.5cm or so square Section, is inoculated on MS culture mediums, culture medium contains 3.0% sucrose, 0.8% agar, pH6.0.
- 5. according to culture materials described in right 4, it is characterised in that：Every bottle of 3~5 pieces of inoculum concentration/piece/section, callus most preferably lure It is MS+2.4-D 0.5mg/L+6-BA 1.0mg/L to lead culture medium prescription.
- 6. the material obtained according to right 5, it is characterised in that：Optimal culture condition is cultivated in 25 1 DEG C of constant incubator of soil, Intensity of illumination 2 000lx, illumination 12h/d, 30~60d of incubation time.
- 7. the material obtained according to right 6, it is characterised in that：The callus of vigorous growth is selected, is induced in aftergrowth 30~60d root inductions are grown on culture medium and are come into leaves, obtain product of the present invention --- mountain orchid regeneration plant.
- 8. according to the technical step described in right 2~7, it is characterised in that：Optimal aftergrowth Fiber differentiation based formulas is MS+ NAA1.0mg/L+6-BA 1.0mg /L。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201711006958.1A CN107736245A (en) | 2017-10-25 | 2017-10-25 | A kind of mountain orchid Fast Asexual Propagation Technique |
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| CN201711006958.1A CN107736245A (en) | 2017-10-25 | 2017-10-25 | A kind of mountain orchid Fast Asexual Propagation Technique |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109699498A (en) * | 2019-03-11 | 2019-05-03 | 成都大学 | A kind of cultural method of Cremastra appendiculata callus |
| CN110178734A (en) * | 2019-07-16 | 2019-08-30 | 中国农业科学院特产研究所 | A kind of the post-directed training base and post-directed training seedling establishment method of mountain orchid germination seed |
-
2017
- 2017-10-25 CN CN201711006958.1A patent/CN107736245A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109699498A (en) * | 2019-03-11 | 2019-05-03 | 成都大学 | A kind of cultural method of Cremastra appendiculata callus |
| CN109699498B (en) * | 2019-03-11 | 2021-09-03 | 成都大学 | Rhododendron callus culture method |
| CN110178734A (en) * | 2019-07-16 | 2019-08-30 | 中国农业科学院特产研究所 | A kind of the post-directed training base and post-directed training seedling establishment method of mountain orchid germination seed |
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Application publication date: 20180227 |