CN106942056A - A kind of Radix Glycyrrhizae cotyledon inducing culture screening technique - Google Patents
A kind of Radix Glycyrrhizae cotyledon inducing culture screening technique Download PDFInfo
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- CN106942056A CN106942056A CN201710171895.9A CN201710171895A CN106942056A CN 106942056 A CN106942056 A CN 106942056A CN 201710171895 A CN201710171895 A CN 201710171895A CN 106942056 A CN106942056 A CN 106942056A
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- cotyledon
- radix glycyrrhizae
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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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- 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Developmental Biology & Embryology (AREA)
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention provides a kind of Radix Glycyrrhizae cotyledon inducing culture screening technique, it carries out Fiber differentiation by unitary variant method to Radix Glycyrrhizae cotyledon, analytic statistics inductivity, melting brown rate and increment, not only successfully to induce as unique evaluation criterion among evaluating, the production of commercial plant is more conformed to as evaluation mark with reference to increment.
Description
Technical field
The invention belongs to Radix Glycyrrhizae technical field of tissue culture, more particularly to a kind of Radix Glycyrrhizae cotyledon inducing culture screening side
Method.
Background technology
Radix Glycyrrhizae,（Scientific name：Glycyrrhiza uralensis Fisch）Alias：State is old, Herba Hedyotis cantonensis, Glycyrrhiza Uralensis, sweet
Son.Pulse family, Glycyrrhiza perennial herb, root are sturdy with root-like stock, are a kind of help Chinese herbal medicines.To a kind of good medicine of human body,
Medicinal part is root and rhizome, and Medicinal Materials Characters root is long 25~100 centimetres in cylinder, 0.6~3.5 centimetre of diameter.Crust is elastic
Differ, surface rufous or taupe brown.Rhizome is in cylinder, and surface, which has in the middle part of bud scar, section, marrow.Gas is micro-, and taste is sweet and special.
Major function is clearing heat and detoxicating, expelling phlegm and arresting coughing, gastral cavity abdomen etc..Happiness is dark moist, the low dry climate of the long temperature of sunshine.Being grown Radix Glycyrrhizae more
In arid, semiarid desert steppe, desert edge and loess hill area.Root and root-like stock hyoscine.Traditional Radix Glycyrrhizae is obtained
It is excavation leaf plant resource to take method, causes soil erosion and serious soil sandification.In addition, Glycyrrhiza Seeds are essentially from open country
Licorice, its variet complexity, seed is rare, and germination percentage is extremely low, the need for wild resource is not suitable with scale and standardized production.
Simultaneously because people devastatingly excavate to leaf licorice, wild plant is set almost to become extinct, the protectiveness utilization to Radix Glycyrrhizae is compeled
The eyebrows and eyelashes, but because licorice planting germination percentage is not high and people are to the demand of its this life metabolin, so that the tissue cultures of Radix Glycyrrhizae
Play an important roll to research and production, but current Screening of Media method is all the screening technique for the purpose of scientific research,
It is unable to the demand of commercial plant metaplasia production.
Rare earth element is the general designation of 17 kinds of special elements, and gaining the name for it is because Sweden scientist is in Extraction of rare earth element
When apply rare earth compound, so rare earth element of gaining the name.But the title that rare earth, which is left over by history, to get off, rare earth is from 18 generation
End beginning of recording finds that water insoluble soild oxide was often called soil by people at that time successively, for example, aluminum oxide is referred to as into " pottery
Soil ", calcium oxide is referred to as " alkaline earth " etc..Rare earth is usually to be separated with oxidation state, at that time than sparser, thus is gained the name
For rare earth（Rare Earth, abbreviation RE or R）, but pass through substantial amounts of scientific research, rare earth is with the addition of among tissue cultures
Element can greatly improve the quality of tissue cultures.
Cerium is periodic system Ι Ι Ι races subgroup lanthanide series, a kind of rare earth element.Atomic number 58.Stable isotope：136、
138、140、142.Grey metal, there is malleability.Density：Quadratic crystal 6.9, cubic crystal 6.7.799 DEG C of fusing point, boiling point 3426
℃.Cerium is a kind of silver-gray active metal, and powder easy spontaneous combustion in atmosphere is soluble in acid.The title of cerium derives from asteroid
The English name of ceres.Content of the cerium in the earth's crust about 0.0046%, is abundance highest in rare earth element.
The content of the invention
It is partial to scientific research in the screening technique of tissue culture and inducement culture medium based on Radix Glycyrrhizae in the prior art, is not suitable for business work
The problem of factory's metaplasia is produced, the present invention provides a kind of Radix Glycyrrhizae cotyledon inducing culture screening technique, and it is by unitary variant method to sweet
Grass seed leaf carries out Fiber differentiation, analytic statistics inductivity, melting brown rate and increment, not only successfully to induce work among evaluating
For unique evaluation criterion, the production of commercial plant is more conformed to as evaluation mark with reference to increment.
A kind of Radix Glycyrrhizae cotyledon inducing culture screening technique, it comprises the following steps：
Step S10 determines preliminary screening scope, and data of literatures is consulted in retrieval, contrasts other plant species tissue culture and inducement process
The amount of middle addition rare earth element simultaneously determines rare earth element preliminary screening scope；
Step S20 selects explant, and selection health, the Radix Glycyrrhizae cotyledon that growing environment is consistent and growth conditions are similar are used as explant
Body, carries out same sterilization to explant and handles；
Step S30 control cultures, the culture medium of various concentrations rare earth element addition is prepared using unitary variant method, is inoculated with respectively
Same amount of Radix Glycyrrhizae cotyledon, and carry out conventional induced tissue culture；
Step S40 record detection parameters, culture counts in each rare earth element concentration gradient after 20 days and induces callus
Quantity and the quantity of browning, and calculate inductivity and melting brown rate；
Step S50 increments are calculated, and are respectively taken out the callus in each parallel laboratory test group of same concentrations gradient, with clear
Water is rinsed well, dries surface moisture and callus is weighed, and records weight；
Step S60 calculate assess factor, by weight amount reduce 10 times, along with germination rate subtract melting brown rate calculate assessment because
Number, contrasts the assessment factor of various concentrations gradient, picks out the maximum corresponding rare earth element concentration gradient of assessment factor of numerical value；
Step S70 reduces screening scope, and step S10-S60 is repeated using the concentration gradient scope picked out in step S60
Reduce screening scope or determine optimum concentration；
Step S80 reversely screens basal medium, the different basal mediums of optimum concentration collocation determined using step S70, weight
Multiple step S40 and step S50, and assessment factor is calculated using step S60 computational methods, it is determined that most suitable basal medium.
Preferably, it is to set 5-15 concentration gradient that various concentrations are prepared in described step S30.
Preferably, the quantity of the culture medium for the Radix Glycyrrhizae cotyledon being inoculated with described step S30 is the training of each concentration gradient
Support base and 10-20 group parallel laboratory test groups are set.
Preferably, described rare earth element is Ce elements.
Preferably, the number of times repeated in described step S70 is 2-3 times.
Preferably, basal medium different in described step S80 include MS culture mediums, N6 culture mediums, B5 medium,
Nitsh culture mediums, White culture mediums and Miller culture mediums.
Embodiment
With reference to specific embodiment, the invention will be further described.
A kind of Radix Glycyrrhizae cotyledon inducing culture screening technique, it comprises the following steps：
Step S10 determines preliminary screening scope, and data of literatures is consulted in retrieval, contrasts other plant species tissue culture and inducement process
The amount of middle addition rare earth element simultaneously determines rare earth element preliminary screening scope；
Step S20 selects explant, and selection health, the Radix Glycyrrhizae cotyledon that growing environment is consistent and growth conditions are similar are used as explant
Body, carries out same sterilization to explant and handles；
Step S30 control cultures, the culture medium of various concentrations rare earth element addition is prepared using unitary variant method, is inoculated with respectively
Same amount of Radix Glycyrrhizae cotyledon, and carry out conventional induced tissue culture；
Step S40 record detection parameters, culture counts in each rare earth element concentration gradient after 20 days and induces callus
Quantity and the quantity of browning, and calculate inductivity and melting brown rate；
Step S50 increments are calculated, and are respectively taken out the callus in each parallel laboratory test group of same concentrations gradient, with clear
Water is rinsed well, dries surface moisture and callus is weighed, and records weight；
Step S60 calculate assess factor, by weight amount reduce 10 times, along with germination rate subtract melting brown rate calculate assessment because
Number, contrasts the assessment factor of various concentrations gradient, picks out the maximum corresponding rare earth element concentration gradient of assessment factor of numerical value；
Step S70 reduces screening scope, and step S10-S60 is repeated using the concentration gradient scope picked out in step S60
Reduce screening scope or determine optimum concentration；
Step S80 reversely screens basal medium, the different basal mediums of optimum concentration collocation determined using step S70, weight
Multiple step S40 and step S50, and assessment factor is calculated using step S60 computational methods, it is determined that most suitable basal medium.
As the presently preferred embodiments, it is to set 10 concentration gradients that various concentrations are prepared in described step S30.
As the presently preferred embodiments, the quantity of the culture medium for the Radix Glycyrrhizae cotyledon being inoculated with described step S30 is each concentration
The culture medium of gradient sets 10 groups of parallel laboratory test groups.
As the presently preferred embodiments, the number of times repeated in described step S70 is 2 times.
As the presently preferred embodiments, basal medium different in described step S80 include MS culture mediums, N6 culture mediums,
B5 medium, Nitsh culture mediums, White culture mediums and Miller culture mediums.
By filtering out the culture medium based on MS culture mediums, addition 6-BA (6- benzyls aminoadenine) concentration is
0.6mg/L、2,4-D（2,4 dichlorophenoxyacetic acid）Concentration be that 3 mg/L, the concentration of activated carbon are 0.1 mg/L, cerous nitrate
Concentration is 3g/L, and the concentration of vitamin is 0.3mg/L, and the concentration of agar powder is that 10g/L, the concentration of white granulated sugar are 30g/L, most suitable
Close Radix Glycyrrhizae cotyledon batch production induction production.
Embodiment described above only expresses one embodiment of the present invention, and it describes more specific and detailed, but simultaneously
Therefore the limitation to the scope of the claims of the present invention can not be interpreted as.It should be pointed out that for one of ordinary skill in the art
For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the guarantor of the present invention
Protect scope.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (6)
1. a kind of Radix Glycyrrhizae cotyledon inducing culture screening technique, it is characterised in that it comprises the following steps：
Step S10 determines preliminary screening scope, and data of literatures is consulted in retrieval, contrasts other plant species tissue culture and inducement process
The amount of middle addition rare earth element simultaneously determines rare earth element preliminary screening scope；
Step S20 selects explant, and selection health, the Radix Glycyrrhizae cotyledon that growing environment is consistent and growth conditions are similar are used as explant
Body, carries out same sterilization to explant and handles；
Step S30 control cultures, the culture medium of various concentrations rare earth element addition is prepared using unitary variant method, is inoculated with respectively
Same amount of Radix Glycyrrhizae cotyledon, and carry out conventional induced tissue culture；
Step S40 record detection parameters, culture counts in each rare earth element concentration gradient after 20 days and induces callus
Quantity and the quantity of browning, and calculate inductivity and melting brown rate；
Step S50 increments are calculated, and are respectively taken out the callus in each parallel laboratory test group of same concentrations gradient, with clear
Water is rinsed well, dries surface moisture and callus is weighed, and records weight；
Step S60 calculate assess factor, by weight amount reduce 10 times, along with germination rate subtract melting brown rate calculate assessment because
Number, contrasts the assessment factor of various concentrations gradient, picks out the maximum corresponding rare earth element concentration gradient of assessment factor of numerical value；
Step S70 reduces screening scope, and step S10-S60 is repeated using the concentration gradient scope picked out in step S60
Reduce screening scope or determine optimum concentration；
Step S80 reversely screens basal medium, the different basal mediums of optimum concentration collocation determined using step S70, weight
Multiple step S40 and step S50, and assessment factor is calculated using step S60 computational methods, it is determined that most suitable basal medium.
2. a kind of Radix Glycyrrhizae cotyledon inducing culture screening technique according to claim 1, it is characterised in that described step
It is to set 5-15 concentration gradient that various concentrations are prepared in S30.
3. a kind of Radix Glycyrrhizae cotyledon inducing culture screening technique according to claim 1, it is characterised in that described step
The quantity of the culture medium for the Radix Glycyrrhizae cotyledon being inoculated with S30 sets 10-20 group parallel laboratory test groups for the culture medium of each concentration gradient.
4. a kind of Radix Glycyrrhizae cotyledon inducing culture screening technique according to claim 1, it is characterised in that described rare earth
Element is Ce elements.
5. a kind of Radix Glycyrrhizae cotyledon inducing culture screening technique according to claim 1, it is characterised in that described step
The number of times repeated in S70 is 2-3 times.
6. a kind of Radix Glycyrrhizae cotyledon inducing culture screening technique according to claim 1, it is characterised in that described step
In S80 different basal mediums include MS culture mediums, N6 culture mediums, B5 medium, Nitsh culture mediums, White culture mediums and
Miller culture mediums.
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| CN201710171895.9A CN106942056A (en) | 2017-03-22 | 2017-03-22 | A kind of Radix Glycyrrhizae cotyledon inducing culture screening technique |
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| CN201710171895.9A CN106942056A (en) | 2017-03-22 | 2017-03-22 | A kind of Radix Glycyrrhizae cotyledon inducing culture screening technique |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1653886A (en) * | 2005-03-08 | 2005-08-17 | 天津大学 | Tissue culturing method for licorice adventitious root |
| CN102150618A (en) * | 2011-01-21 | 2011-08-17 | 甘肃农业大学 | Method for quickly propagating high-quality liquorice seedlings |
| CN102499072A (en) * | 2011-05-18 | 2012-06-20 | 天津大学 | Licorice rapid propagation seedling culture method |
| CN104686345A (en) * | 2015-02-24 | 2015-06-10 | 陈桂容 | Tissue culture rapid propagation method of liquorice |
| CN105340747A (en) * | 2015-11-27 | 2016-02-24 | 中国科学院华南植物园 | Asexual rapid propagation method for radix glycyrrhizae |
-
2017
- 2017-03-22 CN CN201710171895.9A patent/CN106942056A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1653886A (en) * | 2005-03-08 | 2005-08-17 | 天津大学 | Tissue culturing method for licorice adventitious root |
| CN102150618A (en) * | 2011-01-21 | 2011-08-17 | 甘肃农业大学 | Method for quickly propagating high-quality liquorice seedlings |
| CN102499072A (en) * | 2011-05-18 | 2012-06-20 | 天津大学 | Licorice rapid propagation seedling culture method |
| CN104686345A (en) * | 2015-02-24 | 2015-06-10 | 陈桂容 | Tissue culture rapid propagation method of liquorice |
| CN105340747A (en) * | 2015-11-27 | 2016-02-24 | 中国科学院华南植物园 | Asexual rapid propagation method for radix glycyrrhizae |
Non-Patent Citations (7)
| Title |
|---|
| 刘颖等: "稀土元素对甘草细胞生长及甘草酸合成的影响", 《广西植物》 * |
| 李孟楼: "《科研基本方法》", 31 August 2011 * |
| 杨世海等: "不同附加物对甘草愈伤组织培养中黄酮类化合物形成的影响", 《中国药学杂志》 * |
| 杨世海等: "稀土元素对甘草愈伤组织生长及黄酮类化合物含量的影响", 《中药材》 * |
| 王水琦: "《植物组织培养》", 31 March 2007 * |
| 胡海英等: "胀果甘草愈伤组织诱导培养", 《药物生物技术》 * |
| 董春英: "稀土元素在组织培养中的作用研究进展", 《湖南林业科技》 * |
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Application publication date: 20170714 |