CN105746347B - A kind of in-vitro conservation method of meadowrueleaf corydalis root - Google Patents
A kind of in-vitro conservation method of meadowrueleaf corydalis root Download PDFInfo
- Publication number
- CN105746347B CN105746347B CN201610108551.9A CN201610108551A CN105746347B CN 105746347 B CN105746347 B CN 105746347B CN 201610108551 A CN201610108551 A CN 201610108551A CN 105746347 B CN105746347 B CN 105746347B
- Authority
- CN
- China
- Prior art keywords
- culture
- seedling
- meadowrueleaf corydalis
- vitro
- culture medium
- 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
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a kind of in-vitro conservation methods of meadowrueleaf corydalis root, two step seedling of tissue culture including meadowrueleaf corydalis root, the test tube seedling with 4 ~ 6 leaves for recycling two step seedling of tissue culture to obtain is inoculated into Storaged media and carries out Plantlet in vitro, and Storaged media of the present invention is 1/2MS+PPP3331.0mg/l+AC0.3%+ sucrose 4.5%+ agar 0.4%.The pH value of culture medium is 5.8, and temperature is 21 ± 1 °C, 1400 ~ 1600lx of intensity of illumination, and light application time is 8 ~ 10 hours/day.The high quality seedling of a large amount of meadowrueleaf corydalis roots can be provided in a short time using the method for the present invention; reduce the step of preparing culture medium and rolling bottle simultaneously, saves a large amount of manpower and materials, therefore simple and practicable, production cost reduces; also, efficiently solve the problems, such as meadowrueleaf corydalis root Germ-plasma resources protection and scale breeding.
Description
Technical field
The invention belongs to biological tissue's culture technique fields, and in particular, to the in-vitro conservation method of meadowrueleaf corydalis root.
Background technology
Meadowrueleaf corydalis root (Corydalis saxicola Bunting) is bloodroot, is herbaceos perennial, distribution
In provinces such as China Guangxi, Guangdong, Fujian, Hunan.All herbal medicine, is famous strong precious jade drug kind, and herb contains deydrokaividing
(Corydalis Saxicolae) isoreactivity ingredient；With significant antibacterial, anti-inflammatory, analgesia and stable effect by force, and inhibition tumour is observed
Cytosis；It cures mainly the diseases meadowrueleaf corydalis root plant distributions such as sore furuncle poison, hepatitis, hepatic sclerosis, liver cancer and is confined to Limestone Mountain Areas, belong to stone
Mountain endemic species.Since habitat conditions is severe, natural propagation rate is very low, in addition its seed is tiny, breeding is relatively difficult, kind mass-sending
Obstacle is opened up, resource reserves are extremely limited.People's largely excavation and purchase in recent years, resulting in wild resource, supply falls short of demand,
Wild meadowrueleaf corydalis root resource is on the verge of exhaustion, at the same it is more difficult in introducing and planting survive, to preserving seed with it is larger tired using bringing
It is difficult.As a large amount of excavations and purchase cause wild resource to be on the verge of exhaustion, supply falls short of demand in market, and artificial cultivation planting cost is high and not
On a large scale, it is needed for situation to carry out economizing type tissue cultures factorial praluction.Using biotechnology tissue culture technique, Ke Yiyou
Effect rapidly improves the reproduction speed and quality of meadowrueleaf corydalis root seedling, realizes the industrial seedling rearing of meadowrueleaf corydalis root high quality seedling, to meet
Needs in production.
The key technology of tissue cultures is culture medium and its condition of culture.
Also fewer for the research of meadowrueleaf corydalis root Plantlet in vitro at present, applicant is authorizing Publication No. CN
Utilization is also disclosed that in a kind of 201310537770.5 Chinese invention patent " quick breeding method for tissue culture of meadowrueleaf corydalis root "
The method that tissue culture technique quickly breeds meadowrueleaf corydalis root, but in the present invention applicant by the strong sprout during meadowrueleaf corydalis root tissue-culturing rapid propagation
It is carried out at the same time with taking root, reduces a step, can reached and similarly expand numerous effect, therefore manpower will be saved using the present invention
Material resources also disclose meadowrueleaf corydalis root Plantlet in vitro culture medium and condition of culture, therefore, research of the invention and disclose with very
Important meaning.
Invention content
For presently, there are the problem of, the present invention provides a kind of in-vitro conservation method of meadowrueleaf corydalis root, utilizes tissue cultures skill
Art carries out it two step seedling-growing rapid breeding of tissue culture, and Plantlet in vitro is carried out to it by using suitable Storaged media, not only can be with
Effectively save meadowrueleaf corydalis root species in imminent danger, make renewable resource, can also be planted for meadowrueleaf corydalis root provide it is excellent consistent
Seedling technology and indoor Plantlet in vitro provide technical support.
To achieve the goals above, this invention takes following technical solutions：
（1）Adventitious bud induction culture Fiber differentiation：Meadowrueleaf corydalis root stem apex is taken as explant, impregnate 30 in 75% ethyl alcohol ~
45s, aseptic water washing 2 ~ 3 times put into 0.1% mercuric chloride 4min of 2 drop tweens of addition, then with aseptic water washing 2 ~ 3 times, again
0.1% mercuric chloride 4min of 2 drop tweens of input addition are finally seeded in being cut into 0.5 ~ 1.0cm long after aseptic water washing 2 ~ 3 times
It on inducing culture, cultivates 10 ~ 14 days, stem apex is differentiated to form light yellow green Multiple Buds；
（2）Multiple Buds strengthening seedling and rooting culture：Through 25 ~ 35 days, Multiple Buds were grown to 2 ~ 3cm, were transferred to the training of strengthening seedling and rooting culture medium
After supporting 1 month, grow up to the test tube seedling of tool root and 4 ~ 6 leaves, 4 or more radicals；
（3）Test tube seedling Plantlet in vitro：The test tube seedling with 4 ~ 6 leaves, 4 or more radicals that will be generated by differentiation
It is inoculated into Storaged media and carries out Plantlet in vitro.
Step（1）Described in inducing culture be：MS+6-BA1.5mg/l+NAA0.5 mg/l+KT0.4mg/l+ sugarcanes
Sugared 2.5%+agar 0.4%；The pH value of the culture medium is 5.8, and cultivation temperature is 24 ~ 28 °C, 1400 ~ 2000lx of intensity of illumination,
Light application time is 12 ~ 14 hours/day.
Step（2）Described in strengthening seedling and rooting culture medium be：MS+IAA0.6mg/l+6-BA0.3mg/l+ABT 0.5 mg/
L+AC0.5%+ sucrose 3.5%+ agar 0.4%；The pH value of the culture medium is 5.8, and temperature is 22 ~ 25 °C, intensity of illumination 2600 ~
2800lx, light application time are 12 ~ 14 hours/day.
Step（3）Described in Storaged media be 1/2MS+ PPP3331.0mg/l+AC0.3%+ sucrose 4.5%+ agar
0.4%；The pH value of the culture medium is 5.8, and temperature is 21 ± 1 °C, 1200 ~ 1600lx of intensity of illumination, and light application time is 6 ~ 8 small
When/day.
Beneficial effects of the present invention：
A kind of Chinese invention patent " the group of meadowrueleaf corydalis root of the present invention applicant in Publication No. CN 201310537770.5
Knit culture rapid propagation method " on the basis of carried out further improvement, by during meadowrueleaf corydalis root tissue-culturing rapid propagation strong sprout and life
Root is carried out at the same time, and is reduced a step, can be reached and similarly expand numerous effect, use the present invention will save human and material resources with
And the time, therefore simple and practicable, production cost reduces.
The present invention has broken the blank studied meadowrueleaf corydalis root Plantlet in vitro, and the method using the present invention can make germ plasm resource
Long-term preservation is obtained, and obtained seedling is healthy and strong, survival rate is high, can provide high-quality kind of a large amount of meadowrueleaf corydalis roots in a short time
Seedling efficiently solves the problems, such as meadowrueleaf corydalis root category Germ-plasma resources protection and scale breeding, is with a wide range of applications.
Description of the drawings
Influence Fig. 1 shows light intensity of the present invention to meadowrueleaf corydalis root Plantlet in vitro.
Wherein, abscissa indicates intensity of illumination（lx）, ordinate expression preservation number of days（d）.
Specific implementation mode
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
A kind of meadowrueleaf corydalis root category in-vitro conservation method, is realized by the following method：
One, meadowrueleaf corydalis root is through quickly breeding to obtain culture：
1, material to be tested
Meadowrueleaf corydalis root Corydalis saxicola Bunting stem apexs.
2, explant is disinfected
By the stem apex of selection, 30 ~ 45s, aseptic water washing 2 ~ 3 times, input 2% Tween-20 of addition are impregnated in 75% ethyl alcohol
0.1% mercuric chloride 4min, then with aseptic water washing 2 ~ 3 times, 0.1% mercuric chloride 4min of 2% Tween-20 of input addition, is finally used again
Aseptic water washing 2 ~ 3 times.
3, explant induces
It is cut into the segment of 0.5 ~ 1.0cm long after collected meadowrueleaf corydalis root stem apex is sterilized, is seeded in culture medium（MS+6-
2.5%+agar of BA1.5mg/l+NAA0.5 mg/l+KT0.4mg/l+ sucrose 0.4%）On, it cultivates 8 ~ 14 days, stem apex breaks up shape
At Multiple Buds, Multiple Buds are cleavable to be repeated to cultivate.On this culture medium, grow up to complete test tube seedling, for transplanting.The culture medium
PH value is 5.8, and cultivation temperature is 24 ~ 28 °C, 1400 ~ 2000lx of intensity of illumination, and light application time is 12 ~ 14 hours/day.
4, Multiple Buds strengthening seedling and rooting
Meadowrueleaf corydalis root terminal bud culture 16 days or so, can be obtained a large amount of Multiple Buds in previous step（70% or more）, and process 20 ~
Culture in 30 days can obtain high about 2 ~ 3cm sprouts, be transferred to strengthening seedling and rooting culture medium（Strengthening seedling and rooting culture medium is：MS+
0.5 mg/l+AC0.5%+ sucrose 3.5%+ agar 0.4% of IAA0.6mg/l+6-BA0.3mg/l+ABT）, the pH of the culture medium
Value is 5.8, and temperature is 22 ~ 25 °C, 2600 ~ 2800lx of intensity of illumination, and light application time is 12 ~ 14 hours/day.After culture 1 month
Reinforce light（3000Lx or more）, cultivate one month or so, test tube seedling can reach the standard of transplanting seedlings.Whole cycle is 2 ~ 3 months.
5, test tube seedling Plantlet in vitro
Select it is above-mentioned have 4 ~ 6 leaves, 4 or more radical meadowrueleaf corydalis root test tube seedlings for test material carry out Plantlet in vitro.
The screening of type of culture medium, temperature, intensity of illumination, growth inhibitor is carried out to it below, preserves number of days with examination
Pipe seedling survival rate is counted for 60%.
（1）, influence of the culture medium to meadowrueleaf corydalis root Plantlet in vitro
Test material is inoculated on tri- kinds of culture mediums of experimental design MS, 1/2MS, 1/4MS, is inoculated with 10 bottles, every bottle of 5 plants of lists
Bud；The pH value of agar containing 35g/L sucrose and 4g/L in culture medium, culture medium is 5.8,25 ± 1 DEG C of cultivation temperature, light intensity
2000lx, illumination 7h/d；The result shows that meadowrueleaf corydalis root holding time longest on MS culture mediums, is 163 days, but cultivated in 1/2MS
The holding time is 159 days on base, and to select 1/2MS be its Storaged media to be cost-effective, and concrete outcome is shown in Table 1.
Influence of 1 culture medium of table to the meadowrueleaf corydalis root Plantlet in vitro time
| Culture medium | Holding time（d） | Upgrowth situation |
| MS | 163 | Plant is light green, well-grown, starts after 4 months dead |
| 1/2MS | 159 | Plant is light green, well-grown, starts death after 5 months, |
| 1/4MS | 138 | Bud strain is slow-growing, after 3 months material hop to it is withered die, |
（2）, influence of the temperature to meadowrueleaf corydalis root Plantlet in vitro
Test material access 1/2MS culture mediums are respectively placed in 15 DEG C of temperature, 18 DEG C, 21 DEG C, 24 DEG C of illumination cultivation
Case culture is inoculated with 10 bottles, every bottle of 5 plants of simple buds；Agar containing 30g/L sucrose and 4.85g/L, the pH value of culture medium in culture medium
It is 5.8, light intensity 2000lx, illumination 7h/d, the results showed that, meadowrueleaf corydalis root holding time longest in temperature is 21 DEG C of incubators is
187 days；Specifically it is shown in Table 2.
Influence of 2 temperature of table to the meadowrueleaf corydalis root Plantlet in vitro time
（3）, influence of the intensity of illumination to meadowrueleaf corydalis root Plantlet in vitro
Test material is inoculated into 1/2MS culture mediums, design 0,1200lx, 1400lx, 1600lx, five kinds of light of 1800lx
According to intensity, each light intensity is inoculated with 10 bottles, every bottle of 5 plants of simple buds；Agar containing 35g/L sucrose and 4.5g/L in culture medium, culture
The pH value of base is 5.8,21 ± 1 DEG C of cultivation temperature, illumination 7h/d；The result shows that meadowrueleaf corydalis root is protected under the conditions of light intensity is 1600lx
Time longest is deposited, is 251 days, specific comparison result is shown in that Fig. 1, abscissa are to preserve number of days, and ordinate is intensity of illumination.
（4）, influence of the growth inhibitor to meadowrueleaf corydalis root Plantlet in vitro
Test material is inoculated into addition following various concentration CCC, PPP3331/2MS culture mediums on, be inoculated with 10 bottles, often
5 plants of simple buds of bottle；The pH value of agar containing 30g/L sucrose and 4.5g/L in culture medium, culture medium is 5.8, cultivation temperature 21 ± 1
DEG C, light intensity 1600lx, illumination 7h/d, the results showed that, meadowrueleaf corydalis root is in PPP333A concentration of 1.0 mg/L is holding time longest, is
291 days；Concrete condition is shown in Table 3.
Influence of 3 hormone of table to the meadowrueleaf corydalis root holding time
| Hormone | Concentration（mg/L） | Holding time（d） | Upgrowth situation |
| CCC | 0.8 | 209 | Plant is light green, well-grown, starts within 5 months dead |
| CCC | 1.0 | 243 | Plant is light green, well-grown, starts within 5 months dead |
| CCC | 1.2 | 238 | Plant is light green, well-grown, starts within 5 months dead |
| PP333 | 0.8 | 256 | Plant is light green, well-grown, starts within 6 months dead |
| PP333 | 1.0 | 291 | Plant is light green, well-grown, starts within 8 months dead |
| PP333 | 1.2 | 270 | Plant is light green, well-grown, starts within 6 months dead |
By above-mentioned experiment, determine that the best Plantlet in vitro culture medium of meadowrueleaf corydalis root is 1/2MS+ PP3331.0mg/l +AC0.3%
+ sucrose 3.5%+ agar 0.4%；The pH value of the culture medium is 5.8, and temperature is 21 ± 1 °C, 1200 ~ 1600lx of intensity of illumination, light
It is 6 ~ 8 hours/day according to the time.It is 291 days to preserve number of days.
5, upgrowth situation is investigated
The growth recovery situation of the meadowrueleaf corydalis root test tube seedling of Plantlet in vitro is investigated：By the meadowrueleaf corydalis root test tube seedling simple bud of robust growth
It cuts and is inoculated on best Storaged media after preservation 290d, the meadowrueleaf corydalis root test tube seedling of survival is inoculated into MS+6-BA1.5mg/l+
NAA0.3mg/l+KT0.3mg/l adds AC0.1%, adds on 3% sucrose and 0.45% agar medium, per 30d subcultures 1
It is secondary, after subculture 3 times, the recovery feelings of meadowrueleaf corydalis root test tube seedling are investigated using plant height, growth rate, breeding rate, single plant rooting rate as index
Condition is shown in Table 4.It is good from can be seen that the test tube seedling recovery situation after 290d Plantlet in vitro in comparison.
The comparison of test tube seedling before the test tube seedling restored after 4 Plantlet in vitro of table and preservation
| Material | Average life rate/% | Breeding coefficient/times | Rooting rate/% | Plant height/cm |
| Normally | 6.67±0.11 | 7.1±0.51 | 99.1±0.51 | 7.11±0.13 |
| It preserves | 6.29±0.24 | 6.7±0.33 | 98.3±0.34 | 6.67±0.22 |
It is 6-BA that substance used in the present invention annotates respectively（6-benzyl aminopurine）, NAA（Methyl α-naphthyl acetate）,KT（Excitement
Element）, IAA (heteroauxin), AC (activated carbon), ABT (root-inducing powder), CCC（Cycocel）, PPP333（Paclobutrazol）.
Claims (1)
1. a kind of in-vitro conservation method of meadowrueleaf corydalis root, which is characterized in that include the following steps：
（1）Adventitious bud induction culture：It takes meadowrueleaf corydalis root stem apex as explant, 30 ~ 45s, aseptic water washing is impregnated in 75% ethyl alcohol
2 ~ 3 times, 0.1% mercuric chloride 4min of 2 drop tweens of input addition, then with aseptic water washing 2 ~ 3 times, tweens are dripped in input addition 2 again
0.1% mercuric chloride 4min is finally seeded on inducing culture with being cut into 0.5 ~ 1.0cm long after aseptic water washing 2 ~ 3 times, is cultivated
10 ~ 14 days, stem apex was differentiated to form light yellow green Multiple Buds；
（2）Multiple Buds strengthening seedling and rooting culture：Through 25 ~ 35 days, Multiple Buds were grown to 2 ~ 3cm, were transferred to strengthening seedling and rooting medium culture 1
After month, grow up to the test tube seedling of tool root and 4 ~ 6 leaves；
（3）Test tube seedling Plantlet in vitro：The test tube seedling with 4 ~ 6 leaves that will be generated by differentiation is inoculated into preservation culture
Base carries out Plantlet in vitro；
Step（1）Described in inducing culture be：MS+6-BA1.5mg/l+NAA0.5 mg/l+KT0.4mg/l+ sucrose
2.5%+agar 0.4%；The pH value of the culture medium is 5.8, and cultivation temperature is 24 ~ 28 °C, 1400 ~ 2000lx of intensity of illumination, light
It is 12 ~ 14 hours/day according to the time；
Step（2）Described in strengthening seedling and rooting culture medium be：MS+IAA0.6mg/l+6-BA0.3mg/l+ABT 0.5 mg/l +
AC0.5%+ sucrose 3.5%+ agar 0.4%；The pH value of the culture medium is 5.8, and temperature is 22 ~ 25 °C, intensity of illumination 2600 ~
2800lx, light application time are 12 ~ 14 hours/day；
Step（3）Described in Storaged media be 1/2MS+ PPP3331.0mg/l+AC0.3%+ sucrose 4.5%+ agar 0.4%；
The pH value of the culture medium is 5.8, and temperature is 21 ± 1 °C, 1200 ~ 1600lx of intensity of illumination, and light application time is 6 ~ 8 hours/day.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610108551.9A CN105746347B (en) | 2016-02-29 | 2016-02-29 | A kind of in-vitro conservation method of meadowrueleaf corydalis root |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610108551.9A CN105746347B (en) | 2016-02-29 | 2016-02-29 | A kind of in-vitro conservation method of meadowrueleaf corydalis root |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105746347A CN105746347A (en) | 2016-07-13 |
| CN105746347B true CN105746347B (en) | 2018-09-25 |
Family
ID=56330481
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610108551.9A Expired - Fee Related CN105746347B (en) | 2016-02-29 | 2016-02-29 | A kind of in-vitro conservation method of meadowrueleaf corydalis root |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105746347B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106635951A (en) * | 2016-12-28 | 2017-05-10 | 安徽檀鑫科技有限公司 | Culture medium for cultivating corydalis saxicola bunting cell and preparation method of culture medium |
| CN106718923A (en) * | 2016-12-30 | 2017-05-31 | 广西壮族自治区药用植物园 | Meadowrueleaf corydalis root callus highly effective revulsion induction method |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103548695B (en) * | 2013-11-04 | 2015-10-21 | 广西壮族自治区药用植物园 | A kind of meadowrueleaf corydalis root quick breeding method for tissue culture |
-
2016
- 2016-02-29 CN CN201610108551.9A patent/CN105746347B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN105746347A (en) | 2016-07-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101103700B (en) | Tissue culture quick reproduction method of radix tetrastigmae | |
| CN102369881B (en) | Rapid propagation technique of Dendrobium candidum axillary buds | |
| CN102301952B (en) | Method for breeding chamomile | |
| CN103563746A (en) | Method for culturing shoot apical meristem of dendranthema morifolium pamat | |
| CN104996298A (en) | Method for cultivating polygala fallax hemsl tissue culture seedlings based on multiple internodal stem segments integration | |
| CN101953300B (en) | Tissue culture method for Curcuma wenyujin No.1 | |
| CN106538387B (en) | A kind of method for tissue culture of Ku Zhi | |
| CN105746347B (en) | A kind of in-vitro conservation method of meadowrueleaf corydalis root | |
| CN104488723A (en) | Tissue-culture and rapid-propagation method of epimedium koreanum nakai | |
| CN104996304B (en) | Culture medium and culture method for inducing callus differentiation through peony leaves | |
| CN103548695B (en) | A kind of meadowrueleaf corydalis root quick breeding method for tissue culture | |
| CN104285792B (en) | A kind of tissue rapid propagation method of Alnus formosana Plantation | |
| CN105660415B (en) | A kind of in-vitro conservation method of Mongolian oak fern | |
| CN102499082A (en) | Test tube breeding method of lilium oriental hybrid seed ball | |
| CN107711514A (en) | A kind of nonirrigated farmland rose of Sharon purple flower system fine individual plant tissue culture and rapid propagation method | |
| CN104813932A (en) | In vitro preservation method for Dendrobium officinale | |
| CN106922527B (en) | The quick breeding by group culture method of henry clematis | |
| CN114190277B (en) | Method for promoting blooming and fructification of large root orchid test tube | |
| CN106613970B (en) | The quick breeding by group culture method of sealwort leaf elegant jessamine | |
| CN105191795A (en) | Rapid propagation method of gold leaf metasequoia tissue culture | |
| CN102228006B (en) | Method for quickly growing seedling of wild buckwheat rhizome by tissue culture | |
| CN106561453B (en) | The rapid propagation method of Campanumoea javanica | |
| CN108094200A (en) | A kind of be heat-treated combines the breeding method that stem apex stripping acquisition peace ancestral spends detoxic seedling | |
| CN104756863A (en) | In-vitro conservation method for Hemiboea follicularis Clarke | |
| CN104904592B (en) | A kind of in-vitro conservation method of Hemiboea lungzhouensis W. T Wang ex Z. Y. Li. |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180925 Termination date: 20190228 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |