CN106332784A - Open gerbera jamesonii tissue culture method - Google Patents
Open gerbera jamesonii tissue culture method Download PDFInfo
- Publication number
- CN106332784A CN106332784A CN201610988977.8A CN201610988977A CN106332784A CN 106332784 A CN106332784 A CN 106332784A CN 201610988977 A CN201610988977 A CN 201610988977A CN 106332784 A CN106332784 A CN 106332784A
- Authority
- CN
- China
- Prior art keywords
- tissue culture
- sodium hypochlorite
- culture
- gerbera jamesonii
- agar
- 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.)
- Pending
Links
Classifications
-
- 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
-
- 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
Landscapes
- 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
The invention belongs to the field of plant tissue culture and provides an open gerbera jamesonii tissue culture method. The inhibition on growth and reproduction of bacteria in the gerbera jamesonii tissue culture process is achieved by utilizing the bacteriostatic effect of sodium hypochlorite and adding the sodium hypochlorite in a gerbera jamesonii tissue culture medium, and accordingly reproduction and rooting culture of gerbera jamesonii under the nonsterile conditions are achieved. The method has the advantages of being simple in operation, capable of saving time and labor, greatly reducing the costs and the like. The method is a novel rapid and efficient technological means, and an important basis is laid for development of an industrialized gerbera jamesonii seedling raising technology.
Description
Technical field
The invention belongs to field of plant tissue culture, it is related to a kind of method for culturing open type tissue of African Chrysanthemum.
Background technology
African Chrysanthemum (gerbera jamesoniiBolus), Compositae Gerbera, also known as African daisy, are perennial root
Draft ornamental flower.It is one of big Fresh Cutting flower in the world five, for arranging flowers and making the gaily decorated basket, also can make potted plant viewing and admiring.In recent years, non-
Continent chrysanthemum routine division propagation, reproduction speed is slow, and uses division propagation always, and variety deterioration is serious；The gerbera seeds life-span
Extremely short, and adopt seminal propagation, bloom late.Therefore, for meeting the vigorous African Chrysanthemum market demand, all utilize tissue culture's skill
Art produces seedling, and it can be largely overcoming traditional seminal propagation and division propagation breeds slow deficiency.
Though African Chrysanthemum tissue culture can provide the seedling of substantial amounts in a short time, step into commercialization rule when tissue culture produces
After mould produces, the high cost problem that it produces increasingly bursts.And tissue culture cost can be substantially reduced by open tissue culture thus
Realize the scale commodity production of high benefit.Relatively conventional tissue culture, open tissue culture without sterile working inoculation and
Gnotobasiss are cultivated, and only need to the addition of safe and efficient antibacterial in the medium, can inoculate, cultivate under the conditions of having collarium border,
Eliminate the larger high-pressure sterilizing pot of ratio between investments and superclean bench.At present, open tissue is cultivated in tens kinds of plants
On obtain successfully, but African Chrysanthemum open tissue culture yet there are no report.
Before this, by the bacterium test that naturally falls, at 4 kinds of antibacterial (sodium hypochlorite, potassium sorbate, Mancozeb, carbendazim)
In filter out the most obvious sodium hypochlorite of fungistatic effect for follow-up test.To sodium hypochlorite concentration in African Chrysanthemum enrichment culture
It is optimized, comprehensive pollution rate and growth coefficient conclude that the sodium hypochlorite of 0.004-0.010% presses down as African Chrysanthemum tissue culture
The effect of microbial inoculum is best.
Content of the invention
It is an object of the invention to provide a kind of method for culturing open type tissue of African Chrysanthemum, by being added on African Chrysanthemum group
Knit interpolation sodium hypochlorite in culture medium, thus realizing propagation under nonsterile conditionses for the African Chrysanthemum and taking root.Depart from strict aseptic
Condition, without large-scale instruments such as pressure cooker, superclean benches, fundamentally simplifies tissue culture link, substantially reduces tissue culture cost.
For achieving the above object, the present invention adopts the following technical scheme that
A kind of method for culturing open type tissue of African Chrysanthemum, by adding antibacterial in propagation and root media, directly exists
Opening has inoculated and cultured under conditions of bacterium.
Described proliferated culture medium is ms+6-ba 0.5mg/l+naa 0.1mg/l+ sucrose 30g/l+agar 7g/l.
Described root media is 1/2ms+naa 0.2mg/l+ sucrose 30g/l+agar 7g/l.
Described antibacterial is sodium hypochlorite, the final concentration of 0.004-0.010wt.% in culture medium.
It is an advantage of the current invention that:
1st, the present invention passes through systematic study, establishes African Chrysanthemum open tissue cultivating system first；
2nd, the present invention passes through to add certain density sodium hypochlorite, realizes propagation under nonsterile conditionses for the African Chrysanthemum and takes root；
3rd, the present invention saves the use of the large-scale instruments such as pressure cooker and superclean bench, significantly reduces production cost.
Brief description
Fig. 1 falls bacterium test (culture 9 days after) naturally, a. ms+ sucrose 30 g/l+ agar 7 g/l；B. ms+ sucrose 30
G/l+agar 7 g/l+0.02% sodium hypochlorite；C. ms+ sucrose 30 g/l+agar 7 g/l+0.02 g/l Mancozeb；
D. ms+ sucrose 30 g/l+agar 7 g/+ 100 times of l carbendazim；E. ms+ sucrose 30 g/l+agar 7 g/l+0.02g/
L potassium sorbate.
Fig. 2 African Chrysanthemum tissue cultured seedling enrichment culture 20d；A. ms+6-ba 0.5 mg/l+naa 0.1 mg/l+ sucrose 30 g/
L+agar 7 g/l(aseptic condition is inoculated)；B. ms+6-ba 0.5 mg/l+naa 0.1 mg/l+ sucrose 30 g/l+agar 7
G/l+0.004% sodium hypochlorite (open condition inoculation)；C. ms+6-ba 0.5 mg/l+naa 0.1 mg/l+ sucrose 30g+
Agar 7 g/l+0.008% sodium hypochlorite (open condition inoculation)；D. ms+6-ba 0.5 mg/l+naa 0.1 mg/l+ sugarcane
Sugared 30g+ agar 7 g/l+0.010% sodium hypochlorite (open condition inoculation).
Fig. 3 African Chrysanthemum tissue cultured seedling root culture 20d；A. 1/2ms+naa 0.2mg/l+ sucrose 30g/l+agar 7g/l
(aseptic condition inoculation)；B. 1/2ms+naa 0.2mg/l+ sucrose 30g/l+agar 7g/l+0.004% sodium hypochlorite is (open
Condition is inoculated)；C. 1/2ms+naa 0.2mg/l+ sucrose 30g/l+agar 7g/l+0.008% sodium hypochlorite (open condition
Inoculation)；D. 1/2ms+naa 0.2mg/l+ sucrose 30g/l+(open condition connects agar 7g/l+0.010% sodium hypochlorite
Kind).
Specific embodiment
Embodiment 1
1. wash --- culture bottle washs.
Culture medium in culture bottle is rinsed, then in clear water, soaks 1h, scrub dirt in bottle, steep the detergent into 1%
In lysate, then scrubbed, especially bottle mouth position also will conscientiously be scrubbed.Scrub to rinse with flowing water under faucet and done
Only.Finally it is inverted in the vessel basket of cleaning, to drain away the water.
2. join --- culture medium is prepared.
According to designed culture medium prescription, (proliferated culture medium is ms+6-ba 0.5mg/l+naa 0.1mg/l+ sucrose
30g/l+ agar 7g/l, root media are 1/2ms+naa 0.2mg/l+ sucrose 30g/l+ agar 7g/l), draw female in proportion
In liquid injection capacity bottle, add water constant volume, pours in stainless-steel pan plus agar and sugar.Heating while stirring in order to avoid boiling paste, making agar
It is completely dissolved with sugar.Adjust ph value with hydrochloric acid or sodium hydroxide solution, boil, be cooled to when 50-60 DEG C add 0.008%
Sodium hypochlorite.
3. fill --- culture medium subpackage.
Culture medium is uniformly dispensed in culture bottle while hot, in 30 min, covers bottle cap.Culture bottle is set level, can after solidification
Inoculated.
4. kind --- tissue culture plant inoculation.
Including subinoculation and inoculation of taking root.
It is ms+6-ba 0.5mg/l+naa 0.1mg/l+ sucrose 30g/l+ agar 7g/ that subinoculation uses proliferated culture medium
l；Inoculation of taking root is 1/2ms+naa 0.2mg/l+ sucrose 30g/l+ agar 7g/l using root media.
1), working environment: the room of optional neat and tidy, suitable desk, a chair, what a prepares not
The rust little pallet of steel or the glass of one piece of cleaning, two surgical scissorses, two scalpels, two tweezers, one equipped with appropriate 70% wine
Wide mouthed bottle (highly suitably lower, to be easy to inoculating tool immersion, take out), a bag absorbent cotton and a paper basket of essence.
2), seeded process:
1. before inoculating；The necessary washing hand with soap of staff, subculture, bottle seedling of taking root clean the dirt outside culture bottle with clean gauze
Soil.
2. by culture bottle, bottle seedling, scalpel, tweezers, 70% ethanol wide mouthed bottle, absorbent cotton, tool rack, aseptic backing plate etc., neatly
Reasonably it is placed on desk.
3. bottle seedling is taken out, be placed on through, on the aseptic backing plate of 70% alcohol disinfecting, carrying out the cutting of inoculation material, often inoculating
Complete one bottle, residue is poured into paper basket, wiped several times with the absorbent cotton being moistened with 70% ethanol, do not stay blind area, after plate face ethanol is slightly dry,
Carry out the cutting of next bottle of inoculation material.
4. the knife of cutting material, cut, tweezers etc., often inoculated one bottle, with cotton wool clean leaf fragment, agar, be immersed in
In 70% ethanol.
5. cover tightly bottle cap after having connect.
5th, support --- tissue cultured seedling is cultivated.
The indoor temperature control of culture is in 25(± 2) DEG C.Intensity of illumination is 2000lx, the daily 11h of light application time.
6th, plant --- tissue culture transplantation of seedlings.
Before transplanting seedlings, wash clean sticks to the culture medium of Miao Genshang of taking root, and must not damage the young root of tissue cultured seedling.During transplanting, root
Put in the aperture accomplished fluently in advance, so that root system is unfolded, fill up soil and be fully compacted, make root soil contiguity, too deep, nest root of planting will be prevented
Or crab rooting, transplant seedlings one plant in each container, pour permeable after transplanting immediately.
Embodiment 2
Sodium hypochlorite final concentration of 0.004wt.% in the medium.
The indoor temperature control of culture is in 25(± 2) DEG C.Intensity of illumination is 2000lx, the daily 10h of light application time.
Remaining method is same as Example 1.
Embodiment 3
Sodium hypochlorite final concentration of 0.01wt.% in the medium.
The indoor temperature control of culture is in 25(± 2) DEG C.Intensity of illumination is 2000lx, the daily 12h of light application time.
Remaining method is same as Example 1.
Result of implementation is analyzed:
According to Fig. 1, without antibacterial culture medium cultivate 9 days through the bacterium that naturally falls after surface cover with various funguses and bacterial clump
(a)；Add the no any bacterium colony (b) of media surface of 0.02% sodium hypochlorite；Add the culture medium of 0.02 g/l Mancozeb
The no any bacterium colony (c) in surface；The media surface adding 100 times of carbendazim covers with fungus colony, but no bacterial clump growth
(d)；The media surface adding 0.02g/l potassium sorbate covers with fungus colony, but no bacterial clump growth (e).Illustrate four kinds
Antibacterial all can bacteria growing inhibiting, and sodium hypochlorite and Mancozeb suppress the effect of funguses growth be significantly better than carbendazim and
Potassium sorbate.
According to Fig. 2, after African Chrysanthemum tissue cultured seedling enrichment culture 20d, compared with the control, add the tissue culture of 0.010% sodium hypochlorite
Seedling growth is suppressed, and Seedling gesture is significantly less intensive.African Chrysanthemum enrichment culture related data statistics is as follows:
Known by table, sodium hypochlorite concentration is higher, and pollution rate is lower, the lower necrosis rate of growth coefficient is higher.Consider,
0.008% sodium hypochlorite concentration is compared with being suitably applied the open enrichment culture of African Chrysanthemum.
According to Fig. 3, after African Chrysanthemum tissue cultured seedling root culture 20d, compared with the control, add the tissue culture of 0.010% sodium hypochlorite
Seedling rooting is suppressed.African Chrysanthemum root culture related data statistics is as follows:
Known by table, sodium hypochlorite concentration is higher, and rooting rate is lower, and necrosis rate is lower.Consider, 0.008% sodium hypochlorite is dense
Degree is suitably applied the open root culture of African Chrysanthemum.
The foregoing is only presently preferred embodiments of the present invention, all impartial changes done according to scope of the present invention patent with
Modify, all should belong to the covering scope of the present invention.
Claims (4)
1. a kind of method for culturing open type tissue of African Chrysanthemum it is characterised in that: by propagation and root media add
Antibacterial, directly inoculated and cultured under conditions of opening has bacterium.
2. a kind of African Chrysanthemum according to claim 1 method for culturing open type tissue it is characterised in that: described propagation
Culture medium is ms+6-ba 0.5mg/l+naa 0.1mg/l+ sucrose 30g/l+agar 7g/l.
3. a kind of African Chrysanthemum according to claim 1 method for culturing open type tissue it is characterised in that: described takes root
Culture medium is 1/2ms+naa 0.2mg/l+ sucrose 30g/l+agar 7g/l.
4. a kind of African Chrysanthemum according to claim 1 method for culturing open type tissue it is characterised in that: described is antibacterial
Agent is sodium hypochlorite, the final concentration of 0.004-0.010wt.% in culture medium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610988977.8A CN106332784A (en) | 2016-11-10 | 2016-11-10 | Open gerbera jamesonii tissue culture method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610988977.8A CN106332784A (en) | 2016-11-10 | 2016-11-10 | Open gerbera jamesonii tissue culture method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106332784A true CN106332784A (en) | 2017-01-18 |
Family
ID=57841156
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610988977.8A Pending CN106332784A (en) | 2016-11-10 | 2016-11-10 | Open gerbera jamesonii tissue culture method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106332784A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113678737A (en) * | 2021-10-11 | 2021-11-23 | 浙江省亚热带作物研究所 | Open tissue culture method for polygonatum cyrtonema |
| CN114451306A (en) * | 2022-02-23 | 2022-05-10 | 南阳师范学院 | Open tissue culture method for kiwi fruits |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101361456A (en) * | 2008-07-25 | 2009-02-11 | 云南省农业科学院花卉研究所 | Efficient flameray-gerbera propagation production method using excised leaf |
| CN102524073A (en) * | 2012-01-31 | 2012-07-04 | 北京市农林科学院 | Plant tissue culture medium added with sodium hypochlorite for sterilization instead of high temperature and culture method |
| CN102726290A (en) * | 2011-04-12 | 2012-10-17 | 山东省烟台农业学校 | Method for eliminating tissue culture seedling pollution |
| CN104642141A (en) * | 2015-03-13 | 2015-05-27 | 西南林业大学 | Gerbera adventitious bud induction and plant regeneration method by using root as explant |
-
2016
- 2016-11-10 CN CN201610988977.8A patent/CN106332784A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101361456A (en) * | 2008-07-25 | 2009-02-11 | 云南省农业科学院花卉研究所 | Efficient flameray-gerbera propagation production method using excised leaf |
| CN102726290A (en) * | 2011-04-12 | 2012-10-17 | 山东省烟台农业学校 | Method for eliminating tissue culture seedling pollution |
| CN102524073A (en) * | 2012-01-31 | 2012-07-04 | 北京市农林科学院 | Plant tissue culture medium added with sodium hypochlorite for sterilization instead of high temperature and culture method |
| CN104642141A (en) * | 2015-03-13 | 2015-05-27 | 西南林业大学 | Gerbera adventitious bud induction and plant regeneration method by using root as explant |
Non-Patent Citations (1)
| Title |
|---|
| 冯新等: "非洲菊离体快繁条件的优化", 《亚热带农业研究》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113678737A (en) * | 2021-10-11 | 2021-11-23 | 浙江省亚热带作物研究所 | Open tissue culture method for polygonatum cyrtonema |
| CN114451306A (en) * | 2022-02-23 | 2022-05-10 | 南阳师范学院 | Open tissue culture method for kiwi fruits |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102124946B (en) | Method for tissue culture of paeonia lactiflora | |
| CN102577976B (en) | Simple tissue culture method for broussonetia papyrifera | |
| CN103563745B (en) | A kind of method for tissue culture of ilex verticillata | |
| CN102657094B (en) | Ex-vitro soilless cutting rooting method for tissue-cultured and proliferated seedlings of gerbera jamesonii bolus | |
| CN103283598B (en) | Primary fir wood culture bud induction method | |
| CN105638477A (en) | Rapid propagation method for dendrobium hancockii seeds | |
| CN105052738B (en) | A kind of method of Chinese tamarisk tissue-culturing quick-propagation | |
| CN103202233A (en) | Butterfly orchid anti-browning tissue culture method and anti-browning culture medium | |
| CN104396742B (en) | The method that aseptic seedling is differentiated again with five-step approach induction Lilium sulphureum Baker bulbil calluss | |
| CN103931493A (en) | Tissue culture method of pinellian ternate forming seedling through one-step culture and novel pinellia ternate medium | |
| CN102318559A (en) | Culture medium composition for rapid rooting of blueberry tissue culture seedling outside test tube and preparation method | |
| CN102613083A (en) | North American redwood tissue cultivation method | |
| CN104335887A (en) | Method for increasing transplanting survival rate of tissue culture hybrid seedlings of orchid | |
| CN105010147A (en) | Special culture medium for improving tissue culture propagation speed of haworthia succulent plants and tissue culture method | |
| CN103270946B (en) | A kind of seedlings of Dendrobium officinale is produced and hardening synchronous method | |
| CN102823502A (en) | Method for intermediately propagating and culturing vitis quinquangularis in vitro | |
| CN104396759B (en) | The method that ash tree tissue cultures is bred fast | |
| CN103270950A (en) | Chrysanthemum simplified tissue culturing method | |
| CN103975851B (en) | A kind of method of FLOS CHRYSANTHEMI ALBA from Haizhou of China tissue cultures and breeding | |
| CN103798142B (en) | A kind of take stamen as the method that explant sets up lily embryo callus subculture regenerating system | |
| CN104885943B (en) | A kind of oncidiumLuridum chemical disinfection tissue culture method | |
| CN106359107A (en) | Tissue culture method of wild lilium | |
| CN103947548A (en) | Method for establishing agapanthus high-frequency regeneration system | |
| CN106332784A (en) | Open gerbera jamesonii tissue culture method | |
| CN109220789A (en) | The tissue culture and rapid propagation method of apple rootstock M9-T337 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170118 |