CN106386484A - Anaphalis bulleyana cultivation and breeding method - Google Patents
Anaphalis bulleyana cultivation and breeding method Download PDFInfo
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- 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
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Abstract
The invention relates to a plant cultivation and breeding method, in particular to a drug cultivation and breeding method. The method includes the operation steps of: a. Anaphalis bulleyana explant disinfection treatment; b. operation of a sterile ultra-clean workbench; c. cultivation of parenchyma cells; d. propagation culture of Anaphalis bulleyana explant callus; e. culture of Anaphalis bulleyana stem-leaf differentiation seedlings; f. Anaphalis bulleyana rooting culture; and g. seedling hardening and transplanting. Through repeated tests, the seedling culture success rate and the survival rate after transplanting both reach 90% or more, therefore the method achieves stable cultivation and breeding of Anaphalis bulleyana, reaches the purposes of low investment and high economic benefits, and solves the extinction risk of Anaphalis bulleyana due to low seed yield and slow breeding speed.
Description
Technical field
The present invention relates to a kind of culturing and reproducing of plant, particularly to the culturing and reproducing of medicine.
Background technology
Stickyhair pearleverlasting herb Anaphalis bulleyana (J.F.Jaffr.) Chang [Pluchea bulleayana
J.F.Jeffr.], also referred to as：Careless, the wild peppery cigarette of rhizoma cyperi, suchow mosla herb, the Yis is " nest octopus poem ", is that composite family anaphalis plant felt hair is fragrant
Blue or green herb, nature and flavor are pungent, warm, enter stomach, lung channel, have the effect of clearing heat and promoting diuresis, cough-relieving, preventing malaria, anti-inflammatory analgetic, stomach invigorating promoting the circulation of qi,
Cure mainly pharyngitis, anemopyretic cold, tonsillitis, bronchitis, acute gastroenteritis, urinary tract infections, cystitis, urethritis, tracheitis,
Pertussis, infantile malnutrition, the common disease such as malaria, not only widely use in traditional Chinese medicine decoction, or many treatment pharyngitis,
The important source material of the common disease Chinese patent drug such as tonsillitis, flu, infantile malnutrition due to digestive disturbances or intestinalparasites, is Yi nationality, distributed over Yunnan, Sichuan and Guizhou's conventional Chinese medicine, is embodied in《Yunnan Chinese herbal medicine》、
《The southern regions of the Yunnan Province book on Chinese herbal medicine》And《Yunnan Province's drug standards》(Yi nationality, distributed over Yunnan, Sichuan and Guizhou's medicine).Be also Yi nationality's medicine " pharynx relieving capsule ", " pharynx oral liquid for relaxation " mainly former
Material medicine.
Stickyhair pearleverlasting herb be distributed mainly on three of Dayao County, iron lock, San Chahe, the dark and damp ditch of high mountain of bharal, mountain valley with clumps of trees and bamboo side, Cao Po
Ground and low mountain meadow, are Yi nationality, distributed over Yunnan, Sichuan and Guizhou's habituation common medicine, large usage quantity among the people, but due to year after year arid in recent years, wild noon perfume
Grass is endangered, and, all can not find relevant on all of document and website：Yi nationality's medicine stickyhair pearleverlasting herb tissue culture propagating appoint
What technology, production does not have which unit carry out Yi nationality's medicine stickyhair pearleverlasting herb tissue culture propagating yet and produces.
Content of the invention
The purpose of the present invention is intended to overcome the defect of prior art, provides that a kind of reproduction speed is fast, invest little solution medicine
With a plant difficult problem in imminent danger, and ensure the stickyhair pearleverlasting herb culturing and reproducing of medicine resource.
Stickyhair pearleverlasting herb culturing and reproducing of the present invention, its step is：
A. the method for operating that stickyhair pearleverlasting herb explant is disinfected：In fine day afternoon, after cutting stickyhair pearleverlasting herb young leaflet tablet, first use
The debris on spire surface gently removed by the instruments such as writing brush, toothbrush, operating scissors, are put in immersion 5min in saturation detergent liquid, then put
Rinsed 5 times with distilled water to cleaning after little water in clean running water rinses 30min, then move into aseptic ultra-clean work
Platform；
B. the method for operating of aseptic superclean bench：First with 75% alcohol-pickled 10-20s, then the liter with 0.1-0.2%
Mercury (HgCl2) soaking disinfection 3-6min, after then using aseptic water washing 5-10 time, then blots spire surface water with aseptic filter paper
Point, it is cut into about 0.5-1.0 centimetre of length and width, about 1 square centimeter about of blade area with sterile scissors, finally use aseptic inoculation work
Tool is inoculated in superclean bench in the culture medium of callus induction and cultivates；
C. the stickyhair pearleverlasting herb callus described in step b, is aseptically, by organs such as the spire of stickyhair pearleverlasting herb, tender stems
Or a part for tissue scales off, it is placed on and is cultivated on suitable synthetic medium, make these organ or tissues carry out cell
Division, forms a parenchyma cell；
D. stickyhair pearleverlasting herb explant callus proliferation culture, in the callus that step c is induced, tissue cuts into soybean grain
Size, is placed in the culture medium of stickyhair pearleverlasting herb explant callus proliferation, daily illumination 24 hours, and intensity of illumination is 2500LX,
25 ± 2 DEG C of cultivation temperature, inoculation was opened as grown stickyhair pearleverlasting herb callus cell group after 20 50 days；
E. stickyhair pearleverlasting herb cauline leaf differentiation seedling culture, is inoculated into stickyhair pearleverlasting herb callus cell group respectively and is cultivated for basic with MS
Base, and add the 6-BA that concentration proportioning is 0.2-2.0, the IAA of 0.1-0.5,0.2 1.0 NAA, 3.0% sucrose, 6g/L
Agar, carry out cauline leaf differentiation culture, callus differentiation after about 30-50 days in the culture medium of sterilized sterilizing of pH value 5.5
Go out Multiple Buds, the sprout tuber that the Callus material with Multiple Buds is cut into 3-5 Multiple Buds is put in proliferated culture medium,
Transfer 3 pieces for every bottle, cultivate 40-60 days, culture in root media can be proceeded to when crowd shoots grow to 2-5cm；
F. stickyhair pearleverlasting herb culture of rootage, when crowd shoots grow to high 2-5cm, is cut and is proceeded to 1/2MS for basic culture
In base, and add the 6-BA that concentration proportioning is 0.05,0.5 NAA, 20% sucrose, the agar of 6g/L, the taking root of pH value 5.5
In culture medium, start within 6-10 days long root, grow up within 25-30 days the high band offspring of 2-3cm, grew through 40 50 days complete
Article 35, root system；
G. acclimatization and transplantses, the culture band offspring of step f gained are placed under normal temperature and deposit 57 days, then throw off bottle cap,
Take out seedling, the culture medium of clean root after hardening 3-4 days, then by the seeding row spacing of 10cm 30cm, seedling replanting is entered vermiculite etc.
In seedbed, and with covered rearing with plastic film keeping temperature and humidity, remove film after surviving through 7-10 days seedlings and carry out normal clay fertilizer
Water management, its survival rate is up to more than 90%.
Stickyhair pearleverlasting herb explant calli induction media described in step c, it is filled a prescription is with MS as minimal medium, adds dense
Degree proportioning is the 6-BA of 2,4-D, the 0.2 of 2.0, the sucrose of the PVP of 1.0g/L, 5g/L, the agar of 7g/L, its PH=5.8.
Stickyhair pearleverlasting herb explant callus proliferation medium described in step d, it is filled a prescription is to be cultivated with 1/2MS for basic
Base, and 6-BA, the PVP of 1.0g/L, the sucrose 15g/L of add that concentration ratio is 1.0 2,4-D, 0.2, agar 7g/L, PH=5.8.
Acclimatization and transplantses described in step g, are topdressed with the aqueous solution of 1/5MS a great number of elements in the growth period of seedling.
Stickyhair pearleverlasting herb culturing and reproducing of the present invention, the complete stool of stickyhair pearleverlasting herb is yellow, and plant height is up to 100cm, quilt
White wool；Stem is cylindrical, has obvious vertical wrinkle, cane upper end multi-branched on cane.Cane thick diameter 2-8mm, matter are crisp
Easily snap off, section is in that yellow green, center have white flesh.Dan Ye, alternate, rosette-stape, full edge, obovate or lanceolar, along stem
Downward wedgewise width wing, tip point, edge is put down, and there are glandular hairs on two sides.Topmost leaf is narrow and small, close white on wire lanceolar, vein
The long wool of color；Floral white, capitulum, gas delicate fragrance, bitter is cool.Microscopical characters feature：Parenchyma cell similar round or polygonal；Oil
Cell similar round；Conduit reticulate pattern and Kong Wen, 7-30 μm of diameter；Fiber fusiform；The flat shaft type of pore；Glandular hairs are many, more piece.Because the noon is fragrant
By thread white cotton wool, cotton wool length up to 10cm, plant is also long simultaneously the rust more piece glandular hairs with handle for the brown cement, band to careless complete stool
There is substantial amounts of miscellaneous bacteria, lead to explant easily to pollute, the operation sequence of its control pollution is：Explant blade hair
Brush goes debris saturation detergent liquid to soak the clean running water of 5mim cleaning and rinses the rinsing of 30min distilled water
Move into superclean bench sterilization for 5 times from 75% ethanol postincubation 10-15s → rinsed with sterile water 5 times → 0.1%
HgCl2 (dropping Tween-80, every liter plus 20) 3-5min → aseptic washing 8-10 time, the pollution rate of controllable explant.
Therefore, stickyhair pearleverlasting herb explant sterilization method technical system, the method for operating that stickyhair pearleverlasting herb explant is disinfected：Fine
In its afternoon, after cutting stickyhair pearleverlasting herb young leaflet tablet, first gently remove the debris on spire surface with instruments such as writing brush, toothbrush, operating scissors,
It is put in immersion 5min in saturation detergent liquid, then is put into little water in the clean running water of cleaning and use distilled water after rinsing 30min
Rinsing 5 times, then moves into all operationss program that completes in aseptic superclean bench：First with 75% alcohol-pickled 10-20s,
Use mercuric chloride (HgCl2) the soaking disinfection 3-6min of 0.1-0.2% again, after then using aseptic water washing 5-10 time, then with aseptic
Filter paper blots spire surface moisture, is cut into about 0.5-1.0 centimetre of length and width, about 1 square centimeter of left side of blade area with sterile scissors
The right side, is finally inoculated in superclean bench with aseptic inoculation instrument in the culture medium of callus induction and cultivates, outside induction stickyhair pearleverlasting herb
Implant callus.
Complete stickyhair pearleverlasting herb explant calli induction media recipe determination, stickyhair pearleverlasting herb callus is aseptically,
A part for the organ or tissues such as the spire of stickyhair pearleverlasting herb, tender stem is scaled off, is placed on and is cultivated on suitable synthetic medium,
These organ or tissues will carry out cell division, a parenchyma cell of formation.It is in suitable illumination, temperature and certain
Under the conditions of nutriment and hormone etc., callus just starts to break up, and produces various organs and the tissue of plant, and then develops
Become a complete plant, it is the basis of stickyhair pearleverlasting herb Fast-propagation.
Because stickyhair pearleverlasting herb complete stool is carried substantial amounts of body of gland by spider's thread shape white wool, body surface, include rust brown cement juice,
Both carried a large amount of miscellaneous bacterias, easily caused pollution, and easily produced brown stain, lead to callus and Brown, simultaneously to plant
Thing growth regulator is extremely sensitive, and temporarily the investigative technique of no stickyhair pearleverlasting herb fast breeding technique can increase the noon fragrant for reference both at home and abroad
The difficulty of careless tissue cultures.
Researcher's priority with 1/2MS, MS, B5, N6 culture medium as minimal medium, add the 6-BA of variable concentrations, Kt,
2,4-D, the combination matching such as IBA, IAA, NAA, PVP, activated carbon, sucrose, agar, filters out the noon from 121 culture medium prescriptions
Vanilla explant calli induction media is filled a prescription, and can turn out substantial amounts of callus through 30 days about.Its formula is as follows：
MS+2,4-D 2.0+6-BA 0.2+PVP 1.0g/L+ sucrose 15g/L+ agar 7g/L (PH=5.8)
Complete stickyhair pearleverlasting herb explant callus proliferation medium recipe determination, the callus inducing is cut into Huang
Beans size, is inoculated into respectively with 1/2MS, MS, B5, N6 culture medium as minimal medium, add the 6-BA of variable concentrations, Kt,
2,4-D, the combination matching such as IBA, IAA, NAA, PVP, activated carbon, sucrose, agar, filters out the noon from 80 culture medium prescriptions
The culture medium prescription of vanilla explant callus proliferation：1/2MS+2,4-D 1.0+6-BA 0.2+PVP 1.0g/L+ sucrose
15g/L+ agar 7g/L (PH=5.8), daily illumination 24 hours, intensity of illumination is 2500LX, 25 ± 2 DEG C of cultivation temperature, inoculation
Open after 20 50 days as grown substantial amounts of stickyhair pearleverlasting herb callus cell group, thus realizing callus propagation.
Complete stickyhair pearleverlasting herb cauline leaf differentiation seedling screening of medium formula, stickyhair pearleverlasting herb callus cell group is inoculated into respectively：
MS is minimal medium, adds 6-BA 0.2-2.0, IAA0.1-0.5, NAA0.2 1.0, the sucrose of variable concentrations proportioning
3.0%th, carry out cauline leaf differentiation culture in being cultivated in the culture medium of prior preparation sterilization of agar 6g/L, pH value 5.5,
After about 30-50d, callus differentiates Multiple Buds.Callus material with Multiple Buds is cut into 3-5 Multiple Buds
Sprout tuber be put in proliferated culture medium, every bottle transfer 3 pieces, culture 40-60d, life can be proceeded to when crowd shoots grow to 2-5cm
Cultivate in root culture medium.
Complete stickyhair pearleverlasting herb culture of rootage research, as squamous subculture height of seedling 2-5cm, cut and proceed to 1/2MS and substantially train
In foster base, add 6-BA 0.05+NAA 0.5+20% sucrose+agar 6g/L, pH value 5.5.General 6-10d starts long root, 30d
Left and right can grow up to the high band offspring of 2-3cm, can grow complete 35 root system through 40 50d.
Acclimatization and transplantses technology, culture band offspring is placed under normal temperature and deposits 5 7d, then throw off bottle cap, after hardening 3-4d
Take out seedling, the culture medium of clean root, then by the seeding row spacing of 10cm 30cm, seedling replanting is entered in the seedbeds such as vermiculite, be used in combination
Covered rearing with plastic film keeping temperature and humidity, remove film and carry out normal soil fertilizer and water management, it survives after surviving through 7-10d seedling
Rate is up to more than 90%.Growth period can be topdressed with the aqueous solution of 1/5MS a great number of elements, can accelerate the growth of seedling.
Stickyhair pearleverlasting herb culturing and reproducing of the present invention, explores through test of many times, has explored the tissue of stickyhair pearleverlasting herb
Culture test tube seedling and correlation technique system, it is achieved that stickyhair pearleverlasting herb sapling multiplication is not subject to area and season limit, is cultivated, plant through multiple
Survival rate after training, nursery success rate and transplanting it is achieved that stable ectogenesis stickyhair pearleverlasting herb, has reached throwing all more than 90%
Money is few, the purpose of high financial profit；Solve stickyhair pearleverlasting herb and produce the few reproduction speed of seed slowly, face the danger of extinction.
Specific embodiment
With reference to embodiment, the present invention is described further, but is not limited to embodiment.
Embodiment 1
1st, control stickyhair pearleverlasting herb explant pollution method
Control pollution operation sequence be：Explant blade writing brush type brush removes debris saturation detergent liquid
Soak the clean running water of 5mim cleaning and rinse 5 immigration superclean benches of 30min distilled water rinsing
Sterilization (drips Tween-80, every liter adds from the HgCl2 of 75% ethanol postincubation 10-15s → rinsed with sterile water 5 times → 0.1%
20) 3-5min → aseptic washing 8-10 time, the pollution rate of controllable explant.
2nd, stickyhair pearleverlasting herb explant sterilization method technical system
The method of operating that stickyhair pearleverlasting herb explant is disinfected：In fine day afternoon, after cutting stickyhair pearleverlasting herb young leaflet tablet, first use hair
The debris on spire surface gently removed by the instruments such as pen, toothbrush, operating scissors, are put in immersion 5min in saturation detergent liquid, then are put into
Clean after little water in clean running water rinses 30min and rinsed 5 times with distilled water, then move in aseptic superclean bench
Complete all operationss program：First with 75% alcohol-pickled 10-20s, then mercuric chloride (HgCl2) soaking disinfection with 0.1-0.2%
3-6min, after then using aseptic water washing 5-10 time, then blots spire surface moisture with aseptic filter paper, is cut into sterile scissors
About 0.5-1.0 centimetre of length and width, about 1 square centimeter about of blade area, finally connect in superclean bench with aseptic inoculation instrument
Plant in the culture medium of callus induction and cultivate, induce stickyhair pearleverlasting herb explant callus.
3rd, stickyhair pearleverlasting herb explant calli induction media formula
MS+2,4-D 2.0+6-BA 0.2+PVP 1.0g/L+ sucrose 15g/L+ agar 7g/L (PH=5.8)
4th, complete stickyhair pearleverlasting herb explant callus proliferation medium formula
The callus inducing is cut into soybean grain size, is inoculated in 1/2MS+2,4-D 1.0+6-BA0.2+PVP
1.0g/L+ sucrose 15g/L+ agar 7g/L (PH=5.8), daily illumination 24 hours, intensity of illumination is 2500LX, cultivation temperature 25
± 2 DEG C, inoculation was opened as grown substantial amounts of stickyhair pearleverlasting herb callus cell group after 20 50 days, thus realizing callus propagation.
5th, complete stickyhair pearleverlasting herb cauline leaf differentiation seedling screening of medium formula
Stickyhair pearleverlasting herb callus cell group is inoculated into respectively：MS is minimal medium, adds the 6- of variable concentrations proportioning
BA 0.2-2.0, IAA0.1-0.5, NAA 0.2 1.0, sucrose 3.0%, agar 6g/L, the prior preparation sterilization of pH value 5.5 are gone out
Carry out cauline leaf differentiation culture, after about 30-50d, callus differentiates Multiple Buds in being cultivated in the culture medium of bacterium.To carry
The sprout tuber that the Callus material of Multiple Buds is cut into 3-5 Multiple Buds is put in proliferated culture medium, transfers 3 pieces for every bottle, training
Foster 40-60d, culture can be proceeded in root media when crowd shoots grow to 2-5cm.
6th, complete stickyhair pearleverlasting herb culture of rootage research
As squamous subculture height of seedling 2-5cm, cut and proceeded in 1/2MS minimal medium, added 6-BA0.05+NAA
0.5+20% sucrose+agar 6g/L, pH value 5.5.General 6-10d starts long root, 30d about can grow up to the high band root of 2-3cm
Seedling, can grow complete 35 root system through 40 50d.
7th, acclimatization and transplantses technology
Culture band offspring is placed under normal temperature and deposits 5 7d, then throw off bottle cap, take out seedling after hardening 3-4d, clean
The culture medium of root, then by the seeding row spacing of 10cm 30cm, seedling replanting is entered in the seedbeds such as vermiculite, and use covered rearing with plastic film
Keeping temperature and humidity, remove film and carry out normal soil fertilizer and water management after surviving through 7-10d seedling, its survival rate up to 90% with
On.
Claims (4)
1. stickyhair pearleverlasting herb culturing and reproducing is it is characterised in that its operating procedure is：
A. the method for operating that stickyhair pearleverlasting herb explant is disinfected：In fine day afternoon, after cutting stickyhair pearleverlasting herb young leaflet tablet, first use hair
The debris on spire surface gently removed by the instruments such as pen, toothbrush, operating scissors, are put in immersion 5min in saturation detergent liquid, then are put into
Clean after little water in clean running water rinses 30min and rinsed 5 times with distilled water, then move into aseptic superclean bench；
B. the method for operating of aseptic superclean bench：First with 75% alcohol-pickled 10-20s, then the mercuric chloride with 0.1-0.2%
(HgCl2) soaking disinfection 3-6min, after then using aseptic water washing 5-10 time, then blots spire surface water with aseptic filter paper
Point, it is cut into about 0.5-1.0 centimetre of length and width, about 1 square centimeter about of blade area with sterile scissors, finally use aseptic inoculation work
Tool is inoculated in superclean bench in the culture medium of callus induction and cultivates；
C. the stickyhair pearleverlasting herb callus described in step b, is aseptically, by the organs such as the spire of stickyhair pearleverlasting herb, tender stem or group
The part knitted scales off, and is placed on and is cultivated on suitable synthetic medium, makes these organ or tissues carry out cell division,
Form a parenchyma cell；
D. stickyhair pearleverlasting herb explant callus proliferation culture, in the callus that step c is induced, tissue cuts into soybean grain size,
It is placed in the culture medium of stickyhair pearleverlasting herb explant callus proliferation, daily illumination 24 hours, intensity of illumination is 2500LX, culture temperature
25 ± 2 DEG C of degree, inoculation was opened as grown stickyhair pearleverlasting herb callus cell group after 20 50 days；
E. stickyhair pearleverlasting herb cauline leaf differentiation seedling culture, is inoculated into stickyhair pearleverlasting herb callus cell group respectively with MS as minimal medium,
And add the 6-BA that concentration proportioning is 0.2-2.0, the IAA of 0.1-0.5,0.2 1.0 NAA, 3.0% sucrose, the fine jade of 6g/L
Carry out cauline leaf differentiation culture, after about 30-50 days, callus differentiates clump in fat, the culture medium of sterilized sterilizing of pH value 5.5
Sprout, the sprout tuber that the Callus material with Multiple Buds is cut into 3-5 Multiple Buds is put in proliferated culture medium, every bottle
3 pieces of switching, cultivates 40-60 days, can proceed to culture in root media when crowd shoots grow to 2-5cm；
F. stickyhair pearleverlasting herb culture of rootage, when crowd shoots grow to high 2-5cm, cut proceed to 1/2MS be minimal medium in,
And add the 6-BA that concentration proportioning is 0.05,0.5 NAA, 20% sucrose, the agar of 6g/L, the root media of pH value 5.5
In, start within 6-10 days long root, grow up within 25-30 days the high band offspring of 2-3cm, grew complete 35 through 40 50 days
Root system；
G. acclimatization and transplantses, the culture band offspring of step f gained are placed under normal temperature and deposit 57 days, then throw off bottle cap, hardening
Take out seedling, the culture medium of clean root after 3-4 days, then by the seeding row spacing of 10cm 30cm, seedling replanting is entered the seedbeds such as vermiculite
In, and with covered rearing with plastic film keeping temperature and humidity, remove film after surviving through 7-10 days seedlings and carry out normal clay fertilizer water pipe
Reason, its survival rate is up to more than 90%.
2. stickyhair pearleverlasting herb culturing and reproducing according to claim 1 is it is characterised in that stickyhair pearleverlasting herb explant described in step c
Body calli induction media, it is filled a prescription is with MS as minimal medium, and addition concentration proportioning is the 6-BA of 2,4-D, the 0.2 of 2.0,
The sucrose of the PVP of 1.0g/L, 5g/L, the agar of 7g/L, its PH=5.8.
3. stickyhair pearleverlasting herb culturing and reproducing according to claim 1 is it is characterised in that stickyhair pearleverlasting herb explant described in step d
Body callus proliferation medium, it is filled a prescription is with 1/2MS as minimal medium, and add that concentration ratio is 1.0 2,4-D, 0.2
6-BA, the PVP of 1.0g/L, sucrose 15g/L, agar 7g/L, PH=5.8.
4. stickyhair pearleverlasting herb culturing and reproducing according to claim 1 is it is characterised in that acclimatization and transplantses described in step g,
The growth period of seedling is topdressed with the aqueous solution of 1/5MS a great number of elements.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110192527A (en) * | 2019-07-01 | 2019-09-03 | 中央民族大学 | The fragrant green method for tissue culture of fragrant plant bell bell |
| CN112335498A (en) * | 2020-11-05 | 2021-02-09 | 云南金碧制药有限公司 | Planting method of cymbopogon flexuosus and grass pest control method |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102090342A (en) * | 2011-01-10 | 2011-06-15 | 南京农业大学 | Method for establishing high-efficient regeneration system of dendranthema zawadskii |
| CN103229721A (en) * | 2013-05-15 | 2013-08-07 | 聊城大学 | Tissue culture propagation method of Gynura formosana |
| CN103688856A (en) * | 2013-12-13 | 2014-04-02 | 广西桂西制药有限公司 | Rapid propagation method of Blumea riparia (Bl.) DC medicinal material |
| CN103960129A (en) * | 2014-04-24 | 2014-08-06 | 江苏农林职业技术学院 | Atractylis lancea tissue culturing and rapid propagating method |
| CN105052740A (en) * | 2015-08-11 | 2015-11-18 | 中国热带农业科学院橡胶研究所 | Method for regenerating plants by using rubber grass leaves |
-
2016
- 2016-08-30 CN CN201610777414.4A patent/CN106386484A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102090342A (en) * | 2011-01-10 | 2011-06-15 | 南京农业大学 | Method for establishing high-efficient regeneration system of dendranthema zawadskii |
| CN103229721A (en) * | 2013-05-15 | 2013-08-07 | 聊城大学 | Tissue culture propagation method of Gynura formosana |
| CN103688856A (en) * | 2013-12-13 | 2014-04-02 | 广西桂西制药有限公司 | Rapid propagation method of Blumea riparia (Bl.) DC medicinal material |
| CN103960129A (en) * | 2014-04-24 | 2014-08-06 | 江苏农林职业技术学院 | Atractylis lancea tissue culturing and rapid propagating method |
| CN105052740A (en) * | 2015-08-11 | 2015-11-18 | 中国热带农业科学院橡胶研究所 | Method for regenerating plants by using rubber grass leaves |
Non-Patent Citations (1)
| Title |
|---|
| 王跃华等: "川芎幼嫩叶片植株再生的研究", 《西南农业学报》 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110192527A (en) * | 2019-07-01 | 2019-09-03 | 中央民族大学 | The fragrant green method for tissue culture of fragrant plant bell bell |
| CN110192527B (en) * | 2019-07-01 | 2020-11-24 | 中央民族大学 | Tissue culture method of aromatic plant Linglingqing |
| CN112335498A (en) * | 2020-11-05 | 2021-02-09 | 云南金碧制药有限公司 | Planting method of cymbopogon flexuosus and grass pest control method |
| CN112335498B (en) * | 2020-11-05 | 2021-07-02 | 云南金碧制药有限公司 | Planting method for cymbopogon flexuosus |
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