CN102577955A - Banana liquid tissue rapid propagation technology and special culture medium thereof - Google Patents
Banana liquid tissue rapid propagation technology and special culture medium thereof Download PDFInfo
- Publication number
- CN102577955A CN102577955A CN2012100347925A CN201210034792A CN102577955A CN 102577955 A CN102577955 A CN 102577955A CN 2012100347925 A CN2012100347925 A CN 2012100347925A CN 201210034792 A CN201210034792 A CN 201210034792A CN 102577955 A CN102577955 A CN 102577955A
- Authority
- CN
- China
- Prior art keywords
- culture
- culture medium
- medium
- banana
- final concentration
- 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
Images
Abstract
The invention relates to a banana liquid tissue rapid propagation technology, which is characterized by comprising the step of selecting a liquid culture medium taking water as a main solvent and carrying out banana tissue rapid propagation. A culture medium formula comprises a enrichment culture medium and a rooting culture medium, wherein each culture medium is mainly prepared by adding solvents, such as BA (benzyladenine), NNA (Naphthyl Acetic Acid) and GA (Gibberellic acid) into an MS(Murashige Sroog) basal culture medium solution taking water as a main solvent; the final concentration of the BA in the enrichment culture medium is 2.0mg/L, and the final concentration of the NNA in the enrichment culture medium is 1.0mg/L; and the final concentration of the GA in the rooting culture medium is 1.0mg/L, and the final concentration of the NAA in the rooting culture medium is 1.5mg/L. According to the banana liquid tissue rapid propagation technology, because the liquid culture mediums are adopted and a coagulating agent, such as agar does not need to be added, semiautomation of a culturing process is achieved; and because reinoculation is not required in the subculture process and only are the culture mediums changed, the labor is saved. Compared with the traditional solid culture method, the banana liquid tissue rapid propagation technology has the advantages that: the material and labor cost can be reduced by more than 30 percent, and the cost of each seedling can be reduced by 0.075Yuan.
Description
Technical field
The present invention relates to a kind of novel technical method of banana tissue rapid propagation, set up corresponding culture technique and supporting medium system, belong to Plant Tissue Breeding and biological technical field.
Background technology
Banana is that China also is one of main in the world fresh fruit kind, and annual banana sales volume accounts for about 40% of the total fruit sales volume in the whole world, and demand is very big.Along with the continuous expansion of domestic cultivated area, the situation that supply falls short of demand often appears in the also increase day by day of demand to seedling.Present most banana seedling all derives from tissue cultivating seedling, and traditional banana method for tissue culture is a solid culture.In the operating process, from just cultivating successive transfer culture, each cultivation stage all is manual operations, and production process needs a large amount of hand labours, is a kind of labor-intensive technology.The medium that the tradition cultural method is selected for use is solid culture medium, needs to add coagulating agents such as agar, has increased cost of material.The tradition cultural method is because of the restriction of culture vessel, and the growing space of tissue cultivating seedling is limited, and a bottle only can be put 3-5 tissue culture seedlings of bananas, and incubation do not have air interchanger, poor aeration, and growth coefficient is limited, and the quality of seedling is uneven.These all are the common problems that exist in present traditional tissue culture seedlings of bananas production, also are the higher reasons of tissue cultivating seedling production cost.
Summary of the invention
The novel technical method and the supporting medium system that the purpose of this invention is to provide a kind of banana tissue rapid propagation, the propagation multiple of raising tissue cultivating seedling shortens rootage duration; Need not add coagulating agents such as agar; Reduce the input of culture medium raw material, incubation is semi-automatic, saves labour and cost.
The present invention is the semi-automatic tissue rapid propagation new technology of a kind of banana liquid; This technology comprises enrichment culture and required intermittent time in culture of rootage stage; Control such as chopper frequency parameter; Inoculum density, supporting this system that is applicable to carries out the proliferated culture medium and the culture of rootage based formulas of banana liquid culture.
A kind of banana liquid tissue quick-breeding method is characterized in that: selecting for use with water is that the liquid nutrient medium of primary solvent carries out the banana tissue rapid propagation, and culture apparatus is an intermittent immersion bioreactor.
The fast numerous special culture media of banana liquid tissue; Comprise proliferated culture medium and root media; The composition of medium mainly is with water in the MS minimal medium solution that is primary solvent; Adding following solute is made into: 6-benzyladenine (hereinafter to be referred as 6-BA), methyl (hereinafter to be referred as NAA) and gibberellin (hereinafter to be referred as GA);
The enrichment culture based formulas is: MS+6-BA 2.0mg/L+NAA0.1mg/L+30g/L sucrose;
The culture of rootage based formulas is: MS+GA 1.5mg/L+NAA1.0mg/L+30g/L sucrose.
The final concentration of 6-BA is 2.0mg/L in the said proliferated culture medium, and the final concentration of NAA is 0.1mg/L;
The final concentration of GA is 1.0mg/L in the said root media, and the final concentration of NAA is 1.5mg/L.
Each solute concentration is following in the MS minimal medium: potassium nitrate (KNO3) 1900mg/L, ammonium nitrate (NH4NO3) 1650mg/L, potassium dihydrogen phosphate (KH2PO) 170mg/L, magnesium sulfate (MgSO4.7H2O) 370mg/L; Calcium chloride (CaCl2.2H2O) 440mg/L, KI (KI) 0.83mg/L, boric acid (H3BO3) 6.2mg/L, manganese sulphate (MnSO4.4H2O) 22.3mg/L; Zinc sulphate (ZnSO4.7H2O) 8.6mg/L, sodium molybdate (Na2MoO4.2H2O) 0.25mg/L, copper sulphate (CuSO4.5) 0.025mg/L; Cobalt chloride (CoCl2.6H2O) 0.025mg/L, disodium ethylene diamine tetraacetate (Na2.EDTA) 37.3mg/L, ferrous sulfate (FeSO2.7H2O) 27.8mg/L; Inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride (VB1) 0.1mg/L; Puridoxine hydrochloride (VB6) 0.5mg/L, nicotinic acid (VB5) 0.5mg/L, sucrose (sucrose) 30g/L.
Described banana liquid tissue culture technique method is characterized in that:
(1) the subculture seedling should select for use the tissue cultivating seedling of semi-solid cultivation of the 7th generation to be advisable; Inoculum density is every strain/100mL medium,
(2) during enrichment culture, intermittently submergence bioreactor culture condition is: the intermittent time is 3 hours, and chopper frequency is 4min/3h, and the 6-BA concentration of proliferated culture medium is 2.0mg/L; Can make banana tissue culture generation propagation more than 14 times, far exceed conventional method 2-3 doubly.
(3) the culture of rootage stage, intermittently submergence bioreactor culture condition is: chopper frequency is 4min/6h, and root media is suitable for root induction with GA 1.5mg/L+NAA1.0mg/L.
The present invention adopts intermittent immersion bioreactor (hereinafter to be referred as TIBs); Be that a cover massive planting fabric texture is cultivated and the research of Secondary Metabolism of Plant material is the equipment of one; Its principle is to utilize liquid nutrient medium being that power carries out batch (-type) to the plant tissue culture seedling and cultivates through filtered air pressure; For plant provides a good environment in the process of liquid culture, can carry out the gas exchange in the incubator well, comprise the gas componant in the gas and medium in the container.TIBs can control the Immersion time of liquid nutrient medium in the incubation, and the submergence frequency guarantees that incubation is in germ-free condition.The recycle of medium can effectivelyly prevent the accumulation of nutrition deposition and harmful substance, thereby more effectively utilizes the nutrient component of medium.Utilize the TIBs system to carry out the Application Research of banana tissue-culturing rapid propagation, can further promote commercially producing of banana group training health seedling, improve the productivity effect of banana.
Result of the test shows, utilizes technical method of the present invention and supporting medium system, and the propagation multiple of generation tissue cultivating seedling can reach more than 12 times, than the high 2-3 of conventional solid cultural method doubly, sees table 1 for details.The enrichment culture stage is 30 days, and the elongation cultivation stage is 20 days, and the tissue cultivating seedling rootage duration is 20 days, has shortened 10-20 days than conventional method, and the result sees table 2.The present invention adopts liquid nutrient medium, need not add coagulating agents such as agar, reduces the input of culture medium raw material.Incubation is semi-automatic, and successive transfer culture and process of rooting culture do not need artificial infection, only need change medium and get final product, and saves the labour.The production cost of the every strain tissue culture seedlings of bananas of traditional semi-solid cultural method is greatly between 0.25-0.48 unit at present; Banana liquid tissue fast breeding technique of the present invention is compared with the conventional solid cultural method; Can reduce raw material and labor cost more than 30%, the cost that is equivalent to every young plant can reduce more than 0.075 yuan.
Table 1 TIBs cultivates with tradition is semi-solid and cultivates required fate relatively
Generation seedling proliferation rate that table 2TIBs cultivates and the tradition semisolid is cultivated and seedling mass ratio are
Description of drawings
Fig. 1 is an intermittent immersion bioreactor TIBs sketch map.
Among the figure, (1) air compressor (2) magnetic valve and time-controlled switch (3) needle-based air cleaner.
Embodiment
Practical implementation method of the present invention is following:
1, TIBs sets up according to the mentality of designing of Lorenzo (1998), and main building block is air compressor machine, magnetic valve, time-controlled switch, syringe filters, blake bottle and liquid storage bottle, and corresponding pipeline etc.Put into the material that needs cultivation in the blake bottle, tapping body medium in the liquid storage bottle.Blake bottle and liquid storage bottle all are the big spoken parts in an opera look vial of 3L among the TIBs, the high 30cm of bottle, diameter 15cm.The liquid nutrient medium volume is every bottle of 1L/ in the liquid storage bottle.
Accompanying drawing A, B, C are TIBs operation principle sketch map:
A shows air inlet after the compressor operating, and the liquid nutrient medium in the liquid storage bottle flows into blake bottle under pressure.
B shows that banana seedlings is immersed in the liquid nutrient medium in the blake bottle.
After C showed that Immersion time finishes, compressor is task again, and the liquid nutrient medium in the blake bottle flows back to liquid storage bottle under pressure.
2, the composition of medium mainly is with water in the MS minimal medium solution that is primary solvent, adds following solute and is made into: BA, NAA and GA, and the final concentration of BA is 2.0mg/L in the said proliferated culture medium, the final concentration of NAA is 0.1mg/L; The final concentration of GA is 1.0mg/L in the said root media, and the final concentration of NAA is 1.5mg/L; The solvent of said MS basic culture solution is a water.
With preparation 1L medium is example, each composition addition such as following table in the medium:
By the amount of required each composition of last table, good with the flat weighing of electronic scale, be placed in the beaker, be dissolved in water, after having dissolved solution is forwarded in the volumetric flask of 1L, add water to scale, shake all, be the medium of the 1L capacity for preparing.
Prepared culture medium is packed in the liquid storage bottle of 3L, seals bottleneck, and is subsequent use behind the sterilization 20min through 121 ℃, the tweezers that operating process is used, and instruments such as scalpel all use behind high-temperature sterilization.
The concrete operations step is following:
1, learn from else's experience semi-solid cultivate the 7th generation the subculture seedling be material; Cut single bud; Remove the part of blade-section and brown stain blackout, leave and take base portion and be no more than 1cm bud point, the single bud that cuts is inoculated in the blake bottle that 1L liquid proliferated culture medium is housed as inoculation material.
2, with blake bottle and liquid storage bottle, be connected in the TIBs device with silica gel hose, open time-controlled switch, set chopper frequency and intermittent time, regulate culturing room's temperature at 26 ± 2 ℃, intensity of illumination 2000lx, photoperiod 16h illumination/8h is dark.
3, the enrichment culture stage is 30 days, and the elongation stage is 20 days, and the stage of taking root is 20 days.Each cultivation stage is changed corresponding medium and is got final product after finishing.The replacing method of medium is following: the medium that corresponding vegetative stage is needed prepares, and is contained in the liquid storage bottle, and 121 ℃, sterilization 20min is connected among the TIBs then, changes the medium of previous stage.When changing medium, surface, positions such as the pipeline that is connected with liquid storage bottle with 75% alcohol disinfecting earlier, interface.
4, after culture of rootage finishes, can tissue cultivating seedling be taken out the refining seedling, transplant the land for growing field crops.
Claims (3)
1. banana liquid tissue quick-breeding method is characterized in that: selecting for use with water is that the liquid nutrient medium of primary solvent carries out the banana tissue rapid propagation, and culture apparatus is an intermittent immersion bioreactor.
2. fast numerous special culture media of banana liquid tissue; It is characterized in that: comprise proliferated culture medium and root media; The composition of medium mainly is with water in the MS minimal medium solution that is primary solvent; Adding following solute is made into: 6-benzyladenine 6-BA, methyl NAA and gibberellin GA; The final concentration of 6-BA is 2.0mg/L in the said proliferated culture medium, and the final concentration of NAA is 0.1mg/L; The final concentration of GA is 1.0mg/L in the said root media, and the final concentration of NAA is 1.5mg/L;
The enrichment culture based formulas is: MS+6-BA 2.0mg/L+NAA0.1mg/L+30g/L sucrose;
The culture of rootage based formulas is: MS+GA 1.5mg/L+NAA1.0mg/L+30g/L sucrose;
Each solute concentration is following in the minimal medium: potassium nitrate 1900mg/L, ammonium nitrate 1650mg/L, potassium dihydrogen phosphate 170mg/L, magnesium sulfate 370mg/L; Calcium chloride 440mg/L, KI 0.83mg/L, boric acid 6.2mg/L, manganese sulphate 22.3mg/L; Zinc sulphate 8.6mg/L, sodium molybdate 0.25mg/L, copper sulphate 0.025mg/L, cobalt chloride 0.025mg/L; Disodium ethylene diamine tetraacetate 37.3mg/L, ferrous sulfate 27.8mg/L, inositol 100mg/L, glycine 2mg/L; Thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, sucrose 30g/L.
3. banana liquid tissue quick-breeding method according to claim 1 is characterized in that:
(1) the subculture seedling is selected the semi-solid tissue cultivating seedling of cultivating of the 7th generation for use; Inoculum density is every strain/100mL medium;
(2) during enrichment culture, intermittently submergence bioreactor culture condition is: the intermittent time is 3 hours, and chopper frequency is 4min/3h, and the 6-BA concentration of proliferated culture medium is 2.0mg/L;
(3) the culture of rootage stage, intermittently submergence bioreactor culture condition is: chopper frequency is 4min/6h, root media GA 1.5mg/L+NAA1.0mg/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012100347925A CN102577955A (en) | 2012-02-16 | 2012-02-16 | Banana liquid tissue rapid propagation technology and special culture medium thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012100347925A CN102577955A (en) | 2012-02-16 | 2012-02-16 | Banana liquid tissue rapid propagation technology and special culture medium thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102577955A true CN102577955A (en) | 2012-07-18 |
Family
ID=46467645
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012100347925A Pending CN102577955A (en) | 2012-02-16 | 2012-02-16 | Banana liquid tissue rapid propagation technology and special culture medium thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102577955A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103988743A (en) * | 2014-04-21 | 2014-08-20 | 黄少伟 | Banana planting method |
| CN104067937A (en) * | 2014-06-23 | 2014-10-01 | 云南省农业科学院花卉研究所 | Tide type bioreactor and method for culturing lily bulblets thereof |
| CN111543324A (en) * | 2020-06-04 | 2020-08-18 | 云南省农业科学院农业环境资源研究所 | Banana tissue culture method suitable for DNA material requirements of BioNano technology |
-
2012
- 2012-02-16 CN CN2012100347925A patent/CN102577955A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 杨柳等: "利用间歇浸没式生物反应器进行香蕉（Musa AAA Cavendish var. Williams B6）组培快繁研究", 《果树学报》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103988743A (en) * | 2014-04-21 | 2014-08-20 | 黄少伟 | Banana planting method |
| CN104067937A (en) * | 2014-06-23 | 2014-10-01 | 云南省农业科学院花卉研究所 | Tide type bioreactor and method for culturing lily bulblets thereof |
| CN104067937B (en) * | 2014-06-23 | 2016-01-20 | 云南省农业科学院花卉研究所 | A kind of method of tidal type bio-reactor and cultivation lily seed ball thereof |
| CN111543324A (en) * | 2020-06-04 | 2020-08-18 | 云南省农业科学院农业环境资源研究所 | Banana tissue culture method suitable for DNA material requirements of BioNano technology |
| CN111543324B (en) * | 2020-06-04 | 2021-09-03 | 云南省农业科学院农业环境资源研究所 | Banana tissue culture method suitable for DNA material requirements of BioNano technology |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102150624B (en) | Tissue culture and rapid propagation method for pinellia tuber plant | |
| CN104472359B (en) | A kind of ginseng adventitious root proliferative induction method | |
| CN101647393B (en) | Fast tissue culture reproducing method of actinidia eriantha | |
| CN102119655B (en) | Natural light rapid breeding method for dendrobium officinale | |
| CN101933456A (en) | Method for quickly breeding seedlings of dendrobium officinale capsule | |
| CN101720670B (en) | Rapid breeding method for pinellia tuber tissue culture | |
| CN103120126B (en) | Method for carrying out tissue cultivation propagation of anoectochilus roxburghii by intermittent submerged bioreactor | |
| CN103004599B (en) | Method for obtaining crowtoe regeneration plantlet by anther culture | |
| CN103931492A (en) | Tissue-culture rapid seedling growing method for apple rootstock M9 | |
| CN104255524A (en) | Method for quickly producing Chinese fir seedlings through micro-cutting | |
| CN105638455B (en) | A kind of method and culture medium that Drying pepper haplobiont is obtained by Anther Culture | |
| CN103444523A (en) | Method for quickly introducing embryonic callus through anther to regenerate plant | |
| CN102405830B (en) | Induction method of rosa chinensis receptacle callus tissues | |
| CN104381133A (en) | Tissue culture breeding method of phlox subulata | |
| CN100394845C (en) | In-bottle production method of detoxified small seed ball of east lily | |
| CN103688862B (en) | A kind of method of water chestnut plantlet in vitro high efficiently multiplying | |
| CN102577955A (en) | Banana liquid tissue rapid propagation technology and special culture medium thereof | |
| CN102763592B (en) | Peony embryo culturing method | |
| CN104542296A (en) | Open rooting method for sugarcane tissue culture seedlings | |
| CN101658133A (en) | Method for quickly and directly establishing albizzia regeneration plant system | |
| CN112400696B (en) | Tissue culture method of evergreen common selfheal fruit-spike bamboo | |
| CN105494105B (en) | A kind of peony tissue culture vessel seedling technology | |
| KR20080073388A (en) | Mass production of bulblet via somatic embryogenic cell culture in lily | |
| CN103125387A (en) | Method of producing lily bulb by utilizing plant tissue culture technique | |
| CN104782501B (en) | May melon vine seedling cultural method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20120718 |