CN101816288A - Culture medium for establishing embryogenic suspension culture system of garlic - Google Patents
Culture medium for establishing embryogenic suspension culture system of garlic Download PDFInfo
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- CN101816288A CN101816288A CN 201010169619 CN201010169619A CN101816288A CN 101816288 A CN101816288 A CN 101816288A CN 201010169619 CN201010169619 CN 201010169619 CN 201010169619 A CN201010169619 A CN 201010169619A CN 101816288 A CN101816288 A CN 101816288A
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- garlic
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- sucrose
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Abstract
The invention discloses a culture medium for establishing an embryogenic suspension culture system of garlic, which belongs to a tissue culture technology of garlic. 2,4-D, KT, sucrose and agar are dissolved into a basic culture medium B5, and a pH value is adjusted to 5.8-6.0 to prepare the garlic. The culture medium comprises the following components metered by the adding amount of the basic culture medium B5 of 1L: 1L of the B5, 2mg of the 2,4-D, 0.5-2mg of the KT and 30g of the sucrose. The suspension culture system of the garlic is established by utilizing the culture medium, and thus, the technical problems of easy embryogenic performance loss and difficult system formation during suspension culture of embryonic callus induced by the apical meristem of the garlic are solved.
Description
Technical field
The present invention relates to a kind of garlic tissue culture technique, specifically is the medium that a kind of embryogenic suspension culture system of garlic is set up usefulness.Be specially adapted to utilize the embryo callus that induces from the garlic shoot apical meristem to carry out suspension culture.
Background technology
Garlic (Allium sativumL.) is an asexually propagated plant, has higher edible and medical value, utilize biotechnology to improve its quality. proterties such as disease and insect resistance have now caused widely to be paid attention to, and the key link of these biotechnology applications is to set up effective plant regeneration system that a garlic cellular tissure is cultivated, bred.The suspension culture technology can be turned out the test material of a large amount of homogeneities at short notice, increases substantially test efficiency, for the garlic biotechnology provides desirable test material.Zhang Yi etc. utilize the garlic suspension culture to extract the garlic secondary metabolites report (1993); But utilize garlic suspension culture technology, set up suspension culture system and effective plant high-frequency regeneration system and do not report as yet.
Summary of the invention
The invention provides a kind of embryogenic suspension culture system of garlic and set up the medium of usefulness, purpose is an easy forfeiture embryo when solving the garlic shoot apical meristem and inducing the embryo callus suspension culture, be difficult to become the technical barrier that is, set up the cells,primordial of a garlic and cultivate system.
Technical scheme of the present invention is as follows: a kind of embryogenic suspension culture system of garlic is set up the medium of usefulness, be with 2,4 dichlorphenoxyacetic acids 2,4-D, kinetin KT, sucrose and agar are dissolved among the minimal medium B5, pH value is adjusted to 5.8-6.0 and makes, and the consumption of its each component is: in 1L minimal medium B5 addition:
B5 1L
2，4-D 2mg
KT 0.5-2mg
Sucrose 30g.
Be preferably:
B5 1L
2，4-D 2mg
KT 0.5mg
Sucrose 30g.
In the above-mentioned raw materials, B5 is a minimal medium, and cultured tissue grow required inorganic nutritive element and part organic nutrition element are provided; 2,4-D, KT are plant growth regulator, have the effect that promotes that the culture callus forms and grows; Sucrose provide cultured tissue grow carbon nutrition.
The consumption of above-mentioned each component is that the inventor tests in a large number to grope to sum up on the basis of theory analysis and draws, and prepares medium according to the above ratio, has the high-quality of formation embryo callus particle and induces effect.
The preparation method of above-mentioned medium is: with 2,4-D, KT and sucrose dissolved stir in minimal medium B5, and pH value is adjusted into 5.8-6.0 and gets final product.
Good effect of the present invention is: utilize basal culture medium can set up garlic suspension culture system, easy forfeiture embryo when having solved the garlic shoot apical meristem and inducing the embryo callus suspension culture is difficult to become the technical barrier that is.
Embodiment:
Further set forth beneficial effect of the present invention with reference examples by the following examples.
Embodiment 1:
B5 1L
2，4-D 2mg
6-BA 2mg
Sucrose 30g.
Embodiment 2:
B5 1L
2，4-D 2mg
6-BA 0.5mg
NAA 0.2mg
Sucrose 30g.
Embodiment 3:
B5 1L
2，4-D 2mg
KT 0.5mg
Sucrose 30g.
Embodiment 4:
B5 1L
2，4-D 2mg
KT 2mg
Sucrose 30g.
Test method and result:
The medium of the foregoing description is used for the foundation test of embryogenic suspension culture system of garlic:
Material: the embryo callus that garlic Xuzhou white garlic shoot apical meristem induces.
Method: garlic Xuzhou white garlic shoot apical meristem is induced embryo callus be broken into little cell mass, it is transferred in the triangular flask that the foregoing description liquid culture 20ml (PH5.5) is housed, be placed on the horizontal shaking table of 100r/min rotating speed and carry out shaken cultivation, condition of culture is 27 ± 1 ℃, 12h.1000lx illumination every day.Suspension culture every 2 all subcultures once cultivated for 10 weeks.
Result: induce the embryo callus tube that obtains to be seeded in above-mentioned 4 embodiment liquid culture the garlic shoot apical meristem and carry out shaken cultivation.The suspension system formation rate is up to 86.7% in embodiment 3, can obtain propagation rapidly. the cells,primordial suspension culture system that degree of scatter is good.The suspension system formation rate is minimum among the embodiment 1, is 6.6%; The suspension system formation rate is 40% among the embodiment 2; The suspension system formation rate is 63.3% among the embodiment 4.In embodiment 1,2,4, callus bleaches gradually, loses embryo at last, and the part cell mass forms root shape tissue simultaneously.This shows that in the garlic somatic embryo forming process, too high growth hormone is unfavorable for the generation of its body embryo, and cause direction differentiation to root.The cells,primordial that in embodiment 3 medium, obtains cultivate be weekly subculture once, during each subculture, with hard. faint yellow. diameter is broken into small cell cluster below the 0.1mm or unicellular greater than the cell mass of 3mm, by the suspension culture in 2 weeks, the part cell mass can develop into the cell mass of 2-3mm again.By said method, induce the embryo callus that obtains to obtain a large amount of homogeneity embryo callus at short notice by a shoot apical meristem.
Statistics sees Table 1.
Table 1 garlic suspension culture system induces the result to add up (10 week)
Comprehensive above the analysis draws the optimal medium that embryogenic suspension culture system of garlic induces and is:
B5 1L
2，4-D 2mg
KT 0.5mg
Sucrose 30g.
Claims (2)
1. medium of setting up embryogenic suspension culture system of garlic, it is characterized in that: with 2,4 dichlorphenoxyacetic acids 2,4-D, kinetin KT, sucrose, agar are dissolved among the minimal medium B5, pH value is adjusted to 5.8-6.0 and makes, and the consumption of its each component is: in 1L minimal medium B5 addition:
B5 1L
2，4-D 2mg
KT 0.5-2mg
Sucrose 30g.
2. medium of setting up embryogenic suspension culture system of garlic, it is characterized in that: with 2,4 dichlorphenoxyacetic acids 2,4-D, kinetin KT, sucrose, agar are dissolved among the minimal medium B5, pH value is adjusted to 5.8-6.0 and makes, and the consumption of its each component is: in 1L minimal medium B5 addition:
B5 1L
2，4-D 2mg
KT 0.5mg
Sucrose 30g.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010169619 CN101816288A (en) | 2010-05-06 | 2010-05-06 | Culture medium for establishing embryogenic suspension culture system of garlic |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010169619 CN101816288A (en) | 2010-05-06 | 2010-05-06 | Culture medium for establishing embryogenic suspension culture system of garlic |
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| CN101816288A true CN101816288A (en) | 2010-09-01 |
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ID=42651693
Family Applications (1)
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| CN 201010169619 Pending CN101816288A (en) | 2010-05-06 | 2010-05-06 | Culture medium for establishing embryogenic suspension culture system of garlic |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103004596A (en) * | 2012-12-17 | 2013-04-03 | 南京工业大学 | Method for quickly obtaining garlic seedlings by using intermittent immersion cultivation mode |
| CN103598090A (en) * | 2013-10-31 | 2014-02-26 | 李本明 | Method used for induction of garlic calluses from garlic cloves |
| CN110226517A (en) * | 2019-06-26 | 2019-09-13 | 北京市农林科学院 | A kind of method of onion Regeneration in Vitro and its culture medium used |
-
2010
- 2010-05-06 CN CN 201010169619 patent/CN101816288A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 《全国作物细胞工程与分子技术育种学术研讨会论文集》 20031231 张允刚等 大蒜悬浮培养及有效植株再生 1 , 2 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103004596A (en) * | 2012-12-17 | 2013-04-03 | 南京工业大学 | Method for quickly obtaining garlic seedlings by using intermittent immersion cultivation mode |
| CN103004596B (en) * | 2012-12-17 | 2014-07-16 | 南京工业大学 | Method for quickly obtaining garlic seedlings by using intermittent immersion cultivation mode |
| CN103598090A (en) * | 2013-10-31 | 2014-02-26 | 李本明 | Method used for induction of garlic calluses from garlic cloves |
| CN103598090B (en) * | 2013-10-31 | 2016-08-17 | 青岛文创科技有限公司 | A kind of method utilizing garlic clove induction garlic callus |
| CN110226517A (en) * | 2019-06-26 | 2019-09-13 | 北京市农林科学院 | A kind of method of onion Regeneration in Vitro and its culture medium used |
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Application publication date: 20100901 |