CN104263690A - Rapid propagation method for callus and suspension cells of Ficus virens - Google Patents
Rapid propagation method for callus and suspension cells of Ficus virens Download PDFInfo
- Publication number
- CN104263690A CN104263690A CN201410446370.8A CN201410446370A CN104263690A CN 104263690 A CN104263690 A CN 104263690A CN 201410446370 A CN201410446370 A CN 201410446370A CN 104263690 A CN104263690 A CN 104263690A
- Authority
- CN
- China
- Prior art keywords
- callus
- pueraria lobota
- suspension cell
- culture
- yellow
- 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
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a rapid propagation method for a callus and suspension cells of Ficus virens. The rapid propagation method comprises the following steps of obtaining sterile explants, inducing the callus, proliferating the callus and then culturing the suspension cells, and the like. According to the rapid propagation method, Ficus virens cells with stable inheritable characters can be rapidly obtained by virtue of the cell suspension culture method, the reproductive rate is high, a large number of pharmaceutical raw materials are provided in a short term and the production cost can be effectively reduced.
Description
Technical field
Under the present invention relates to condition of tissue culture, the cultivation of yellow Pueraria lobota suspension cell, belongs to plant technology field.
Background technology
Huang Geshu, latin name
ficus virens, another name Ammonium alum method, great Ye banyan, horse hair banyan, sparrow tree, Moraceae Ficus fallen leaves megaphanerophyte.The cultivation of the ground such as Chongqing, Sichuan, Hubei is best, and by residence, bridge side, trackside be seen everywhere, and after young leaves unfolding, cherry stipule lands one after another, very attractive in appearance.One of conventional shade tree, shade tree.It is now the city tree of Chongqing in China.Huang Geshu likes light, drought-enduring, barren-resistant, has aerial root, and adaptive faculty is strong especially.Ecotope: be born in sparse woods or small stream limit wetland, winter hardiness comparatively banyan is slightly strong.Yellow its root of Ge Shu, leaf are used as medicine.Root: dispel rheumatism is clearing heat and detoxicating.For treating rheumatic ostealgia, flu, tonsillitis, eye conjunctivitis.Leaf: swelling and pain relieving.Treating swelling and pain by traumatic injury is controlled in external application.Modes of reproduction main is at present cottage propagation and seed propagation.Tissue culture breeding method there is no people's research, and utilization group training suspension cell culture can shorten culture cycle, keeps genetic stability, obtains a large amount of pharmaceutical raw materials in a short time.
Summary of the invention
Technical problem to be solved by this invention is to provide the rapid breeding method of yellow Pueraria lobota cell suspension culture, and obtain the yellow Pueraria lobota cell of stabilization characteristics of genetics fast, breeding potential is high, provides a large amount of pharmaceutical raw materials in a short time, and effectively can reduce production cost.
For solving the problems of the technologies described above, the present invention adopts following technical proposal:
Get the blade of yellow Pueraria lobota, first in chlorinated lime, soak 3min, running water is 25min, clorox process 10min on Bechtop, aseptic water washing 5-7 time, induction of callus is carried out in the vast mycin inducing culture of yellow Pueraria lobota blade access substratum MS+IAA0.2mg/L+2ip1mg/l+15mg/L of disinfecting, additional saccharose 30g/L, agar 6.5g/L, PH5.8, dark phase 16h, photophase 8 h, the callus derived puts into substratum MS+2, 4-D 0.2mg/L+TDZ0.1mg/L+ citric acid 100mg/L carries out callus proliferation cultivation, additional saccharose 30g/L, agar 6.5g/L, PH5.8, illumination 3000lx, callus after propagation is put into liquid nutrient medium MS+NAA 0.1mg/L+ZT0.3mg/L+ABA2mg/L+ lanthanum nitrate 2mg/L and is carried out suspension cell culture, additional saccharose 30g/L, PH5.8, illumination 4000lx, culture condition is culture temperature 25 DEG C, illumination 11 ~ 13 h/d, rotating speed 100 ~ 120 r/min.
Adopt yellow Pueraria lobota suspension cell prepared by the present invention, fast growth, proliferation rate is high, and be beneficial to large production operation, energy consumption is little, pollutes few.
Below in conjunction with embodiment, the present invention is further elaborated, but the scope of protection of present invention is not limited to following embodiments.
Embodiment
Embodiment 1
Get the blade of yellow Pueraria lobota, first in chlorinated lime, soak 3min, running water is 25min, clorox process 10min on Bechtop, aseptic water washing 5-7 time, the yellow Pueraria lobota blade access culture medium prescription of disinfecting is carry out induction of callus in the vast mycin inducing culture of MS+IAA0.2mg/L+2ip1mg/l+15mg/L, additional saccharose 30g/L, agar 6.5g/L, PH5.8, dark phase 16h, photophase 8 h, the callus derived puts into substratum MS+2, 4-D 0.2mg/L+TDZ0.1mg/L+ citric acid 100mg/L carries out callus proliferation cultivation, additional saccharose 30g/L, agar 6.5g/L, PH5.8, illumination 3000lx, callus after propagation is put into liquid nutrient medium MS+NAA 0.05mg/L+ZT0.2mg/L+ABA1mg/L+ lanthanum nitrate 1mg/L and is carried out suspension cell culture, additional saccharose 30g/L, PH5.8, illumination 4000lx.
Embodiment 2
Get the blade of yellow Pueraria lobota, first in chlorinated lime, soak 3min, running water is 25min, clorox process 10min on Bechtop, aseptic water washing 5-7 time, the yellow Pueraria lobota blade access culture medium prescription of disinfecting is carry out induction of callus in the vast mycin inducing culture of MS+IAA0.2mg/L+2ip1mg/l+15mg/L, additional saccharose 30g/L, agar 6.5g/L, PH5.8, dark phase 16h, photophase 8 h, the callus derived puts into substratum MS+2, 4-D0.2mg/L+TDZ0.1mg/L+ citric acid 100mg/L carries out callus proliferation cultivation, additional saccharose 30g/L, agar 6.5g/L, PH5.8, illumination 3000lx, callus after propagation is put into liquid nutrient medium MS+NAA 0.1mg/L+ZT0.4 mg/L+ABA3mg/L+ lanthanum nitrate 3 mg/L and is carried out suspension cell culture, additional saccharose 30g/L, PH5.8, illumination 4000lx.
Embodiment 3
Get the blade of yellow Pueraria lobota, first in chlorinated lime, soak 3min, running water is 25min, clorox process 10min on Bechtop, aseptic water washing 5-7 time, the yellow Pueraria lobota blade access culture medium prescription of disinfecting is carry out induction of callus in the vast mycin inducing culture of MS+IAA0.2mg/L+2ip1mg/l+15mg/L, additional saccharose 30g/L, agar 6.5g/L, PH5.8, dark phase 16h, photophase 8 h, the callus derived puts into substratum MS+2, 4-D0.2mg/L+TDZ0.1mg/L+ citric acid 100mg/L carries out callus proliferation cultivation, additional saccharose 30g/L, agar 6.5g/L, PH5.8, illumination 3000lx, callus after propagation is put into liquid nutrient medium MS+NAA 0.1mg/L+ZT0.4 mg/L+ABA3mg/L+ lanthanum nitrate 2mg/L and is carried out suspension cell culture, additional saccharose 30g/L, PH5.8, illumination 4000lx.
Embodiment 4
Get the blade of yellow Pueraria lobota, first in chlorinated lime, soak 3min, running water is 25min, clorox process 10min on Bechtop, aseptic water washing 5-7 time, the yellow Pueraria lobota blade access culture medium prescription of disinfecting is carry out induction of callus in the vast mycin inducing culture of MS+IAA0.2mg/L+2ip1mg/l+15mg/L, additional saccharose 30g/L, agar 6.5g/L, PH5.8, dark phase 16h, photophase 8 h, the callus derived puts into substratum MS+2, 4-D0.2mg/L+TDZ0.1mg/L+ citric acid 100mg/L carries out callus proliferation cultivation, additional saccharose 30g/L, agar 6.5g/L, PH5.8, illumination 3000lx, callus after propagation is put into liquid nutrient medium MS+NAA0.05mg/L+ZT0.3 mg/L+ABA2mg/L+ lanthanum nitrate 2mg/L and is carried out suspension cell culture, additional saccharose 30g/L, PH5.8, illumination 4000lx.
Claims (5)
1. a rapid breeding method for yellow Pueraria lobota callus and suspension cell, comprise the acquisition of aseptic explant, the induction of callus, the propagation, suspension cell culture etc. of callus, key step is as follows:
(1) get yellow Pueraria lobota young leaflet tablet, sterilization method is conveniently to its disinfection;
(2) induction of callus is carried out in the vast mycin inducing culture of yellow Pueraria lobota blade access substratum MS+IAA0.2mg/L+2ip1mg/l+15mg/L of step (1) being disinfected, additional saccharose 30g/L, agar 6.5g/L, PH5.8, dark phase 16h, photophase 8h;
(3) get the callus that step (2) derives and put into substratum MS+2,4-D 0.2mg/L+TDZ0.1mg/L+ citric acid 100mg/L carries out callus proliferation cultivation, additional saccharose 30g/L, agar 6.5g/L, PH5.8, illumination 3000lx;
(4) callus got after step (3) propagation is put into liquid nutrient medium MS+NAA 0.05-0.1mg/L+ZT0.2-0.4 mg/L+ABA1-3mg/L+ lanthanum nitrate 1-3 mg/L and is carried out suspension cell culture, additional saccharose 30g/L, PH5.8, illumination 4000lx;
According to the rapid breeding method of a kind of yellow Pueraria lobota callus according to claim 1 and suspension cell, it is characterized in that: the acquisition of the aseptic blade of yellow Pueraria lobota described in step (1) is, get the blade of yellow Pueraria lobota, first in chlorinated lime, soak 3min, running water is 25min, clorox process 10min, aseptic water washing 5-7 time on Bechtop.
2., according to the rapid breeding method of a kind of yellow Pueraria lobota callus according to claim 1 and suspension cell, it is characterized in that: the induction of callus described in step (2) addition of the vast mycin of 15mg/L in the medium, can eliminate endophyte.
3., according to the rapid breeding method of a kind of yellow Pueraria lobota callus according to claim 1 and suspension cell, it is characterized in that: addition of citric acid in the propagation of callus described in step (3), effectively can prevent the browning of callus.
4. according to the rapid breeding method of a kind of yellow Pueraria lobota callus according to claim 1 and suspension cell, it is characterized in that: the auspicious wooden suspension cell of the Huang described in step (4) is according to obtaining by the following method: select free-running property good, healthy callus, after subculture 3 times, aseptically put into the Erlenmeyer flask that 25 mL liquid nutrient mediums are housed respectively, after adding the granulated glass sphere jolting of sterilization, be placed in shaking culture case and carry out horizontal oscillations cultivation, culture condition is culture temperature 25 DEG C, illumination 11 ~ 13 h/d, rotating speed 100 ~ 120 r/min.
5., according to the rapid breeding method of the auspicious wooden callus of a kind of Huang according to claim 1 and suspension cell, it is characterized in that: step adds rare earth lanthanum nitrate in (4), has promoter action to the growth of cell suspension culture.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410446370.8A CN104263690A (en) | 2014-09-04 | 2014-09-04 | Rapid propagation method for callus and suspension cells of Ficus virens |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410446370.8A CN104263690A (en) | 2014-09-04 | 2014-09-04 | Rapid propagation method for callus and suspension cells of Ficus virens |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104263690A true CN104263690A (en) | 2015-01-07 |
Family
ID=52155274
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410446370.8A Pending CN104263690A (en) | 2014-09-04 | 2014-09-04 | Rapid propagation method for callus and suspension cells of Ficus virens |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104263690A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105900741A (en) * | 2016-05-05 | 2016-08-31 | 广西壮族自治区农业科学院生物技术研究所 | Method for cultivating pachyrhizua angulatus tissue culture seedlings by means of plant growth lamps |
| CN110923190A (en) * | 2019-11-28 | 2020-03-27 | 大连普瑞康生物技术有限公司 | Method for improving flavone phenylpropanoid compounds of saussurea involucrate cell culture |
-
2014
- 2014-09-04 CN CN201410446370.8A patent/CN104263690A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105900741A (en) * | 2016-05-05 | 2016-08-31 | 广西壮族自治区农业科学院生物技术研究所 | Method for cultivating pachyrhizua angulatus tissue culture seedlings by means of plant growth lamps |
| CN110923190A (en) * | 2019-11-28 | 2020-03-27 | 大连普瑞康生物技术有限公司 | Method for improving flavone phenylpropanoid compounds of saussurea involucrate cell culture |
| WO2021103863A1 (en) * | 2019-11-28 | 2021-06-03 | 大连普瑞康生物技术有限公司 | Method for increasing the content of flavonoid phenylpropanoid compounds in saussurea involucrata cell culture |
| EP4067479A4 (en) * | 2019-11-28 | 2023-01-11 | Dalian Practical Biotechnology Co., Ltd. | Method for increasing the content of flavonoid phenylpropanoid compounds in saussurea involucrata cell culture |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106386511B (en) | The rapid propagation method of beautiful millettia root suspension cell culture | |
| CN102668989B (en) | Shallow culture method for sugarcane liquid | |
| CN102577976B (en) | Simple tissue culture method for broussonetia papyrifera | |
| CN101766122B (en) | Method for cultivating sweet sorghum tissue and special culture medium thereof | |
| CN103548694B (en) | Tissue culture and rapid propagation method for dracaena cochinchinensis | |
| CN103688865A (en) | Inducing medium and method for improving survival rate of butterfly orchid pedicel | |
| CN105165626A (en) | Culture medium formula of plant callus induction and subculture multiplication of astragalus membranaceus (fisch.) bunge and culture method of astragalus membranaceus (fisch.) bunge | |
| CN106577291B (en) | The method that radix stemonae sessilifoliae callus efficiently induces seedling | |
| CN104263690A (en) | Rapid propagation method for callus and suspension cells of Ficus virens | |
| CN102640710B (en) | Method for induction and subculture of taxus chinensis leaf callus and culture medium used by method | |
| CN104285792B (en) | A kind of tissue rapid propagation method of Alnus formosana Plantation | |
| CN105850729A (en) | Verbena bonariensis cultivation method | |
| CN106577272B (en) | A kind of method for tissue culture of water shield | |
| CN104255478A (en) | Method for quickly reproducing Murraya exotica L. suspension cells | |
| CN107125136A (en) | A kind of Camellia nitidissima tissue culture mating system | |
| CN106172002A (en) | A kind of production method of Folium Artemisiae Argyi tissue cultured seedling | |
| CN103314859A (en) | Stem tip tissue culture propagation method of dendrobium officinale | |
| CN102511393A (en) | Method for establishing racomitrium japonicum gametophyte regeneration system | |
| CN105660415A (en) | In-vitro preservation method of drynaria roosii nakaike | |
| CN109730010A (en) | A kind of American shad stereo ecological cultivating system | |
| CN104304007B (en) | A kind of in-vitro culture method of berchemia lineata | |
| CN101243773B (en) | Induction method for Camptotheca acuminata Decne polyploid | |
| CN102805032B (en) | Method for preventing daemonorops margaritae callus browning phenomena from occurring | |
| CN105746347A (en) | In-vitro preservation method of corydalis thalictrifolia | |
| CN107114241A (en) | A kind of method for culturing seedlings of passion fruit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150107 |
|
| WD01 | Invention patent application deemed withdrawn after publication |