CN104823853A - Method for rapidly proliferating tissue culture sprout of dendrobium - Google Patents
Method for rapidly proliferating tissue culture sprout of dendrobium Download PDFInfo
- Publication number
- CN104823853A CN104823853A CN201510234922.3A CN201510234922A CN104823853A CN 104823853 A CN104823853 A CN 104823853A CN 201510234922 A CN201510234922 A CN 201510234922A CN 104823853 A CN104823853 A CN 104823853A
- Authority
- CN
- China
- Prior art keywords
- tissue culture
- bud
- dendrobium
- dendrobe tissue
- medium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Abstract
The invention discloses a method for rapidly proliferating a tissue culture sprout of dendrobium. The method comprises the steps of carrying out short-time soaking stimulation on the tissue culture sprout of dendrobium by using negative ion water; then, carrying out dark starvation treatment in a sugar-free culture medium; and finally, transferring and inoculating to an eutrophic culture medium, and recovering the illumination condition to promote the rapid proliferation of the tissue culture sprout of dendrobium. The method concretely comprises the following operation steps of (1) negative ion water soaking pretreatment; (2) dark starvation treatment of the tissue culture sprout of dendrobium; and (3) proliferation culture of the tissue culture sprout of dendrobium. The tissue culture sprout of dendrobium is rapidly proliferated through combining short-time negative ion water, dark starvation stress treatment and later eutrophic culture. The method is simple and easy to realize, low in cost and suitable for large-scale industrial production; and the proliferation efficiency of the tissue culture sprout of dendrobium is high.
Description
Technical field
Patent of the present invention relates to plant tissue culture technical field, particularly relates to a kind of method of dendrobe tissue culture bud fast breeding.
Background technology
Dendrobe tissue culture bud is the intermediary of the stem of noble dendrobium by embryo seedling differentiation, is also to cultivate the key link forming test-tube plantlet.Under common condition of culture, dendrobe tissue culture Shoot propagation differentiation speed is slow, and proliferate efficiency is low.Conventional means of adding hormone impel the Proliferation, Differentiation of group training bud at present, but the interpolation of exogenous hormone, can, along with the carrying out of the growth of plant, be enriched in plant part, hidden danger is existed to edible safety.The problems such as the method seeking to promote dendrobe tissue culture bud fast breeding to break up safely and effectively is slow to stem of noble dendrobium reproduction restraint in the production of solution existing plant layoutization, hormone residues are significant.
Summary of the invention
The object of the invention is the method in order to solve a kind of dendrobe tissue culture bud fast breeding that the problems such as the growing multiplication run in existing dendrobe tissue culture Shoot propagation production process is slow, exogenous hormone uses, production life cycle is long provide, simple to operate, lower cost.
The technical solution realizing above-mentioned purpose is as follows:
A method for dendrobe tissue culture bud fast breeding, is characterized in that carrying out according to following step:
(1) anion water dipping pretreatment: taken out from blake bottle by dendrobe tissue culture bud, is added into anion culture fluid to submergence, under 25 DEG C of conditions, soaks 4-8 hour.
(2) the hungry dark treatment of dendrobe tissue culture bud: the dendrobe tissue culture bud after soaking is taken out, is transferred in desaccharification medium, under 25 DEG C of conditions, light culture 48-96 hour.
(3) Multiplying culture of dendrobe tissue culture bud: take out dendrobe tissue culture bud from desaccharification medium, transfers in proliferated culture medium, at temperature 25 ± 2 DEG C, 50 μm of ol/ (m
2s), illumination every day 14 hours, intensity of illumination 1500-2000LX condition under cultivate 30-40 days, in this period, dendrobe tissue culture bud can obtain fast breeding.
The described stem of noble dendrobium is the one in Dendrobidium huoshanness, dendrobium candidum, Dendrobium Moniliforme.
The budlet that described dendrobe tissue culture bud is Dendrobidium huoshanness, dendrobium candidum or Dendrobium Moniliforme seed are sprouted in basic MS culture medium.
Anion concentration in anion water described in step (1) is not less than 200/cm
3.
Desaccharification medium described in step (2) is the liquid MS medium removing sucrose.
Proliferated culture medium in step (3), for containing 30g/L sucrose: maltose ratio is the disaccharide solids MS medium of 1:0.5 ~ 1:1.5.
Principle summary:
Under anion refers to external condition, atoms outermost electronics breaks away from atomic nucleus, discharges from atom, and the free electron of effusion can be attached on some molecule or atom, forms new electronegative molecule.The anion obtained by ionization part water can increase ion concentration, promotes that plant is to the absorption of moisture and nutriment, strengthens the matter transportation function of plant, the equilibrium osmotic pressure of regulating plant body.In anion forming process, meeting release portion oxygen, strengthens the absorption of root, thus reaches the effect promoting plant metabolism.
The Hunger stress of short-term can not produce damage to body, and plant can be impelled on the contrary to occur compensatory growth phenomenon.After coercing releasing, the photosynthetic efficiency of plant and being increased Nutrient Absorption ability, growth rate can increase substantially.
Compared with prior art, method of the present invention has following beneficial effect: the present invention produces to sprout by the anion process of short-term and coerces to impel dendrobe tissue culture bud to strengthen its metabolic vigor in conjunction with light culture, improve the speed to photosynthetic response, reach synchronous, growth and propagation fast object.
1, the present invention adopts the anion of short-term to sprout to coerce in conjunction with light culture and to instead of tradition with eutrophy cultural method and use hormone to impel dendrobe tissue culture Shoot propagation, avoids exogenous hormone and uses harm to health.
2, the present invention adopts the anion of short-term to sprout to coerce and eutrophy cultural method in conjunction with light culture, significantly improve the rate of increase, within 30-40 days, the dendrobe tissue culture bud fresh weight rate of increase is up to more than 400%, and the rate of increase of bud reaches more than 110%, is about 4 times of untreated control group.Before the rate of increase=(before processing rear-process)/process × 100%.
3, the present invention adopts short-term anion to sprout to coerce and eutrophy cultural method in conjunction with light culture, and simple to operate, lower cost, for enterprise's batch production, the large-scale production stem of noble dendrobium provide new method.
Embodiment
Below in conjunction with concrete embodiment, the present invention is further described.The features and advantages of the invention will be clearer along with description, but these exemplary embodiments are only used for the present invention is described, do not form any restriction to scope of the present invention.
In this patent, involved Dendrobidium huoshanness group training bud, candidum tissue culturing bud, Dendrobium Moniliforme group training bud are Anhui Kang Erxin Biology Pharmacy Co., Ltd and produce; It is pure that MS medium medicine is analysis, all buys from the bright bio tech ltd of Pood; Anion water machine is bought in Ge Hao brine electrolysis Co., Ltd, and anion concentration is not less than 200/cm
3.
(1) anion water dipping pretreatment: taken out from blake bottle by dendrobe tissue culture bud, is added into anion culture fluid to submergence, under 25 DEG C of conditions, soaks 4-8 hour.
(2) the hungry dark treatment of dendrobe tissue culture bud: the dendrobe tissue culture bud after soaking is taken out, is transferred in desaccharification medium, under 25 DEG C of conditions, light culture 48-96 hour.
(3) Multiplying culture of dendrobe tissue culture bud: the dendrobe tissue culture bud taken out from light culture, transfers in proliferated culture medium, at temperature 25 ± 2 DEG C, 50 μm of ol/ (m
2s), illumination every day 14 hours, intensity of illumination 1500-2000LX condition under cultivate 30-40 days, in this period, dendrobe tissue culture bud can obtain fast breeding.The group training bud switch over operation more than related to all should carry out in superclean bench.
embodiment 1
A method for Dendrobidium huoshanness group training bud fast breeding, comprises following operating procedure:
Step 1: anion water dipping pretreatment
From blake bottle, take out Dendrobidium huoshanness group training bud 20g, average bud number 32/g, puts into anion concentration and is not less than 200/cm
3anion water 200 mL in submergence, under 25 DEG C of conditions, immersion treatment 4 hours.
Step 2: the hungry dark treatment of Dendrobidium huoshanness group training bud
Dendrobidium huoshanness group training bud after soaking is taken out, is transferred in desaccharification MS medium, the every bottle of about 4g that transfers, at temperature 25 ± 2 DEG C, light culture 96 hours.
Step 3: the Multiplying culture of Dendrobidium huoshanness group training bud
From light culture take out Dendrobidium huoshanness group training bud, transfer containing 30g/L(sucrose: maltose ratio is 1:0.5) disaccharide MS solid multiplication medium in, every bottle switching about 4 g, at temperature 25 ± 2 DEG C, 50 μm of ol/ (m
2s), illumination every day 14 hours, cultivate 35 days under the condition of intensity of illumination 1500LX.Within 35 days, take out the Dendrobidium huoshanness group training bud of propagation afterwards, every bottle of weight average is 21.47 g, and rate of body weight gain is 436.75%, and the number that sprouts on average can reach 77/g, and bud ratio is 140%.
embodiment 2
A method for candidum tissue culturing bud fast breeding, comprises following operating procedure:
Step 1: anion water dipping pretreatment
From blake bottle, take out candidum tissue culturing bud 20g, average bud number 41/g, puts into anion concentration and is not less than 200/cm
3anion water 200 mL in submergence, under 25 DEG C of conditions, immersion treatment 6 h.
Step 2: the hungry dark treatment of candidum tissue culturing bud
Candidum tissue culturing bud after soaking is taken out, is transferred in the liquid MS medium removing sucrose, the every bottle of about 4g that transfers, at temperature 25 ± 2 DEG C, light culture 72 h.
Step 3: the Multiplying culture of candidum tissue culturing bud
From light culture take out candidum tissue culturing bud, transfer containing 30 g/L(sucrose: maltose ratio is 1:1) disaccharide MS solid multiplication medium in, the every bottle of about 4g that transfers, at temperature 25 ± 2 DEG C, 50 μm of ol/ (m
2cultivate 30 days under the condition of s), illumination every day 14 hours, intensity of illumination 1800LX.Within 30 days, take out the candidum tissue culturing bud of propagation afterwards, every bottle of weight average is 22.12 g, and rate of body weight gain is 453.00%, and bud number on average can reach 87/g, and bud ratio is 112.20%.
embodiment 3
A method for Dendrobium Moniliforme group training bud fast breeding, comprises following operating procedure:
Step 1: anion water dipping pretreatment
From blake bottle, take out Dendrobium Moniliforme group training bud 20g, average bud number 38/g, puts into anion concentration and is not less than 200/cm
3anion water 200 mL in submergence, under 25 DEG C of conditions, immersion treatment 4 h.
Step 2: the hungry dark treatment of Dendrobium Moniliforme group training bud
Dendrobium Moniliforme group training bud after soaking is taken out, is transferred in desaccharification MS medium, every bottle of switching about 4 g, at temperature 25 ± 2 DEG C, light culture 48 h.
Step 3: the Multiplying culture of Dendrobium Moniliforme group training bud
From light culture take out Dendrobium Moniliforme group training bud, transfer containing 30g/L(sucrose: maltose ratio is 1:1.5) disaccharide MS solid multiplication medium in, every bottle switching about 4 g, at temperature 25 ± 2 DEG C, 50 μm of ol/ (m
2cultivate 30 days under the condition of s), illumination every day 14 hours, intensity of illumination 2000LX.Within 30 days, take out the Dendrobium Moniliforme group training bud of propagation afterwards, every bottle of weight average is 20.37g, and rate of body weight gain is 409.25%, and bud number on average can reach 82/g, and bud ratio is 115.79%.
Claims (6)
1. a method for dendrobe tissue culture bud fast breeding, is characterized in that carrying out according to following step:
(1) anion water dipping pretreatment: taken out from blake bottle by dendrobe tissue culture bud, is added into anion culture fluid to submergence, under 25 DEG C of conditions, soaks 4-8 hour;
(2) the hungry dark treatment of dendrobe tissue culture bud: the dendrobe tissue culture bud after soaking is taken out, is transferred in desaccharification medium, under 25 DEG C of conditions, light culture 48-96 hour;
(3) Multiplying culture of dendrobe tissue culture bud: take out dendrobe tissue culture bud from desaccharification medium, transfers in proliferated culture medium, at temperature 25 ± 2 DEG C, 50 μm of ol/ (m
2s), illumination every day 14 hours, intensity of illumination 1500-2000LX condition under cultivate 30-40 days, in this period, dendrobe tissue culture bud can obtain fast breeding.
2. the method for a kind of dendrobe tissue culture bud fast breeding according to claim 1, is characterized in that the described stem of noble dendrobium is the one in Dendrobidium huoshanness, dendrobium candidum, Dendrobium Moniliforme.
3. the method for a kind of dendrobe tissue culture bud fast breeding according to claim 1, is characterized in that described dendrobe tissue culture bud is Dendrobidium huoshanness, budlet that dendrobium candidum or Dendrobium Moniliforme seed are sprouted in basic MS culture medium.
4. the method for a kind of dendrobe tissue culture bud fast breeding according to claim 1, is characterized in that the anion concentration in the anion water described in step (1) is not less than 200/cm
3.
5. the method for a kind of dendrobe tissue culture bud fast breeding according to claim 1, is characterized in that desaccharification medium described in step (2) is the liquid MS medium removing sucrose.
6. the method for a kind of dendrobe tissue culture bud fast breeding according to claim 1, is characterized in that the proliferated culture medium in step (3), for containing 30g/L sucrose: maltose ratio is the disaccharide solids MS medium of 1:0.5 ~ 1:1.5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510234922.3A CN104823853B (en) | 2015-05-11 | 2015-05-11 | A kind of method of dendrobe tissue culture bud fast breeding |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510234922.3A CN104823853B (en) | 2015-05-11 | 2015-05-11 | A kind of method of dendrobe tissue culture bud fast breeding |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104823853A true CN104823853A (en) | 2015-08-12 |
| CN104823853B CN104823853B (en) | 2017-06-06 |
Family
ID=53802389
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510234922.3A Active CN104823853B (en) | 2015-05-11 | 2015-05-11 | A kind of method of dendrobe tissue culture bud fast breeding |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104823853B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0523070A (en) * | 1991-07-17 | 1993-02-02 | P C Shiyu Technol:Kk | Production of seedling of saffron |
| CN104429961A (en) * | 2014-12-08 | 2015-03-25 | 河源市裕森农林发展有限公司 | Rapid propagation tissue culture method for dendrobium officinale |
| CN104542270A (en) * | 2013-10-14 | 2015-04-29 | 厦门塔斯曼生物工程有限公司 | Rapid propagation method for medicinal dendrobium nobile |
-
2015
- 2015-05-11 CN CN201510234922.3A patent/CN104823853B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0523070A (en) * | 1991-07-17 | 1993-02-02 | P C Shiyu Technol:Kk | Production of seedling of saffron |
| CN104542270A (en) * | 2013-10-14 | 2015-04-29 | 厦门塔斯曼生物工程有限公司 | Rapid propagation method for medicinal dendrobium nobile |
| CN104429961A (en) * | 2014-12-08 | 2015-03-25 | 河源市裕森农林发展有限公司 | Rapid propagation tissue culture method for dendrobium officinale |
Non-Patent Citations (3)
| Title |
|---|
| ROXAS, ET AL.: "Electrogenic machine and Nitrazyme soil conditioner: their influence on plants", 《CANOPY INTERNATIONAL》 * |
| 丁玉梅等: "植物组织培养电刺激效应研究进展", 《生物学通报》 * |
| 卢文芸等: "五种药用石斛快速繁殖的研究", 《种子》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104823853B (en) | 2017-06-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104472359B (en) | A kind of ginseng adventitious root proliferative induction method | |
| Salimi et al. | Effects of gibberellic acid and carbon disulphide on sprouting of potato minitubers | |
| CN105010109B (en) | The ciltivating process of Momordica grosvenori | |
| CN103202127A (en) | Method for stopping Taxus chinensis var. mairei seed dormancy | |
| CN105010147A (en) | Special culture medium for improving tissue culture propagation speed of haworthia succulent plants and tissue culture method | |
| CN102884981B (en) | Zanthoxylum nitidum tissue culture medium | |
| CN103305456B (en) | Construction method and ultralow temperature freezing and storing method of sinocyclocheilus grahami saccus olfactorius cell line | |
| WO2012061950A1 (en) | Thermo-photo-bioreactor and method for the culture and mass micropropagation of deschampsia antarctica in vitro | |
| CN105613291A (en) | Fast breeding method for dendrobium candidum protocorm tissue culture | |
| CN107047319A (en) | The method for tissue culture and root media of a kind of ginkgo | |
| CN109220810A (en) | A kind of method of tree peony embryo high-effective root-growing under aseptic condition | |
| CN106106147B (en) | A kind of renovation process of lotus unmature subleaf somatic embryo development ways | |
| CN103766039B (en) | Method for breaking dormancy of nervilis fordii schlecht bulb | |
| CN102067818B (en) | Inducing technology of test tube lotus root | |
| CN104823853A (en) | Method for rapidly proliferating tissue culture sprout of dendrobium | |
| CN105409773A (en) | Lophophora williamsii sterile seeding and regeneration system establishing method | |
| CN102499082A (en) | Test tube breeding method of lilium oriental hybrid seed ball | |
| CN108703068B (en) | Method for removing endophyte in arrowhead culture process, culture method and application | |
| RU2731061C2 (en) | Method of storing rubus plants in vitro | |
| Yamagishi | Effects of cold treatment, BA and GA3 on enlargement and leaf emergence of in vitro cultured bulblets of Lilium japonicum Thunb. | |
| CN104255453A (en) | Preservation method of stevia rebaudiana tissue culture seedlings | |
| CN107306794B (en) | A method of extending the time-to-live of duckweed germplasm Preservation in sterile condition | |
| CN104170741B (en) | A kind of Storaged media of chrysanthemum tissue cultured seedling | |
| Abdulmalik et al. | Micropropagation of banana (Musa spp) using temporary immersion bioreactor system | |
| CN110720393A (en) | Method for tissue culture and rapid propagation of ficus microcarpa |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |