CN108012926A - A kind of proliferated culture medium of pseudo-ginseng anther callus and preparation method thereof - Google Patents
A kind of proliferated culture medium of pseudo-ginseng anther callus and preparation method thereof Download PDFInfo
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- CN108012926A CN108012926A CN201711447128.2A CN201711447128A CN108012926A CN 108012926 A CN108012926 A CN 108012926A CN 201711447128 A CN201711447128 A CN 201711447128A CN 108012926 A CN108012926 A CN 108012926A
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- ginseng
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- Developmental Biology & Embryology (AREA)
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a kind of proliferated culture medium of pseudo-ginseng anther callus and preparation method thereof, proliferated culture medium includes active ingredient by weight：4,050 4130 parts of a great number of elements；378.88 393.12 parts of trace element；330 350 parts of calcium salt；63 67 parts of molysite；100.895 105.305 parts of organic component；2,4 0.5 1.2 parts of D；0.25 0.4 parts of kinetin；30,000 50000 parts of sucrose；Preparation method includes：The prewired step of solution：A great number of elements solution, trace element solution, calcium salt soln, iron salt solutions, organic component solution, 2,4 solution Ds and kinetin solution are prepared respectively；Blend step：Each solution, sucrose and agar are mixed and dissolved, it is constant volume after 5.4 5.8 to adjust pH, to obtain the final product；The proliferated culture medium improves the growth coefficient in pseudo-ginseng flower pesticide callus tissue culture.
Description
Technical field
The present invention relates to a kind of proliferated culture medium of pseudo-ginseng anther callus and preparation method thereof, belongs to Vitro Plant training
Support reagent technique field.
Background technology
Pseudo-ginseng Panax notoginseng (Burk.) F.H.Chen is Araliaceae Araliacea e Panaxes Panax
Linn. herbaceos perennial, is the distinctive rare traditional Chinese medicine in China, main product is in Yunnan mountain of papers.Stop with hemostasis scattered silt, detumescence
Bitterly, Xu-tonic and other effects.Pseudo-ginseng has a long history as daily health caring, can relate earliest to the Tang Dynasty.In recent years due to
Pseudo-ginseng is increasingly approved that demand is also increasing in unique curative effect for the treatment of cardiac and cerebral vascular diseases by people.Pseudo-ginseng gives birth to growth
State environment has particular/special requirement, its suitable cultivation region is small.In addition the problems such as growth cycle is long, pest and disease damage is serious so that pseudo-ginseng
Breed breeding is very difficult.
The important channel that vegetative propagation is preservation and the utilization species resource is carried out to it using plant tissue culture technique,
And the breeding cycle is short, efficient.Anther Culture can produce the haplobiont from pollen, have on crop breeding important
Value.Not only homozygous speed is fast, optional efficient, and can improve breeding efficiency, overcomes distant hybridization sterile, creates novel species
Matter etc..Each plant has its corresponding callus induction culture medium, and the Fiber differentiation of some plants is hormone kind and proportioning
Difference, even minimal medium is all different for some, and so far, the research of tissue culture medium (TCM) in relation to pseudo-ginseng flower pesticide does not appear in the newspapers
Road.
The content of the invention
For overcome the deficiencies in the prior art, of the invention first purpose is to provide a kind of pseudo-ginseng anther callus
Proliferated culture medium, which is studied for the tissue cultures of pseudo-ginseng flower pesticide, improves plant in culture medium and give birth to
Long modifier concentration proportioning, is conducive to improve the growth coefficient of callus.
Second object of the present invention is to provide a kind of preparation of the proliferated culture medium of above-mentioned pseudo-ginseng anther callus
Method.
Realizing first purpose of the present invention can be reached by adopting the following technical scheme that：A kind of pseudo-ginseng antherderived callus group
The proliferated culture medium knitted, the proliferated culture medium include following active ingredient by weight：
Further, proliferated culture medium includes following active ingredient by weight：
Further, a great number of elements includes following component by weight：
Further, the trace element includes following component by weight：
Further, the calcium salt is calcium chloride.
Further, the molysite is prepared by ferrous sulfate and EDTA-2Na.Further, the organic component bag
Include following component by weight：
Realize that second object of the present invention can reach by adopting the following technical scheme that：A kind of pseudo-ginseng antherderived callus group
The preparation method for the proliferated culture medium knitted, including：
The prewired step of solution：A great number of elements solution, trace element solution, calcium salt soln, iron salt solutions, organic is prepared respectively
Component solution, 2,4-D solution and kinetin solution；
Blend step：By a great number of elements solution, trace element solution, calcium salt soln, iron salt solutions, organic component solution,
2,4-D solution, kinetin solution, sucrose and agar are mixed and dissolved, and it is constant volume after 5.4-5.8 to adjust pH, to obtain the final product.
Further, in blend step, it is 5.6 to adjust pH.
Further, the active ingredient concentration in the proliferated culture medium obtained is 34.5-55.5g/L；The active ingredient
For a great number of elements, trace element, calcium salt, molysite, organic component, 2,4-D, kinetin and sucrose.
Compared with prior art, the beneficial effects of the present invention are：
The present invention proliferated culture medium be for pseudo-ginseng flower pesticide tissue cultures feature it is for further research, improve
Plant growth regulator concentration proportioning in culture medium, substantially increases the growth coefficient of pseudo-ginseng anther callus, utilizes this hair
Growth coefficient that is bright and cultivating pseudo-ginseng anther callus reaches more than 9.
Brief description of the drawings
Fig. 1 is used for tissue cultures 45d callus growth situations for 1 culture medium of embodiment；
Fig. 2 is used for tissue cultures 45d callus growth situations for 2 culture medium of embodiment.
Embodiment
In the following, with reference to embodiment, the present invention is described further：
A kind of proliferated culture medium of pseudo-ginseng anther callus, including following active ingredient by weight：
The active ingredient of the proliferated culture medium of 1 pseudo-ginseng anther callus of form
The preparation method of the proliferated culture medium of pseudo-ginseng anther callus, including：
The prewired step of solution：A great number of elements solution, trace element solution, calcium salt soln, iron salt solutions, organic is prepared respectively
Component solution, 2,4-D solution and kinetin solution；
Wherein in a great number of elements solution, the concentration of potassium nitrate is 164-166g/L, and the concentration of ammonium nitrate is 189-191g/L,
The concentration of magnesium sulfate is 36-38g/L, and the concentration of potassium dihydrogen phosphate is 16-18g/L；
Wherein in trace element solution, the concentration of potassium iodide is 8-9g/L, and the concentration of boric acid is 60-63g/L, manganese sulfate
Concentration is 223-230g/L, and sulfuric acid zinc concentration is 85-88g/L, and the concentration of sodium molybdate is 2.4-2.6g/L, the concentration of copper sulphate
For 0.24-0.26g/L, the concentration of cobalt chloride is 0.24-0.26g/L；
Wherein the concentration of calcium salt soln is 33-35g/L；
Wherein iron salt solutions are dissolved in water by ferrous sulfate and EDTA-2Na and obtain, containing chelating ferrous in iron salt solutions
(EDTA-Fe), molysite total concentration is 6.3-6.7g/L；
Wherein in organic component solution, the concentration of inositol is 9.8-10.2g/L, and the concentration of nicotinic acid is 0.045-0.055g/
L, the concentration of puridoxine hydrochloride are 0.045-0.055g/L, and the concentration of vitamin B1 is 9.5-10.5mg/L, the concentration of glycine
For 0.19-0.21g/L；
The concentration of 2,4-D solution is 0.05-0.12g/L, and the concentration of kinetin solution is 0.025-0.04g/L；
Blend step：Take a great number of elements solution 10mL, trace element solution 1mL, calcium salt soln 10mL, iron salt solutions
10mL, organic component solution 10mL, 2,4-D solution 10mL, kinetin solution 10mL, sucrose 30-50g and agar 5g mixing are simultaneously molten
Solution, adjusts after pH is 5.6 and is settled to 1L；
Sterilization steps：Temperature is placed in as the 22-24min that sterilizes under the conditions of 121-125 DEG C, pressure 0.10-0.12MPa, finally
Obtained proliferated culture medium.
Embodiment 1-3：
Embodiment 1-3 is the proliferated culture medium of the pseudo-ginseng anther callus of configuration 1L, is comprised the following steps：
The prewired step of solution：A great number of elements solution, trace element solution, calcium salt soln, iron salt solutions, organic is prepared respectively
Component solution, 2,4-D solution and kinetin solution；Each component in solution concentration is as shown in Table 2；
Constituent concentration in 2 prewired solution of form
Blend step：Take a great number of elements solution 10mL, trace element solution 1mL, calcium salt soln 10mL, iron salt solutions
10mL, organic component solution 10mL, 2,4-D solution 10mL, kinetin solution 10mL, sucrose 30g and agar 5g are mixed and dissolved,
Adjust after pH is 5.6 and be settled to 1L；
Sterilization steps：Temperature is placed in as the 22-24min that sterilizes under the conditions of 121-125 DEG C, pressure 0.10-0.12MPa, finally
Obtained proliferated culture medium.
In obtained culture medium, each constituent concentration is as shown in Table 3：
Constituent concentration in 3 culture medium of form
The culture medium of embodiment 1-3 is respectively used to tissue cultures, culture pseudo-ginseng anther callus promotes to sprout, with one
The effective bud number formed in a blake bottle/inoculation callus block number, is calculated growth coefficient.The growth coefficient of embodiment 1 is 9,
Fig. 1 show the growing state that embodiment 1 cultivates 45d callus；The growth coefficient of embodiment 2 show embodiment for 11, Fig. 2
The growing state of 2 culture 45d callus, the growth coefficient of embodiment 3 is 10；The growth coefficient of 3 embodiments is more than 9.
For those skilled in the art, technical solution that can be as described above and design, make other each
Kind is corresponding to be changed and deforms, and all these change and deform the protection model that should all belong to the claims in the present invention
Within enclosing.
Claims (10)
1. a kind of proliferated culture medium of pseudo-ginseng anther callus, it is characterised in that proliferated culture medium is included by weight
Following active ingredient：
2. the proliferated culture medium of pseudo-ginseng anther callus as claimed in claim 1, it is characterised in that proliferated culture medium includes
Following active ingredient by weight：
3. the proliferated culture medium of pseudo-ginseng anther callus as claimed in claim 1, it is characterised in that a great number of elements bag
Include following component by weight：
4. the proliferated culture medium of pseudo-ginseng anther callus as claimed in claim 1, it is characterised in that the trace element bag
Include following component by weight：
5. the proliferated culture medium of pseudo-ginseng anther callus as claimed in claim 1, it is characterised in that the calcium salt is chlorination
Calcium.
6. the proliferated culture medium of pseudo-ginseng anther callus as claimed in claim 1, it is characterised in that the molysite is by sulfuric acid
Ferrous and EDTA-2Na is prepared.
7. the proliferated culture medium of pseudo-ginseng anther callus as claimed in claim 1, it is characterised in that the organic component bag
Include following component by weight：
A kind of 8. preparation method of the proliferated culture medium of pseudo-ginseng anther callus, it is characterised in that including：
The prewired step of solution：A great number of elements solution, trace element solution, calcium salt soln, iron salt solutions, organic component are prepared respectively
Solution, 2,4-D solution and kinetin solution；
Blend step：By a great number of elements solution, trace element solution, calcium salt soln, iron salt solutions, organic component solution, 2,4-D
Solution, kinetin solution, sucrose and agar are mixed and dissolved, and it is constant volume after 5.4-5.8 to adjust pH, to obtain the final product.
9. preparation method as claimed in claim 8, it is characterised in that in blend step, it is 5.6 to adjust pH.
10. preparation method as claimed in claim 8, it is characterised in that the active ingredient concentration in obtained proliferated culture medium
For 34.5-55.5g/L；The active ingredient is a great number of elements, trace element, calcium salt, molysite, organic component, 2,4-D, excitement
Element and sucrose.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711447128.2A CN108012926A (en) | 2017-12-27 | 2017-12-27 | A kind of proliferated culture medium of pseudo-ginseng anther callus and preparation method thereof |
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| CN201711447128.2A CN108012926A (en) | 2017-12-27 | 2017-12-27 | A kind of proliferated culture medium of pseudo-ginseng anther callus and preparation method thereof |
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| CN201711447128.2A Pending CN108012926A (en) | 2017-12-27 | 2017-12-27 | A kind of proliferated culture medium of pseudo-ginseng anther callus and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114158366A (en) * | 2021-11-10 | 2022-03-11 | 内蒙古蒙草生态环境（集团）股份有限公司 | Method for cutting propagation of chenopodium camellinum |
-
2017
- 2017-12-27 CN CN201711447128.2A patent/CN108012926A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| 张传海等: "《植物生物技术》", 30 September 2015 * |
| 张成国: "西洋参花药愈伤组织无性系建立及高产无性系的筛选", 《中国优秀硕士学位论文全文数据库，农业科技辑》 * |
| 段承俐等: "三七花药培养的研究（Ⅰ）愈伤组织的诱导", 《云南农业大学学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114158366A (en) * | 2021-11-10 | 2022-03-11 | 内蒙古蒙草生态环境（集团）股份有限公司 | Method for cutting propagation of chenopodium camellinum |
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Effective date of registration: 20190403 Address after: 650000 6th Floor, Block A, Sunshine Building, 999 Kegao Road, Wuhua District, Kunming City, Yunnan Province Applicant after: Kangmei Pharmaceutical (Kunming) Germplasm Resources Co., Ltd. Address before: 663099 Wenshan Zhong and Miao Autonomous Prefecture Wenshan City, Yunnan Province, Kaihua South Road Wenshan Biovalley of Chinese Traditional Medicine 4-21 Applicant before: Kangmei Pharmaceutical (Wenshan) medicinal material planting Management Co., Ltd. |
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Application publication date: 20180511 |