CN106359082A - Tissue culture method with effect of improving general flavone content of gynura divaricata test-tube plantlet - Google Patents
Tissue culture method with effect of improving general flavone content of gynura divaricata test-tube plantlet Download PDFInfo
- Publication number
- CN106359082A CN106359082A CN201610694142.1A CN201610694142A CN106359082A CN 106359082 A CN106359082 A CN 106359082A CN 201610694142 A CN201610694142 A CN 201610694142A CN 106359082 A CN106359082 A CN 106359082A
- Authority
- CN
- China
- Prior art keywords
- general flavone
- test tube
- culture
- radix
- flavone content
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a tissue culture method with the effect of improving general flavone content of a gynura divaricata test-tube plantlet. The tissue culture method comprises 1, taking an explant and carrying out disinfection, 2, carrying out induced culture through inoculating a MS induced culture medium with the disinfected explant and carrying out induced culture to obtain callus, and 3, inoculating a culture medium of test-tube plantlet induction with the callus and carrying out induced culture to obtain a high general flavone content test-tube plantlet. The test-tube plantlet can be directly used as a general flavone extraction plant, can be used as a seedling for traditional drug cultivation production, can also be used for later development of high-general flavone content gynura divaricata and can provide technical support for high-quality gynura divaricata seedling cultivation. The tissue culture method has substantial effects, utilizes a simple and convenient medium formula, can be used for batch culture of gynura divaricata and is safe and reliable.
Description
Technical field
The invention belongs to field of plant tissue culture technique, it is related to a kind of promotion Radix et Rhizoma Gynurae divaricatae test tube seedling general flavone content and carries
High method for tissue culture.
Background technology
Radix et Rhizoma Gynurae divaricatae (gynura divaricata (l.) dc), perennial herb, is that Compositae Radix Gynurae belongs to perennial herb
Plant, is one kind of Chinese traditional herbs, widely distributed, is a kind of edible and the medicinal improved seeds having concurrently, mainly in South China
Portion, the west and south and Taiwan.Radix et Rhizoma Gynurae divaricatae contains abundant polysaccharide, flavone and alkaloid, has medicinal and health care.Its each
Part can be used as medicine.There are some researches show that it contains abundant total flavonoid etc., flavonoid belongs to Secondary metabolites, be many
Plant the material base that Chinese medicine plays physiological action.Its extract has been proved to reduce blood pressure and drop hypoglycemic in zoopery
Effect.Its tender stem and leaf can also directly eat, and because having health care, become a kind of plant being becoming increasingly popular.
But its wild resource is seldom, there is the features such as fecundity is poor, seed amount is few simultaneously, and traditional cutting propagation quantity also has very much
Limit, thus conventional research is concentrated mainly on it and quickly breeds, and has been set up with low cost, cycle is short quickly numerous at present
Grow system.
But the content for effective medicinal components total flavones in Radix et Rhizoma Gynurae divaricatae, the especially content in the test tube seedling stage are not closed
Note.
Content of the invention
For at present with regard to Radix et Rhizoma Gynurae divaricatae research situation, the present invention is shown by substantial amounts of experimentation, changes the white back of the body three
The culture medium prescription of seven test tube seedlings has the effect of raising for general flavone content in Radix et Rhizoma Gynurae divaricatae test tube seedling.The purpose of the present invention
It is to obtain the test tube seedling of high general flavone content, so that the seedling of the high general flavone content of large-scale production is it is adaptable to Radix et Rhizoma Gynurae divaricatae
Large-scale production, the defect that existing reproduction technique exists can be overcome simultaneously, and then provide and a kind of improve Radix et Rhizoma Gynurae divaricatae test tube
The method for tissue culture of general flavone content in Seedling.
The purpose of the present invention is achieved through the following technical solutions:
A kind of method for tissue culture improving general flavone content in Radix et Rhizoma Gynurae divaricatae test tube seedling, comprises the steps:
First, the sterilization of drawing materials of explant: outer value body is first cleaned up with tap water, then sterilized water is used on super-clean bench
Cleaning 3 times, using liquor natrii hypochloritises' sterilization of 20g/l, then with aseptic water washing 3 times, is then cut into 0.5 centimeter square, standby
With.
2nd, inducing culture: the explant disinfecting is inoculated in inducing culture in ms inducing culture and obtains calluss,
Described condition of culture is: intensity of illumination 2000lx, daily illumination 16 hours, cultivation temperature be 25 ± 2 DEG C, incubation time be 15～
20 days, in described ms inducing culture in addition to ms medium component, also include 30g/l sucrose, agar content is 7%, ph=5.5
～6, naa concentration is 0.5mg/l, and 6-ba concentration is 2.0mg/l.
3rd, calluss are inoculated into the test tube that inducing culture in the culture medium of induction test tube seedling obtains high general flavone content
Seedling, described condition of culture is intensity of illumination 2000lx, daily illumination 16 hours, and cultivation temperature is 25 ± 2 DEG C, and incubation time is 45
～50 days, the culture medium of described induction test tube seedling was ms+ carbon source sucrose 30g/l, trace element compares mn2+: zn2+For 44.6mg/l:
13.2mg/l, nitrogen source kno3: nh4no3It is 0.1mg/l:1.5mg/l for 2020mg/l:1600mg/l, hormone than naa:6-ba.
In the present invention, described outer value body is selected from the October Radix et Rhizoma Gynurae divaricatae top young leaflet tablet given birth to then or stem and leaf.
In the present invention, sterile working to be strictly observed in all steps.
The present invention has the advantage that
1st, the present invention improves the content of total flavones in the test tube seedling stage, without going past generation variation in step afterwards
Step, therefore hereditary stability are good, can obtain stable seedling.
2nd, the plant obtained by the present invention both can extract plant directly as total flavones, is supplied to tradition but also as seedling
Drug cultivation uses on producing, and can be also Radix et Rhizoma Gynurae divaricatae good seed as the subsequent development of high general flavone content Radix et Rhizoma Gynurae divaricatae
Cultivate and technical support is provided.
3rd, effect is significant of the present invention, culture medium prescription is simple, convenient, can be used for the Batch Culture of Radix et Rhizoma Gynurae divaricatae, and safety
Reliable.
Brief description
Fig. 1 is the picture obtaining each stage Radix et Rhizoma Gynurae divaricatae in the present invention, wherein: a blade inducing culture；What b obtained heals
Injured tissue；C: subsequent experimental calluss.
Specific embodiment
Below in conjunction with the accompanying drawings technical scheme is further described, but is not limited thereto, every to this
Inventive technique scheme is modified or equivalent, without deviating from the spirit and scope of technical solution of the present invention, all should cover
In protection scope of the present invention.
Embodiment 1
Present embodiments provide a kind of method for tissue culture improving general flavone content in Radix et Rhizoma Gynurae divaricatae test tube seedling, specifically real
Apply step as follows:
First, explant is drawn materials and is sterilized: chooses October Radix et Rhizoma Gynurae divaricatae top young leaflet tablet, is first cleaned with tap water dry
Only, aseptic water washing 3 times in superclean bench according to sterile working's code, then sterilization 10 in 20g/l liquor natrii hypochloritises
Minute, then take out aseptic water washing 3 times, be cut into 0.5cm about fritter, standby.
2nd, the explant cutting is accessed in ms inducing culture (Fig. 1 a), condition of culture is: intensity of illumination 2000lx,
Light application time 16 hours, cultivation temperature is 25 ± 2 DEG C, obtains calluss (Fig. 1 b) within about 15～20 days about, described ms lures
Lead in culture medium in addition to ms medium component, also include 30g/l sucrose, agar content is 7%, ph=5.5～6, naa concentration is
0.5mg/l, 6-ba concentration is 2.0mg/l.
3rd, the well-grown calluss obtaining are required to be inoculated in the culture medium of induction test tube seedling according to sterile working
In (ms+ sucrose 30g/l, trace element compare mn2+: zn2+For 44.6mg/l:13.2mg/l, nitrogen source kno3: nh4no3For 2020mg/
L:1600mg/l, hormone are 0.1mg/l:1.5mg/l than naa:6-ba) induction obtain test tube seedling (Fig. 1 c).Incubation time is 45
～50 days about, condition of culture is intensity of illumination 2000lx, light application time 16 hours, and cultivation temperature is 25 ± 2 DEG C.
The test tube seedling obtaining is cut, removes culture medium and calluss, clean up, measure always yellow in whole strain test tube seedling
The content of ketone, test result indicate that, in test tube seedling, the content of total flavones improves than the content of total flavones in original plant
15%.
Embodiment 2
First, explant is drawn materials and is sterilized: chooses October Radix et Rhizoma Gynurae divaricatae top children young stem and leaf, is first cleaned with tap water dry
Only, aseptic water washing 3 times in superclean bench according to sterile working's code, then sterilization 10 in 20g/l liquor natrii hypochloritises
Minute, then take out aseptic water washing 3 times, be cut into 0.5cm about fritter, standby.
2nd, the explant cutting is accessed in ms inducing culture (Fig. 1 a), condition of culture is: intensity of illumination 2000lx,
Light application time 16 hours, cultivation temperature is 25 ± 2 DEG C, obtains within about 15～20 days calluss (Fig. 1 b), described ms induction training
In foster base in addition to ms medium component, also include 30g/l sucrose, agar content is 7%, ph=5.5～6, naa concentration is
0.5mg/l, 6-ba concentration is 2.0mg/l.
3rd, the well-grown calluss obtaining are required to be inoculated in the culture medium of induction test tube seedling according to sterile working
In (ms+ sucrose 30g/l, trace element compare mn2+: zn2+For 44.6mg/l:13.2mg/l, nitrogen source kno3: nh4no3For 2020mg/
L:1600mg/l, hormone are 0.1mg/l:1.5mg/l than naa:6-ba) induction obtain test tube seedling (Fig. 1 c).Incubation time is 45
～50 days about, condition of culture is intensity of illumination 2000lx, light application time 16 hours, and cultivation temperature is 25 ± 2 DEG C.
The test tube seedling obtaining is cut, removes culture medium and calluss, clean up, measure always yellow in whole strain test tube seedling
The content of ketone, test result indicate that, in test tube seedling, the content of total flavones improves than the content of total flavones in original plant
14.8%.
Claims (8)
1. a kind of method for tissue culture improving general flavone content in Radix et Rhizoma Gynurae divaricatae test tube seedling is it is characterised in that methods described step
As follows:
First, the sterilization of drawing materials of explant；
2nd, inducing culture: the explant disinfecting is inoculated in inducing culture in ms inducing culture and obtains calluss；
3rd, calluss are inoculated into the test tube seedling that inducing culture in the culture medium of induction test tube seedling obtains high general flavone content.
2. the method for tissue culture improving general flavone content in Radix et Rhizoma Gynurae divaricatae test tube seedling according to claim 1, its feature
It is in described step one, the sterilization method of drawing materials of explant is as follows: outer value body is first cleaned up with tap water, then super
With sterile water wash 3 times on net platform, using liquor natrii hypochloritises' sterilization of 20g/l, then with aseptic water washing 3 times, it is then cut into
0.5 centimeter square, standby.
3. the method for tissue culture improving general flavone content in Radix et Rhizoma Gynurae divaricatae test tube seedling according to claim 1 and 2, it is special
Levy and be that described outer value body is selected from the October Radix et Rhizoma Gynurae divaricatae top young leaflet tablet given birth to then or stem and leaf.
4. the method for tissue culture improving general flavone content in Radix et Rhizoma Gynurae divaricatae test tube seedling according to claim 1, its feature
It is in described step 2, condition of culture is: intensity of illumination 2000lx, daily illumination 16 hours, cultivation temperature is 25 ± 2 DEG C,
Incubation time is 15～20 days.
5. the method for tissue culture improving general flavone content in Radix et Rhizoma Gynurae divaricatae test tube seedling according to claim 1, its feature
It is in described step 2, in ms inducing culture in addition to ms medium component, also include 30g/l sucrose, agar content is
7%, ph=5.5～6, naa concentration is 0.5mg/l, and 6-ba concentration is 2.0mg/l.
6. the method for tissue culture improving general flavone content in Radix et Rhizoma Gynurae divaricatae test tube seedling according to claim 1, its feature
It is in described step 3, condition of culture is intensity of illumination 2000lx, daily illumination 16 hours, and cultivation temperature is 25 ± 2 DEG C, training
The foster time is 45～50 days.
7. the method for tissue culture improving general flavone content in Radix et Rhizoma Gynurae divaricatae test tube seedling according to claim 1, its feature
It is in described step 3, the culture medium of induction test tube seedling is ms+ carbon source sucrose 30g/l, trace element compares mn2+: zn2+For
44.6mg/l:13.2mg/l, nitrogen source kno3: nh4no3It is 0.1mg/l for 2020mg/l:1600mg/l, hormone than naa:6-ba:
1.5mg/l.
8. the method for tissue culture improving general flavone content in Radix et Rhizoma Gynurae divaricatae test tube seedling according to claim 1, its feature
It is described step one, sterile working to be strictly observed in two, three.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610694142.1A CN106359082A (en) | 2016-08-19 | 2016-08-19 | Tissue culture method with effect of improving general flavone content of gynura divaricata test-tube plantlet |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610694142.1A CN106359082A (en) | 2016-08-19 | 2016-08-19 | Tissue culture method with effect of improving general flavone content of gynura divaricata test-tube plantlet |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106359082A true CN106359082A (en) | 2017-02-01 |
Family
ID=57879525
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610694142.1A Pending CN106359082A (en) | 2016-08-19 | 2016-08-19 | Tissue culture method with effect of improving general flavone content of gynura divaricata test-tube plantlet |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106359082A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107823233A (en) * | 2017-10-31 | 2018-03-23 | 安徽新华学院 | A kind of extracting method of the flavones based on angelica keiskei koidz |
| CN107853174A (en) * | 2017-10-31 | 2018-03-30 | 安徽新华学院 | A kind of method for improving angelica keiskei koidz lateral bud survival rate |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105850749A (en) * | 2016-06-09 | 2016-08-17 | 泉州正和堂生物科技有限公司 | Culture method for detoxified gynura divaricata tissue cells |
-
2016
- 2016-08-19 CN CN201610694142.1A patent/CN106359082A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105850749A (en) * | 2016-06-09 | 2016-08-17 | 泉州正和堂生物科技有限公司 | Culture method for detoxified gynura divaricata tissue cells |
Non-Patent Citations (4)
| Title |
|---|
| 李琰等: "微量元素对雷公藤不定根生长和次生代谢产物含量的影响", 《林业科学》 * |
| 杨世海等: "培养基中碳源和氮源对甘草愈伤组织生长和黄酮类化合物合成的影响", 《中国中药杂志》 * |
| 范三微等: "植物激素对灵菊七愈伤组织中黄酮含量的影响", 《湖北农业科学》 * |
| 覃兰琼等: "白背三七总黄酮含量季节性变化规律研究", 《中国药师》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107823233A (en) * | 2017-10-31 | 2018-03-23 | 安徽新华学院 | A kind of extracting method of the flavones based on angelica keiskei koidz |
| CN107853174A (en) * | 2017-10-31 | 2018-03-30 | 安徽新华学院 | A kind of method for improving angelica keiskei koidz lateral bud survival rate |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101946705B (en) | Method for propagating cochinchnese asparagus root simply, efficiently and quickly | |
| CN105104190B (en) | Herba Anoectochili roxburghii tissue culture seedling establishment method | |
| CN105165617B (en) | The tissue cultures and fast seedling-cultivating method of blood aspidistra seed | |
| CN102907323A (en) | Method for aseptically producing miniature seed stems of common bletilla pseudobulb seeds | |
| CN103798145A (en) | Culture medium for tissue culture of vernonia amygdalina del. | |
| CN106358821A (en) | Cuttage treatment method in cuttage seedling culturing process of honeysuckles | |
| CN106172016A (en) | A kind of Cortex Acanthopancis tissue culture and rapid propagation method | |
| CN103548694A (en) | Tissue culture and rapid propagation method for dracaena cochinchinensis | |
| CN109220790A (en) | A kind of in vitro outer breeding method of red fruit ginseng | |
| CN104094845B (en) | A kind of in-vitro culture method of Dendranthema indicum | |
| CN103392593B (en) | A kind of quickly tissue culture propagation method of Dendrobidium huoshanness seedling | |
| CN106508661A (en) | Method for constructing anoectochilus formosanus polyploidy resource | |
| CN106386507A (en) | Quick tissue-culture propagation method of dendrobium huoshanense seed seedlings | |
| CN106359082A (en) | Tissue culture method with effect of improving general flavone content of gynura divaricata test-tube plantlet | |
| CN106106190A (en) | A kind of tissue cultivation rapid breeding method of Radix Zanthoxyli | |
| CN101715733A (en) | Tissue culturing method of protocorms of dendrobium candidum | |
| CN102783420B (en) | Method for quickly breeding cordate houttuynia | |
| CN106134987A (en) | Exocarpium Citri Rubrum tissue-culturing quick-propagation and method for culturing seedlings | |
| Anburaj et al. | In vitro regeneration of Cleome viscosa–an important medicinal herb | |
| CN106386494B (en) | A kind of sweet potato stem tip detoxification and breeding method | |
| CN107801635A (en) | A kind of dendrobium candidum protocorm largely breaks up and rooting method | |
| CN107155881B (en) | One kind being suitable for polytype plant rapid propagation method | |
| CN106417012B (en) | A kind of tissue culture method for promoting bamboo shoot orchid efficiently to breed | |
| CN104160965A (en) | Rapid propagation method for litsea rotundifolia | |
| CN108541594A (en) | A kind of tissue culture and rapid propagation method of polygonatum cirrhifolium Royle |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170201 |
|
| WD01 | Invention patent application deemed withdrawn after publication |