CN100566558C - Radix bupleuri tissue culturing fast seedling-cultivating method - Google Patents
Radix bupleuri tissue culturing fast seedling-cultivating method Download PDFInfo
- Publication number
- CN100566558C CN100566558C CNB2007101396792A CN200710139679A CN100566558C CN 100566558 C CN100566558 C CN 100566558C CN B2007101396792 A CNB2007101396792 A CN B2007101396792A CN 200710139679 A CN200710139679 A CN 200710139679A CN 100566558 C CN100566558 C CN 100566558C
- Authority
- CN
- China
- Prior art keywords
- radix bupleuri
- medium
- bud
- tissue culturing
- cultivating method
- 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.)
- Expired - Fee Related
Links
Images
Classifications
-
- Y02P60/216—
Landscapes
- Cultivation Of Plants (AREA)
Abstract
The invention provides a kind of tissue culturing fast seedling-cultivating method of medicinal plant radix bupleuri.This method selects the stem with bud of radix bupleuri as explant, is inoculated in the B of additional 6-BA 1.0mg/L and KT 0.2mg/L after the sterilization
5The fast breeding of induced bud in the medium; At B
5To not have offspring in the minimal medium after strong sprout and change root induction in the 1/2MS medium that adds NAA 0.1mg/L, IBA 0.5mg/L and DSC 1.0mg/L over to; Transplant after hardening, the transplanting survival rate of test-tube plantlet can reach 94%～97%.Method of the present invention is quick, efficient, and cost is low, and the transplanting survival rate height is convenient to promote, and genetic stability is good, is applicable to the cultivar or the type of various radix bupleuri.Can be according to market situation, the radix bupleuri of a large amount of at short notice breedings and production high-quality is cultivated plant or high quality seed is provided.
Description
Technical field
The present invention relates to plant regeneration, specifically belong to a kind of tissue culturing fast seedling-cultivating method of medicinal plant radix bupleuri by tissue culture technique.
Background technology
Radix bupleuri is important medicinal plant, also is the employed large medicinal materials of state such as China, Korea S and Japan.Chinese Pharmacopoeia (version in 2000) is included samphire Bupleurum Chinese Bupleurum Chinese DC. (claim not only hard seedling radix bupleuri) and radix bupleuri scorzoneraefolii B.scorzonerifolium Willd. (but also claiming soft seedling radix bupleuri or RADIX BUPLEURI SCORZONERIFOLII) is the former plant of medicinal radix bupleuri.Radix bupleuri is used as medicine with root, contains saikoside and volatilization wet goods composition in the root, has antiviral, antibacterium endotoxin, anti-inflammatory, antitumor, anticonvulsion, analgesic, reducing blood lipid and promotes to secrete, protect the liver, improve effect such as immunity, and medical value is very high.
In recent years, the market demand of radix bupleuri increases year by year, traditional method of obtaining radix bupleuri is to be cost to gather and to consume wild resource, owing to unplanned coyoting is for many years dug excessively and the natural resources regeneration capacity that destruction caused of natural environment is descended the degradation reason greatly, make natural wild medicinal radix bupleuri resource wretched insufficiency, can only the amount of satisfying the demands about 50%, remaining 50% need replenish by artificial cultivation, and therefore the emphasis of radix bupleuri supply is in recent years progressively transferred to the artificial cultivation plant and come up.China's cultivated area maximum is the Bupleurum falcatum that originates in Japan at present, but Bupleurum falcatum is not the certified products medicinal material, and relevant department of Chinese government explicit order requires plantation to originate in the radix bupleuri kind of China and forbid planting Bupleurum falcatum.Yet, because various radix bupleuri plant appearance similars, and usually being a plurality of kinds and mixing and be born in the same place of production that morphology is identified relatively difficulty, academia handles also disputable to its classification.Through our investigation, all there is variet complexity phenomenon in various degree in each place of production of Bupleurum Chinese, different modification under the existing Bupleurum Chinese kind, the sibling species that also has form easily to mix, the various morphosiss that occur behind the introducing and planting are more arranged, cause the germplasm of Bupleurum Chinese medicinal material impure thus, variet complexity, quality be uneven, causes oeverall quality to descend, have influence on the accuracy of medication and the stability of drug quality, also had influence on the foreign exchange earning of radix bupleuri.Therefore, press for a kind of science more, system, effective, the practical technology of adopting, on the basis of the pollination type of the hereditary feature of investigating thoroughly the main variation type of Bupleurum Chinese and radix bupleuri, carry out the screening and the cultivation of radix bupleuri improved seeds, particularly to existing, undertaken by the artificial cultivation kind that numerous medicinal herb growers accepted that germplasm is purified, rejuvenation, cultivating high yield best in quality, that variation is little, high-quality radix bupleuri plant (seed) and batch process and supply the market, is present problem demanding prompt solution.
Plant tissue culture technique is the effective ways of fast seedling growing, has obtained in fields such as gardening, forests using widely.The application organizes cultural method is bred the medicinal radix bupleuri of high-quality fast, is to solve the quick of radix bupleuri disparities between supply and demand, raising plantation level and radix bupleuri quality and valid approach.Yet, the kind difference of plant, kind difference, even the selected explant difference of same kind, also there is very big difference in required medium component, condition of culture and the transplanting and management conditions etc. of tissue culture.
Summary of the invention
The objective of the invention is to set up the tissue culturing fast seedling-cultivating method of medicinal radix bupleuri, large-scale production high-quality radix bupleuri seedling and seed.This method is simple and practical, and is rapidly efficient, suitability for scale production.
The present invention cultivates through the multiplex screening optimum experimental and breeds condition, has proposed the method that suitable medicinal radix bupleuri tissue-culturing quick-propagation is grown seedlings.It is explant that the present invention selects the stem section of high-quality Bupleurum Chinese (Bupleurum chineseDC.) or radix bupleuri scorzoneraefolii (B.scorzonerifoliumWilld); Screened the medium constituent of bud fast breeding; Screened the condition of strong seedling culture; Having screened the culture of rootage medium forms; Screen and set up the condition and the measure of test-tube seedling transplanting and field management.
Radix bupleuri tissue culturing fast seedling-cultivating method of the present invention, concrete steps comprise:
(1) sterilization of explant: the radix bupleuri that will pick up from the field washes away surface dust with running water, and the young stem of getting the band axillalry bud is cut into the long segment of 3cm～5cm, through 75% alcohol immersion 30s, 0.1%HgCl as explant
2Aqueous solution sterilization 10min, aseptic water washing 4 times;
(2) inducing and breed of bud: will be cut into stem-segment with single bud through the explant of above-mentioned processing, and be inoculated in the B that contains exogenous plant hormones respectively
5In the medium, under 25 ± 2 ℃, 1500lux～2500lux, 12～14h/d illumination, cultivate 30d～40d and obtain not having offspring, every 30d～the 40d successive transfer culture once; Culture medium prescription is: B
5Medium+0.2mg/L～1.5mg/L6-BA+0.1mg/L～0.2mg/L KT+0mg/L～0.2mg/L NAA+2.5%～3.5% sucrose, pH 5.8～6.0.In above medium, the axillalry bud on the explant all can normally be sprouted.
Above-mentioned medium is preferably: B
5Medium+1.0mg/L6-BA+0.2mg/L KT+3.0% sucrose, pH 5.8, and at this inoculation of medium 1 all backs axillary bud sprouting, after 2～3 weeks of inoculation, bud is bred rapidly and is formed the bud of growing thickly, no offspring robust growth, height of seedling 3cm～4cm.But successive transfer culture behind 30d～40d.
(3) strong sprout: through behind the successive transfer culture repeatedly, no offspring growth conditions can weaken, and the no offspring that propagation need be obtained is inoculated into and contains 2.5%～3.5% sucrose, the B of pH 5.8～6.0
5In the medium, under 25 ± 2 ℃, 1500lux～2500lux, 12～14h/d illumination, cultivate 15d～35d strong sprout.
(4) root induction: choosing the no offspring of growing thickly healthy and strong, that 1cm～3cm is high, is simple bud with its cutting, goes to root induction in the root media, obtains complete test tube plantlet; The culture of rootage based formulas is: 1/2MS medium+0.1mg/L～1.0mg/L NAA+0.1mg/L～1.0mg/LIBA+0mg/L～1.0mg/LDSC+2.5%～3.5% sucrose, and pH 5.8～6.0; Under 25 ± 2 ℃, 1500lux～2500lux, 12～14h/d illumination, cultivate 35d～45d.Radix bupleuri does not have offspring and almost can both take root in above medium.
Preferred root media is: 1/2MS medium+NAA 0.1mg/L+IBA 0.5mg/L+DSC 1.0mg/L+3.0% sucrose, and pH 5.8, and this medium does not have taking root of offspring to radix bupleuri good facilitation is arranged, and rooting rate is 100%.
(5) hardening and transplanting: choose and take root many and healthy and strong plant is opened and seals film hardening 3d～5d; clean medium then; immigration is equipped with in the nutritive cube of turfy soil and perlite (volume ratio 1: 1～3: 1) mixed-matrix and is placed the greenhouse; should note protecting the tip of a root of test-tube plantlet during transplanting; water permeable after the transplanting; cover to preserve moisture with plastic film; throwing off film behind the 3d gradually ventilates; to avoid mould contamination, add intense light irradiation, in time supply nutritive cube moisture; after 1～3 week; place the field to adapt to the land for growing field crops environment nutritive cube, root system grows up to after 2～4 weeks, transplants to the big Tanaka who imposes phosphate fertilizer.The transplanting initial stage will be noted the control of damage by disease and insect and replenishing of moisture.The survival rate of transplanted seedling can reach 94%～97%.
Compared with prior art, the present invention has the following advantages and effect:
(1) shortened the cycle that radix bupleuri selects the good child-rearing seedling.Radix bupleuri tradition plantation breeding cycle is longer, is difficult to satisfy requirement of large-scale production.The present invention can shorten the growing-seedling period of radix bupleuri greatly, and can select fine quality or type to carry out large-scale industrialized production.
(2) improved reproduction coefficient, the survival rate height of transplanted seedling.The common radix bupleuri cultivated form breeding cycle is long.Reproduction speed of the present invention is fast, and the seedling growth is neat, and simple bud breeds the 30d subculture once, and the propagation multiple is 5～10 times; Filter out suitable radix bupleuri test-tube plantlet hardening condition and cultivation matrix, make transplanting survival rate reach 94%～97%; Have the low characteristics of cost.
(3) with the stem section as explant, breed fast by the propagation of bud, guaranteed hereditary stability, and drawn materials easily that it is convenient to upgrade.
(4) can rationally arrange and adjust production scale according to relation between market supply and demand, in time tissue is produced, and can produce in the realization anniversary.
(5) has general applicability.The present invention is based on the Bupleurum Chinese of 8 area plantations in the main planting site Shanxi of radix bupleuri, Shaanxi, Gansu and the quick breeding research of radix bupleuri scorzoneraefolii cultivated form, the radix bupleuri of various places all can be tamed and be carried out test tube rapid propagation, helps to screen from now on the radix bupleuri fine quality that obtains or the quick breeding and the popularization of type.
Description of drawings
Fig. 1 shows that Bupleurum Chinese does not have the propagation of offspring bud: the Bupleurum Chinese stem section that will take from the field inserts medium through after the surface sterilization, after axillalry bud grows, is the B of 6-BA 1.0mg/L+KT 0.2mg/L at hormone combination
5Carry out the quick expansion propagation of bud in the medium, obtain the bud of growing thickly after 3 weeks.
Fig. 2 shows that Bupleurum Chinese does not have offspring and induces adventive root: through B
5Minimal medium is after strong sprout, and the bud branch of will growing thickly is cut to simple bud, and inserting hormone combination is the 1/2MS medium root induction of NAA 0.1mg/L, IBA 0.5mg/L and DSC 1.0mg/L, and 35
The complete test-tube plantlet that obtains behind the d.
Fig. 3 shows that the Bupleurum Chinese test-tube seedling transplanting is to the nutritive cube that matrix is arranged: test-tube plantlet is cleaned the medium of root after hardening, be transplanted in the nutritive cube that contains turfy soil and perlite mixed-matrix.
Fig. 4 shows Bupleurum Chinese test-tube seedling transplanting land for growing field crops: the test-tube plantlet in the nutritive cube adapts to the land for growing field crops environment, and root system development is transplanted in the soil growth situation of doing well after improving.
Fig. 5 shows that the Bupleurum Chinese test-tube plantlet after the transplanting blooms: test-tube seedling transplanting is gone into the land for growing field crops and is begun after 2 months to bloom, and the florescence reaches 4 months.
Fig. 6 shows that the Bupleurum Chinese test-tube plantlet after the transplanting is solid: the test-tube plantlet that moves into the land for growing field crops is normally solid, and full seed.
Embodiment
Embodiment 1
The young stem section of Bupleurum Chinese axillalry bud that to take from field, Lingchuan, Shanxi is as explant, and flowing water washes away surface dust, is cut into the long segment of 3cm～5cm.Through 75% alcohol immersion 30s, 0.1%HgCl
2Behind the aqueous solution sterilization 10min, aseptic water washing 4 times, be cut into 20 sections of stem-segment with single bud about 0.5cm, be inoculated in and contain 6-BA 1.0mg/L, KT 0.2mg/L, sucrose 3.0%, the B of pH5.8
5On the medium, cultivate under 25 ± 2 ℃, 2000lux, 14h/d illumination, carry out inducing and fast breeding of bud, bud (as Fig. 1) obtained growing thickly after 3 weeks.Successive transfer culture after 4 weeks, subculture is not had offspring later on 3 times in a large number, it is changed over to contain 3.0% sucrose, and pH is 5.8 B
5Minimal medium strong sprout.
Cultivate about 15d, offspring not had grows to about 2cm when high, chooses the seedling of growing thickly of robust growth, changes the root induction medium after the cutting over to.Consisting of of root induction medium: 1/2MS+NAA 0.1mg/L+IBA 0.5mg/L+DSC 1.0mg/L+3.0% sucrose, pH 5.8.Obtain about 700 strains of complete test-tube plantlet (as Fig. 2) behind the 35d.Remaining no offspring is cultivated 30d in the strong seedling culture base after, continue cutting, change the fast propagating culture medium shoot proliferation over to.
Take root behind the 40d, test-tube plantlet hardening 3d～5d cleans the medium of root, is transplanted in the nutritive cube of the mixed-matrix that contains turfy soil and perlite (1: 1) (as Fig. 3).Water permeablely after the transplanting immediately, cover to preserve moisture, place the greenhouse with plastic film.Ventilate gradually behind the 3d and throw off film, add intense light irradiation.
2 weeks of test-tube plantlet behind the transplant survival were placed on the land for growing field crops hardening about 1 month, treat that root system grows up to after, move into the land for growing field crops.The tissue cultivating seedling form is identical with former cultivation plant, and growth is than common cultivation robust plant (as Fig. 4), and the transplanting average plant height of plant then is more than 1.2m.Bloom after 2 months, the florescence reaches 4 months (as Fig. 5), and mid or late September begins solid, full seed (as Fig. 6).
Embodiment 2
To take from the stem section of the Bupleurum Chinese band axillalry bud in field, Taigu, Shanxi (originating in Zuoquan, Shanxi), flowing water washes away surface dust, is cut into the long segment of long 3cm～5cm.Through 75% alcohol immersion 30s, 0.1%HgCl
2Behind the aqueous solution sterilization 10min, aseptic water washing 4 times, be cut into the stem-segment with single bud about 0.5cm, be inoculated in additional 6-BA 0.5mg/L, KT 0.1mg/L, the B of NAA 0.1mg/L
5In the medium, under 25 ± 2 ℃, 2000lux, 14h/d illumination, cultivate the sprouting of inducing axillalry bud, 2 week the back axillary bud sproutings, forms no offspring, 4 all after buds bred about 3 times.
The no offspring of gained is continued to cut into stem-segment with single bud, change the bud proliferated culture medium that screening obtains over to.Consisting of of medium: B
5+ 6-BA 1.0mg/L+KT 0.2mg/L+3.0% sucrose, pH 5.8.In the successive transfer culture process, base portion at no offspring forms the bud of growing thickly, the bud multiplication rate is increased to about 6,30d was 1 generation, subculture obtains a large amount of Bupleurum Chineses after does not repeatedly have offspring, but growth conditions weakens to some extent, and the blastogenesis of growing thickly that has is long closely is difficult for cutting apart, and changes it in the strong seedling culture base (with embodiment 1) strong sprout.
Select the bud of growing thickly of robust growth behind the strong sprout 20d, cutting is the simple bud plant, changes over to carry out root induction and hardening, transplanting in the root media (with embodiment 1).
Remaining no offspring is cultivated 35d in the strong seedling culture base after, continue cutting, change subculture propagation in the bud proliferated culture medium over to.
Embodiment 3
To take from the stem section of the radix bupleuri scorzoneraefolii in field, Taigu, Shanxi, through the sterilization after be inoculated into bud induce with proliferated culture medium in, condition of culture and medium obtain the test-tube plantlet plant of radix bupleuri scorzoneraefolii with embodiment 1.
Claims (6)
1, a kind of radix bupleuri tissue culturing fast seedling-cultivating method is characterized in that, comprises the steps:
(1) sterilization of explant: with the young stem of radix bupleuri band axillalry bud as explant, segment, sterilization, aseptic water washing;
(2) inducing and breed of bud: will be cut into stem-segment with single bud through the explant of above-mentioned processing, and be inoculated in the B that contains exogenous plant hormones respectively
5In the medium, at 25 ± 2 ℃, 1500lux～2500lux, 12～14h/d illumination is cultivated down 30d～40d and is obtained not having offspring, every 30d～40d successive transfer culture 1 time; Culture medium prescription is: B
5Medium+0.2mg/L～1.5mg/L6-BA+0.1mg/L～0.2mg/L KT+0mg/L～0.2mg/L NAA+2.5%～3.5% sucrose, pH 5.8～6.0;
(3) strong sprout: through behind the successive transfer culture, the no offspring that propagation is obtained is transferred to and contains 2.5%～3.5% sucrose, the B of pH 5.8～6.0
5In the medium, under 25 ± 2 ℃, 1500lux～2500lux, 12～14h/d illumination, cultivate 15d～35d;
(4) root induction: choosing the no offspring of growing thickly healthy and strong, that 1cm～3cm is high, is simple bud with its cutting, goes to root induction in the root media, obtains complete test tube plantlet; The culture of rootage based formulas is: 1/2MS medium+0.1mg/L～1.0mg/L NAA+0.1mg/L～1.0mg/L IBA+0mg/L～1.0mg/L DSC+2.5%～3.5% sucrose, and pH 5.8～6.0; Under 25 ± 2 ℃, 1500lux～2500lux, 12～14h/d illumination, cultivate 35d～45d;
(5) hardening and transplanting: choose and take root many and healthy and strong plant is opened and seals the film hardening, clean medium then, immigration contains the nutritive cube of turfy soil and perlite mixed-matrix and places the greenhouse, cover with plastic film, throw off film behind the 3d gradually and ventilate, increase illumination, in time supply nutritive cube moisture, after 1～3 week, nutritive cube is placed the field, the back transplanting of 2～4 weeks is to big Tanaka.
2, radix bupleuri tissue culturing fast seedling-cultivating method according to claim 1 is characterized in that bud is induced with proliferated culture medium in the described step (2) and is: B
5Medium+1.0mg/L 6-BA+0.2mg/L KT+3.0% sucrose, pH 5.8.
3, radix bupleuri tissue culturing fast seedling-cultivating method according to claim 1, it is characterized in that root media is in the described step (4): 1/2MS medium+NAA 0.1mg/L+IBA 0.5mg/L+DSC 1.0mg/L+3.0% sucrose, pH 5.8.
4, radix bupleuri tissue culturing fast seedling-cultivating method according to claim 1 is characterized in that turfy soil and perlite volume ratio are 1: 1～3: 1 in described step (5) mixed-matrix.
5, radix bupleuri tissue culturing fast seedling-cultivating method according to claim 1 is characterized in that explant sterilization in the described step (1), is through 75% alcohol immersion 30s, 0.1%HgCl with explant
2Aqueous solution sterilization 10min, aseptic water washing 4 times.
6, radix bupleuri tissue culturing fast seedling-cultivating method according to claim 1 is characterized in that described radix bupleuri is Bupleurum Chinese or radix bupleuri scorzoneraefolii.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2007101396792A CN100566558C (en) | 2007-11-02 | 2007-11-02 | Radix bupleuri tissue culturing fast seedling-cultivating method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2007101396792A CN100566558C (en) | 2007-11-02 | 2007-11-02 | Radix bupleuri tissue culturing fast seedling-cultivating method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101142898A CN101142898A (en) | 2008-03-19 |
| CN100566558C true CN100566558C (en) | 2009-12-09 |
Family
ID=39205771
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2007101396792A Expired - Fee Related CN100566558C (en) | 2007-11-02 | 2007-11-02 | Radix bupleuri tissue culturing fast seedling-cultivating method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100566558C (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105706729B (en) * | 2016-03-31 | 2018-08-31 | 西南科技大学 | A kind of radix bupleuri method for culturing seedlings rapidly and efficiently |
| CN106718898A (en) * | 2016-12-08 | 2017-05-31 | 大连民族大学 | A kind of high yield cultivating method of Laoshan celery |
| CN115380830B (en) * | 2022-10-08 | 2023-08-01 | 浙江大学 | Propagation method of adventitious buds of peucedanum praeruptorum dunn |
-
2007
- 2007-11-02 CN CNB2007101396792A patent/CN100566558C/en not_active Expired - Fee Related
Non-Patent Citations (8)
| Title |
|---|
| 三岛柴胡体细胞胚胎发生与植株再生的研究. 赵胜利等.中国中药杂志，第18卷第9期. 1993 |
| 三岛柴胡体细胞胚胎发生与植株再生的研究. 赵胜利等.中国中药杂志，第18卷第9期. 1993 * |
| 柴胡属(Bupleurum L)植物研究进展. 杜景红等.东北农业大学学报，第34卷第3期. 2003 |
| 柴胡属(Bupleurum L)植物研究进展. 杜景红等.东北农业大学学报，第34卷第3期. 2003 * |
| 柴胡细胞固定化培养. 郝建平等.山西大学学报(自然科学版)，第18卷第2期. 1995 |
| 柴胡细胞固定化培养. 郝建平等.山西大学学报(自然科学版)，第18卷第2期. 1995 * |
| 药用植物柴胡种子萌发特性与组织培养研究. 徐丽霞.山西大学2006届硕士学位论文. 2006 |
| 药用植物柴胡种子萌发特性与组织培养研究. 徐丽霞.山西大学2006届硕士学位论文. 2006 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101142898A (en) | 2008-03-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104719152B (en) | Rhizoma bletillae industrialized seedling method | |
| CN100444722C (en) | Cyrtopterin tissue culturing method and fast reproduction thereof | |
| CN102301951B (en) | Method for rapidly propagating roots of subprostrate sophora by tissue culture | |
| CN104067939A (en) | Tissue culture rapid propagation method of radix gentianae | |
| CN101116425A (en) | Method for preparing thick leaf rhizoma bergeniae by using biology tissue culture process | |
| CN112335549A (en) | Method for obtaining larch regeneration plant through tissue in-vitro culture | |
| CN103190344A (en) | Tissue culture method of fargesii | |
| CN107027627B (en) | Microtuber propagation method for young embryo culture of polygonatum cyrtonema | |
| CN105309317A (en) | Method for propagating moghania macrophylla by tissue culture | |
| CN105594438A (en) | Azalea seedling culture method | |
| CN101176431A (en) | Method for cultivating rapidly white potato breeder's seed | |
| CN105010142B (en) | The method of the strange nanmu agalloch eaglewood tissue cultures of Vietnam | |
| CN104542307A (en) | Culturing method of momordica cochinchinensis | |
| CN106818468A (en) | A kind of shellflower seed asepsis sprouting and rapid propagation method | |
| CN100566558C (en) | Radix bupleuri tissue culturing fast seedling-cultivating method | |
| CN101816286B (en) | Method for tissue culture and rapid propagation of narcissus pseudonarcissus by using ramentum | |
| CN101731149A (en) | Method for rapid propagation of rauwolfia yunnanensis by tissue culture technology | |
| CN101015280B (en) | Tissue culture method for fast propagation of primula denticulata ssp.sino-denticulata | |
| CN103548695B (en) | A kind of meadowrueleaf corydalis root quick breeding method for tissue culture | |
| CN108377911A (en) | The tissue culture mating system of manglietia glauca | |
| CN105900564B (en) | A kind of method that rare or endangered species beet seeds is promoted efficiently to sprout | |
| CN104686344A (en) | Tissue culture method of liriope muscari | |
| CN101743908A (en) | Tissue culture, rapid propagation and cultivation method of grevillea banksii | |
| CN101015279B (en) | Tissue culture method for fast propagation of primula poissonii | |
| CN114568307B (en) | Method for rapid breeding of seedlings by using stem tips of semen katsumadai |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20091209 Termination date: 20121102 |