CN103141385A - Culture medium for efficient induction of garlic calli - Google Patents
Culture medium for efficient induction of garlic calli Download PDFInfo
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- CN103141385A CN103141385A CN 201310062350 CN201310062350A CN103141385A CN 103141385 A CN103141385 A CN 103141385A CN 201310062350 CN201310062350 CN 201310062350 CN 201310062350 A CN201310062350 A CN 201310062350A CN 103141385 A CN103141385 A CN 103141385A
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Abstract
The invention discloses a culture medium for efficient induction of garlic calli, relating to a technology for garlic tissue culture, and belongs to the technical field of agriculture. The culture medium is characterized in that the dosage of every component on the basis of the added amount of MS in 1L of a basic culture medium of is as follows: 1L of MS, 1.0-2.0mg of forchlorfenuron, 0.05-0.5mg of compound sodium nitrophenolate, 200mg of casein hydrolysate, 20g of sucrose and 6g of agar. The culture medium has the beneficial effects that the culture medium has the functions of promoting cell division and expansion, the organofaction and synthesis of protein, improving the efficiency of photosynthesis, enhancing stress resistance, delaying senescence, promoting flower bud differentiation on melon and fruit plants, protecting flowers and fruits, improving the fruit setting rate, promoting fruit expansion, and the like; and the use of a compound plant growth regulator ensures that the culture medium can quickly permeate into plant bodies after contacting with plants.
Description
Technical field
The present invention relates to the Garlic Tissue culture technique, belong to agricultural technology field, specifically a kind of garlic callus medium of efficiently inducing.
Background technology
Garlic, herbaceos perennial, Liliaceae allium.Underground bulb distinguish is divided into purple skin kind and white skin kind by the color of the leather difference.Pungent, irritant smell, edible or confession seasoning also can be used as medicine.Garlic nutrient is abundant: every 100 gram moisture content 69.8 grams, protein 4.4 grams, fatty 0.2 gram, carbohydrate 23.6 grams, 5 milligrams of calcium, 44 milligrams, phosphorus, 0.4 milligram of iron, 3 milligrams of vitamin Cs.In addition, also contain the trace elements such as thiamine, vitamin b3, niacin, allicin, citral and selenium and germanium.Contain volatile oil approximately 0.2%, in oil, main component is allicin, has bactericidal action, is that alliin contained in garlic is produced by the effect hydrolysis of large allinase.Still contain the sulfide compound of multiple pi-allyl, propyl group and methyl composition etc.Contain " garlic amine " in garlic, this material to the benefit of brain than Cobastab, strong manyfold also.
After plant is subjected to wound, be called callus at the tissue of hindering face new life.The callus of inducing the explant somatocyte division to form in Plant Tissue Breeding, it not mostly the result of damage, but the plant hormone substance inducing cell that utilizes external source begins to divide formation, commonly used have auxin substance as 2,4-=chlorophenoxyacetic acid, methyl α-naphthyl acetate, heteroauxin and the basic element of cell division as 6 benzyl purines etc.Growth hormone and the basic element of cell division are used in conjunction with in finite concentration and ratio, are easy to induce callus.
At present, by tissue and Organ culture techniques, only set up vitro Regeneration System on a few allium.Garlic belongs to asexually propagated crop, and most garlic kinds show very strong sterility.All adopt the garlic cultivar of traditional landrace and natural variation to cultivate in production, be difficult to obtain or create the garlic material that to educate and carry out the sexual hybridization breeding.But along with new breeding technique development, the new means such as the separation of induced mutations, protoplast and fusion and tissue cultivation are emerged in large numbers in succession, and the breeding work of garlic is made great progress.
Summary of the invention
The medium that the object of the present invention is to provide a kind of garlic callus efficiently to induce overcomes the garlic bulb point and organizes the low problem of inductivity, and the medium that the inventive method utilization is efficiently induced is used for the research and production of garlic.
For achieving the above object, the medium that a kind of garlic callus of the present invention is efficiently induced, it is characterized in that: the medium that this garlic callus is induced is dissolved in CPPU, compound sodium nitrophenolate, caseinhydrolysate, sucrose, agar in minimal medium MS and forms, and the consumption of its each component is: in 1L minimal medium MS addition: MS 1L, CPPU 1.0～2.Omg, compound sodium nitrophenolate 0.05～0.5mg, caseinhydrolysate 200mg, sucrose 20g, agar 6g.
The medium that a kind of garlic callus of the present invention is efficiently induced, its implementation is as follows:
One, the test plant material adopts the common white skin garlic of abolishing resting stage that market is bought, and the multiplex CPPU of this experiment, compound sodium nitrophenolate, caseinhydrolysate are Beijing Suo Laibao Science and Technology Ltd. product, and testing other reagent used, to be domestic analysis pure;
Two, go white skin garlic the garlic clove strip off to be placed in the culture dish of diameter 12cm, 5 layers of gauze of underlay are used water-soaked, and the garlic clove that strips is positioned on gauze, and 25 ℃ of water cultures are taken root, and sooner or later respectively change water 1 time every day;
Three, go the fresh tip of a root 3～5mm of robust growth to rinse with a large amount of clear water, use afterwards 75% alcohol-pickled 10～15s, then put into 0.1% mercury chloride 7～8min, afterwards with a large amount of aseptic water washings 8～10 times to remove residual disinfectancy liquid;
Four, the above-mentioned explant after sterilizing is connected to and contains CPPU, compound sodium nitrophenolate, caseinhydrolysate, sucrose, agar and be dissolved on the medium of MS, and described medium is in 1L minimal medium MS addition: MS 1L, CPPU 1.0～2.Omg, compound sodium nitrophenolate 0.05～0.5mg, caseinhydrolysate 200mg, sucrose 20g, agar 6g; The pH value is adjusted into 5.8～6.0.
In described raw material, MS is minimal medium, and cultured tissue grow required inorganic nutrients and part organic nutrition element are provided: CPPU and compound sodium nitrophenolate are plant growth regulator, has to promote cultured tissue to form the effect of embryo callus; Caseinhydrolysate provides organic polypeptide class nutritive element; Sucrose provide cultured tissue grow carbon nutrition: the agar Main Function is to solidify medium, plays a supportive role.
The preparation method of described medium is that CPPU, compound sodium nitrophenolate, caseinhydrolysate, sucrose, agar are dissolved in MS, stirs, and the pH value gets final product between being adjusted into 5.8～6.0.
Described CPPU is a kind of highly active phenyl urea derivatives, has cytokine activity.Have promote cell division and expansion, organ to form and protein synthetic, improve photosynthetic efficiency, strengthen resistance, delay senility, can promote flower bud differentiation on the melon and fruit plant, flower and fruit protecting improves fruit-setting rate, promotes the effect such as Fruit.
Described compound sodium nitrophenolate is a kind of powerful cell-activating agent, in plant corpus, promotes the protoplasm of cell to flow with energy rapid permeability after plant contact, improves cell viability.
The medium that a kind of garlic callus of the present invention is efficiently induced, its beneficial effect is: medium of the present invention have promote cell division and expansion, organ to form and protein synthetic, improve photosynthetic efficiency, strengthen resistance, delay senility, can promote flower bud differentiation on the melon and fruit plant, flower and fruit protecting improves the effects such as fruit-setting rate, promotion Fruit; After the use of composite plant growth regulator makes medium and plant contact, the energy rapid permeability is in plant corpus, promote the protoplasm of cell to flow, improve cell viability, utilize the inductivity of callus in the cultivation of basal culture medium Allium fistulosum stem tips tissue all greater than 34%, and can reach 82% left and right.
Embodiment
Following examples are that the present invention further goes out to set forth.
Embodiment 1
One, the test plant material adopts the common white skin garlic of abolishing resting stage that market is bought.The multiplex CPPU of this experiment, compound sodium nitrophenolate, caseinhydrolysate are Beijing Suo Laibao Science and Technology Ltd. product, and testing other reagent used, to be domestic analysis pure;
Two, go white skin garlic the garlic clove strip off to be placed in the culture dish of diameter 12cm, 5 layers of gauze of underlay are used water-soaked, and the garlic clove that strips is positioned on gauze, and 25 ℃ of water cultures are taken root, and sooner or later respectively change water 1 time every day;
Three, go the fresh tip of a root 3mm of robust growth to rinse with a large amount of clear water, use afterwards 75% alcohol-pickled 10s, then put into 0.1% mercury chloride 7min, afterwards with a large amount of aseptic water washings 8 times to remove residual disinfectancy liquid;
Four, the above-mentioned explant after sterilizing is connected to and contains CPPU, compound sodium nitrophenolate, caseinhydrolysate, sucrose, agar and be dissolved on the medium of MS, and described medium is in 1L minimal medium MS addition: MS 1L, CPPU 1.0mg, compound sodium nitrophenolate 0.05mg, caseinhydrolysate 200mg, sucrose 20g, agar 6g pH value are adjusted into 5.8.
Embodiment 2
One, the test plant material adopts the common white skin garlic of abolishing resting stage that market is bought.The multiplex CPPU of this experiment, compound sodium nitrophenolate, caseinhydrolysate are Beijing Suo Laibao Science and Technology Ltd. product, and testing other reagent used, to be domestic analysis pure;
Two, go white skin garlic the garlic clove strip off to be placed in the culture dish of diameter 12cm, 5 layers of gauze of underlay are used water-soaked, and the garlic clove that strips is positioned on gauze, and 25 ℃ of water cultures are taken root, and sooner or later respectively change water 1 time every day;
Three, go the fresh tip of a root 4mm of robust growth to rinse with a large amount of clear water, use afterwards 75% alcohol-pickled 12s, then put into 0.1% mercury chloride 7min, afterwards with a large amount of aseptic water washings 9 times to remove residual disinfectancy liquid;
Four, the above-mentioned explant after sterilizing is connected to and contains CPPU, compound sodium nitrophenolate, caseinhydrolysate, sucrose, agar and be dissolved on the medium of MS, and described medium is in 1L minimal medium MS addition: MS 1L, CPPU 1.5mg, compound sodium nitrophenolate 0.3mg, caseinhydrolysate 200mg, sucrose 20g, agar 6g pH value are adjusted into 5.9.
Embodiment 3
One, the test plant material adopts the common white skin garlic of abolishing resting stage that market is bought.The multiplex CPPU of this experiment, compound sodium nitrophenolate, caseinhydrolysate are Beijing Suo Laibao Science and Technology Ltd. product, and testing other reagent used, to be domestic analysis pure;
Two, go white skin garlic the garlic clove strip off to be placed in the culture dish of diameter 12cm, 5 layers of gauze of underlay are used water-soaked, and the garlic clove that strips is positioned on gauze, and 25 ℃ of water cultures are taken root, and sooner or later respectively change water 1 time every day;
Three, go the fresh tip of a root 5mm of robust growth to rinse with a large amount of clear water, use afterwards 75% alcohol-pickled 15s, then put into 0.1% mercury chloride 8min, afterwards with a large amount of aseptic water washings 10 times to remove residual disinfectancy liquid;
Four, the above-mentioned explant after sterilizing is connected to and contains CPPU, compound sodium nitrophenolate, caseinhydrolysate, sucrose, agar and be dissolved on the medium of MS, and described medium is in 1L minimal medium MS addition: MS 1L, CPPU 2.Omg, compound sodium nitrophenolate 0.5mg, caseinhydrolysate 200mg, sucrose 20g, agar 6g pH value are adjusted into 6.0.
Claims (3)
1. medium that garlic callus is efficiently induced, it is characterized in that: the medium of this garlic embryonic callus induction is dissolved in CPPU, compound sodium nitrophenolate, caseinhydrolysate, sucrose, agar in minimal medium MS and forms, and the consumption of its each component is: in 1L minimal medium MS addition: MS 1L, CPPU 1.0～2.Omg, compound sodium nitrophenolate 0.05～0.5mg, caseinhydrolysate 200mg, sucrose 20g, agar 6g.
2. a kind of garlic callus medium of efficiently inducing as claimed in claim 1, it is characterized in that: the preparation method of described medium is that CPPU, compound sodium nitrophenolate, caseinhydrolysate, sucrose, agar are dissolved in MS, stir, the pH value gets final product between being adjusted into 5.8～6.0.
3. a kind of garlic callus medium of efficiently inducing as claimed in claim 1, it is characterized in that: using method is as follows:
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| CN 201310062350 CN103141385A (en) | 2013-02-28 | 2013-02-28 | Culture medium for efficient induction of garlic calli |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104082355A (en) * | 2014-07-24 | 2014-10-08 | 宜宾云辰乔木园林有限责任公司 | Natural extract-containing plant growth accelerator |
| CN104642109A (en) * | 2015-02-04 | 2015-05-27 | 天津师范大学 | Method for constructing garlic micropropagation by taking bulb sheets as explants |
| CN106912320A (en) * | 2017-05-09 | 2017-07-04 | 中科天华生物科技有限公司 | A kind of culture medium for paper mulberry nursery and preparation method thereof |
-
2013
- 2013-02-28 CN CN 201310062350 patent/CN103141385A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104082355A (en) * | 2014-07-24 | 2014-10-08 | 宜宾云辰乔木园林有限责任公司 | Natural extract-containing plant growth accelerator |
| CN104642109A (en) * | 2015-02-04 | 2015-05-27 | 天津师范大学 | Method for constructing garlic micropropagation by taking bulb sheets as explants |
| CN104642109B (en) * | 2015-02-04 | 2016-11-02 | 天津师范大学 | A kind of is the method that outer implant builds Bulbus Allii micro-propagation with bulb sheet |
| CN106912320A (en) * | 2017-05-09 | 2017-07-04 | 中科天华生物科技有限公司 | A kind of culture medium for paper mulberry nursery and preparation method thereof |
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Application publication date: 20130612 |