CN105746347A - In-vitro preservation method of corydalis thalictrifolia - Google Patents

In-vitro preservation method of corydalis thalictrifolia Download PDF

Info

Publication number
CN105746347A
CN105746347A CN201610108551.9A CN201610108551A CN105746347A CN 105746347 A CN105746347 A CN 105746347A CN 201610108551 A CN201610108551 A CN 201610108551A CN 105746347 A CN105746347 A CN 105746347A
Authority
CN
China
Prior art keywords
corydalis
culture medium
thalictrifolia franch
corydalis thalictrifolia
thalictrifoliae
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.)
Granted
Application number
CN201610108551.9A
Other languages
Chinese (zh)
Other versions
CN105746347B (en
Inventor
张占江
李翠
郭晓云
赵以民
李林轩
缪剑华
韦莹
韦坤华
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Guangxi Botanical Garden of Medicinal Plants
Original Assignee
Guangxi Botanical Garden of Medicinal Plants
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Guangxi Botanical Garden of Medicinal Plants filed Critical Guangxi Botanical Garden of Medicinal Plants
Priority to CN201610108551.9A priority Critical patent/CN105746347B/en
Publication of CN105746347A publication Critical patent/CN105746347A/en
Application granted granted Critical
Publication of CN105746347B publication Critical patent/CN105746347B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

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 an in-vitro preservation method of corydalis thalictrifolia. The method comprises the following steps: performing tissue culture two-step seedling development on corydalis thalictrifolia, and inoculating test-tube seedlings with 4-6 leaves obtained by utilizing tissue culture two-step seedling development into a preservation medium for performing in-vitro preservation. The preservation medium refers to 1/2MS+PPP3331.0mg/l+0.3% of AC+ 4.5% of cane sugar+0.4 % of agar. The pH value of the medium is 5.8, the temperature is 21+/-1 DEG C, the illumination intensity is 1400-1600lx, and the illumination time is 8-10 hours per day. According to the method disclosed by the invention, lots of high quality seedlings of corydalis thalictrifolia can be provided in a short time, the steps of preparing the medium and spinning the bottle are reduced, and lots of manpower and material resources are saved; and therefore, the method is simple and feasible, the production cost is reduced, and the purpose of preservation and large-scale seedling growing of corydalis thalictrifolia germplasm resources can be effectively achieved.

Description

A kind of in-vitro conservation method of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae)
Technical field
The invention belongs to biological tissue's culture technique field, in particular it relates to the in-vitro conservation method of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae).
Background technology
Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) (CorydalissaxicolaBunting) is bloodroot, for herbaceos perennial, is distributed in China Guangxi, Guangdong, Fujian, Hunan etc. and economizes.All herbal medicine, is famous strong precious jade medicine kind, and herb is containing deydrokaividing (Corydalis Saxicolae) isoreactivity composition；There is antibacterial, antiinflammatory, analgesia and strong stable effect significantly, and observed suppression tumor cell effect；Cure mainly the disease Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) plant distributions such as furuncle toxic swelling, hepatitis, liver cirrhosis, hepatocarcinoma and be confined to Limestone Mountain Areas, belong to tor endemic species.Owing to habitat conditions is severe, its natural propagation rate is very low, adds that its seed is tiny, breeds relatively difficult, and population development obstacles, resource reserves are extremely limited.People excavate in a large number and purchase in recent years, and supply falls short of demand to result in wild resource, and wild Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) resource is on the verge of exhaustion, simultaneously more difficult when introducing and planting survive, and bring bigger difficulty to preserving seed with utilizing.Along with excavating in a large number and purchase causes that wild resource is on the verge of exhaustion, supply falls short of demand in market, and artificial culture planting cost is high and not on a large scale, and carrying out economizing type tissue culture factorial praluction is needed for situation.Adopt biotechnology tissue culture technique, it is possible to effectively and rapidly improve reproduction speed and the quality of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) seedling, it is achieved the industrial seedling rearing of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) high quality seedling, with the needs in satisfied production.
The key technology of tissue culture is culture medium and condition of culture thereof.
Also fewer for the research of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) in vitro conservation at present, applicant also discloses that, in the Chinese invention patent " quick breeding method for tissue culture of a kind of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) " authorizing publication number to be CN201310537770.5, the method utilizing tissue culture technique Fast-propagation Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae), but the strong sprout in Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) tissue-culturing rapid propagation process is carried out by applicant with taking root in the present invention simultaneously, decrease a step, same expanding propagation effect can be reached, therefore the present invention is adopted to use manpower and material resources sparingly, also disclose Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) in vitro conservation culture medium and condition of culture, therefore, research and the open tool of the present invention are of great significance.
Summary of the invention
For the problem that presently, there are, the present invention provides the in-vitro conservation method of a kind of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae), utilize tissue culture technique that it is carried out group training two step seedling-growing rapid breedings, by adopting suitable Storaged media that it is carried out in vitro conservation, it is possible not only to effectively save Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) species in imminent danger, make Renewable resource, it is also possible to provide excellent consistent seedling technology and indoor in vitro conservation to provide technical support for Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) plantation.
To achieve these goals, the present invention has taken following technical scheme:
(1) adventitious bud induction culture inducing culture: take Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) stem apex as outer implant, 75% ethanol soaks 30 ~ 45s, aseptic water washing 2 ~ 3 times, put into the 0.1% mercuric chloride 4min adding 2 tweens, again with aseptic water washing 2 ~ 3 times, again put into the 0.1% mercuric chloride 4min adding 2 tweens, be finally seeded on inducing culture with being cut into 0.5 ~ 1.0cm length after aseptic water washing 2 ~ 3 times, cultivating 10 ~ 14 days, stem apex is differentiated to form light yellow green Multiple Buds；
(2) Multiple Buds strengthening seedling and rooting is cultivated: through 25 ~ 35 days, Multiple Buds length, to 2 ~ 3cm, proceeded to strengthening seedling and rooting culture medium culturing after 1 month, grows up to the test tube Seedling of tool root and 4 ~ 6 leaves, more than 4 radicals；
(3) test tube Seedling in vitro conservation: by produced by differentiation there are 4 ~ 6 leaves, the test tube Seedling of more than 4 radicals is inoculated into Storaged media and carries out in vitro conservation.
Inducing culture described in step (1) is: MS+6-BA1.5mg/l+NAA0.5mg/l+KT0.4mg/l+ sucrose 2.5%+ agar 0.4%；The pH value of described culture medium is 5.8, and cultivation temperature is 24 ~ 28 ° of C, intensity of illumination 1400 ~ 2000lx, and light application time is 12 ~ 14 hours/day.
Strengthening seedling and rooting culture medium described in step (2) is: MS+IAA0.6mg/l+6-BA0.3mg/l+ABT0.5mg/l+AC0.5%+ sucrose 3.5%+ agar 0.4%；The pH value of described culture medium is 5.8, and temperature is 22 ~ 25 ° of C, intensity of illumination 2600 ~ 2800lx, and light application time is 12 ~ 14 hours/day.
Storaged media described in step (3) is 1/2MS+PPP3331.0mg/l+AC0.3%+ sucrose 4.5%+ agar 0.4%；The pH value of described culture medium is 5.8, and temperature is 21 ± 1 ° of C, intensity of illumination 1200 ~ 1600lx, and light application time is 6 ~ 8 hours/day.
Beneficial effects of the present invention:
Present invention applicant has carried out further improvement on Chinese invention patent " quick breeding method for tissue culture of a kind of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) " basis that publication number is CN201310537770.5, strong sprout in Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) tissue-culturing rapid propagation process is carried out with taking root simultaneously, decrease a step, same expanding propagation effect can be reached, adopting the present invention will save human and material resources and time, therefore simple, production cost reduces.
The present invention has broken the blank to the research of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) in vitro conservation; the method utilizing the present invention can make germ plasm resource obtain long-term preservation; and the seedling obtained is healthy and strong, survival rate is high; the high quality seedling of a large amount of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) can be provided in a short time; efficiently solve Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) and belong to Germ-plasma resources protection and scale breeding problem, be with a wide range of applications.
Accompanying drawing explanation
Fig. 1 represents the light intensity of the present invention impact on Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) in vitro conservation.
Wherein, abscissa represents intensity of illumination (lx), and vertical coordinate represents preservation natural law (d).
Detailed description of the invention
The experimental technique used in following embodiment if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, all commercially obtain.
A kind of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) belongs to in-vitro conservation method, is realized by the following method:
One, Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) obtains culture through Fast-propagation:
1, material to be tested
Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) CorydalissaxicolaBunting stem apex.
2, the disinfecting of outer implant
The stem apex that will select, soaks 30 ~ 45s, aseptic water washing 2 ~ 3 times in 75% ethanol, put into the 0.1% mercuric chloride 4min adding 2% tween 20, use aseptic water washing 2 ~ 3 times again, again put into the 0.1% mercuric chloride 4min adding 2% tween 20, finally use aseptic water washing 2 ~ 3 times.
3, outer implant induction
The segment of 0.5 ~ 1.0cm length will be cut into after the Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) stem apex sterilizing collected, it is seeded in culture medium (MS+6-BA1.5mg/l+NAA0.5mg/l+KT0.4mg/l+ sucrose 2.5%+ agar 0.4%), cultivating 8 ~ 14 days, stem apex is differentiated to form Multiple Buds, and Multiple Buds can cut and repeat to cultivate.In this culture medium, grow up to complete test tube Seedling, for transplanting.The pH value of described culture medium is 5.8, and cultivation temperature is 24 ~ 28 ° of C, intensity of illumination 1400 ~ 2000lx, and light application time is 12 ~ 14 hours/day.
4, Multiple Buds strengthening seedling and rooting
In previous step, Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) terminal bud is cultivated about 16 days, a large amount of Multiple Buds (more than 70%) can be obtained, high about 2 ~ 3cm sprout can be obtained again through the cultivation of 20 ~ 30 days, proceed to strengthening seedling and rooting culture medium (strengthening seedling and rooting culture medium is: MS+IAA0.6mg/l+6-BA0.3mg/l+ABT0.5mg/l+AC0.5%+ sucrose 3.5%+ agar 0.4%), the pH value of described culture medium is 5.8, temperature is 22 ~ 25 ° of C, intensity of illumination 2600 ~ 2800lx, and light application time is 12 ~ 14 hours/day.Adding high light (3000Lx and more than) after cultivating 1 month, cultivate about one month, test tube Seedling can reach the standard of transplanting seedlings.The whole cycle is 2 ~ 3 months.
5, test tube Seedling in vitro conservation
Select above-mentioned have 4 ~ 6 leaves, more than 4 radical Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) test tube Seedlings are that test material carries out in vitro conservation.
Below it is carried out the screening of type of culture medium, temperature, intensity of illumination, growth inhibitor, preserve natural law and all add up for 60% with test tube Seedling survival rate.
(1), the culture medium impact on Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) in vitro conservation
Test material is inoculated in EXPERIMENTAL DESIGN MS, tri-kinds of culture medium of 1/2MS, 1/4MS, inoculates 10 bottles, every bottle of 5 strain simple buds；Containing the agar of 35g/L sucrose and 4g/L in culture medium, the pH value of culture medium is 5.8, cultivation temperature 25 ± 1 DEG C, light intensity 2000lx, illumination 7h/d；It is shown that Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) holding time in MS culture medium is the longest, be 163 days, but in 1/2MS culture medium the holding time be 159 days, for save cost select 1/2MS to be its Storaged media, concrete outcome is in Table 1.
The impact on the Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) in vitro conservation time of table 1 culture medium
Culture medium Holding time (d) Upgrowth situation
MS 163 Plant is light green, well-grown, starts death after 4 months
1/2MS 159 Plant is light green, well-grown, starts death after 5 months,
1/4MS 138 Bud strain poor growth, after 3 months, material hops to it withered dying,
(2), the temperature impact on Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) in vitro conservation
Test material accessing 1/2MS culture medium and is respectively placed in temperature 15 DEG C, 18 DEG C, the illumination box of 21 DEG C, 24 DEG C is cultivated, and inoculates 10 bottles, every bottle of 5 strain simple buds；Agar containing 30g/L sucrose and 4.85g/L in culture medium, the pH value of culture medium is 5.8, light intensity 2000lx, illumination 7h/d, it is shown that Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) temperature be in 21 DEG C of incubators the holding time the longest, be 187 days；It is specifically shown in table 2.
The impact on the Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) in vitro conservation time of table 2 temperature
(3), the intensity of illumination impact on Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) in vitro conservation
Test material is inoculated into 1/2MS culture medium, design 0,1200lx, 1400lx, five kinds of intensities of illumination of 1600lx, 1800lx, and every kind of light intensity inoculates 10 bottles, every bottle of 5 strain simple buds；Containing the agar of 35g/L sucrose and 4.5g/L in culture medium, the pH value of culture medium is 5.8, cultivation temperature 21 ± 1 DEG C, illumination 7h/d；It is shown that Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) holding time when light intensity is 1600lx is the longest, being 251 days, concrete comparative result is shown in Fig. 1, and abscissa is for preserving natural law, and vertical coordinate is intensity of illumination.
(4), the growth inhibitor impact on Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) in vitro conservation
Test material is inoculated into interpolation following variable concentrations CCC, PPP3331/2MS culture medium on, inoculate 10 bottles, every bottle of 5 strain simple buds；Containing the agar of 30g/L sucrose and 4.5g/L in culture medium, the pH value of culture medium is 5.8, cultivation temperature 21 ± 1 DEG C, light intensity 1600lx, and illumination 7h/d, it is shown that Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) is at PPP333Concentration is 1.0mg/L is that the holding time is the longest, is 291 days；Concrete condition is in Table 3.
The impact on the Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) holding time of table 3 hormone
Hormone Concentration (mg/L) Holding time (d) Upgrowth situation
CCC 0.8 209 Plant is light green, well-grown, and 5 months start death
CCC 1.0 243 Plant is light green, well-grown, and 5 months start death
CCC 1.2 238 Plant is light green, well-grown, and 5 months start death
PP333 0.8 256 Plant is light green, well-grown, and 6 months start death
PP333 1.0 291 Plant is light green, well-grown, and 8 months start death
PP333 1.2 270 Plant is light green, well-grown, and 6 months start death
By above-mentioned experiment, it is determined that Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) the best in vitro conservation culture medium is 1/2MS+PP3331.0mg/l+AC0.3%+ sucrose 3.5%+ agar 0.4%；The pH value of described culture medium is 5.8, and temperature is 21 ± 1 ° of C, intensity of illumination 1200 ~ 1600lx, and light application time is 6 ~ 8 hours/day.Preserving natural law is 291 days.
5, upgrowth situation is investigated
The growth recovery situation of the Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) test tube Seedling of in vitro conservation is investigated: is cut by the Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) test tube Seedling simple bud of robust growth and is inoculated on best Storaged media after preservation 290d, the Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) test tube Seedling of survival is inoculated into MS+6-BA1.5mg/l+NAA0.3mg/l+KT0.3mg/l, add AC0.1%, add on the sucrose of 3% and the agar culture medium of 0.45%, every 30d subculture 1 time, after subculture 3 times, the recovery situation of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) test tube Seedling is investigated, in Table 4 with plant height, rate of growth, breeding rate, individual plant rooting rate for index.It can be seen that the test tube Seedling recovery situation after 290d in vitro conservation is good from contrast.
The test tube Seedling recovered after table 4 in vitro conservation and the comparison preserving front test tube Seedling
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
Preserve 6.29±0.24 6.7±0.33 98.3±0.34 6.67±0.22
Material used in the present invention annotates respectively as 6-BA(6-benayl aminopurine), NAA(naphthalene acetic acid), KT(kinetins), IAA (heteroauxing), AC (activated carbon), ABT (root-inducing powder), CCC(chlorocholine chloride), PPP333(paclobutrazol).

Claims (4)

1. the in-vitro conservation method of a Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae), it is characterised in that comprise the steps:
(1) adventitious bud induction culture: take Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) stem apex as outer implant, 75% ethanol soaks 30 ~ 45s, aseptic water washing 2 ~ 3 times, put into the 0.1% mercuric chloride 4min adding 2 tweens, again with aseptic water washing 2 ~ 3 times, again put into the 0.1% mercuric chloride 4min adding 2 tweens, be finally seeded on inducing culture with being cut into 0.5 ~ 1.0cm length after aseptic water washing 2 ~ 3 times, cultivating 10 ~ 14 days, stem apex is differentiated to form light yellow green Multiple Buds；
(2) Multiple Buds strengthening seedling and rooting is cultivated: through 25 ~ 35 days, Multiple Buds length, to 2 ~ 3cm, proceeded to strengthening seedling and rooting culture medium culturing after 1 month, grows up to the test tube Seedling of tool root and 4 ~ 6 leaves；
(3) test tube Seedling in vitro conservation: the test tube Seedling with 4 ~ 6 leaves that will be produced by differentiation, is inoculated into Storaged media and carries out in vitro conservation.
2. the in-vitro conservation method of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) as claimed in claim 1, it is characterised in that the inducing culture described in step (1) is: MS+6-BA1.5mg/l+NAA0.5mg/l+KT0.4mg/l+ sucrose 2.5%+ agar 0.4%；The pH value of described culture medium is 5.8, and cultivation temperature is 24 ~ 28 ° of C, intensity of illumination 1400 ~ 2000lx, and light application time is 12 ~ 14 hours/day.
3. the in-vitro conservation method of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) as claimed in claim 1, it is characterised in that the strengthening seedling and rooting culture medium described in step (2) is: MS+IAA0.6mg/l+6-BA0.3mg/l+ABT0.5mg/l+AC0.5%+ sucrose 3.5%+ agar 0.4%；The pH value of described culture medium is 5.8, and temperature is 22 ~ 25 ° of C, intensity of illumination 2600 ~ 2800lx, and light application time is 12 ~ 14 hours/day.
4. the in-vitro conservation method of Corydalis thalictrifolia Franch. (Radix Corydalis Thalictrifoliae) as claimed in claim 1, it is characterised in that the Storaged media described in step (3) is 1/2MS+PPP3331.0mg/l+AC0.3%+ sucrose 4.5%+ agar 0.4%；The pH value of described culture medium is 5.8, and temperature is 21 ± 1 ° of C, intensity of illumination 1200 ~ 1600lx, and light application time is 6 ~ 8 hours/day.
CN201610108551.9A 2016-02-29 2016-02-29 A kind of in-vitro conservation method of meadowrueleaf corydalis root Expired - Fee Related CN105746347B (en)

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 true CN105746347A (en) 2016-07-13
CN105746347B 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)

Cited By (2)

* Cited by examiner, † Cited by third party
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

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103548695A (en) * 2013-11-04 2014-02-05 广西壮族自治区药用植物园 Tissue culture and rapid propagation method for corydalis saxicola bunting

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103548695A (en) * 2013-11-04 2014-02-05 广西壮族自治区药用植物园 Tissue culture and rapid propagation method for corydalis saxicola bunting

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
张娟: "柑橘试管苗种质保存及其内源激素变化研究", 《中国优秀硕士学位论文全文数据库农业科技辑》 *
张明海等: "《魔芋高产栽培与加工技术》", 31 December 2007, 湖北科学技术出版社 *

Cited By (2)

* Cited by examiner, † Cited by third party
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

Also Published As

Publication number Publication date
CN105746347B (en) 2018-09-25

Similar Documents

Publication Publication Date Title
CN101258835B (en) Fast reproducing method for high quality seedling of dendrobium officinale
CN101531991B (en) Rhodida plant callus and suspension cell granule culture method
CN102150624A (en) Tissue culture and rapid propagation method for pinellia tuber plant
CN106538387B (en) A kind of method for tissue culture of Ku Zhi
CN102405830B (en) Induction method of rosa chinensis receptacle callus tissues
CN101953300B (en) Tissue culture method for Curcuma wenyujin No.1
CN104488723A (en) Tissue-culture and rapid-propagation method of epimedium koreanum nakai
CN106973796A (en) A kind of tissue cultivating and seedling method of Idesia polycarpa
CN104285792B (en) A kind of tissue rapid propagation method of Alnus formosana Plantation
CN103548695B (en) A kind of meadowrueleaf corydalis root quick breeding method for tissue culture
CN105746352A (en) Tissue culture method of succulent Haworthia emelyae v.comptoniana 'KYODAI AKASEN' HO1
CN105746347B (en) A kind of in-vitro conservation method of meadowrueleaf corydalis root
CN104813932A (en) In vitro preservation method for Dendrobium officinale
CN107711514A (en) A kind of nonirrigated farmland rose of Sharon purple flower system fine individual plant tissue culture and rapid propagation method
CN110663555B (en) Rapid propagation method of odorous jasminum grandiflorum
CN105660415B (en) A kind of in-vitro conservation method of Mongolian oak fern
CN100420741C (en) Agrobacterium rhizogenes mediated aconitum coreanum and its obtaining method
CN102499082A (en) Test tube breeding method of lilium oriental hybrid seed ball
CN102511393A (en) Method for establishing racomitrium japonicum gametophyte regeneration system
CN106613970B (en) The quick breeding by group culture method of sealwort leaf elegant jessamine
CN108094200A (en) A kind of be heat-treated combines the breeding method that stem apex stripping acquisition peace ancestral spends detoxic seedling
CN104756864B (en) In-vitro conservation method for Hemiboea cavaleriei var. paucinervis
KR20080073388A (en) Mass production of bulblet via somatic embryogenic cell culture in lily
CN104756863B (en) A kind of in-vitro conservation method of south China half capsule lettuce tongue
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