CN1653886A - Tissue culturing method for licorice adventitious root - Google Patents
Tissue culturing method for licorice adventitious root Download PDFInfo
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- CN1653886A CN1653886A CN 200510016274 CN200510016274A CN1653886A CN 1653886 A CN1653886 A CN 1653886A CN 200510016274 CN200510016274 CN 200510016274 CN 200510016274 A CN200510016274 A CN 200510016274A CN 1653886 A CN1653886 A CN 1653886A
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Abstract
The present invention discloses the process of culturing adventitious root tissue of licorice, and belongs to the tissue culture of licorice as one kind of Chinese medicinal materials. The process of the present invention includes soaking licorice seed in solution with ethanol in 70-75 vol% and sodium hypochlorite in 2 vol% to sterilize and planting in MS culture medium to induce bacteria-free bud; inoculating the cut cotyledon and bud in MS culture medium containing different plant growth regulator to induce callus; transferring the callus growing for 20 days in MS culture medium with indolebutyric acid and kinin to induce adventitious root; and finally transferring the adventitious root to liquid culture medium containing the same plant growth regulator for fast growth. The present invention has simple process, high inducing rate, fast adventitious root proliferating speed and other features.
Description
Technical field:
The invention discloses a kind of tissue culturing method for licorice adventitious root, belong to Chinese medicine Radix Glycyrrhizae field of tissue culture.
Background technology:
Radix Glycyrrhizae is a pulse family Glycyrrhiza herbaceos perennial, is used as medicine with root and rhizome.Modern pharmacological research shows that Radix Glycyrrhizae has pharmacological actions such as antiulcer, anti-inflammatory, antiallergy, kobadrin.Simultaneously, Radix Glycyrrhizae still is the western desert of China, the important sand-fixation plant in half-desert area.Licorice medicinal materials mainly relies on wild resource for many years, but in recent years because the uncontrolled of Radix Glycyrrhizae excavated, makes China's Radix Glycyrrhizae resource suffer heavy damage, the reserves of present wild Radix Glycyrrhizae only be before the founding of the state 50%.And artificial cultivation is from being seeded into results, generally needs 3～4 years and is difficult for emerging; And its content of effective is compared the very big gap of existence with wild Radix Glycyrrhizae; Residual the exceeding standard of farming also limited the development prospect of artificial cultivation Radix Glycyrrhizae simultaneously.The Radix Glycyrrhizae method for tissue culture has in unlimited time, place, season; Can provide the culture of effective component content homogeneous; Help characteristics such as large-scale industrialization production, have very big development prospect.But up to the present fewer about the report of Radix Glycyrrhizae tissue culture, and mainly concentrate on callus culture, the cultivation of adventive root does not appear in the newspapers as yet.
Summary of the invention:
The object of the present invention is to provide a kind of method of licorice adventitious root tissue culture.This method is feasible fast, and the culture of homogeneous can be provided.
The present invention is realized by following technical proposals.By the set out method of evoking liquorice adventive root of Radix Glycyrrhizae seed, it is characterized in that comprising following process: with the seed of Radix Glycyrrhizae with 70%～75% (V/V) alcohol immersion after 10～15 seconds, with aseptic water washing 2～4 times, soaked 15～20 minutes with 2% liquor natrii hypochloritis again, use aseptic water washing then 2～4 times, it is inoculated in the MS medium cultivates, when treating its bud length to 1.1～1.3cm bud and cotyledon are downcut respectively, be transferred to and contain heteroauxin (IAA) 0.1mg/L～4mg/L, 2,4-dichlorphenoxyacetic acid (2,4-D) 1mg/L～4mg/L, cultivate in the MS medium of kinetin (KT) 0mg/L～2mg/L, cultivate after 15～20 days the callus that forms is cut into small pieces, be transferred to and contain indolebutyric acid (IBA) 1mg/L～4mg/L, in the MS solid culture medium of KT 0mg/L～1mg/L, through 15～20 days cultivations was the adventive root generation of villous shape, adventive root continued growth afterwards, 20～23 days adventive root of growth is transferred in the identical liquid nutrient medium of composition cultivates, its growth rate is faster.
The present invention utilizes method for tissue culture to cultivate licorice adventitious root, have more practical significance than cultivating licorice cell, its growth rate is more faster than primitive root, and the productive rate of active secondary metabolite is high and more stable, is not subjected to the influence of natural environments such as natural disaster simultaneously again.
Embodiment:
Example one:
The seed of Ural Radix Glycyrrhizae (available from Beijing time precious Study of Medicinal Herbs institute) is rinsed well with running water, after using 75% alcohol immersion 10s then, with aseptic water washing 2～3 times, soak 15min with 2% liquor natrii hypochloritis again, behind aseptic water washing 3～4 times, seed is inoculated in the MS medium, cultivate 4d, treat that bud length is behind 1.1cm, the segment that will its root middle part be cut into 0.5cm, be transferred to additional 2, on the MS solid culture medium of 4-D 2mg/L and KT 0.7mg/L, behind the 20d callus that forms is cut into small pieces again, be transferred in the MS solid culture medium that adds IBA 2mg/L and KT 0.2mg/L and cultivate, the cultivation of process 20d is that the adventive root of villous shape generates adventive root continued growth afterwards, the adventive root of growth 20d is transferred in the identical liquid nutrient medium of composition and cultivates, and its growth rate is faster.
Example two:
The seed of Ural Radix Glycyrrhizae is rinsed well with running water, after using 72% alcohol immersion 14s then, with aseptic water washing 2～3 times, soak 20min with 2% liquor natrii hypochloritis again, behind aseptic water washing 3～4 times, seed is inoculated in the MS medium, cultivate 4d, treat that bud length is behind 1.3cm, its tip of a root is cut into the segment of 0.7cm, be transferred to additional 2, on the MS solid culture medium of 4-D 1mg/L and KT 1mg/L, behind the 18d callus that forms is cut into small pieces again, be transferred in the MS solid culture medium that adds IBA 1mg/L and KT 0.1mg/L and cultivate, the cultivation of process 17d is that the adventive root of villous shape generates adventive root continued growth afterwards, the adventive root of growth 23d is transferred in the identical liquid nutrient medium of composition and cultivates, and its growth rate is faster.
Example three:
The seed of Ural Radix Glycyrrhizae is rinsed well with running water, after using 71% alcohol immersion 11s then, with aseptic water washing 2～3 times, soak 18min with 2% liquor natrii hypochloritis again, behind aseptic water washing 3～4 times, seed is inoculated in the MS medium, cultivate 4d, after treating bud length to 1.1～1.3cm, its roots and tops is cut into the segment of 0.3cm, be transferred on the MS solid culture medium of additional IAA 3mg/L and KT 1mg/L, behind the 15d callus that forms be cut into small pieces again, be transferred in the MS solid culture medium that adds IBA 3mg/L and KT 0.5mg/L and cultivate, cultivation through 20d is the adventive root generation of villous shape, adventive root continued growth afterwards is transferred to the adventive root of growth 22d in the identical liquid nutrient medium of composition and cultivates, and its growth rate is faster.
Example four:
The seed of Ural Radix Glycyrrhizae is rinsed well with running water, after using 73% alcohol immersion 17s then, with aseptic water washing 2～3 times, soak 16min with 2% liquor natrii hypochloritis again, behind aseptic water washing 3～4 times, seed is inoculated in the MS medium, cultivate 4d, after treating bud length to 1.1～1.3cm, its cotyledon is downcut, be transferred to and add 2, on the MS solid culture medium of 4-D 2mg/L, behind the 16d callus that forms is cut into small pieces again, be transferred in the MS solid culture medium that adds IBA 2mg/L and cultivate, the cultivation of process 20d is that the adventive root of villous shape generates adventive root continued growth afterwards, the adventive root of growth 20d is transferred in the identical liquid nutrient medium of composition and cultivates, and its growth rate is faster.
Example five:
The seed of Radix Glycyrrhizae is rinsed well with running water, after using 70% alcohol immersion 15s then, with aseptic water washing 2～3 times, soak 20min with 2% liquor natrii hypochloritis again, behind aseptic water washing 3～4 times, seed is inoculated in the MS medium, cultivate 4d, after treating bud length to 1.1～1.3cm, its root middle part is cut into the segment of 0.7cm, be transferred on the MS solid culture medium of additional IAA 4mg/L and KT 1mg/L, behind the 15d callus that forms be cut into small pieces again, be transferred in the MS solid culture medium that contains IBA 4mg/L and KT 1mg/L and cultivate, through the cultivation about 16d is the adventive root generation of villous shape, adventive root continued growth afterwards is transferred to the adventive root of growth 21d in the identical liquid nutrient medium of composition and cultivates, and its growth rate is faster.
Claims (1)
1. the method for a licorice adventitious root tissue culture is characterized in that comprising following process:
The seed of Radix Glycyrrhizae after 10～15 seconds, is used aseptic water washing 2～4 times with 70%～75% alcohol immersion, soaked 15～20 minutes with 2% liquor natrii hypochloritis again, use aseptic water washing then 2～4 times, it is inoculated in the MS medium cultivates; When treating its bud length to 1.1～1.3cm bud and cotyledon are downcut respectively, be transferred in the MS medium that contains heteroauxin 0.1mg/L～4mg/L, 2,4 dichlorophenoxyacetic acid 1mg/L～4mg/L, kinetin 0mg/L～2mg/L and cultivate; Cultivate after 15～20 days the callus that forms is cut into small pieces, be transferred in the MS solid culture medium that contains indolebutyric acid 1mg/L～4mg/L, kinetin 0mg/L～1mg/L, through 15～20 days cultivations was the adventive root generation of villous shape, adventive root continued growth afterwards, 20～23 days adventive root of growth is transferred in the identical liquid nutrient medium of composition cultivates, its growth rate is faster.
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Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN103120085A (en) * | 2011-11-18 | 2013-05-29 | 陈芳 | Seedling transplantation method of ecological restoration type liquorices |
| CN104620987A (en) * | 2015-02-12 | 2015-05-20 | 天津大学 | Tissue-culturing method for improving content of flavone compounds in adventitious roots of glycyrrhiza uralensis |
| CN106614000A (en) * | 2017-03-22 | 2017-05-10 | 陈志育 | Screening method of glycyrrhiza uralensis rooting medium containing aspirin |
| CN106614001A (en) * | 2017-03-22 | 2017-05-10 | 陈志育 | Screening method of light source conditions for induction culture of liquorice cotyledon |
| CN106665369A (en) * | 2017-03-22 | 2017-05-17 | 陈志育 | High-inductivity induction culture medium for cotyledons of radix glycyrrhizae |
| CN106665356A (en) * | 2016-12-27 | 2017-05-17 | 天津大学 | Tissue culture method for promoting accumulation of glycyrrhizic acid in liquorice adventitious roots |
| CN106718948A (en) * | 2017-03-22 | 2017-05-31 | 陈志育 | Improve the screening technique of the aseptic seeding method of Glycyrrhiza Seeds germination rate |
| CN106797891A (en) * | 2017-03-22 | 2017-06-06 | 陈志育 | A kind of Radix Glycyrrhizae method for tissue culture for promoting root |
| CN106888975A (en) * | 2017-03-22 | 2017-06-27 | 陈志育 | A kind of Radix Glycyrrhizae cotyledon induced tissue cultural method |
| CN106888973A (en) * | 2017-03-22 | 2017-06-27 | 陈志育 | A kind of Radix Glycyrrhizae root media containing aspirin |
| CN106888972A (en) * | 2017-03-22 | 2017-06-27 | 陈志育 | A kind of Radix Glycyrrhizae aseptic seeding culture medium of germination rate high |
| CN106900556A (en) * | 2017-03-22 | 2017-06-30 | 陈志育 | A kind of Radix Glycyrrhizae aseptic seeding method of germination rate high |
| CN106912382A (en) * | 2017-03-22 | 2017-07-04 | 陈志育 | A kind of screening technique of the condition of tissue culture for promoting Radix Glycyrrhizae to go out root |
| CN106912381A (en) * | 2017-03-22 | 2017-07-04 | 陈志育 | A kind of germination rate Radix Glycyrrhizae aseptic seeding Screening of Media method high |
| CN106942056A (en) * | 2017-03-22 | 2017-07-14 | 陈志育 | A kind of Radix Glycyrrhizae cotyledon inducing culture screening technique |
-
2005
- 2005-03-08 CN CNB2005100162740A patent/CN1319438C/en not_active Expired - Fee Related
Cited By (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN103120085A (en) * | 2011-11-18 | 2013-05-29 | 陈芳 | Seedling transplantation method of ecological restoration type liquorices |
| CN103120085B (en) * | 2011-11-18 | 2015-05-06 | 陈芳 | Seedling transplantation method of ecological restoration type liquorices |
| CN104620987A (en) * | 2015-02-12 | 2015-05-20 | 天津大学 | Tissue-culturing method for improving content of flavone compounds in adventitious roots of glycyrrhiza uralensis |
| CN106665356A (en) * | 2016-12-27 | 2017-05-17 | 天津大学 | Tissue culture method for promoting accumulation of glycyrrhizic acid in liquorice adventitious roots |
| CN106614001A (en) * | 2017-03-22 | 2017-05-10 | 陈志育 | Screening method of light source conditions for induction culture of liquorice cotyledon |
| CN106665369A (en) * | 2017-03-22 | 2017-05-17 | 陈志育 | High-inductivity induction culture medium for cotyledons of radix glycyrrhizae |
| CN106614000A (en) * | 2017-03-22 | 2017-05-10 | 陈志育 | Screening method of glycyrrhiza uralensis rooting medium containing aspirin |
| CN106718948A (en) * | 2017-03-22 | 2017-05-31 | 陈志育 | Improve the screening technique of the aseptic seeding method of Glycyrrhiza Seeds germination rate |
| CN106797891A (en) * | 2017-03-22 | 2017-06-06 | 陈志育 | A kind of Radix Glycyrrhizae method for tissue culture for promoting root |
| CN106888975A (en) * | 2017-03-22 | 2017-06-27 | 陈志育 | A kind of Radix Glycyrrhizae cotyledon induced tissue cultural method |
| CN106888973A (en) * | 2017-03-22 | 2017-06-27 | 陈志育 | A kind of Radix Glycyrrhizae root media containing aspirin |
| CN106888972A (en) * | 2017-03-22 | 2017-06-27 | 陈志育 | A kind of Radix Glycyrrhizae aseptic seeding culture medium of germination rate high |
| CN106900556A (en) * | 2017-03-22 | 2017-06-30 | 陈志育 | A kind of Radix Glycyrrhizae aseptic seeding method of germination rate high |
| CN106912382A (en) * | 2017-03-22 | 2017-07-04 | 陈志育 | A kind of screening technique of the condition of tissue culture for promoting Radix Glycyrrhizae to go out root |
| CN106912381A (en) * | 2017-03-22 | 2017-07-04 | 陈志育 | A kind of germination rate Radix Glycyrrhizae aseptic seeding Screening of Media method high |
| CN106942056A (en) * | 2017-03-22 | 2017-07-14 | 陈志育 | A kind of Radix Glycyrrhizae cotyledon inducing culture screening technique |
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