CN110226518A - A kind of tissue culture and rapid propagation method of brook anemone - Google Patents
A kind of tissue culture and rapid propagation method of brook anemone Download PDFInfo
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- CN110226518A CN110226518A CN201910641302.XA CN201910641302A CN110226518A CN 110226518 A CN110226518 A CN 110226518A CN 201910641302 A CN201910641302 A CN 201910641302A CN 110226518 A CN110226518 A CN 110226518A
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- Prior art keywords
- brook anemone
- tissue culture
- rapid propagation
- culture
- anemone
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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
Abstract
The invention discloses a kind of tissue culture and rapid propagation method of brook anemone, 1) it sterilizes；2) by the explant after surface sterilization along the crosscutting square at 2~6mm in vertical middle arteries direction, the blade back of the explant is inoculated in downward in the induced medium of culture bottle, 10 plants of explants of every bottle of inoculation；3) adventitious bud induced；4) adventitious bud of the induction is inoculated into root media, culture can take root for 60~70 days, by to brook anemone in vitro culture, set up brook anemone tissue culture rapid propagation system, make survival rate up to 95% or more, the foundation of brook anemone tissue culture and rapid propagation method solves the problems, such as that brook anemone seed is rare, germination rate is low, reproduction speed is slow, is suitable for brook anemone industrialization nursery；Reproduction speed is significantly improved, and rare Yi nationality's medicine brook anemone is made to measure increase with seedling, expands the germ plasm resource quantity of brook anemone, while can reduce cost, easy to operate, high financial profit.
Description
Technical field
The present invention relates to Chinese medicine plant technical field more particularly to a kind of tissue culture and rapid propagation methods of brook anemone.
Background technique
Brook anemone Anemones rivularis Buch.-ham.ex Dc is called anemone rivularis, sees that wind is green, sees wind indigo plant, is hair
Gelsemium section perennial herb medicinal plant, has clearing heat and detoxicating, activates blood and relax tendons, antibechic, eliminating the phlegm, antibacterial, anticancer and other effects.Brook anemone
It is often born between the mountainous region of Yunnan height above sea level 1750m~3300m, grass, by small stream or waits wetlands or border.Secondly in Sichuan, Guizhou, Gansu
The ground such as the west and south, southeast Qinghai, the Hubei west and south, West Guangxi, southern Tibet and east are also distributed, but with environment item
Part changes, and current wild brook anemone is endangered, causes shortage of resources, supply falls short of demand.
Setting percentage is very low under field conditions (factors) and seed longeivity is short, easily loses germination for brook anemone, while its seed progeny
Breeding is easy to lose original kind of property, morphs, causes bennet to attenuate, softens, flower pattern becomes smaller.Division propagation can keep former
Some kind of property feature, does not morph, but seedling is easy to be infected by disease pest in reproductive process, and growth rate is slow, it is difficult to
Meet the urgent need that market produces brook anemone.Therefore, brook anemone seeling industry technical system rapidly and efficiently is established to treasure
Dilute Yi nationality's medicine brook anemone seedling amount and the germ plasm resource quantity for expanding brook anemone all have urgent realistic meaning.
Summary of the invention
Of the invention provides a kind of tissue culture and rapid propagation method of brook anemone.
The scheme of the invention is:
A kind of tissue culture and rapid propagation method of brook anemone, including the following steps:
1) for clip brook anemone spire as explant, the sodium hypochlorite with 75% alcohol and 5% is molten on superclean bench
Surface sterilization is carried out to the explant；
2) by the explant after surface sterilization along the crosscutting square at 2~6mm in vertical middle arteries direction, the explant
The blade back of body is inoculated in downward in the induced medium of culture bottle, 10 plants of explants of every bottle of inoculation；
3) explant being seeded in culture bottle is first subjected to 3 days dark treatments, then intensity of illumination be 800~
2000Lux, 12~14h/d of light application time, temperature be 20~28 DEG C under conditions of cultivate 20~35 days be induced it is indefinite
Bud；
4) adventitious bud of the induction is inoculated into root media, inoculation number is 10 in every bottle of root media cultivation
Strain, under conditions of intensity of illumination is 800~2000Lux, 12~14h/d of light application time, temperature are 20~28 DEG C cultivate 60~
It can take root within 70 days.
As a preferred technical solution, the induced medium be containing 1.0~6.0mg/L of 6-BA, IAA 0.1~
The MS culture medium of 0.4mg/L.
The root media is the MS culture medium containing 0.1~0.3mg/L of IAA as a preferred technical solution,.
Intensity of illumination is 1500Lux in the step 4) as a preferred technical solution,.
Light application time 12h/d in the step 4) as a preferred technical solution,.
Temperature is 24 DEG C in the step 4) as a preferred technical solution,.
Culture 60 days in the step 4) as a preferred technical solution,.
The root that the length taken root is 9~11cm as a preferred technical solution,.
The every plant of brook anemone to take root in the step 4) as a preferred technical solution, takes root 7~9, in the chicken of tubbiness
Claw-like, plant is tall and straight, and for every plant of brook anemone height in 11~13cm, blade is light green.
By adopting the above-described technical solution, a kind of tissue culture and rapid propagation method of brook anemone, 1) clip on superclean bench
Brook anemone spire carries out surface sterilization to the explant as explant, with the sodium hypochlorite of 75% alcohol and 5% is molten；2) will
The explant after surface sterilization is along the crosscutting square at 2~6mm in vertical middle arteries direction, and the blade back of the explant is downward
It is inoculated in the induced medium of culture bottle, 10 plants of explants of every bottle of inoculation；3) explant that will be seeded in culture bottle
3 days dark treatments are first carried out, then intensity of illumination is 800~2000Lux, 12~14h/d of light application time, temperature are 20~28 DEG C
Under conditions of cultivate 20~35 days i.e. adventitious bud that is induced；4) adventitious bud by the induction is inoculated into root media
In, every bottle of root media plant in inoculation number be 10 plants, intensity of illumination be 800~2000Lux, 12~14h/d of light application time,
Temperature is cultivated 60~70 days and can be taken root under conditions of being 20~28 DEG C.
Advantages of the present invention:
Firstly, by brook anemone in vitro culture, it is established that brook anemone tissue culture rapid propagation system, make survival rate up to 95% with
On, the foundation of brook anemone tissue culture and rapid propagation method solves the problems, such as that brook anemone seed is rare, germination rate is low, reproduction speed is slow, is applicable in
In brook anemone industrialization nursery；
Reproduction speed is significantly improved, and so that rare Yi nationality's medicine brook anemone is measured increase with seedling, expands brook anemone
Germ plasm resource quantity, while can reduce cost, easy to operate, high financial profit.
Specific embodiment
In order to make up the above deficiency, the present invention provides a kind of tissue culture and rapid propagation methods of brook anemone to solve above-mentioned background skill
The problems in art.
A kind of tissue culture and rapid propagation method of brook anemone, including the following steps:
1) for clip brook anemone spire as explant, the sodium hypochlorite with 75% alcohol and 5% is molten on superclean bench
Surface sterilization is carried out to the explant；
2) by the explant after surface sterilization along the crosscutting square at 2~6mm in vertical middle arteries direction, the explant
The blade back of body is inoculated in downward in the induced medium of culture bottle, 10 plants of explants of every bottle of inoculation；
3) explant being seeded in culture bottle is first subjected to 3 days dark treatments, then intensity of illumination be 800~
2000Lux, 12~14h/d of light application time, temperature be 20~28 DEG C under conditions of cultivate 20~35 days be induced it is indefinite
Bud；
4) adventitious bud of the induction is inoculated into root media, inoculation number is 10 in every bottle of root media cultivation
Strain, under conditions of intensity of illumination is 800~2000Lux, 12~14h/d of light application time, temperature are 20~28 DEG C cultivate 60~
It can take root within 70 days.
The induced medium is the MS culture medium containing 1.0~6.0mg/L of 6-BA, 0.1~0.4mg/L of IAA.
The root media is the MS culture medium containing 0.1~0.3mg/L of IAA.
Intensity of illumination is 1500Lux in the step 4).
Light application time 12h/d in the step 4).
Temperature is 24 DEG C in the step 4).
Culture 60 days in the step 4).
The length taken root is the root of 9~11cm.
The every plant of brook anemone to take root in the step 4) takes root 7~9, and in the chicken feet shape of tubbiness, plant is tall and straight, every plant of tiger
Natural plant height degree is slapped in 11~13cm, blade is light green.
In order to be easy to understand the technical means, the creative features, the aims and the efficiencies achieved by the present invention, tie below
Specific embodiment is closed, the present invention is further explained.
Embodiment 1:
1) for clip brook anemone spire as explant, the sodium hypochlorite with 75% alcohol and 5% is molten on superclean bench
Surface sterilization is carried out to the explant；
2) by the explant after surface sterilization along the crosscutting square at 2mm in vertical middle arteries direction, the explant
Blade back be inoculated in the induced medium of culture bottle downward, 10 plants of explants of every bottle of inoculation；
3) explant being seeded in culture bottle is first subjected to 3 days dark treatments, is then 800Lux, illumination in intensity of illumination
Time 12h/d, temperature cultivate 20 days adventitious buds induced under conditions of being 20 DEG C；
4) adventitious bud of the induction is inoculated into root media, inoculation number is 10 in every bottle of root media cultivation
Strain, cultivating 60 days under conditions of intensity of illumination is 800Lux, light application time 12h/d, temperature are 20 DEG C can take root.
The induced medium is the MS culture medium containing 6-BA 1.0mg/L, IAA 0.1mg/L.
The root media is the MS culture medium containing IAA 0.1mg/L.
Embodiment 2:
1) for clip brook anemone spire as explant, the sodium hypochlorite with 75% alcohol and 5% is molten on superclean bench
Surface sterilization is carried out to the explant；
2) by the explant after surface sterilization along the crosscutting square at 6mm in vertical middle arteries direction, the explant
Blade back be inoculated in the induced medium of culture bottle downward, 10 plants of explants of every bottle of inoculation；
3) explant being seeded in culture bottle is first subjected to 3 days dark treatments, is then 2000Lux, light in intensity of illumination
35 days adventitious buds induced are cultivated under conditions of being 28 DEG C according to time 14h/d, temperature；
4) adventitious bud of the induction is inoculated into root media, inoculation number is 10 in every bottle of root media cultivation
Strain, cultivating 70 days under conditions of intensity of illumination is 2000Lux, light application time 14h/d, temperature are 28 DEG C can take root.
The induced medium is the MS culture medium containing 6-BA 6.0mg/L, IAA 0.4mg/L.
The root media is the MS culture medium containing IAA 0.3mg/L.
Embodiment 3:
1) for clip brook anemone spire as explant, the sodium hypochlorite with 75% alcohol and 5% is molten on superclean bench
Surface sterilization is carried out to the explant；
2) by the explant after surface sterilization along the crosscutting square at 4mm in vertical middle arteries direction, the explant
Blade back be inoculated in the induced medium of culture bottle downward, 10 plants of explants of every bottle of inoculation；
3) explant being seeded in culture bottle is first subjected to 3 days dark treatments, is then 1500ux, illumination in intensity of illumination
Time 12h/d, temperature cultivate 30 days adventitious buds induced under conditions of being 25 DEG C；
4) adventitious bud of the induction is inoculated into root media, inoculation number is 10 in every bottle of root media cultivation
Strain, cultivating 60 days under conditions of intensity of illumination is 1500Lux, light application time 12h/d, temperature are 24 DEG C can take root.
The induced medium is the MS culture medium containing 6-BA 5.0mg/L, IAA 0.3mg/L.
The root media is the MS culture medium containing IAA 0.2mg/L.
The length taken root is the root of 10cm.
The every plant of brook anemone to take root in the step 4) takes root 8, and in the chicken feet shape of tubbiness, plant is tall and straight, every plant of tiger palm
For natural plant height degree in 12cm, blade is light green.
Rooting rate has reached 100% through this embodiment.
The above shows and describes the basic principle, main features and advantages of the invention.The technology of the industry
Personnel are it should be appreciated that the present invention is not limited to the above embodiments, and the above embodiments and description only describe this
The principle of invention, without departing from the spirit and scope of the present invention, various changes and improvements may be made to the invention, these changes
Change and improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and its
Equivalent circle.
Claims (9)
1. a kind of tissue culture and rapid propagation method of brook anemone, characterized in that it comprises the following steps:
1) clip brook anemone spire is molten to institute with the sodium hypochlorite of 75% alcohol and 5% as explant on superclean bench
It states explant and carries out surface sterilization；
2) by the explant after surface sterilization along the crosscutting square at 2~6mm in vertical middle arteries direction, the explant
Blade back is inoculated in downward in the induced medium of culture bottle, 10 plants of explants of every bottle of inoculation；
3) explant being seeded in culture bottle is first subjected to 3 days dark treatments, is then 800~2000Lux, light in intensity of illumination
20~35 days adventitious buds induced are cultivated under conditions of being 20~28 DEG C according to 12~14h/d of time, temperature；
4) adventitious bud of the induction being inoculated into root media, inoculation number is 10 plants in every bottle of root media cultivation,
60~70 days are cultivated under conditions of intensity of illumination is 800~2000Lux, 12~14h/d of light application time, temperature are 20~28 DEG C i.e.
It can take root.
2. a kind of tissue culture and rapid propagation method of brook anemone as described in claim 1, it is characterised in that: the induced medium be containing
There are the MS culture medium of 1.0~6.0mg/L of 6-BA, 0.1~0.4mg/L of IAA.
3. a kind of tissue culture and rapid propagation method of brook anemone as described in claim 1, it is characterised in that: the root media is
MS culture medium containing 0.1~0.3mg/L of IAA.
4. a kind of tissue culture and rapid propagation method of brook anemone as claimed in claim 3, it is characterised in that: illumination is strong in the step 4)
Degree is 1500Lux.
5. a kind of tissue culture and rapid propagation method of brook anemone as described in claim 1, it is characterised in that: in the step 4) when illumination
Between 12h/d.
6. a kind of tissue culture and rapid propagation method of brook anemone as described in claim 1, it is characterised in that: temperature is in the step 4)
24℃。
7. a kind of tissue culture and rapid propagation method of brook anemone as described in claim 1, it is characterised in that: culture 60 in the step 4)
It.
8. a kind of tissue culture and rapid propagation method of brook anemone as described in claim 1, it is characterised in that: the length taken root is 9
The root of~11cm.
9. a kind of tissue culture and rapid propagation method of brook anemone as described in claim 1, it is characterised in that: take root in the step 4)
Every plant of brook anemone takes root 7~9, and in the chicken feet shape of tubbiness, plant is tall and straight, and in 11~13cm, blade is every plant of brook anemone height
Light green.
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| CN201910641302.XA CN110226518B (en) | 2019-07-16 | 2019-07-16 | Tissue culture and rapid propagation method of saxifrage |
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Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6264998B1 (en) * | 2000-03-01 | 2001-07-24 | Dabur Research Foundation | Extracting betulinic acid from Ziziphus jujuba |
| CN101390496A (en) * | 2008-09-28 | 2009-03-25 | 华中农业大学 | Vitro quick di-wu breeding method |
| US20090104295A1 (en) * | 2005-08-12 | 2009-04-23 | Kenji Kohno | Agent for Hair Growth |
| CN101810143A (en) * | 2010-04-29 | 2010-08-25 | 华中农业大学 | Rapid in-vitro propagation method and application of anemone flaccida fr.schmidt |
| CN104450748A (en) * | 2014-11-18 | 2015-03-25 | 西南大学 | Asperugo procumbens delta6-fatty acid desaturase ApD6D gene family as well as recombinant expression vector and application thereof |
| CN104784331A (en) * | 2015-04-30 | 2015-07-22 | 宋彬 | Traditional Chinese medicine for treating dilated cardiomyopathy and preparation method thereof |
| CN105168610A (en) * | 2015-08-26 | 2015-12-23 | 朱小芬 | Composition for cold cough and preparation method thereof |
| CN108451141A (en) * | 2018-04-22 | 2018-08-28 | 南京正宽医药科技有限公司 | A kind of facial mask with moisture-keeping functions |
| CN109608263A (en) * | 2018-12-27 | 2019-04-12 | 天津亚德尔生物质科技股份有限公司 | A kind of preparation method of charcoal selenium fertilizer |
| CN110214561A (en) * | 2019-07-22 | 2019-09-10 | 楚雄师范学院 | A kind of quick division propagation method of brook anemone |
-
2019
- 2019-07-16 CN CN201910641302.XA patent/CN110226518B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6264998B1 (en) * | 2000-03-01 | 2001-07-24 | Dabur Research Foundation | Extracting betulinic acid from Ziziphus jujuba |
| US20090104295A1 (en) * | 2005-08-12 | 2009-04-23 | Kenji Kohno | Agent for Hair Growth |
| CN101390496A (en) * | 2008-09-28 | 2009-03-25 | 华中农业大学 | Vitro quick di-wu breeding method |
| CN101810143A (en) * | 2010-04-29 | 2010-08-25 | 华中农业大学 | Rapid in-vitro propagation method and application of anemone flaccida fr.schmidt |
| CN104450748A (en) * | 2014-11-18 | 2015-03-25 | 西南大学 | Asperugo procumbens delta6-fatty acid desaturase ApD6D gene family as well as recombinant expression vector and application thereof |
| CN104784331A (en) * | 2015-04-30 | 2015-07-22 | 宋彬 | Traditional Chinese medicine for treating dilated cardiomyopathy and preparation method thereof |
| CN105168610A (en) * | 2015-08-26 | 2015-12-23 | 朱小芬 | Composition for cold cough and preparation method thereof |
| CN108451141A (en) * | 2018-04-22 | 2018-08-28 | 南京正宽医药科技有限公司 | A kind of facial mask with moisture-keeping functions |
| CN109608263A (en) * | 2018-12-27 | 2019-04-12 | 天津亚德尔生物质科技股份有限公司 | A kind of preparation method of charcoal selenium fertilizer |
| CN110214561A (en) * | 2019-07-22 | 2019-09-10 | 楚雄师范学院 | A kind of quick division propagation method of brook anemone |
Non-Patent Citations (2)
| Title |
|---|
| SEDIVA, J等: ""An efficient in vitro propagation protocol for snowdrop anemone (Anemone sylvestris L.)"", 《HORTICULTURAL SCIENCE 》 * |
| 中国植物学会系统与进化植物学专业委员会、云南省植物学会: ""中国—喜马拉雅草玉梅繁殖生物学及生态适应研究"", 《2011年全国系统与进化植物学暨第十届青年学术研讨会论文集》 * |
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