CN106912382A - A kind of screening technique of the condition of tissue culture for promoting Radix Glycyrrhizae to go out root - Google Patents
A kind of screening technique of the condition of tissue culture for promoting Radix Glycyrrhizae to go out root Download PDFInfo
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- CN106912382A CN106912382A CN201710171879.XA CN201710171879A CN106912382A CN 106912382 A CN106912382 A CN 106912382A CN 201710171879 A CN201710171879 A CN 201710171879A CN 106912382 A CN106912382 A CN 106912382A
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- culture
- radix glycyrrhizae
- root
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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
- A01H4/008—Methods for regeneration to complete plants
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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
Abstract
The present invention provides a kind of screening technique of the condition of tissue culture for promoting Radix Glycyrrhizae to go out root, strengthen the physiological effect of the wavelength to Radix Glycyrrhizae tissue culturing seedling of light source by using the adjustment test of unitary variant method, it is determined that most suitable enhancing optical source wavelength, and the influence according to light sources with different wavelengths to Radix Glycyrrhizae tissue-cultured seedling, determine that light sources with different wavelengths goes out combination, and by keeping enhancing light source constant reverse determination basal medium, while introducing tissue-cultured seedling actually goes out root rate as screening criteria, it is more suitable for commercial plant metaplasia product.
Description
Technical field
The invention belongs to Radix Glycyrrhizae technical field of tissue culture, more particularly to a kind of sieve of the condition of tissue culture for promoting Radix Glycyrrhizae to go out root
Choosing 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.In cylinder, there is bud scar on surface to rhizome, and there is marrow at section middle part.Gas is micro-, and taste is sweet and special.
Indication is clearing heat and detoxicating, expelling phlegm and arresting coughing, gastral cavity abdomen etc..Happiness is dark moist, the low dry climate of sunshine temperature long.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
The method of taking is excavation leaf plant resource, 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 rarely growth of Glycyrrhiza uralensis is adjusted by strengthening light source in existing Radix Glycyrrhizae culture medium
Technology.
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
Above mentioned problem is had based on prior art, the present invention provides a kind of screening side of the condition of tissue culture for promoting Radix Glycyrrhizae to go out root
Method, the physiological effect of the wavelength to Radix Glycyrrhizae tissue culturing seedling of light source is strengthened by using the adjustment test of unitary variant method, it is determined that most
Suitable enhancing optical source wavelength, and the influence according to light sources with different wavelengths to Radix Glycyrrhizae tissue-cultured seedling, determine that light sources with different wavelengths goes out combination, and
By keeping the enhancing constant reverse determination basal medium of light source, while the root rate that actually goes out for introducing tissue-cultured seedling is marked as screening
Standard, is more suitable for commercial plant metaplasia product.
A kind of screening technique of the condition of tissue culture for promoting Radix Glycyrrhizae to go out root, it is comprised the following steps：
Step S10 determines preliminary screening scope, and data of literatures is consulted in retrieval, contrasts other plant species tissue culture rooting process
Middle addition strengthens light source category and determines the preliminary screening scope of optical source wavelength；
Step S20 selects explant, selection health, the Radix Glycyrrhizae Multiple Buds that growing environment is consistent and growth conditions are similar, by clump
Sprout and be divided into simple bud；
Step S30 control culture, the culture medium based on 1/2MS culture mediums, add aspirin solution volume fraction be for
0.03%、NAA（Methyl α-naphthyl acetate）Concentration for the concentration of 0.05mg/L, ZT (zeatin) be that 2.0 mg/L, the concentration of lanthanum nitrate are
3g/L, vitamin B12Concentration be 0.3mg/L, the concentration of agar powder is 10g/L, the concentration of white granulated sugar is 7g/L, is configured to survey
Culture medium on probation, is inoculated with same amount of Radix Glycyrrhizae Multiple Buds respectively, and carries out conventional root tissues culture as a control group, in routine
Used as experimental group, control group all sets multiple parallel condition different-waveband enhancing enhancing light source with experimental group on the basis of culture
Group；
Step S40 record detection parameters, culture counts the survival Radix Glycyrrhizae bud that goes out root in each experimental group and control group after 20 days
Quantity, and calculating effectively goes out root rate；
Step S50 increments are calculated, and respectively take out the bud in each parallel laboratory test group of same light source wave-length coverage, use clear water
Rinse well, whole strain plant is weighed after drying surface moisture, then root is cut out, and root is weighed, record
Weight；
Step S60 calculates assessment factor, will effectively go out root rate be multiplied by ten point seven of root weight and plant overall weight ten/
Four sum, draws assessment factor, contrasts the assessment factor of various concentrations gradient, picks out the maximum assessment factor of numerical value corresponding
Optical source wavelength scope；
Step S70 reduces screening scope, and step S10- is repeated using the enhancing optical source wavelength scope picked out in step S60
S60 reduces screening scope or determines optimum wavelength scope；
Step S80 reversely screens basal medium, the different basis cultures of optimum wavelength scope collocation determined using step S70
Base, repeat step S40 and step S50, and using the computational methods calculating assessment factor of step S60, it is determined that most suitable basis culture
Base；
Step S90 different wave length combined sortings, determine that different wave length is joined to the physiology of Radix Glycyrrhizae adventitious buds proliferation by single factor
Number influence, adjustment is best suitable for wavelength combination.
Preferably, each experimental group or control group set 5-15 parallel group in described step S30.
Preferably, the number of times repeated in described step S70 is 2-3 times.
Preferably, different basal mediums includes 1/2MS culture mediums, MS culture mediums, N6 cultures in described step S80
Base, B5 medium, Nitsh culture mediums, White culture mediums and Miller culture mediums.
Specific embodiment
With reference to specific embodiment, the invention will be further described.
A kind of screening technique of the condition of tissue culture for promoting Radix Glycyrrhizae to go out root, it is comprised the following steps：
Step S10 determines preliminary screening scope, and data of literatures is consulted in retrieval, contrasts other plant species tissue culture rooting process
Middle addition strengthens light source category and determines the preliminary screening scope of optical source wavelength；
Step S20 selects explant, selection health, the Radix Glycyrrhizae Multiple Buds that growing environment is consistent and growth conditions are similar, by clump
Sprout and be divided into simple bud；
Step S30 control culture, the culture medium based on 1/2MS culture mediums, add aspirin solution volume fraction be for
0.03%、NAA（Methyl α-naphthyl acetate）Concentration for the concentration of 0.05mg/L, ZT (zeatin) be that 2.0 mg/L, the concentration of lanthanum nitrate are
3g/L, vitamin B12Concentration be 0.3mg/L, the concentration of agar powder is 10g/L, the concentration of white granulated sugar is 7g/L, is configured to survey
Culture medium on probation, is inoculated with same amount of Radix Glycyrrhizae Multiple Buds respectively, and carries out conventional root tissues culture as a control group, in routine
Used as experimental group, control group all sets multiple parallel condition different-waveband enhancing enhancing light source with experimental group on the basis of culture
Group；
Step S40 record detection parameters, culture counts the survival Radix Glycyrrhizae bud that goes out root in each experimental group and control group after 20 days
Quantity, and calculating effectively goes out root rate；
Step S50 increments are calculated, and respectively take out the bud in each parallel laboratory test group of same light source wave-length coverage, use clear water
Rinse well, whole strain plant is weighed after drying surface moisture, then root is cut out, and root is weighed, record
Weight；
Step S60 calculates assessment factor, will effectively go out root rate be multiplied by ten point seven of root weight and plant overall weight ten/
Four sum, draws assessment factor, contrasts the assessment factor of various concentrations gradient, picks out the maximum assessment factor of numerical value corresponding
Optical source wavelength scope；
Step S70 reduces screening scope, and step S10- is repeated using the enhancing optical source wavelength scope picked out in step S60
S60 reduces screening scope or determines optimum wavelength scope；
Step S80 reversely screens basal medium, the different basis cultures of optimum wavelength scope collocation determined using step S70
Base, repeat step S40 and step S50, and using the computational methods calculating assessment factor of step S60, it is determined that most suitable basis culture
Base；
Step S90 different wave length combined sortings, determine that different wave length is joined to the physiology of Radix Glycyrrhizae adventitious buds proliferation by single factor
Number influence, adjustment is best suitable for wavelength combination.
As the presently preferred embodiments, each experimental group or control group set 10 parallel groups in described step S30.
As the presently preferred embodiments, the number of times repeated in described step S70 is 3 times.
As the presently preferred embodiments, different basal mediums includes that 1/2MS culture mediums, MS are cultivated in described step S80
Base, N6 culture mediums, B5 medium, Nitsh culture mediums, White culture mediums and Miller culture mediums.
Being defined below enhancing light source by screening has the quality for actually going out root rate and root higher：Use natural light, indigo plant
Light strengthens light source and feux rouges enhancing light source irradiation Multiple Buds, and red light source is the feux rouges that wave band is 660nm, and blue-light source is wave band
It is the blue light of 46nm, light application time is illumination 16 hours/dark 8 hours, feux rouges 70%, blue light 30%.
Claims (4)
1. a kind of promotion Radix Glycyrrhizae goes out the screening technique of the condition of tissue culture of root, it is characterised in that it is comprised the following steps：
Step S10 determines preliminary screening scope, and data of literatures is consulted in retrieval, contrasts other plant species tissue culture rooting process
Middle addition strengthens light source category and determines the preliminary screening scope of optical source wavelength；
Step S20 selects explant, selection health, the Radix Glycyrrhizae Multiple Buds that growing environment is consistent and growth conditions are similar, by clump
Sprout and be divided into simple bud；
Step S30 control culture, the culture medium based on 1/2MS culture mediums, add aspirin solution volume fraction be for
0.03%、NAA（Methyl α-naphthyl acetate）Concentration for the concentration of 0.05mg/L, ZT (zeatin) be that 2.0 mg/L, the concentration of lanthanum nitrate are
3g/L, vitamin B12Concentration be 0.3mg/L, the concentration of agar powder is 10g/L, the concentration of white granulated sugar is 7g/L, is configured to survey
Culture medium on probation, is inoculated with same amount of Radix Glycyrrhizae Multiple Buds respectively, and carries out conventional root tissues culture as a control group, in routine
Used as experimental group, control group all sets multiple parallel condition different-waveband enhancing enhancing light source with experimental group on the basis of culture
Group；
Step S40 record detection parameters, culture counts the survival Radix Glycyrrhizae bud that goes out root in each experimental group and control group after 20 days
Quantity, and calculating effectively goes out root rate；
Step S50 increments are calculated, and respectively take out the bud in each parallel laboratory test group of same light source wave-length coverage, use clear water
Rinse well, whole strain plant is weighed after drying surface moisture, then root is cut out, and root is weighed, record
Weight；
Step S60 calculates assessment factor, will effectively go out root rate be multiplied by ten point seven of root weight and plant overall weight ten/
Four sum, draws assessment factor, contrasts the assessment factor of various concentrations gradient, picks out the maximum assessment factor of numerical value corresponding
Optical source wavelength scope；
Step S70 reduces screening scope, and step S10- is repeated using the enhancing optical source wavelength scope picked out in step S60
S60 reduces screening scope or determines optimum wavelength scope；
Step S80 reversely screens basal medium, the different basis cultures of optimum wavelength scope collocation determined using step S70
Base, repeat step S40 and step S50, and using the computational methods calculating assessment factor of step S60, it is determined that most suitable basis culture
Base；
Step S90 different wave length combined sortings, determine that different wave length is joined to the physiology of Radix Glycyrrhizae adventitious buds proliferation by single factor
Number influence, adjustment is best suitable for wavelength combination.
2. a kind of promotion Radix Glycyrrhizae according to claim 1 goes out the screening technique of the condition of tissue culture of root, it is characterised in that described
The step of S30 in each experimental group or control group 5-15 parallel group is set.
3. a kind of promotion Radix Glycyrrhizae according to claim 1 goes out the screening technique of the condition of tissue culture of root, it is characterised in that described
The step of S70 in repeat number of times be 2-3 times.
4. a kind of promotion Radix Glycyrrhizae according to claim 1 goes out the screening technique of the condition of tissue culture of root, it is characterised in that described
The step of S80 in different basal medium include 1/2MS culture mediums, MS culture mediums, N6 culture mediums, B5 medium, Nitsh trainings
Support base, White culture mediums and Miller culture mediums.
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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 CN201710171879.XA patent/CN106912382A/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 (4)
| Title |
|---|
| 李孟楼: "《科研基本方法》", 31 August 2011, 西北农林科技大学出版社 * |
| 李胜等: "《植物组织培养》", 31 July 2015, 中国轻工业出版社 * |
| 梁玉勇等: ""甘草组培苗的培养基优化及其有效成分分析"", 《贵州农业科学》 * |
| 霍云谦等: ""中国甘草的组织培养研究进展"", 《中国农学通报》 * |
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