CN107484660A - A kind of plant tissue culture method using incubator - Google Patents
A kind of plant tissue culture method using incubator Download PDFInfo
- Publication number
- CN107484660A CN107484660A CN201710835784.3A CN201710835784A CN107484660A CN 107484660 A CN107484660 A CN 107484660A CN 201710835784 A CN201710835784 A CN 201710835784A CN 107484660 A CN107484660 A CN 107484660A
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- China
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- stage
- nutrient solution
- box body
- tissue culture
- absorbent substrate
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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
- A01H4/008—Methods for regeneration to complete plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
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- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention belongs to field of plant tissue culture technique, is related to a kind of plant tissue culture method using incubator.It is divided into five stages：First stage is that aseptic process and nutrient solution enter the box body stage, and absorbent substrate is poured on pallet with holes；Second stage is the sterile culture stage, opens protecgulum, plant tissue materials are placed on absorbent substrate and cultivated；Phase III is to change the nutrient solution stage, and the liquid-leaking nozzle of incubator first is opened into drained nutrient solution, closes liquid-leaking nozzle, opens protecgulum, new nutrient solution is poured into, closing lid；Fourth stage is into the seedling rooting stage, and the young plant for growing up to strain is cut out, changes culture of rootage liquid into, pallet with holes before is substituted for into fixed tray, the young plant of strain is put into fixed cover；5th stage was the hardening stage, and gland and adjustable gland are opened, and treated can transplanting in 3 days or so or transport.The plant that the inventive method is cultivated is shorter than the cycle of traditional tissue culture method, and young plant is healthy and strong, and survival rate is high.
Description
Technical field
The invention belongs to field of plant tissue culture technique, is related to a kind of plant tissue culture method using incubator.
Background technology
In traditional plant tissue culture work, using it is more be blake bottle and culture dish, young plant is placed in blake bottle, empty
Between it is small, be unfavorable for young plant and grow tall to become strong.Blade is placed in culture dish, aqueous vapor weight in ware, influences the growing way of callus.Traditional tissue culture
Process it is excessively cumbersome, often, cause it is artificial waste it is more；Root media is agar, needs to wash agar off during transplanting, easily
Hinder its root system, but also bad transport, survival rate are low.
The content of the invention
The present invention provides a kind of method using incubator tissue culture plant to solve the above problems.
The inventive method uses a kind of incubator in sterile tissue incubation, from explant callus induction, to seedling,
Hardening may be by the incubator.The incubator, including enclosed box body, there is protecgulum before box body, pass through sealing ring and box
Body combines, and has two air-vents at the top of box body, and one of air-vent loads sterile film and the gland of hollow out, and another is ventilative
There is a semicircle dividing plate in hole, loads sterile film and adjustable gland, the pressure-adjustable be stamped it is semicircular turn dividing plate, the box body
There is carbon dioxide gas port at top；There is liquid-leaking nozzle the side of the box body close to bottom.
The cultural method of the present invention, is divided into five stages, each stage can be individually operated.
First stage is that aseptic process and nutrient solution enter the box body stage, and all accessories of incubator are first sterilized,
Nutrient solution is poured into box body, then absorbent substrate is poured on support pallet, the bottom of the support pallet and four wall holes；It is logical
Cross absorbent substrate suction nutrient solution；
Second stage is the sterile tissue culture stage, and vegetable material is put on absorbent substrate, is sucked and cultivated by absorbent substrate
Liquid, finally protecgulum is sealed.
Phase III is to change the nutrient solution stage, after nutrient consumption in nutrient solution, changes nutrient solution.First by incubator
Liquid-leaking nozzle is opened, and after nutrient solution is drained, is closed liquid-leaking nozzle, is opened protecgulum, new nutrient solution is poured into, closing lid sealing.Behaviour
Pallet need not be taken out when making, it is simple and convenient.
Fourth stage is into the seedling rooting stage, and the young plant for growing up to strain is cut out, changes culture of rootage liquid into, by branch before
Bracketing disk is substituted for fixed tray, as shown in Fig. 2 the fixed tray is used together with fixed cover, is equipped with the fixed cover
Absorbent substrate, then the young plant of strain is put into each fixed cover；
5th stage was the hardening stage, and gland and adjustable gland are opened, and young plant has been applicable extraneous bad border, it is possible to transplants
Or transport.
The inventive method survival rate is high, simple to operate.Week of the plant that the inventive method is cultivated than traditional tissue culture method
Phase is short, and young plant is healthy and strong, and survival rate is high.
The advantages of the inventive method, has：
1st, the tissue culture cycle is short, tissue culture efficiency high；
2nd, it is convenient to change nutrient solution, it is simple to operate, it is not easy microbiological contamination；
3rd, subculture is few, artificial to waste less；
4th, without condensate problem, can be controlled by adjusting gland；
5th, hardening is convenient, and fixed cover is degradable, can be planted together with young plant in soil, and survival rate is high；
6th, convenient transportation, it is not easily broken.
Brief description of the drawings
Fig. 1 is the culture box structural representation containing pallet with holes.
Fig. 2 is fixed tray schematic diagram.
Wherein, 1 protecgulum, 2 sealing rings, 3 support pallets, 4 absorbent substrates, 5 liquid-leaking nozzles, 6 carbon dioxide gas ports, 7 every bacterium
Film, 8 glands, 9 nutrient solutions, 10 box bodys, 11 adjustable glands, 12 fixed trays, 13 fixed covers, 14 air-vents.
Fig. 3 is to break up figure after 6 days.
Fig. 4 is figure of being taken root after 15 days.
Embodiment
Incubator used in the present invention, including box body, there is protecgulum before box body, combined by sealing ring and box body,
There are two air-vents at the top of box body, one of air-vent loads sterile film and the gland of hollow out, and another air-vent has half
Circular bulkheads, load sterile film and adjustable gland, the pressure-adjustable be stamped it is semicircular turn dividing plate, the top of the box body is also
There is carbon dioxide gas port；There is liquid-leaking nozzle the side of the box body close to bottom.
The accessory of the incubator of the present invention can use chemical agent to sterilize, physical sterilization.Plant is by being filled nutrient solution
Absorbent substrate 4 required nutrition is provided.The condensate and humidity cultivated in box can be controlled by adjusting adjustable gland 11.
Periodically to filling CO 2 in box, in case needed for plant.
When needing to change nutrient solution 9 in implementation process, liquid-leaking nozzle 5 need to only be opened, nutrient solution 9 will be automatically drained out,
Pallet need not be taken out, after drained, liquid-leaking nozzle 5 is blocked, then protecgulum 1 is opened and pours into new nutrient solution, finally closes protecgulum 1
Lid sealing.
The cultural method of the present invention, is divided into five stages：
First stage is the germination stage, MS nutrient solutions is poured into box body, then water suction base is poured on pallet with holes
Matter perlite, the bottom of the pallet with holes and four wall holes；Seed is placed on absorbent substrate, sucked by absorbent substrate
MS nutrient solutions, for needed for seed, finally sealing protecgulum；
Can germination in 4 days or so is only needed with the seed of the inventive method, the cycle is short, and germination percentage is high.Second stage is
In the blade tissue culture stage, after germination, first liquid-leaking nozzle is opened and discharges MS nutrient solutions, protecgulum is opened, induction broth is fallen
Enter, then blade is cut and (such as needs to convert, blade need to be contaminated to dark training three days), without changing pallet, can add on pallet
A little absorbent substrates, then the blade cut is put on absorbent substrate again, blade by absorbent substrate absorb needed for nutrition.Treat 10
After it or so, there is obvious differentiation sign, differentiation is soon.Phase III is to change the nutrient solution stage, and this stage treats that blade is grown into
During the young plant of strain, a nutrient solution need to be changed, only liquid-leaking nozzle need to be opened, then pour into new nutrient solution, it is simple to operate, reduce after
Generation.
Fourth stage is to change culture of rootage liquid, it is necessary to the young plant for growing up to strain is cut out into the seedling rooting stage, will before
Pallet be substituted for fixed tray, as shown in Fig. 2 being equipped with multiple fixed covers on this pallet, load absorbent substrate in fixed cover, then
The young plant of strain is put into each fixed cover, this fixed cover is fibrous material, can be degraded in soil, without taking out during transplanting,
Operation is very easy.
In 5th hardening stage in stage, two glands need to be only opened during the inventive method hardening, treat that 3 days or so cans are moved
Plant, survival rate is high, simple to operate.The plant that the inventive method is cultivated is shorter than the cycle of traditional tissue culture method, and young plant will
Stalwartness, survival rate are high.
The present invention is illustrated with reference to specific case study on implementation.
Tobacco K326 tissue cultures are tested：
The germination test of first stage seed, about 200, tobacco K326 seeds are selected, are put into 5ml in sterile centrifugation tube,
In superclean bench, first with 70% alcohol washes 2min, sterile washing 2 times, 8min then is soaked with 2.5% sodium hypochlorite,
It is stand-by after washing 4 times.Will culture box and its accessory with physically or chemically sterilizing, MS nutrient solutions and sterile film high-temperature sterilization, then
Nutrient solution is poured into culture box, then the pallet equipped with absorbent substrate is put into box, the seed disinfected is equably layered on
On absorbent substrate, closing lid sealing.25 DEG C, the illumination cultivation of 16h illumination/dark 8h.It will germinate within 4 days or so, after 1 month or so
Can seedling, into the second stage blade tissue culture stage, the liquid-leaking nozzle that will cultivate box is opened, and MS nutrient solutions are discharged, pour into cigarette
Careless induction broth then cuts blade, about 0.5cm × 0.5cm big or small slice, the blade cut is put into water suction base in culture box
In matter, such as need to convert, also need blade contaminating dark training three days, treat just to have differentiation sign in 6 days, as shown in Figure 3.Phase III is changed
In the nutrient solution stage, after the young plant that one month or so can grow up to strain, a nutrient solution is changed in this middle of the month, only need by
Liquid-leaking nozzle is opened, and treats that nutrient solution is drained, then pours into new nutrient solution, simple to operate, is not easy microbiological contamination.The 4th can be entered afterwards
Stage takes root the stage, it can be seen that the tobacco plantlet for having stem to extend, it is stand-by to cut these plantlets, in sterile culture box added with
Tobacco culture of rootage liquid.Change support pallet into fixed tray combination, absorbent substrate is added in fixed cover, then it is small by what is cut
Plant is inserted into absorbent substrate, closing lid sealing.Can be by adjusting the gland on culture box come the condensate in control box.In observation box
Plant growth state, whenever necessary can be to supplementing carbon dioxide gas in box.Such as Fig. 4, young plant grows tall within 15 days or so, well developed root system.
After 25 days or so, root system can penetrate fixed cover, at this moment can carry out for the 5th hardening stage in stage, and two glands for cultivating box are beaten
Open, can is taken and transplanted in soil after 3 days, directly plants fixed cover in soil together with young plant, and fixed cover is degradable, operation
Convenient, survival rate is high.
Claims (7)
- A kind of 1. plant tissue culture method using incubator, it is characterised in thatDescribed incubator, it is a kind of enclosed box body, box body has protecgulum, and protecgulum is combined by sealing ring and box body, the top of box body There are two air-vents in portion, and one of air-vent loads sterile film and the gland of hollow out, and another air-vent has semicircle dividing plate, Load every mycoderm and adjustable gland, pressure-adjustable, which is stamped, semicircular turns dividing plate；There is carbon dioxide gas port at the top of box body； There are support pallet or fixed tray in box body, support hole on pallet or fixed tray；Liquid-leaking nozzle is arranged at box body bottom；Cultural method, it is divided into five stages：First stage is that aseptic process and nutrient solution enter the box body stage, and all accessories of incubator are first sterilized, in box body Nutrient solution is inside poured into, then absorbent substrate is poured on support pallet, the bottom of the support pallet and four wall holes；Pass through suction Water-based sucks nutrient solution；Second stage is the sterile tissue culture stage, and vegetable material is put on absorbent substrate, sucks nutrient solution by absorbent substrate, most Protecgulum is sealed afterwards；Phase III is to change the nutrient solution stage, after nutrient consumption in nutrient solution, changes nutrient solution；First by the leakage of incubator Hole is opened, and after nutrient solution is drained, is closed liquid-leaking nozzle, is opened protecgulum, new nutrient solution is poured into, closing lid sealing；Fourth stage is into the seedling rooting stage, and the young plant for growing up to strain is cut out, changes culture of rootage liquid into, by supporting bracket before Disk is substituted for fixed tray, and the fixed tray is used together with fixed cover, and absorbent substrate is housed in the fixed cover, then into The young plant of strain is put into each fixed cover；5th stage was the hardening stage, and gland and adjustable gland are opened, and young plant has adapted to extraneous bad border, it is possible to transplanting or Transport.
- 2. tissue culture method according to claim 1, it is characterised in that described incubator is transparent confinement box body.
- 3. tissue culture method according to claim 1, it is characterised in that the support pallet is by hollow footing and tray set Into.
- 4. hollow footing according to claim 4 and pallet, it is characterised in that suction hole is distributed with footing and pallet Hole.
- 5. tissue culture method according to claim 1, it is characterised in that absorbent substrate is put in pallet and footing, in footing Absorbent substrate sucks nutrient solution by intensive hole, nutrient solution is conducted into top absorbent substrate, then will be cultivated by absorbent substrate Liquid is supplied to clay thing.
- 6. tissue culture method according to claim 1, it is characterised in that the fixed tray combines with fixed cover, fixed cover It is placed in the positioning hole of fixed tray.
- 7. tissue culture method according to claim 6, it is characterised in that fixed cover uses fibrous material, has suction in fixed cover Water-based, absorbent substrate absorb nutrient solution, clay thing nutrient solution being supplied to above matrix.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710835784.3A CN107484660A (en) | 2017-09-16 | 2017-09-16 | A kind of plant tissue culture method using incubator |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710835784.3A CN107484660A (en) | 2017-09-16 | 2017-09-16 | A kind of plant tissue culture method using incubator |
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| CN107484660A true CN107484660A (en) | 2017-12-19 |
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| CN201710835784.3A Pending CN107484660A (en) | 2017-09-16 | 2017-09-16 | A kind of plant tissue culture method using incubator |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112753576A (en) * | 2020-12-31 | 2021-05-07 | 武汉叶动力生物科技股份有限公司 | Plant tissue culture method and plant tissue culture container thereof |
| CN114916422A (en) * | 2022-06-07 | 2022-08-19 | 贵州大学 | Plant growth sterile experiment culture apparatus |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN204482615U (en) * | 2015-03-16 | 2015-07-22 | 武汉叶动力生物科技有限公司 | One Plants aseptic culture device |
-
2017
- 2017-09-16 CN CN201710835784.3A patent/CN107484660A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN204482615U (en) * | 2015-03-16 | 2015-07-22 | 武汉叶动力生物科技有限公司 | One Plants aseptic culture device |
Non-Patent Citations (2)
| Title |
|---|
| 刘勇: "《绿色技术创新在我国绿洲区支柱产业中的实践与管理研究》", 31 December 2013 * |
| 胡一鸿等: "《农业生物技术教程》", 31 August 2015 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112753576A (en) * | 2020-12-31 | 2021-05-07 | 武汉叶动力生物科技股份有限公司 | Plant tissue culture method and plant tissue culture container thereof |
| CN114916422A (en) * | 2022-06-07 | 2022-08-19 | 贵州大学 | Plant growth sterile experiment culture apparatus |
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Application publication date: 20171219 |