CN103283605A - Preparation method of tissue culture medium without agar - Google Patents
Preparation method of tissue culture medium without agar Download PDFInfo
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- CN103283605A CN103283605A CN2013102388430A CN201310238843A CN103283605A CN 103283605 A CN103283605 A CN 103283605A CN 2013102388430 A CN2013102388430 A CN 2013102388430A CN 201310238843 A CN201310238843 A CN 201310238843A CN 103283605 A CN103283605 A CN 103283605A
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Abstract
The invention belongs to plant tissue culture techniques in the field of biotechnology and particularly relates to a preparation method of a tissue culture medium without agar. The preparation method specifically comprises the steps of: firstly, preparing a nutrient solution according to needs, then, absorbing the nutrient solution by super absorbent resin for curing and forming, successively, sub-packaging the cured nutrient solution in tissue culture vessels, and finally, carrying out disinfection and sterilization on a culture medium according to a conventional method to obtain the tissue culture medium for production. By implementing the preparation method, the super absorbent resin is utilized for replacing the agar, and the prepared nutrient solution is directly absorbed by the absorbent resin to be cured and formed, so that the steps of heating and dissolving the nutrient solution, and then sub-packaging the solution into the tissue culture vessels in a conventional preparation method of the culture medium with the agar are omitted; the cost also can be substantially reduced since the curing and forming of 1000ml of culture medium only need 2g of super absorbent resin while 1000ml of culture medium needs 10g of agar in the conventional preparation of the culture medium. By implementing the preparation method, the factorization, large-scale and intensive production of plant tissue culture is facilitated.
Description
Technical field
The invention belongs to the plant tissue culture technique in the biological technical field, be specifically related to a kind of group training medium preparation method without agar.
Technical background
It is to have this theory of totipotency according to plant cell that the tissue of plant is cultivated, a vegetative new technology that grows up in recent decades.Plant Tissue Breeding is plant aseptic culture technology, claim cultured in vitro again, be to have totipotent theory according to plant cell, the organ (as root, stem, leaf, stem apex, flower, fruit etc.), tissue (as forming layer, epidermis, cortex, marrow cell, endosperm etc.) or cell (as megaspore, microspore, somatic cell etc.) and the protoplast that utilize plant corpus to exsomatize, under artificial conditions such as aseptic and suitable synthetic medium and illumination, temperature, can induce callus, indefinite bud, adventive root, form the subject of complete plant at last.
Traditional plant tissue culture is in implementation process, medium preparation is basically: the different materials of component during by MS medium+plant tissue culture formed with hormone+agar that different cultivation stage is added correspondence, then the medium for preparing is added the agar heating for dissolving, transfer to pH5.6-5.8, be sub-packed in the 100mL tissue culture bottle every bottle of about 30mL while hot.After treating the medium cooled and solidified, cover group training lid, or cover and seal film and fasten with cotton thread, then 121 ℃ (1kg/cm2) sterilization 20min down in high-pressure sterilizing pot.
Summary of the invention
The purpose of this invention is to provide brand-new a kind of group training medium preparation method without agar.Concrete is: concrete is: prepare nutrient solution at first as required, with high hydroscopic resin nutrient solution is absorbed curing molding then, then be sub-packed in the tissue culture bottle, according to a conventional method medium is carried out disinfection sterilization at last and must produce with group training medium.By implementing the present invention, utilize high hydroscopic resin to replace agar, after preparing, nutrient solution directly absorbed and curing molding by water-absorbing resin, and having omitted when traditional medium uses agar also needs the heating for dissolving inborn ability to install in the tissue culture bottle.Also can reduce cost significantly, the medium curing molding of 1000ml only needs 2 gram high hydroscopic resins, about 0.8～1 mao of cost, and during traditional medium preparation, the medium of 1000ml needs 10 gram agar, about 2 yuans of cost.Implement the present invention, more be conducive to plant tissue culture batch production, scale, intensive production.
(Super Absorbent Polymer SAP) is a kind of new functional macromolecule material to described high hydroscopic resin.Has hydrophilic radical, moisture can be absorbed in a large number and swelling can maintain the synthetic resin that moisture does not outflow, as the starch grafted acrylate class, graft acrylamide, the high substituted degree cross-linked carboxymethyl cellulose, the cross-linked carboxymethyl cellulose graft acrylamide, cross-linking type hydroxyethylcellulose graft acrylamide polymer etc., generally can absorb and be equivalent to the moisture of resin volume more than 100 times, the highest water absorption rate can reach more than 1000%, super absorbent polymer has nontoxic, to the human body nonirritant, no side reaction, do not cause characteristics such as blood coagulation, in recent years, be widely used in field of medicaments.Generally as medical material, as diaper, sanitary napkin etc., industrial also as loss circulation material.
The present invention is achieved through the following technical solutions:
A kind of group training medium preparation method without agar comprises the steps: in the technical scheme
1, selecting the suction multiple is that high hydroscopic resin more than 500 times is made curing agent.
Different materials during 2, according to plant tissue culture and different cultivation stages are mixed with nutrient solution with the hormone of MS medium prescription+correspondence.
3, according to quantity high hydroscopic resin is added in the nutrient solution, nutrient solution is absorbed curing molding.
4, according to quantity the nutrient solution of curing molding is sub-packed in the tissue culture bottle.
5, according to a conventional method medium is carried out disinfection sterilization and must producing with group training medium.
Compared with prior art, the existing following advantage of the present invention:
1, group proposed by the invention is trained medium preparation method and measure uniqueness, implements easily.
2, by implementing the present invention, utilize high hydroscopic resin to replace agar, after preparing, nutrient solution directly absorbed and curing molding by water-absorbing resin, and having omitted when traditional medium uses agar also needs heating for dissolving, and inborn ability installs in the tissue culture bottle.
3, by implementing the present invention, the medium curing molding of 1000ml only needs 2 gram high hydroscopic resins, about 0.8～1 mao of cost, and during traditional medium preparation, the medium of 1000ml needs 10 gram agar, about 2 yuans of cost; Therefore, adopt the present invention to reduce cost significantly.
4, by enforcement the present invention, after the medium moulding, because its institutional framework is granular, thus the compactness that unlike traditional medium, hardens, but loose, at more favourable root growth of culture of rootage stage.
5, implement the present invention, more be conducive to plant tissue culture batch production, scale, intensive production.
Embodiment
Below in conjunction with embodiment method of the present invention is further specified.
A kind of group training medium preparation method without agar comprises the steps: in the technical scheme
1, selecting the suction multiple is that high hydroscopic resin more than 500 times is made curing agent.
Different materials during 2, according to plant tissue culture and different cultivation stages are mixed with nutrient solution with the hormone of MS medium prescription+correspondence.
3, according to quantity high hydroscopic resin is added in the nutrient solution, nutrient solution is absorbed curing molding.
4, according to quantity the nutrient solution of curing molding is sub-packed in the tissue culture bottle.
5, according to a conventional method medium is carried out disinfection sterilization and must producing with group training medium.
Embodiment is as follows:
1, selecting the suction multiple is that 500 times high hydroscopic resin is made curing agent.
Different materials during 2, according to plant tissue culture and different cultivation stages are mixed with the 1000ml nutrient solution with the hormone of MS medium prescription+correspondence.
3,2 gram high hydroscopic resins are added in the 1000ml nutrient solution, nutrient solution is absorbed curing molding.
4, the nutrient solution with curing molding is sub-packed in the tissue culture bottle, every bottled 30 grams.
5, tissue culture bottle is sealed, 121 ℃ (1kg/cm2) sterilizes 20min down and must produce with group training medium in high-pressure sterilizing pot then.
Claims (1)
1. the group training medium preparation method without agar is characterized in that comprising the steps:
(1) selecting the suction multiple is that high hydroscopic resin more than 500 times is made curing agent;
Different materials during (2) according to plant tissue culture and different cultivation stages are mixed with nutrient solution with the hormone of MS medium prescription+correspondence;
(3) according to quantity high hydroscopic resin is added in the nutrient solution, nutrient solution is absorbed curing molding;
(4) according to quantity the nutrient solution of curing molding is sub-packed in the tissue culture bottle;
(5) according to a conventional method medium is carried out disinfection sterilization and must producing with group training medium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013102388430A CN103283605A (en) | 2013-06-17 | 2013-06-17 | Preparation method of tissue culture medium without agar |
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| Application Number | Priority Date | Filing Date | Title |
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| CN2013102388430A CN103283605A (en) | 2013-06-17 | 2013-06-17 | Preparation method of tissue culture medium without agar |
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| CN103283605A true CN103283605A (en) | 2013-09-11 |
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| CN2013102388430A Pending CN103283605A (en) | 2013-06-17 | 2013-06-17 | Preparation method of tissue culture medium without agar |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104293765A (en) * | 2014-09-17 | 2015-01-21 | 黄秀英 | Preparation method of solid photosynthetic bacterium |
| CN104871980A (en) * | 2015-06-26 | 2015-09-02 | 黄旭胜 | Culture medium for tissue culture |
| CN105104193A (en) * | 2015-08-19 | 2015-12-02 | 单县绿丰种业有限公司 | Method for raising salt-tolerant tomato plants |
| CN109076959A (en) * | 2018-08-31 | 2018-12-25 | 湖州德清玖沐农业科技有限公司 | A kind of generic plant tissue culture culture medium and preparation method |
| CN109673587A (en) * | 2018-12-04 | 2019-04-26 | 上海交通大学 | A kind of preparation method for the transparent earth materials can be used for soil animal movable observing |
Citations (3)
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| CN101955562A (en) * | 2010-10-09 | 2011-01-26 | 内蒙古大学 | Method for preparing super absorbent resin containing dehydrated MS culture medium |
| CN101985482A (en) * | 2010-10-09 | 2011-03-16 | 内蒙古大学 | Method for preparing super absorbent resin containing fulvic acid sodium and MS dry medium |
| CN101985507A (en) * | 2010-10-09 | 2011-03-16 | 内蒙古大学 | Method for preparing super absorbent resin containing composite plant growth regulator and dehydrated Murashige and Skoog's (MS) culture medium |
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2013
- 2013-06-17 CN CN2013102388430A patent/CN103283605A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101955562A (en) * | 2010-10-09 | 2011-01-26 | 内蒙古大学 | Method for preparing super absorbent resin containing dehydrated MS culture medium |
| CN101985482A (en) * | 2010-10-09 | 2011-03-16 | 内蒙古大学 | Method for preparing super absorbent resin containing fulvic acid sodium and MS dry medium |
| CN101985507A (en) * | 2010-10-09 | 2011-03-16 | 内蒙古大学 | Method for preparing super absorbent resin containing composite plant growth regulator and dehydrated Murashige and Skoog's (MS) culture medium |
Non-Patent Citations (1)
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| 涂艺声等: "《经济植物大规模快速繁殖技术》", 31 March 2009, 化学工业出版社 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104293765A (en) * | 2014-09-17 | 2015-01-21 | 黄秀英 | Preparation method of solid photosynthetic bacterium |
| CN104871980A (en) * | 2015-06-26 | 2015-09-02 | 黄旭胜 | Culture medium for tissue culture |
| CN105104193A (en) * | 2015-08-19 | 2015-12-02 | 单县绿丰种业有限公司 | Method for raising salt-tolerant tomato plants |
| CN109076959A (en) * | 2018-08-31 | 2018-12-25 | 湖州德清玖沐农业科技有限公司 | A kind of generic plant tissue culture culture medium and preparation method |
| CN109673587A (en) * | 2018-12-04 | 2019-04-26 | 上海交通大学 | A kind of preparation method for the transparent earth materials can be used for soil animal movable observing |
| CN109673587B (en) * | 2018-12-04 | 2020-07-24 | 上海交通大学 | Preparation method of transparent soil material for soil animal activity observation |
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Application publication date: 20130911 |
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