CN101265462A - Industrial culture method for liquorice cell - Google Patents
Industrial culture method for liquorice cell Download PDFInfo
- Publication number
- CN101265462A CN101265462A CNA2008100530144A CN200810053014A CN101265462A CN 101265462 A CN101265462 A CN 101265462A CN A2008100530144 A CNA2008100530144 A CN A2008100530144A CN 200810053014 A CN200810053014 A CN 200810053014A CN 101265462 A CN101265462 A CN 101265462A
- Authority
- CN
- China
- Prior art keywords
- cell
- culture
- culture medium
- suspending liquid
- liquorice
- 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
Abstract
The invention discloses a method for culturing industrial liquorice cells, and belongs to the liquorice cell culture technology. The method comprises the steps of inoculating liquorice seeds in an MS culture medium after sterilization for inducing aseptic sprouts of the liquorice; transferring cut liquorice sprout segments to a callus culture medium for solid culture so as to generate a liquorice callus; transferring the callus to a cell culture medium for suspension culture and then subculture so as to obtain liquorice cell sap with stable cell output; and transferring the liquorice cell sap to a stirring paddle type reactor for scale suspension culture of the liquorice cells to obtain liquorice cell suspension with cell output of 15-40g/l by cell dry weight. The method is characterized by simple process, quick cell growth, stable output, etc.
Description
Technical field
The present invention relates to a kind of suspension culture method of licorice cell, belong to the licorice cell culture technique.
Background technology
Radix Glycyrrhizae (Glycyrrhiza uralensis Fisch.) is the pulse family per nnial herb, its root and rhizome have invigorate the spleen and benefit qi, effect such as cough-relieving apophlegmatic, clearing heat and detoxicating, coordinating the actions of various ingredients in a prescription, be one of the most frequently used Chinese medicine, saying of " ten sides, nine grass " always just arranged.Modern pharmacological research shows that Radix Glycyrrhizae has pharmacologically active widely, as detoxifcation, anti-oxidant, anti-inflammatory etc.Main activeconstituents is flavonoid and triterpenes in the Radix Glycyrrhizae.Its main triterpenes components Potenlini has multiple pharmacologically actives such as anti-inflammatory, antiulcer agent, antiviral and immunomodulatory, and other has the report Potenlini to have the effect of anti-SARS virus, and immunizing power that can enhanced SAR S the infected.
Radix Glycyrrhizae not only has very high pharmaceutical use, and be the western arid of China, one of important vegetation resources of semiarid zone has important effect for checking winds and fixing drifting sand, improve Soil structure, and the over-ground part that studies show that Radix Glycyrrhizae in addition is a kind of medium herbage.Market is very big to the Radix Glycyrrhizae demand, and domestic annual requirement is about 30,000 tons, and annual export volume is about 50,000 tons, and is growing trend.In the market, 80% Radix Glycyrrhizae is wild, excavates a large amount of degenerations that Radix Glycyrrhizae has been caused the grassland, the Northwest, makes wild Radix Glycyrrhizae resource be on the verge of exhaustion, and has caused serious Desertification phenomenon.Radix Glycyrrhizae has become the second class protection plant of country at present.Radix Glycyrrhizae has become specially control medicinal material of country, and wild Radix Glycyrrhizae is under an embargo and is used in the middle of the healthcare products.And the content of effective constituent such as Potenlini is stable inadequately in the tame Radix Glycyrrhizae, and the artificial culture growth cycle is long, need take a large amount of land resourcess,, alleviate the Radix Glycyrrhizae resource problem to a certain extent so expectation is produced useful secondary metabolite with the method for plant tissue culture.
Cell suspension culture has plurality of advantages, and is controlled such as growth conditions; Save the soil, the production system standardization; The effective constituent synthetic route is carried out genetic manipulation, improve secondary metabolite content etc., had the cell suspension culture of multiple medicinal plant to realize industrialization at present, as Asian puccoon, genseng etc.As seen utilize the medicinal plant cell suspension culture to produce secondary metabolite and have feasibility.
The report of Radix Glycyrrhizae tissue culture aspect mainly concentrates in the research of callus culture and differentiation and hairly root cultivation aspect at present, and less to the research of licorice cell cultivation; Simultaneously, systematic Study and the industrialization cultivation to the licorice cell suspension culture do not appear in the newspapers.The present invention has optimized the condition of licorice cell suspension culture, has obtained growth rapidly, the licorice cell of stable yield, and carried out amplification culture with the reactor of 5L, obtained success, the cell growth is rapid, the stable content of main components such as Potenlini, liquirtin, licorice polysaccharide.
Summary of the invention
The object of the present invention is to provide a kind of cultural method of licorice cell industrialization, this method can realize the suitability for industrialized production of licorice cell.
The present invention is realized by the following technical programs: a kind of cultural method of licorice cell industrialization is characterized in that comprising following process:
1. the seed mass concentration with Radix Glycyrrhizae is that 70%～75% alcohol immersion is after 10～15 seconds, deionized water rinsing through sterilizing 2～4 times, be that 2% chlorine bleach liquor soaked 15～20 minutes with mass concentration again, deionized water rinsing through sterilizing is 2～4 times then, and it is inoculated in Murashige﹠amp; Solid culture in Skoog (MS) substratum, in culture temperature is 23 ℃～28 ℃, and intensity of illumination is 800～2000lux, and light application time is that seed bore bud after the culture condition of 12h/d was cultivated 3～5d down, bud is cut into segment, is transferred in the callus culture base and cultivates.Described callus culture base is by being the MS substratum and pressing 2 of every liter of MS substratum adding 1mg～4mg, 4-dichlorphenoxyacetic acid (2,4-D), add 0mg～2mg kinetin (KT), add 30～60g sucrose and add 8～10g agar and form, with concentration is that the NaOH of 1mol/L or mass concentration are that the pH that 10% HCl regulates cell culture medium is 4.5～7.5, be that 23 ℃～28 ℃ following lucifuges are cultivated in temperature afterwards, after cultivating 20～25d, the Radix Glycyrrhizae callus is covered with culture dish.
2. in 1: 10～20 ratio of the ml volumes amount of every gram callus and cell culture medium, the callus fragment changed over to carry out fluid suspension culture in the substratum.Described cell culture medium adds 2 of 1mg～4mg by the MS substratum and by every liter of substratum, 4-dichlorphenoxyacetic acid (2,4-D), add the kinetin (KT) of 0mg～2mg and 30～60g sucrose of adding is formed, be that the NaOH of 1mol/L or mass concentration are that the pH of 10% HCl adjusting cell culture medium is 4.5～7.5 with concentration.In temperature is 23 ℃～28 ℃, and intensity of illumination is 800～2000lux, and light application time is 12h/d, and shaking speed is 90～120rmin
-1The condition low suspension cultivate 4～7d after, suspension is filtered, the cell mass that filtering is bigger reaches not the tissue of callusization fully, in the dispersiveness that obtains preferably in the cell suspending liquid, according to the ratio of cell suspending liquid volume and the volume of the cell culture medium of adding is that 1: 2～5 ratio adds cell culture medium, behind succeeding transfer culture 20～25d, in the subculture cell suspending liquid, according to the volume ratio of subculture cell suspending liquid volume and the cell culture medium of adding is that 1: 2～5 ratio adds the cell suspending liquid that cell culture medium obtains twice of subculture, carry out succeeding transfer culture 5 times with this subculture method, obtain cell yield is counted 3～10g/L by dry cell weight cell suspending liquid.
3. getting every liter of cell suspending liquid 1～1.5L that contains by dry weight basis 3～10g cell that step 2 obtains and transfer in the 5L stirring rake formula reactor, is 1: 1～3 to add cell culture medium in reactor according to the ratio of the volume of the volume of cell suspending liquid and cell culture medium.In temperature is 23 ℃～28 ℃, intensity of illumination is 800～2000lux, and light application time is 12h/d, and stir speed (S.S.) is 80～130r/min, air flow is to gather in the crops licorice cell after cultivating 20～30d under the culture condition of 1.5～4L/min, and cell yield is counted 15～40g/L by dry cell weight.
Advantage of the present invention: present method has realized the large-scale production of licorice cell; in callus culture, use 2,4-D and KT are as plant-growth regulator; make Radix Glycyrrhizae bud callusization rapid, callus growth is very fast, in good condition in callus succeeding transfer culture process.In cell cultivation process, utilize 2,4-D makes the cell growth rapidly with the excellent fit ratio of KT, in large-scale production, the present invention utilizes stirring reactor to cultivate first, reaches culture effect preferably.Whole culturing process is convenient and easy, and rapid, the stable yield of cell growth is for industrial mass production provides effective way.
Embodiment
Example one:
With the seed of Radix Glycyrrhizae (available from Beijing time precious Study of Medicinal Herbs institute) 3g, with mass concentration is that 70% alcohol immersion is after 10～15 seconds, deionized water rinsing through sterilizing 2 times, be that 2% chlorine bleach liquor soaked 15 minutes with mass concentration again, deionized water rinsing through sterilizing is 2 times then, it is inoculated in solid culture in the 20mL MS substratum, culture condition is: 23 ℃ of temperature, intensity of illumination are that 800lux, light application time are 12h/d, cultivate about 3d seed and bear bud, bud is cut into the segment of 3～4mm.Get 25mL MS substratum to wherein adding 2 of 0.025mg, 4-D adds the KT of 0.05mg, adds 0.75g sucrose, adds 0.25g agar, is made into the callus culture base, is that 10% HCl adjusting pH is 4.5 with mass concentration.Place above-mentioned cultivation to cultivate the budlet segment.Culture condition is: behind 23 ℃ of temperature, lucifuge cultivation, the cultivation 20d, be covered with the Radix Glycyrrhizae callus on culture dish.
By fresh weight the 5g callus is changed in the 100mL cell culture medium, this cell culture medium is the MS substratum by 100mL, and adds 0.1mg 2,4-D, 0.2mg KT adds 3g sucrose and forms, and is that the pH that 10% HCl regulates this cell culture medium is 4.5 with mass concentration again.Culture condition is: temperature is 23 ℃, shaking speed 90rmin
-1, intensity of illumination is 800lux, light application time is 12hd
-1Behind the suspension culture 7d, suspension is filtered, the cell mass that filtering is bigger reaches not fully, and the tissue of callusization obtains dispersiveness cell suspending liquid 100mL preferably, cell suspending liquid is divided equally equal portions, every part of 20mL, add the 40mL cell culture medium to every part of suspension in also again, at culture condition be: temperature is 23 ℃, shaking speed 90rmin
-1, intensity of illumination is 800lux, light application time is 12hd
-1Culture condition under carry out the succeeding transfer culture first time.After continuing to cultivate 20d, subculture suspension is once divided equally equal portions, every part of 20mL adds the 40mL cell culture medium and carries out the succeeding transfer culture second time.Carry out succeeding transfer culture 5 times with this way, obtain cell yield is counted 3g/L by dry cell weight cell suspending liquid.
The cell suspending liquid 1.5L that the cell yield that obtains is counted 3g/L by dry cell weight transfers in the 5L stirring rake formula reactor, adds cell culture medium 1.5L.This cell culture medium is by 1.5LMS minimum medium and the 1.5mg 2 that added, 4-D, and 3mg KT, 45g sucrose is formed, and is that the pH that 10% HCl regulates this cell culture medium is 4.5 with mass concentration.In culture temperature is 23 ℃, and intensity of illumination is 800lux, and light application time is 12h/d, and stir speed (S.S.) is 80r/min, and bubbling air, air flow are harvested cell behind the following 20d of cultivation of the culture condition of 1.5L/min.
Example two:
Seed 4g with Radix Glycyrrhizae, with mass concentration is that 70% alcohol immersion is after 10～15 seconds, deionized water rinsing through sterilizing 2 times, be that 2% chlorine bleach liquor soaked 16 minutes with mass concentration again, deionized water rinsing through sterilizing is 2 times then, and it is inoculated in solid culture in the 20mL MS substratum, and culture condition is: 25 ℃ of temperature, intensity of illumination are that 1000lux, light application time are 12h/d, cultivate about 3d seed and bear bud, bud is cut into the segment of 3～4mm.Get 25mL MS substratum to wherein adding 2 of 0.05mg, 4-D adds the KT of 0.0125mg, adds 0.75g sucrose, adds 0.25g agar, is made into the callus culture base, is that the NaOH adjusting pH of 1mol/L is 5.8 with concentration.Place above-mentioned cultivation to cultivate the budlet segment.Culture condition is: behind 25 ℃ of temperature, lucifuge cultivation, the cultivation 20d, be covered with the Radix Glycyrrhizae callus on culture dish.
By fresh weight the 10g callus is changed in the 100mL cell culture medium, this cell culture medium is the MS substratum by 100mL, and adds 0.2mg 2,4-D, 0.05mg KT adds 3g sucrose and forms, concentration is that the pH that the NaOH of 1mol/L regulates this cell culture medium is 5.8 again.Culture condition is: temperature is 25 ℃, shaking speed 120rmin
-1, intensity of illumination is 1000lux, light application time is 12hd
-1Behind the suspension culture 7d, suspension is filtered, the cell mass that filtering is bigger reaches not fully, and the tissue of callusization obtains dispersiveness cell suspending liquid 100mL preferably, cell suspending liquid is divided equally equal portions, every part of 20mL, add the 60mL cell culture medium to every part of suspension in also again, at culture condition be: temperature is 25 ℃, shaking speed 120rmin
-1, intensity of illumination is 1000lux, light application time is 12hd
-1Culture condition under carry out the succeeding transfer culture first time.After continuing to cultivate 20d, subculture suspension is once divided equally equal portions, every part of 20mL adds the 60mL cell culture medium and carries out the succeeding transfer culture second time.Carry out succeeding transfer culture 5 times with this way, obtain cell yield is counted 10g/L by dry cell weight cell suspending liquid.
The cell suspending liquid 1L that the cell yield that obtains is counted 10g/L by dry cell weight transfers in the 5L stirring rake formula reactor, adds cell culture medium 3L.This cell culture medium is by 3L MS minimum medium and the 6mg 2 that added, 4-D, and 1.5mg KT, 90g sucrose is formed, and is that the pH that the NaOH of 1mol/L regulates this cell culture medium is 5.8 with concentration.In culture temperature is 25 ℃, and intensity of illumination is 1000lux, and light application time is 90r/min for the 12h/d stir speed (S.S.), and bubbling air, air flow are harvested cell behind the following 25d of cultivation of the culture condition of 2L/min.
Example three:
Seed 5g with Radix Glycyrrhizae, with mass concentration is that 75% alcohol immersion is after 10～15 seconds, deionized water rinsing through sterilizing 2 times, be that 2% chlorine bleach liquor soaked 20 minutes with mass concentration again, deionized water rinsing through sterilizing is 2 times then, and it is inoculated in solid culture in the 20mL MS substratum, and culture condition is: 26 ℃ of temperature, intensity of illumination are that 1500lux, light application time are 12h/d, cultivate about 3d seed and bear bud, bud is cut into the segment of 3～4mm.Get 25mL MS substratum to wherein adding 2 of 0.1mg, 4-D adds the KT of 0.0025mg, adds 0.75g sucrose, adds 0.25g agar, is made into the callus culture base, is that the NaOH adjusting pH of 1mol/L is 6.5 with concentration.Place above-mentioned cultivation to cultivate the budlet segment.Culture condition is: behind 25 ℃ of temperature, lucifuge cultivation, the cultivation 20d, be covered with the Radix Glycyrrhizae callus on culture dish.
By fresh weight the 6g callus is changed in the 60mL cell culture medium, this cell culture medium is the MS substratum by 60mL, and adds 0.24mg 2,4-D, 0.006mg KT adds 1.8g sucrose and forms, concentration is that the pH that the NaOH of 1mol/L regulates this cell culture medium is 6.5 again.Culture condition is: temperature is 26 ℃, shaking speed 100rmin
-1, intensity of illumination is 1500lux, light application time is 12hd
-1Behind the suspension culture 7d, suspension is filtered, the cell mass that filtering is bigger reaches not fully, and the tissue of callusization obtains dispersiveness cell suspending liquid 60mL preferably, cell suspending liquid is divided equally equal portions, every part of 20mL, add the 60mL cell culture medium to every part of suspension in also again, at culture condition be: temperature is 26 ℃, shaking speed 100rmin
-1, intensity of illumination is 1500lux, light application time is 12hd
-1Culture condition under carry out the succeeding transfer culture first time.After continuing to cultivate 20d, subculture suspension is once divided equally equal portions, every part of 20mL adds the 60mL cell culture medium and carries out the succeeding transfer culture second time.Carry out succeeding transfer culture 5 times with this way, obtain cell yield is counted 7g/L by dry cell weight cell suspending liquid.
The cell suspending liquid 1L that the cell yield that obtains is counted 7g/L by dry cell weight transfers in the 5L stirring rake formula reactor, adds cell culture medium 2L.This cell culture medium is by 2L MS minimum medium and the 8mg 2 that added, 4-D, and 0.2mg KT, 60g sucrose is formed, and is that the pH that the NaOH of 1mol/L regulates this cell culture medium is 5.8 with concentration.In culture temperature is 25 ℃, and intensity of illumination is 1500lux, and light application time is 12h/d, and stir speed (S.S.) is 100r/min, and bubbling air, air flow are harvested cell behind the following 30d of cultivation of the culture condition of 2.5L/min.
Example four:
Seed 4g with Radix Glycyrrhizae, with mass concentration is that 75% alcohol immersion is after 10～15 seconds, deionized water rinsing through sterilizing 2 times, be that 2% chlorine bleach liquor soaked 15 minutes with mass concentration again, deionized water rinsing through sterilizing is 2 times then, and it is inoculated in solid culture in the 20mL MS substratum, and culture condition is: 28 ℃ of temperature, intensity of illumination are that 2000lux, light application time are 12h/d, cultivate about 3d seed and bear bud, bud is cut into the segment of 3～4mm.Get 25mL MS substratum to wherein adding 2 of 0.075mg, 4-D adds the KT of 0mg, adds 0.75g sucrose, adds 0.25g agar, is made into the callus culture base, is that the NaOH adjusting pH of 1mol/L is 7.5 with concentration.Place above-mentioned cultivation to cultivate the budlet segment.Culture condition is: behind 28 ℃ of temperature, lucifuge cultivation, the cultivation 20d, be covered with the Radix Glycyrrhizae callus on culture dish.
By fresh weight the 6g callus is changed in the 80mL cell culture medium, this cell culture medium is the MS substratum by 60mL, and adds 0.24mg 2,4-D, 0mg KT adds 2.4g sucrose and forms, and concentration is that the pH that the NaOH of 1mol/L regulates this cell culture medium is 7.5 again.Culture condition is: temperature is 28 ℃, shaking speed 110rmin
-1, intensity of illumination is 2000lux, light application time is 12hd
-1Behind the suspension culture 7d, suspension is filtered, the cell mass that filtering is bigger reaches not fully, and the tissue of callusization obtains dispersiveness cell suspending liquid 80mL preferably, cell suspending liquid is divided equally equal portions, every part of 20mL, add the 80mL cell culture medium to every part of suspension in also again, at culture condition be: temperature is 28 ℃, shaking speed 110rmin
-1, intensity of illumination is 2000lux, light application time is 12hd
-1Culture condition under carry out the succeeding transfer culture first time.After continuing to cultivate 20d, subculture suspension is once divided equally equal portions, every part of 20mL adds the 80mL cell culture medium and carries out the succeeding transfer culture second time.Carry out succeeding transfer culture 5 times with this way, obtain cell yield is counted 9g/L by dry cell weight cell suspending liquid.
The cell suspending liquid 1L that the cell yield that obtains is counted 9g/L by dry cell weight transfers in the 5L stirring rake formula reactor, adds cell culture medium 2L.This cell culture medium is by 2L MS minimum medium and the 6mg 2 that added, 4-D, and 0mg KT, 60g sucrose is formed, and is that the pH that the NaOH of 1mol/L regulates this cell culture medium is 7.5 with concentration.In culture temperature is 28 ℃, and intensity of illumination is 2000lux, and light application time is 12h/d, and stir speed (S.S.) is 130r/min, and bubbling air, air flow are harvested cell behind the following 30d of cultivation of the culture condition of 4L/min.
Claims (1)
1. the cultural method of a licorice cell industrialization is characterized in that comprising following process:
1). with the seed mass concentration of Radix Glycyrrhizae is that 70%～75% alcohol immersion is after 10～15 seconds, deionized water rinsing through sterilizing 2～4 times, be that 2% chlorine bleach liquor soaked 15～20 minutes with mass concentration again, deionized water rinsing through sterilizing is 2～4 times then, and it is inoculated in Murashige﹠amp; Solid culture in the Skoog substratum, in culture temperature is that 23 ℃～28 ℃ intensities of illumination are 800～2000lux, light application time is that seed bore bud after the culture condition of 12h/d was cultivated 3～5d down, bud is cut into segment, be transferred in the callus culture base and cultivate, described callus culture base is by being the MS substratum and pressing 2 of every liter of MS substratum adding 1～4mg, the 4-dichlorphenoxyacetic acid, add 0～2mg kinetin, add 30～60g sucrose and add 8～10g agar and form, with concentration is that the NaOH of 1mol/L or mass concentration are that the pH that 10% HCl regulates cell culture medium is 4.5～7.5, be that 23 ℃～28 ℃ following lucifuges are cultivated in temperature afterwards, after cultivating 20～25d, the Radix Glycyrrhizae callus is covered with culture dish;
2). in the ratio of the ml volumes amount 1: 10～20 of every gram callus and cell culture medium, the callus fragment changed over to carry out fluid suspension culture in the substratum, described cell culture medium adds 2 of 1～4mg by the MS substratum and by every liter of substratum, the 4-dichlorphenoxyacetic acid, adding the kinetin of 0～2mg and 30～60g sucrose of adding forms, with concentration is that the NaOH of 1mol/L or mass concentration are that the pH that 10% HCl regulates cell culture medium is 4.5～7.5, in temperature is 23 ℃～28 ℃, intensity of illumination is 800～2000lux, light application time is 12h/d, and shaking speed is 90～120rmin
-1The condition low suspension cultivate 4～7d after, suspension is filtered, the cell mass that filtering is bigger reaches not the tissue of callusization fully, in the dispersiveness that obtains preferably in the cell suspending liquid, according to the ratio of cell suspending liquid volume and the volume of the cell culture medium of adding is that 1: 2～5 ratio adds cell culture medium, behind succeeding transfer culture 20～25d, in the subculture cell suspending liquid, according to the volume ratio of subculture cell suspending liquid volume and the cell culture medium of adding is that 1: 2～5 ratio adds the cell suspending liquid that cell culture medium obtains twice of subculture, carry out subculture 5 times with this subculture method, obtain cell yield is counted 3～10g/L by dry cell weight cell suspending liquid;
3). get step 2) every liter of cell suspending liquid 1～1.5L that contains by dry weight basis 3～10g cell obtaining transfers in the 5L stirring rake formula reactor, according to the ratio of the volume of the volume of cell suspending liquid and cell culture medium is 1: 1～3 to add cell culture medium in reactor, in temperature is 23 ℃～28 ℃, intensity of illumination is 800～2000lux, light application time is 12h/d, stir speed (S.S.) is 80～130r/min, bubbling air, air flow is to cultivate results licorice cell suspension behind 20～30d under the culture condition of 1.5～4L/min, and cell yield is counted 15～40g/L by dry cell weight in the cell suspending liquid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100530144A CN101265462A (en) | 2008-05-07 | 2008-05-07 | Industrial culture method for liquorice cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100530144A CN101265462A (en) | 2008-05-07 | 2008-05-07 | Industrial culture method for liquorice cell |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101265462A true CN101265462A (en) | 2008-09-17 |
Family
ID=39988154
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008100530144A Pending CN101265462A (en) | 2008-05-07 | 2008-05-07 | Industrial culture method for liquorice cell |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101265462A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102532337A (en) * | 2011-12-29 | 2012-07-04 | 天津大学 | Method for producing glycyrrhizia polysaccharide by utilizing tissue culture method |
| CN103789253A (en) * | 2014-03-06 | 2014-05-14 | 高宁 | Wintergreen cell suspension culture method |
| CN104521760A (en) * | 2015-01-14 | 2015-04-22 | 甘肃农业大学 | Method for doubling glycyrrhiza pallidiflora maxim chromosomes through tissue culture |
| CN106367378A (en) * | 2016-08-26 | 2017-02-01 | 珀莱雅化妆品股份有限公司 | Glycyrrhiza glabra callus cell culture method capable of improving content of licoflavone |
| CN111718888A (en) * | 2020-06-23 | 2020-09-29 | 中国医学科学院药用植物研究所 | Culture method for improving glycyrrhizic acid content in suspension culture cells of liquorice |
-
2008
- 2008-05-07 CN CNA2008100530144A patent/CN101265462A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102532337A (en) * | 2011-12-29 | 2012-07-04 | 天津大学 | Method for producing glycyrrhizia polysaccharide by utilizing tissue culture method |
| CN103789253A (en) * | 2014-03-06 | 2014-05-14 | 高宁 | Wintergreen cell suspension culture method |
| CN104521760A (en) * | 2015-01-14 | 2015-04-22 | 甘肃农业大学 | Method for doubling glycyrrhiza pallidiflora maxim chromosomes through tissue culture |
| CN106367378A (en) * | 2016-08-26 | 2017-02-01 | 珀莱雅化妆品股份有限公司 | Glycyrrhiza glabra callus cell culture method capable of improving content of licoflavone |
| CN106367378B (en) * | 2016-08-26 | 2019-05-21 | 珀莱雅化妆品股份有限公司 | The glycyrrhiza glabra callus cell cultural method of licoflavone content can be improved |
| CN111718888A (en) * | 2020-06-23 | 2020-09-29 | 中国医学科学院药用植物研究所 | Culture method for improving glycyrrhizic acid content in suspension culture cells of liquorice |
| CN111718888B (en) * | 2020-06-23 | 2022-08-19 | 中国医学科学院药用植物研究所 | Culture method for improving glycyrrhizic acid content in suspension culture cells of liquorice |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104335903B (en) | It is a kind of to promote Pseudobulbus Bletillae (Rhizoma Bletillae) rapid propagation method | |
| CN101180949B (en) | Stem tip tissue rapid cultivating and seedling method of algam dendrobium nobile | |
| CN104082138B (en) | A kind of tissue cultivation rapid breeding method of Radix Aristolochiae Tagalae | |
| CN104920214A (en) | Method for improving yield of saponin in panax notoginseng tissue culture seedlings | |
| CN102860262A (en) | Propagation method of cyrtomium fortunei | |
| Zhou et al. | Effects of temperature, light, nutrients and smoke-water on seed germination and seedling growth of Astragalus membranaceus, Panax notoginseng and Magnolia officinalis—Highly traded Chinese medicinal plants | |
| CN101265462A (en) | Industrial culture method for liquorice cell | |
| CN1328951C (en) | Dendrobe protocorm hormoneless cultivation method | |
| CN102771397B (en) | Method for establishing adventitious root cultivation system of Psammosilene tuniceoides W. C. Wu et C. Y. Wu and expanding cultivation method of Psammosilene tuniceoides W. C. Wu et C. Y. Wu | |
| CN107027611A (en) | It is a kind of to improve the roxburgh anoectochilus terminal bud tissue-cultured seedling speed of growth and the method for active component content | |
| CN105409771A (en) | Rapid propagation method for Gymnadenia conopsea (L.) R. Br. | |
| CN104488723A (en) | Tissue-culture and rapid-propagation method of epimedium koreanum nakai | |
| CN104604678B (en) | A kind of wild panax japonicus sugar-free open tissue is cultivated fast seedling-cultivating method | |
| CN110604049B (en) | Wild-returning ecological planting method for dendrobium officinale | |
| CN102599065A (en) | Quick propagation method for humulus scandens | |
| CN105123174A (en) | Cultivation method for baphicanthus cusia bremek | |
| CN103430852A (en) | Method for controlling increasing of content of polysaccharide in dendrobium officinale suspension cell | |
| CN103992952B (en) | One strain improves Radix Salviae Miltiorrhizae yield and the Cladosporium fungus of total phenolics content | |
| CN101475929B (en) | Method for producing oleanolic acid by white birch suspension culture | |
| CN100366144C (en) | Method for producing podophyllotoxin induced from sino podophyllum hexandrum callus | |
| CN102511716A (en) | Glycyrrhiza polysaccharide granules and preparation method thereof | |
| CN102499044A (en) | Cultivation method capable of increasing biomass of selfheal clusters and yield of active ingredients | |
| CN105432470A (en) | Tissue culture method for dendrobium moniliforme | |
| CN103444547A (en) | Culture method of aralis suspension cells | |
| CN103992961B (en) | One strain improves Radix Salviae Miltiorrhizae yield and the paecilomyces fungus of total phenolics content |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20080917 |