CN109169271A - A method of use Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant - Google Patents
A method of use Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant Download PDFInfo
- Publication number
- CN109169271A CN109169271A CN201811021459.4A CN201811021459A CN109169271A CN 109169271 A CN109169271 A CN 109169271A CN 201811021459 A CN201811021459 A CN 201811021459A CN 109169271 A CN109169271 A CN 109169271A
- Authority
- CN
- China
- Prior art keywords
- stem
- segment
- node
- plant
- explant
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
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
- A01H4/001—Culture apparatus for tissue culture
-
- 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 methods for using Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant, comprising the following steps: the Yunnan rough gentian plant for choosing robust growth takes its stem-segment with node, cleans, impregnates, rinsing and is after disinfection to reserve；Stem-segment with node is inoculated into pre-culture medium and obtains preculture stem-segment with node；Preculture stem-segment with node is taken out and obtains evoked callus in access induced medium；Evoked callus is accessed in proliferated culture medium and obtains proliferation callus；Proliferation callus is accessed in differential medium, is to carry out obtaining seedling in illumination cultivation 40~60 days under 3500~4500Lx in 25~30 DEG C of temperature, illumination；Seedling is accessed in root media, is to carry out obtaining rooted seedling in low light culture 20~25 days under 1500~2500Lx in 25~30 DEG C of temperature, illumination.The test tube seedling that the method for the present invention is produced, yield 2 times or more at least high compared with prior seed type of seeding.
Description
Technical field
The invention belongs to technical field of cultivation, and in particular to a kind of to use Yunnan rough gentian stem-segment with node for explant regeneration plant
Method.
Background technique
Yunnan rough gentian is Gentianaceae Gentiana perennial herb plant also known as Gentiana rigescens, four leaf gallbladders, radix gentianae, Yunnan dragon
Gallbladder grass, orchid root etc., are distributed mainly on the ground such as Yunnan, Sichuan, Guangxi and Guizhou, are born in 1350~3500m of height above sea level famine on the sunny side more
Ground, careless slope and sparse woods, wherein Yunnan is its Genuine producing area.As social economy rapidly develops, living standard is gradually increased, people
Specific gravity on health investment is continuously increased, to meet the market demand, develop it is a kind of can Yunnan rough gentian Different Organs be used for material
The method for establishing induction and breeding system is very important.
Summary of the invention
The purpose of the present invention is to provide a kind of methods for using Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant.
The object of the present invention is achieved like this, and described uses Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant
Method the following steps are included:
1) the Yunnan rough gentian plant for choosing robust growth, takes its stem-segment with node, cleans, impregnates, rinsing and is after disinfection to reserve；
2) stem-segment with node is inoculated into pre-culture medium, obtains preculture band in 20~25 DEG C of temperature dark culture 5~10 days
Stem segment；
3) by preculture stem-segment with node take out access induced medium on, at 25~30 DEG C of temperature dark culture 25~
Obtain evoked callus within 35 days；
4) evoked callus is accessed in proliferated culture medium, in 25~30 DEG C of temperature, illumination under 1500~2500Lx
It carries out obtaining within low light culture 35~40 days proliferation callus；
5) proliferation callus is accessed in differential medium, in 25~30 DEG C of temperature, illumination under 3500~4500Lx
It carries out obtaining seedling in illumination cultivation 40~60 days；
6) seedling is accessed in root media, is to carry out dim light under 1500~2500Lx in 25~30 DEG C of temperature, illumination
Culture obtains rooted seedling in 20~25 days；
Wherein, the pre-culture medium are as follows: MS+ 2.5~3.0g/L+ of plant gel sucrose 30%, pH5.0~7.0；
The induced medium are as follows: MS+2,4-D 4.0~6.0mg/L+ plant gel 2.5~3.0g/L+ sucrose
30%, pH5.0~7.0；
The proliferated culture medium are as follows: MS+2,4-D 5.0~7.0mg/L+ plant gel 2.5~3.0g/L+ sucrose
30%, pH5.0~7.0；
The differential medium are as follows: MS+6-BA 6mg/L+NAA 0.4~0.6mg/L+KT, 1~3mg/L+ plant is solidifying
2.5~3.0g/L+ of glue sucrose 30%, pH5.0~7.0；
The root media are as follows: MS+IAA 0.2~0.4mg/L+TDZ, 0.5~1.5mg/L+ plant gel 2.5~
3.0g/L+ sucrose 30%, pH5.0~7.0.
The present invention using stem-segment with node be used as explant, in specific culture medium culture after, value-added coefficient up to 65.0 with
On, general breeding coefficient is mostly 5~10 in Gentianaceae, and the present invention adds the PP333 of high concentration in continuing culture medium
Proliferation is synchronous with rejuvenation in (10-20mg/L) carries out, although growth coefficient is declined (20.0 or more), indefinite clump bud is thick
It is strong, branch is few, the selection for the material that is conducive to take root；The PP333 (0.5-1.0mg/L) of low quality concentration is added in root media
Middle inducing adventitious root occurs.When culture of rootage, root induction is carried out only with the stem apex saved in indefinite clump bud with 2-3,
He is partially used as fertile material to continue to cultivate.In this way, no callus and adventitious bud occur after rooting of vitro seedling, the short strong, blade of single seedling
In blackish green, root system more than cauline leaf prosperity；Since the medicinal part of Yunnan rough gentian is exactly its root, produced by the method for the present invention
Test tube seedling, under spring, autumn and winter excavation when, yield 2 times or more at least high compared with prior seed type of seeding.
Specific embodiment
It is of the present invention use Yunnan rough gentian stem-segment with node for explant cultivate regeneration plant method the following steps are included:
1) the Yunnan rough gentian plant for choosing robust growth, takes its stem-segment with node, cleans, impregnates, rinsing and is after disinfection to reserve；
2) stem-segment with node is inoculated into pre-culture medium, obtains preculture band in 20~25 DEG C of temperature dark culture 5~10 days
Stem segment；
3) by preculture stem-segment with node take out access induced medium on, at 25~30 DEG C of temperature dark culture 25~
Obtain evoked callus within 35 days；
4) evoked callus is accessed in proliferated culture medium, in 25~30 DEG C of temperature, illumination under 1500~2500Lx
It carries out obtaining within low light culture 35~40 days proliferation callus；
5) proliferation callus is accessed in differential medium, in 25~30 DEG C of temperature, illumination under 3500~4500Lx
It carries out obtaining seedling in illumination cultivation 40~60 days；
6) seedling is accessed in root media, is to carry out dim light under 1500~2500Lx in 25~30 DEG C of temperature, illumination
Culture obtains rooted seedling in 20~25 days；
Wherein, the pre-culture medium are as follows: MS+ 2.5~3.0g/L+ of plant gel sucrose 30%, pH5.0~7.0；
The induced medium are as follows: MS+2,4-D 4.0~6.0mg/L+ plant gel 2.5~3.0g/L+ sucrose
30%, pH5.0~7.0；
The proliferated culture medium are as follows: MS+2,4-D 5.0~7.0mg/L+ plant gel 2.5~3.0g/L+ sucrose
30%, pH5.0~7.0；
The differential medium are as follows: MS+6-BA 6mg/L+NAA 0.4~0.6mg/L+KT, 1~3mg/L+ plant is solidifying
2.5~3.0g/L+ of glue sucrose 30%, pH5.0~7.0；
The root media are as follows: MS+IAA 0.2~0.4mg/L+TDZ, 0.5~1.5mg/L+ plant gel 2.5~
3.0g/L+ sucrose 30%, pH5.0~7.0.
The cleaning is circulating water rinsing.
The immersion is to impregnate 8~12min with 10% washing powder solution of mass percent.
The flushing is to rinse 20~40min using flowing water.
The disinfection is to handle 3~5s using 75% ethanol solution, then again through the water-soluble liquid disinfectant of 0.1% mercuric chloride 7~
9min。
The pre-culture medium is MS+ plant gel 2.8g/L+ sucrose 30%, pH6.0.
The induced medium is MS+2,4-D 5.0mg/L+ plant gel 2.8g/L+ sucrose 30%, pH6.0；
The proliferated culture medium is MS+2,4-D 6.0mg/L+ plant gel 2.8g/L+ sucrose 30%, pH6.0.
The differential medium is MS+6-BA 6mg/L+NAA 0.5mg/L+KT 2mg/L+ plant gel 2.8g/L+
Sucrose 30%, pH6.0.
The root media is MS+IAA 0.3mg/L+TDZ 1.0mg/L+ plant gel 2.8g/L+ sucrose 30%,
pH6.0。
Embodiment 1
1) the Yunnan rough gentian plant for choosing robust growth, takes its stem-segment with node, cleans, impregnates, rinsing and is after disinfection to reserve；
2) stem-segment with node is inoculated into pre-culture medium, obtains preculture band in 20~25 DEG C of temperature dark culture 5~10 days
Stem segment；
3) by preculture stem-segment with node take out access induced medium on, at 25~30 DEG C of temperature dark culture 25~
Obtain evoked callus within 35 days；
4) evoked callus is accessed in proliferated culture medium, in 25~30 DEG C of temperature, illumination under 1500~2500Lx
It carries out obtaining within low light culture 35~40 days proliferation callus；
5) proliferation callus is accessed in differential medium, in 25~30 DEG C of temperature, illumination under 3500~4500Lx
It carries out obtaining seedling in illumination cultivation 40~60 days；
6) seedling is accessed in root media, is to carry out dim light under 1500~2500Lx in 25~30 DEG C of temperature, illumination
Culture obtains rooted seedling in 20~25 days；
Wherein, pre-culture medium are as follows: MS+ plant gel 2.5g/L+ sucrose 30%, pH5.0；
Induced medium are as follows: MS+2,4-D 4.0mg/L+ plant gel 2.5g/L+ sucrose 30%, pH5.0；
Proliferated culture medium are as follows: MS+2,4-D 5.0mg/L+ plant gel 2.5g/L+ sucrose 30%, pH5.0；
Differential medium are as follows: MS+6-BA 6mg/L+NAA 0.4mg/L+KT 1mg/L+ plant gel 2.5g/L+ sucrose
30%, pH5.0；
Root media are as follows: MS+IAA 0.2mg/L+TDZ 0.5mg/L+ plant gel 2.5g/L+ sucrose 30%,
pH5.0。
Embodiment 2
With embodiment 1, the difference is that pre-culture medium are as follows: MS+ plant gel 3.0g/L+ sucrose 30%, pH7.0；
Induced medium are as follows: MS+2,4-D 6.0mg/L+ plant gel 3.0g/L+ sucrose 30%, pH7.0；
Proliferated culture medium are as follows: MS+2,4-D 7.0mg/L+ plant gel 3.0g/L+ sucrose 30%, pH7.0；
Differential medium are as follows: MS+6-BA 6mg/L+NAA 0.6mg/L+KT 3mg/L+ plant gel 3.0g/L+ sucrose
30%, pH7.0；
Root media are as follows: MS+IAA 0.4mg/L+TDZ 1.5mg/L+ plant gel 3.0g/L+ sucrose 30%,
pH7.0。
Embodiment 3
With embodiment 1, the difference is that pre-culture medium are as follows: MS+ plant gel 2.8g/L+ sucrose 30%, pH6.0；
Induced medium are as follows: MS+2,4-D 5.0mg/L+ plant gel 2.8g/L+ sucrose 30%, pH5.8；
Proliferated culture medium are as follows: MS+2,4-D 6.0mg/L+ plant gel 2.8g/L+ sucrose 30%, pH6.0；
Differential medium are as follows: MS+6-BA 6mg/L+NAA 0.45mg/L+KT 2mg/L+ plant gel 2.8g/L+ sucrose
30%, pH6.0；
Root media are as follows: MS+IAA 0.3mg/L+TDZ 1.0mg/L+ plant gel 2.8g/L+ sucrose 30%,
pH6.0。
Claims (10)
1. a kind of method for using Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant, it is characterized in that: described uses Yunnan
Rough gentian stem-segment with node be explant cultivate regeneration plant method the following steps are included:
1) the Yunnan rough gentian plant for choosing robust growth, takes its stem-segment with node, cleans, impregnates, rinsing and is after disinfection to reserve；
2) stem-segment with node is inoculated into pre-culture medium, obtains preculture belt segment stem in 20~25 DEG C of temperature dark culture 5~10 days
Section；
3) preculture stem-segment with node is taken out in access induced medium, the dark culture 25~35 days at 25~30 DEG C of temperature
Obtain evoked callus；
4) evoked callus is accessed in proliferated culture medium, is to be carried out under 1500~2500Lx in 25~30 DEG C of temperature, illumination
Obtain within low light culture 35~40 days proliferation callus；
5) proliferation callus is accessed in differential medium, is to be carried out under 3500~4500Lx in 25~30 DEG C of temperature, illumination
Obtain seedling within illumination cultivation 40~60 days；
6) seedling is accessed in root media, is to carry out low light culture under 1500~2500Lx in 25~30 DEG C of temperature, illumination
Obtain rooted seedling within 20~25 days；
Wherein, the pre-culture medium are as follows: MS+ 2.5~3.0g/L+ of plant gel sucrose 30%, pH5.0~7.0；
The induced medium are as follows: MS+2,4-D 4.0~6.0mg/L+, 2.5~3.0g/L+ of plant gel sucrose 30%,
PH5.0~7.0；
The proliferated culture medium are as follows: MS+2,4-D 5.0~7.0mg/L+, 2.5~3.0g/L+ of plant gel sucrose 30%,
PH5.0~7.0；
The differential medium are as follows: MS+6-BA 6mg/L+NAA 0.4~0.6mg/L+KT, 1~3mg/L+ plant gel 2.5
~3.0g/L+ sucrose 30%, pH5.0~7.0；
The root media are as follows: MS+IAA 0.2~0.4mg/L+TDZ, 0.5~1.5mg/L+ plant gel 2.5~
3.0g/L+ sucrose 30%, pH5.0~7.0.
2. the method according to claim 1 for using Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant, feature
Be: the cleaning is circulating water rinsing.
3. the method according to claim 1 for using Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant, feature
Be: the immersion is to impregnate 8~12min with 10% washing powder solution of mass percent.
4. the method according to claim 1 for using Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant, feature
Be: the flushing is to rinse 20~40min using flowing water.
5. the method according to claim 1 for using Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant, feature
Be: the disinfection is to handle 3~5s using 75% ethanol solution, then again through the water-soluble 7~9min of liquid disinfectant of 0.1% mercuric chloride.
6. the method according to claim 1 for using Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant, feature
Be: the pre-culture medium is MS+ plant gel 2.8g/L+ sucrose 30%, pH6.0.
7. the method according to claim 1 for using Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant, feature
Be: the induced medium is MS+2,4-D 5.0mg/L+ plant gel 2.8g/L+ sucrose 30%, pH6.0.
8. the method according to claim 1 for using Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant, feature
Be: the proliferated culture medium is MS+2,4-D 6.0mg/L+ plant gel 2.8g/L+ sucrose 30%, pH6.0.
9. the method according to claim 1 for using Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant, feature
Be: the differential medium is MS+6-BA 6mg/L+NAA 0.5mg/L+KT 2mg/L+ plant gel 2.8g/L+ sucrose
30%, pH6.0.
10. the method according to claim 1 for using Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant, feature
Be: the root media is MS+IAA 0.3mg/L+TDZ 1.0mg/L+ plant gel 2.8g/L+ sucrose 30%,
pH6.0。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811021459.4A CN109169271A (en) | 2018-09-03 | 2018-09-03 | A method of use Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811021459.4A CN109169271A (en) | 2018-09-03 | 2018-09-03 | A method of use Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109169271A true CN109169271A (en) | 2019-01-11 |
Family
ID=64914269
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811021459.4A Pending CN109169271A (en) | 2018-09-03 | 2018-09-03 | A method of use Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109169271A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109479720A (en) * | 2018-12-29 | 2019-03-19 | 云南农业大学 | A kind of tissue culture and rapid propagation method of river east rough gentian |
| CN111165352A (en) * | 2020-01-14 | 2020-05-19 | 云南省农业科学院药用植物研究所 | Novel space breeding tissue culture seedling raising method for gentiana rigescens |
| CN115606504A (en) * | 2022-12-19 | 2023-01-17 | 云南省农业科学院药用植物研究所 | Cultivation method of gentiana rigescens polyploidy |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070111955A1 (en) * | 2002-09-11 | 2007-05-17 | Kwak Wie-Jong | Extraction and purification method of active constituents from stem of Lonicera japonica Thunb., its usage for anti-inflammatory and analgesic drug |
| CN102630569A (en) * | 2012-05-10 | 2012-08-15 | 中国科学院西北高原生物研究所 | Tissue culture method of Gentiana dahurica Fisch for rapid in vitro reproduction |
| CN104478894A (en) * | 2014-12-30 | 2015-04-01 | 云南中医学院 | Gnetiolactone compound, as well as preparation method and application thereof |
| CN105766657A (en) * | 2016-04-28 | 2016-07-20 | 四川农业大学 | Tissue culture method of conyza blinii |
| CN107333658A (en) * | 2017-09-13 | 2017-11-10 | 中冶华天工程技术有限公司 | A kind of quick-breeding method of the close thorn eel grass engineering seedling of hormone induction |
-
2018
- 2018-09-03 CN CN201811021459.4A patent/CN109169271A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070111955A1 (en) * | 2002-09-11 | 2007-05-17 | Kwak Wie-Jong | Extraction and purification method of active constituents from stem of Lonicera japonica Thunb., its usage for anti-inflammatory and analgesic drug |
| CN102630569A (en) * | 2012-05-10 | 2012-08-15 | 中国科学院西北高原生物研究所 | Tissue culture method of Gentiana dahurica Fisch for rapid in vitro reproduction |
| CN104478894A (en) * | 2014-12-30 | 2015-04-01 | 云南中医学院 | Gnetiolactone compound, as well as preparation method and application thereof |
| CN105766657A (en) * | 2016-04-28 | 2016-07-20 | 四川农业大学 | Tissue culture method of conyza blinii |
| CN107333658A (en) * | 2017-09-13 | 2017-11-10 | 中冶华天工程技术有限公司 | A kind of quick-breeding method of the close thorn eel grass engineering seedling of hormone induction |
Non-Patent Citations (3)
| Title |
|---|
| 席银凯等: ""滇龙胆丛芽高效诱导与植株再生体系的建立"", 《中草药》 * |
| 李宝平等: "《植物组织培养技术》", 31 May 2000, 中国农业科技出版社 * |
| 罗春梅等: ""不同激素组合对诱导滇龙胆顶芽及腋芽产生不定芽的研究"", 《楚雄师范学院学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109479720A (en) * | 2018-12-29 | 2019-03-19 | 云南农业大学 | A kind of tissue culture and rapid propagation method of river east rough gentian |
| CN111165352A (en) * | 2020-01-14 | 2020-05-19 | 云南省农业科学院药用植物研究所 | Novel space breeding tissue culture seedling raising method for gentiana rigescens |
| CN115606504A (en) * | 2022-12-19 | 2023-01-17 | 云南省农业科学院药用植物研究所 | Cultivation method of gentiana rigescens polyploidy |
| CN115606504B (en) * | 2022-12-19 | 2023-05-05 | 云南省农业科学院药用植物研究所 | Polyploid cultivation method for gentiana rigescens |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105340747B (en) | A kind of Radix Glycyrrhizae asexual rapid propagation method | |
| Pierik et al. | Plantlet formation in callus tissues of Anthurium andraeanum Lind. | |
| CN109169271A (en) | A method of use Yunnan rough gentian stem-segment with node to cultivate regeneration plant for explant | |
| CN101422132A (en) | China rose salt-resistance strain vitro quick-screening method | |
| Jayasree et al. | Somatic embryogenesis and plant regeneration from immature anthers of Hevea brasiliensis (Muell.) Arg. | |
| CN106332777A (en) | Tissue culture method of acer rubrum | |
| CN105766654B (en) | A kind of Nanchuan jackfruit method for tissue culture | |
| CN103404438B (en) | Acer palmatum seed tissue culture method | |
| Cerović et al. | Walnut (Juglans regia L.) selection in Serbia | |
| Naz et al. | In vitro clonal multiplication and acclimatization of different varieties of turmeric (Curcuma longa L.) | |
| CN104082096A (en) | Method for increasing wild Tilia miqueliana transplantation survival rate | |
| CN109819892B (en) | Tissue culture method of good single plant of tsaoko | |
| CN113748987A (en) | Method for producing selenium-rich konjac mutants through mutagenesis | |
| CN109362566A (en) | A kind of Rabdosia amethystoides tissue culture and rapid propagation method | |
| CN108834894A (en) | A kind of method for tissue culture of uncaria | |
| CN107114237A (en) | A kind of production of No. 25 tissue culture detoxic seedlings of cigarette potato and rooting culture method | |
| CN103636506A (en) | Shepherdia argentea caulicle regenerated plant induction culture medium and method for performing plant culture by utilizing culture medium | |
| CN104026017B (en) | The breeding method of Semen Maydiss haplobiont | |
| Kozak et al. | Effect of cytokinins on in vitro multiplication of rhubarb (Rheum rhaponticum L.)‘Karpow lipskiego’shoots and ex vitro acclimatization and growth | |
| CN110199753A (en) | A kind of nuisanceless Chinese prickly ash implantation methods of green ecological | |
| CN109496858A (en) | The quick breeding method for tissue culture of mountain tsaoko | |
| CN107041310B (en) | A kind of tissue culture and rapid propagation method of ethnic drug fevervine | |
| ReyesCastro et al. | Agronomic performance of three cocoyam (Xanthosoma violaceum Schott) genotypes grown in Nicaragua | |
| CN107258536A (en) | A kind of eucalyptus breeding tail eucalyptus camaldulensis DH191 7 tissue culture and rapid propagation method | |
| CN108271687A (en) | A kind of method for tissue culture of butterfly cherry |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190111 |
|
| RJ01 | Rejection of invention patent application after publication |