CN1565171A - Notoginseng adventitious root tissue culture and its production method - Google Patents
Notoginseng adventitious root tissue culture and its production method Download PDFInfo
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- CN1565171A CN1565171A CN 03130265 CN03130265A CN1565171A CN 1565171 A CN1565171 A CN 1565171A CN 03130265 CN03130265 CN 03130265 CN 03130265 A CN03130265 A CN 03130265A CN 1565171 A CN1565171 A CN 1565171A
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Abstract
The invention discloses a notoginseng adventitious root tissue culture and its production method wherein the method comprises, notoginseng callus inducement, notoginseng callus expansion and successive transfer culturing, notoginseng adventitious root inducement, notoginseng adventitious root expansion and successive transfer culturing, screen selecting and active component determination and expansion for different root systems of notoginseng adventitious roots, and reactor culturing of notoginseng adventitious root.
Description
Technical field
The present invention relates to the adventive root tissue culture and the production method of a plant species, particularly relate to a kind of tissue culture and production method of pseudo-ginseng adventive root.
Background technology
Pseudo-ginseng is one of the most frequently used traditional Chinese medicine of China, also is simultaneously the situation of selling well autonomic drug on the international market.It is directly edible that it can be used as invigorant, the still important source material of many health products and medicine,, Yunnan Baiyao logical as thromboembolism and Danshen Root dropping ball etc.Because this medicinal material is subjected to the restriction of geographical conditions,, mainly is distributed in the area of Yunnan Province of China province Wenshan Prefecture one band narrow range, but also is subjected to " influence that the soil of cultivation can not reuse " so its area under cultivation is limited.Because " old seven ground " problem can not solve, so the soil of cultivation pseudo-ginseng can only lean on continuous felling mountain forest, to occupy the farmland and expand, this will cause the local a large amount of water and soil loss and the heavy damage of bio-diversity.At present, the soil that China is suitable for cultivating pseudo-ginseng reduces year by year, and along with the continuous expansion of Chinese pharmaceutical enterprises, the demand of people's lives health care medication increases year by year, and the field production of pseudo-ginseng more and more is difficult to satisfy the needs in market.In addition, the cultivation pseudo-ginseng needs the time in 3 years, all will use a large amount of agricultural chemicals, chemical fertilizer every year during this period, but also to drop into lot of manpower and material resources, will cause agricultural chemicals and heavy-metal residual and cost than problems such as height like this, thereby this just makes the supply of pseudo-ginseng still all can not satisfy the growing market demand qualitatively in quantity, so the pollution-free and pseudo-ginseng that active constituent content is high of a large amount of breedings just becomes the task of top priority of natural resources of Chinese medicinal materials development and utilization in the short time.The medicinal plant tissue culture of industrialized scale can address these problems.Tissue culture is organ or cell and protoplast of a cultured in vitro plant on synthetic medium and the technology that makes its growth, propagation, differentiation and regeneration plant.Wherein the adventive root cultivation is a new technology that grows up the eighties.Adventive root is the root that the callus induction by plant comes out, and compares the stable content of its secondary metabolites with the suspension cell of medicinal plant; Compare with the plant roots of grown in field, its growth rate is wanted fast thousands of hundred times.Therefore, the tissue culture that relatively is fit to medicinal plant with obtain active ingredient or as an alternative the agents area of raw material, particularly most of medicinal plants be its root.China is engaged in the work of medicinal plant tissue culture and rests on culture dish mostly and shake a bottle stage, up to the present, the in-vitro inducing of pseudo-ginseng adventive root, cultivate and utilize bioreactor technology to come the technology of the pseudo-ginseng adventive root aspect that culture of ex vivo induces not appear in the newspapers yet at home and abroad.
Summary of the invention
In order to address the above problem, the object of the present invention is to provide that a kind of technological operation is simple, good reproducibility, large-scale production fast, the tissue culture and the production method of the pseudo-ginseng adventive root of convenient and low production cost.
In order to achieve the above object, the tissue culture of pseudo-ginseng adventive root provided by the invention and production method comprise the step of carrying out in the following order: step 1, inducing of pseudo-ginseng callus: will cut into pieces after the sterilization of pseudo-ginseng root, and be inoculated on the preprepared solid culture medium, under aseptic, room temperature and lucifuge condition, cultivate the formation of 3-5 visible callus after week; Step 2, the amplification of pseudo-ginseng callus and successive transfer culture: the callus that induces is inoculated on the ready solid culture medium, adds plant growth regulator, under aseptic, room temperature and lucifuge condition, cultivate 3-5 week; Step 3, the inducing of pseudo-ginseng adventive root: the callus that amplifies is inoculated on the ready solid culture medium, under aseptic, room temperature and lucifuge condition, cultivates the formation that 3-4 Zhou Houke sees adventive root; Step 4, the numerous and successive transfer culture of the expansion of pseudo-ginseng adventive root: established adventive root is inoculated on the ready solid culture medium, adds plant growth regulator, under aseptic, room temperature and lucifuge condition, cultivate 3-4 week; Step 5, the screening of the different root systems of pseudo-ginseng adventive root and the mensuration of active component and expand numerous: the adventive root to different growing states, different colours and the different thicknesses of turning out is classified, expand numerous and successive transfer culture, after treating that strain system is stable, carry out the mensuration of active component, and set up the finger-print of corresponding composition, filter out optimum adventive root root system according to testing result then, and be inoculated in and expand numerously in the triangular flask of different volumes, cultivation cycle is 3-4 week; Step 6, the reactor of pseudo-ginseng adventive root is cultivated: the pseudo-ginseng adventive root that will expand in step 5 after numerous is inoculated in the liquid nutrient medium of airlift reactor, and adding plant growth regulator, under air feed speed is 0.1vvm, 18-25 ℃ and lucifuge or illumination condition, cultivate, cultivate 6-12 and promptly make pseudo-ginseng adventive root of the present invention after week.
Pseudo-ginseng adventive root provided by the invention is that the adventive root that utilizes reactor to carry out pseudo-ginseng is cultivated, its propagation multiple can reach more than 100 times, has the short and rate of increase advantages of higher of restriction, growth time that is not subjected to geographical position, season and time, be very suitable for the growth of pseudo-ginseng adventive root, therefore opened up the new way of an industrialization for the large-scale production of this rare traditional Chinese medicine resource of pseudo-ginseng, thereby can fundamentally solve the problem that pseudo-ginseng medicine source is subjected to the natural conditions restriction, so economic and social benefit is remarkable.
Embodiment
Be elaborated below in conjunction with the tissue culture and the production method of specific embodiment to pseudo-ginseng adventive root of the present invention.
Embodiment 1
With HCl and NaOH solution 100mL MS solid culture medium is transferred to pH6.0, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 15 minutes is standby; Select the pseudo-ginseng root of well-grown, no damage by disease and insect, use running water, distilled water and washing agent irrigation and disinfection 10 minutes respectively, clean with distilled water then, and with 70% alcohol immersion 3 minutes, adopt 1% clorox (adding several surfactant Tween-20) sterilization 30 minutes afterwards, use distilled water wash 3-5 time at last, can obtain aseptic pseudo-ginseng root.Pseudo-ginseng root after the sterilization is cut into pieces with the scalpel of sterilize, is inoculated on the preprepared MS solid culture medium, under 25 ℃ and lucifuge condition, carry out inducing of pseudo-ginseng callus, cultivate 4 all after the as seen formation of callus.With HCl and NaOH solution 100mL MS solid culture medium is transferred to pH6.0, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 15 minutes is standby; Select well-grown callus, scalpel and tweezers with prior sterilization are inoculated on the ready MS solid culture medium, and adding plant growth regulator 2,4-D, concentration is 1mg/L, carry out the amplification and the successive transfer culture of pseudo-ginseng callus under 23 ℃ and lucifuge condition, cultivation cycle was 4 weeks.With HCl and NaOH solution 100mL MS solid culture medium is transferred to pH6.0, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 15 minutes is standby; Select well-grown callus, be inoculated on the ready MS solid culture medium, under 25 ℃ and lucifuge condition, carry out inducing of pseudo-ginseng adventive root, cultivate the formation that 4 Zhou Houke see adventive root with the scalpel and the tweezers of sterilization in advance.With HCl and NaOH solution 100mL MS solid culture medium is transferred to pH6.0, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 15 minutes is standby; Choose well-grown adventive root, scalpel and tweezers with prior sterilization are inoculated on the ready MS solid culture medium, and add plant growth regulator IBA, and concentration is 1mg/L, carry out the numerous and successive transfer culture of expansion of pseudo-ginseng adventive root under 25 ℃ and lucifuge condition, cultivation cycle was 4 weeks.Adventive root to different growing states, different colours and the different thicknesses of turning out is classified, expand numerous and successive transfer culture, after treating that strain system is stable, adopt the HPLC method to carry out the mensuration of active component, detect the content of ginsenoside Rb1, Rg1 in the pseudo-ginseng, the content of check polysaccharide, and set up the finger-print of corresponding composition, filter out optimum adventive root root system according to testing result then, and be inoculated in expand in the triangular flask of 200mL numerous, interior dress 50mL contains the MS liquid nutrient medium of 3% sucrose, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 20 minutes is standby; Be inoculated in this triangular flask filtering out optimum adventive root root system, and add plant growth regulator IBA, concentration is 1mg/L, and on the shaking table of 100rpm speed, cultivate under 25 ℃ and the lucifuge condition, the cycle was 3 weeks; Adopt the 1000L airlift reactor, interior dress 500L contains the MS liquid nutrient medium of 3% sucrose, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 20 minutes is standby; The pseudo-ginseng adventive root that expands after numerous is inoculated in this reactor, and adds plant growth regulator IBA, concentration is 1mg/L, cultivates under the air feed speed of 0.1vvm, 25 ℃ and lucifuge condition, can obtain pseudo-ginseng adventive root of the present invention after 8 weeks.
Embodiment 2
With HCl and NaOH solution 100mL 1/2MS solid culture medium is transferred to pH6.0, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 15 minutes is standby; Select the pseudo-ginseng root of well-grown, no damage by disease and insect, use running water, distilled water and washing agent irrigation and disinfection 10 minutes respectively, clean with distilled water then, and with 70% alcohol immersion 3 minutes, adopt 1% clorox (adding several surfactant Tween-20) sterilization 30 minutes afterwards, use distilled water wash 3-5 time at last, can obtain aseptic pseudo-ginseng root.Pseudo-ginseng root after the sterilization is cut into pieces with the scalpel of sterilize, is inoculated on the preprepared 1/2MS solid culture medium, under 24 ℃ and lucifuge condition, carry out inducing of pseudo-ginseng callus, cultivate 5 all after the as seen formation of callus.With HCl and NaOH solution 100mL 1/2MS solid culture medium is transferred to pH6.0, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 15 minutes is standby; Select well-grown callus, scalpel and tweezers with prior sterilization are inoculated on the ready 1/2MS solid culture medium, and adding plant growth regulator IAA, concentration is 1mg/L, carry out the amplification and the successive transfer culture of pseudo-ginseng callus under 25 ℃ and lucifuge condition, cultivation cycle was 5 weeks.With HCl and NaOH solution 100mL1/2 MS solid culture medium is transferred to pH6.0, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 15 minutes is standby; Select well-grown callus, be inoculated on the ready 1/2MS solid culture medium, under 24 ℃ and lucifuge condition, carry out inducing of pseudo-ginseng adventive root, cultivate the formation that 4 Zhou Houke see adventive root with the scalpel and the tweezers of sterilization in advance.With HCl and NaOH solution the 100mL1/2MS solid culture medium is transferred to pH6.0, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 15 minutes is standby; Choose well-grown adventive root, scalpel and tweezers with prior sterilization are inoculated on the ready 1/2MS solid culture medium, and add plant growth regulator NAA, and concentration is 1mg/L, carry out the numerous and successive transfer culture of expansion of pseudo-ginseng adventive root under 24 ℃ and lucifuge condition, cultivation cycle was 4 weeks.Adventive root to different growing states, different colours and the different thicknesses of turning out is classified, expand numerous and successive transfer culture, after treating that strain system is stable, adopt the HPLC method to carry out the mensuration of active component, detect the content of ginsenoside Rb1, Rg1 in the pseudo-ginseng, the content of check polysaccharide, and set up the finger-print of corresponding composition, filter out optimum adventive root root system according to testing result then, and be inoculated in expand in the triangular flask of 200mL numerous, interior dress 50mL contains the MS liquid nutrient medium of 3% sucrose, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 20 minutes is standby; Be inoculated in this triangular flask filtering out optimum adventive root root system, and add plant growth regulator IBA, concentration is 1mg/L, and on the shaking table of 100rpm speed, cultivate under 25 ℃ and the lucifuge condition, the cycle was 4 weeks; Adopt the 1000L airlift reactor, interior dress 500L contains the 1/2MS liquid nutrient medium of 4% sucrose, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 20 minutes is standby; The pseudo-ginseng adventive root that expands after numerous is inoculated in this reactor, and adds plant growth regulator IBA, concentration is 1mg/L, cultivates under the air feed speed of 0.1vvm, 18 ℃ and illumination condition, can obtain pseudo-ginseng adventive root of the present invention after 12 weeks.
Embodiment 3
With HCl and NaOH solution 100mL 3/4MS solid culture medium is transferred to pH6.0, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 15 minutes is standby; Select the pseudo-ginseng root of well-grown, no damage by disease and insect, use running water, distilled water and washing agent irrigation and disinfection 10 minutes respectively, clean with distilled water then, and with 70% alcohol immersion 3 minutes, adopt 1% clorox (adding several surfactant Tween-20) sterilization 30 minutes afterwards, use distilled water wash 3-5 time at last, can obtain aseptic pseudo-ginseng root.Pseudo-ginseng root after the sterilization is cut into pieces with the scalpel of sterilize, is inoculated on the preprepared 3/4MS solid culture medium, under 25 ℃ and lucifuge condition, carry out inducing of pseudo-ginseng callus, cultivate 5 all after the as seen formation of callus.With HCl and NaOH solution 100mL 3/4MS solid culture medium is transferred to pH6.0, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 15 minutes is standby; Select well-grown callus, scalpel and tweezers with prior sterilization are inoculated on the ready 3/4MS solid culture medium, and adding plant growth regulator kinetin, concentration is 1mg/L, carry out the amplification and the successive transfer culture of pseudo-ginseng callus under 25 ℃ and lucifuge condition, cultivation cycle was 4 weeks.With HCl and NaOH solution 100mL 3/4MS solid culture medium is transferred to pH6.0, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 15 minutes is standby; Select well-grown callus, be inoculated on the ready 3/4MS solid culture medium, under 25 ℃ and lucifuge condition, carry out inducing of pseudo-ginseng adventive root, cultivate the formation that 4 Zhou Houke see adventive root with the scalpel and the tweezers of sterilization in advance.With HCl and NaOH solution the 100mL3/4MS solid culture medium is transferred to pH6.0, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 15 minutes is standby; Choose well-grown adventive root, scalpel and tweezers with prior sterilization are inoculated on the ready 3/4MS solid culture medium, and adding plant growth regulator kinetin, concentration is 1mg/L, carry out the numerous and successive transfer culture of expansion of pseudo-ginseng adventive root under 24 ℃ and lucifuge condition, cultivation cycle was 4 weeks.Adventive root to different growing states, different colours and the different thicknesses of turning out is classified, expand numerous and successive transfer culture, after treating that strain system is stable, adopt the HPLC method to carry out the mensuration of active component, detect ginsenoside Rb1 in the pseudo-ginseng, the content of Rg1, the content of check polysaccharide, and set up the finger-print of corresponding composition, filter out optimum adventive root root system according to testing result then, and be inoculated in and expand numerously in the triangular flask of 200mL, interior dress 50mL contains the MS liquid nutrient medium of 3% sucrose, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 20 minutes is standby; Be inoculated in this triangular flask filtering out optimum adventive root root system, and add plant growth regulator IBA, concentration is 1mg/L, and on the shaking table of 100rpm speed, cultivate under 25 ℃ and the lucifuge condition, cultivation cycle was 3 weeks; Adopt the 1000L airlift reactor, interior dress 500L contains the 3/4MS liquid nutrient medium of 5% sucrose, at 121 ℃, 1.2kg/cm
-2Pressure down sterilization 20 minutes is standby; The pseudo-ginseng adventive root that expands after numerous is inoculated in this reactor, and adds plant growth regulator IBA, concentration is 1mg/L, cultivates under the air feed speed of 0.1vvm, 20 ℃ and illumination condition, can obtain pseudo-ginseng adventive root of the present invention after 10 weeks.
Claims (6)
1, a kind of tissue culture of pseudo-ginseng adventive root and production method, it is characterized in that: the tissue culture of described pseudo-ginseng adventive root and production method comprise the step of carrying out in the following order: step 1, inducing of pseudo-ginseng callus: will cut into pieces after the sterilization of pseudo-ginseng root, and be inoculated on the preprepared solid culture medium, under aseptic, room temperature and lucifuge condition, cultivate the formation of 3-5 visible callus after week; Step 2, the amplification of pseudo-ginseng callus and successive transfer culture: the callus that induces is inoculated on the ready solid culture medium, adds plant growth regulator, under aseptic, room temperature and lucifuge condition, cultivate 3-5 week; Step 3, the inducing of pseudo-ginseng adventive root: the callus that amplifies is inoculated on the ready solid culture medium, under aseptic, room temperature and lucifuge condition, cultivates the formation that 3-4 Zhou Houke sees adventive root; Step 4, the numerous and successive transfer culture of the expansion of pseudo-ginseng adventive root: established adventive root is inoculated on the ready solid culture medium, adds plant growth regulator, under aseptic, room temperature and lucifuge condition, cultivate 3-4 week; Step 5, the screening of the different root systems of pseudo-ginseng adventive root and the mensuration of active component and expand numerous: the adventive root to different growing states, different colours and the different thicknesses of turning out is classified, expand numerous and successive transfer culture, after treating that strain system is stable, carry out the mensuration of active component, and set up the finger-print of corresponding composition, filter out optimum adventive root root system according to testing result then, and be inoculated in and expand numerously in the triangular flask of different volumes, cultivation cycle is 3-4 week; Step 6, the reactor of pseudo-ginseng adventive root is cultivated: the pseudo-ginseng adventive root that will expand in step 5 after numerous is inoculated in the liquid nutrient medium of airlift reactor, and adding plant growth regulator, under air feed speed is 0.1vvm, 18-25 ℃ and lucifuge or illumination condition, cultivate, cultivate 6-12 and promptly make pseudo-ginseng adventive root of the present invention after week.
2, the tissue culture of pseudo-ginseng adventive root according to claim 1 and production method is characterized in that: described solid and liquid nutrient medium are the MS base.
3, the tissue culture of pseudo-ginseng adventive root according to claim 1 and production method is characterized in that: in the amplification and successive transfer culture of pseudo-ginseng adventive root, described plant growth regulator is 2,4-D, any in IAA and the kinetin.
4, the tissue culture of pseudo-ginseng adventive root according to claim 1 and production method is characterized in that: the expansion of pseudo-ginseng adventive root numerous with successive transfer culture in, described plant growth regulator is any in IBA, NAA and the kinetin.
5, the tissue culture of pseudo-ginseng adventive root according to claim 1 and production method is characterized in that: in the reactor of pseudo-ginseng adventive root was cultivated, described liquid nutrient medium contained the sucrose of 3%-5%.
6, the tissue culture of pseudo-ginseng adventive root according to claim 1 and production method is characterized in that: the concentration of described plant growth regulator is 1mg/L.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100407906C (en) * | 2006-07-10 | 2008-08-06 | 浙江省农业科学院 | Tissue-culturing quick propagation method of wild rhizoma panacis japonici |
| CN1947498B (en) * | 2006-08-31 | 2010-05-12 | 上海交通大学 | Method for induction and tissue-culture of adventitous root of pseudo-ginseng |
| CN102771397A (en) * | 2012-08-17 | 2012-11-14 | 成都市三禾田生物技术有限公司 | 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 |
| CN102907318A (en) * | 2011-08-01 | 2013-02-06 | 东北林业大学 | Rapid propagation of notoginseng regenerated plant by using bioreactor to cultivate notoginseng somatic embryos |
| CN103923874A (en) * | 2014-03-25 | 2014-07-16 | 天津大学 | Pseudo-ginseng cell suspension culture method |
| CN104620986A (en) * | 2015-02-12 | 2015-05-20 | 天津大学 | Induction and tissue culture method of pseudo-ginseng adventitious root |
| CN104920214A (en) * | 2015-06-03 | 2015-09-23 | 中国科学院昆明植物研究所 | Method for improving yield of saponin in panax notoginseng tissue culture seedlings |
| CN110585267A (en) * | 2019-09-06 | 2019-12-20 | 天津大学 | Pharmaceutical composition with thrombin activity inhibition function and application thereof |
| CN113349053A (en) * | 2020-03-04 | 2021-09-07 | 东北林业大学 | Method for producing three main ginsenosides by rapidly proliferating adventitious roots of panax notoginseng |
-
2003
- 2003-06-27 CN CN 03130265 patent/CN1256870C/en not_active Expired - Fee Related
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100407906C (en) * | 2006-07-10 | 2008-08-06 | 浙江省农业科学院 | Tissue-culturing quick propagation method of wild rhizoma panacis japonici |
| CN1947498B (en) * | 2006-08-31 | 2010-05-12 | 上海交通大学 | Method for induction and tissue-culture of adventitous root of pseudo-ginseng |
| CN102907318A (en) * | 2011-08-01 | 2013-02-06 | 东北林业大学 | Rapid propagation of notoginseng regenerated plant by using bioreactor to cultivate notoginseng somatic embryos |
| CN102771397A (en) * | 2012-08-17 | 2012-11-14 | 成都市三禾田生物技术有限公司 | 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 |
| CN102771397B (en) * | 2012-08-17 | 2013-11-27 | 成都市三禾田生物技术有限公司 | 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 |
| CN103923874A (en) * | 2014-03-25 | 2014-07-16 | 天津大学 | Pseudo-ginseng cell suspension culture method |
| CN104620986A (en) * | 2015-02-12 | 2015-05-20 | 天津大学 | Induction and tissue culture method of pseudo-ginseng adventitious root |
| CN104920214A (en) * | 2015-06-03 | 2015-09-23 | 中国科学院昆明植物研究所 | Method for improving yield of saponin in panax notoginseng tissue culture seedlings |
| CN104920214B (en) * | 2015-06-03 | 2017-06-20 | 中国科学院昆明植物研究所 | A kind of method for improving pseudo-ginseng tissue-cultured seedling saponin(e yield |
| CN110585267A (en) * | 2019-09-06 | 2019-12-20 | 天津大学 | Pharmaceutical composition with thrombin activity inhibition function and application thereof |
| CN113349053A (en) * | 2020-03-04 | 2021-09-07 | 东北林业大学 | Method for producing three main ginsenosides by rapidly proliferating adventitious roots of panax notoginseng |
| CN113349053B (en) * | 2020-03-04 | 2022-03-22 | 东北林业大学 | Method for producing three main ginsenosides by rapidly proliferating adventitious roots of panax notoginseng |
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| CN1256870C (en) | 2006-05-24 |
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