CN109430058A - A kind of axenic germination method of adenophora tetraphylla seed - Google Patents
A kind of axenic germination method of adenophora tetraphylla seed Download PDFInfo
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- CN109430058A CN109430058A CN201811587817.8A CN201811587817A CN109430058A CN 109430058 A CN109430058 A CN 109430058A CN 201811587817 A CN201811587817 A CN 201811587817A CN 109430058 A CN109430058 A CN 109430058A
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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
- A01H4/001—Culture apparatus for tissue culture
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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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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
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- Pretreatment Of Seeds And Plants (AREA)
Abstract
The invention discloses a kind of axenic germination methods of adenophora tetraphylla seed, it is related to agricultural plantation technology field, it handles, culture medium configuration, inoculated and cultured, take root including seed asepsis, the Nansha seedling budding time that transplanting breeding goes out shortens, germination percentage greatly improves, seedling physically well develops after being transplanted in soil, seedling is healthy and strong, is a kind of simple, efficient axenic germination cultural method.
Description
Technical field
The present invention relates to agricultural plantation technology field, especially a kind of axenic germination method of adenophora tetraphylla seed.
Background technique
Adenophora tetraphylla is adenophora tetraphylla Adenophora tetraphylla (Thunb.) Fisch. and the root of straight ladybell Adenophora
The dry root of stricta Miq. is recorded in going through an edition Pharmacopoeia of the People's Republic of China.Adenophora tetraphylla as it is clinically used simply
Yin tonics has good nourishiing yin to clear away the lung-heat, resolving sputum, QI invigorating, the function of reinforcing stomach reg fluid.Chinese medicine is commonly used in treatment lung-heat type cough,
Deficiency of yin and overstrain cough, dry cough phlegm is glutinous, deficiency of stomach-Yin, deficiency of food vomiting, deficiency of both vital energy and Yin, restlessness and fever with thirst etc..Modern research shows that in adenophora tetraphylla
Generally containing the chemical components such as polysaccharide, terpene, phenolic acid class, cumarin, sterol, volatile oil, there is kobadrin, anti-oxidant, anti-
The effect of the modern pharmacologies such as aging, immunological regulation, anti-radiation.
The economic value of adenophora tetraphylla is mainly reflected in two aspects: one is edible value, and the root of the root of straight ladybell platymiscium is rich in more
Sugar, steroidal and triterpene compound, in good taste, nutritive value is high, can be used as nutraceutical development and utilization；The root of straight ladybell root is sweet fresh
It is crisp, there is the habit removed the peel and eaten it raw Guizhou is civil；It is also nourished with its stewed chicken in some areas.Two be medical value, is China
One of highly important medicinal material is used as medicine with the root of carrot shape；Script adenophora tetraphylla is pure wild plant medicinal material, however as market
Demand increases year by year, and price rises year by year, and economic value constantly rises, and keeps its wild resource increasingly depleted, it would be highly desirable to artificial
Cultivation.
Adenophora tetraphylla is mainly bred with underground stem tuber as main means at present, discloses one as patent 201711116974.6 is bright
Kind adenophora tetraphylla Regeneration in Vitro tissue cultures technology, using adenophora tetraphylla stem with bud as explant, by bud induction, bud proliferation training
The processes such as feeding, culture of rootage, hardening and transplanting realize adenophora tetraphylla Regeneration in Vitro, but input cost is high, and cultivation technique is immature,
Lead to low output, income is very few；Another common planting technology is seed sowing, and point spring sowing and autumn sowing are sowed in spring in April is upper
In ten days, autumn sowing is before November is freeze-up；Shallow ridges is opened by 30-40 centimetres of line-spacing, seed is uniformly sprinkling upon in ditch, is covered with fine earth flat, gently
It waters after compacting, one layer of grass of lid inspires bud with heat and moisture preserving.It emerges within spring sowing 15-20 days, autumn sowing is emerged in spring in next year；Nansha
The seed sowing technology of ginseng specifically can refer to " Liu Caihong, the cultivation technique of adenophora tetraphylla, base's J Chinese, 2000,73,26 ".
But the cultural method germination percentage that seed is broadcast live is lower, brings difficulty to artificial large-scale planting, serious to restrict adenophora tetraphylla industrialization
Development.
Also there is the tissue culture technique of research adenophora tetraphylla in recent years, sprouted using aseptic culture medium culture adenophora tetraphylla seed,
Clump bud proliferation is taken root, and last hardening is simultaneously transplanted in fertile soil: in the seedbed of perlite (3:1), survival rate is up to 89% or more
(auspicious friend of dragon etc., the tissue culture and rapid proliferation of adenophora tetraphylla, Plant Physiology Communications, in April, 2007, volume 43, the 2nd phase).
The present invention have bred a large amount of test tube seedlings using group culturation rapid propagating technology, can significantly improve the germination rate of adenophora tetraphylla seed, are Nansha
Ginseng carries out artificial development and utilization and provides technical support with wild resource effective protection.
Summary of the invention
In order to solve the above technical problems existing in the prior art, the present invention provides a kind of the sterile of adenophora tetraphylla seed and sprouts
Forwarding method, can rapid, high volume breeding, method is reproducible, and the period is short.
The first aspect of the present invention provides a kind of axenic germination method of adenophora tetraphylla seed, the method includes the steps:
(a) seed is subjected to axenic germination and obtains aseptic seedling；With
(b) aseptic seedling for obtaining the step a) carries out culture of rootage, obtains rooted seedling；
In another preferred example, the method also includes carrying out disinfection to the seed.
In another preferred example, adenophora tetraphylla fruit pod is handled with the following method, and adenophora tetraphylla maturation is taken not crack fruit pod in certainly
Water rinses 30min, impregnates 5-10min with dish washing liquid, is cleaned up, be placed on spare on superclean bench with tap water；
In another preferred example, adenophora tetraphylla fruit pod processing method are as follows: impregnate 10-30S with 75% alcohol, then use mercuric chloride
(0.1%) it impregnates 5-7 minutes, with aseptic water washing 3-5 times, blots fruit pod surface moisture with aseptic filter paper, with solution planing tool solution plane
Fruit is opened, gently picks up seed with tweezers, can be seeded in aseptic culture medium.
In another preferred example, in the step a), the seed is placed in containing NAA (0.4-0.7mg/L), 6-BA
On the 1/2MS solid medium of (0.1-0.4mg/L), sucrose (30g/L) and agar 6.5g/L, in 23-26 DEG C of sterile culture, institute
It states seed and sprouts the acquisition aseptic seedling.
In another preferred example, in the step a), the seed is placed in containing NAA (0.4-0.6mg/L), 6-BA
On the 1/2MS solid medium of (0.1-0.3mg/L), sucrose (30g/L) and agar 6.5g/L, in 23-26 DEG C of sterile culture, institute
It states seed and sprouts the acquisition aseptic seedling.
In another preferred example, in the step a), the seed is placed in containing NAA (0.5-0.7mg/L), 6-BA
On the 1/2MS solid medium of (0.2-0.4mg/L), sucrose (30g/L) and agar 6.5g/L, in 23-26 DEG C of sterile culture, institute
It states seed and sprouts the acquisition aseptic seedling.
In another preferred example, in the step a), Aseptic seedling culture humidity is 30%-50%, intensity of illumination 1200-
2000lx, light application time 8-12h/d.
In another preferred example, in the step a), seed starts to sprout after culture 8-10 days, and seed is sprouted within 25-30 days
Terminate, germination percentage 93-99%.
In another preferred example, in the step b), the culture of rootage refers in the aminoadenine of benzyl containing 0.1-1mg/L
It is cultivated in the MS culture medium of 1-10mg/L gibberellin.
In another preferred example, in the step b), the culture of rootage refers to fast in the amino gland of benzyl containing 0.3-0.8mg/L
It is cultivated in the MS culture medium of purine and 2-7mg/L gibberellin.
In another preferred example, it in the step b), takes root naturally within 28-32 days.
In another preferred example, rooted seedling culture humidity is 30%-50%, intensity of illumination 1200- in the step b)
2000lx, light application time 8-12h/d.
The second aspect of the present invention provides a kind of propagation method of adenophora tetraphylla seed, comprising:
(a) seed is subjected to axenic germination and obtains aseptic seedling；
(b) aseptic seedling for obtaining the step a) carries out culture of rootage, obtains rooted seedling；With
(c) the step b) rooted seedling obtained is cultivated, takes root and obtains plant.
In another preferred example, the method also includes carrying out disinfection to the seed.
In another preferred example, adenophora tetraphylla fruit pod is handled with the following method, and adenophora tetraphylla maturation is taken not crack fruit pod in certainly
Water rinses 30min, impregnates 5-10min with dish washing liquid, is cleaned up, be placed on spare on superclean bench with tap water；
In another preferred example, adenophora tetraphylla fruit pod processing method are as follows: impregnate 10-30S with 75% alcohol, then use mercuric chloride
(0.1%) it impregnates 5-7 minutes, with aseptic water washing 3-5 times, blots fruit pod surface moisture with aseptic filter paper, with solution planing tool solution plane
Fruit is opened, gently picks up seed with tweezers, can be seeded in aseptic culture medium.
In another preferred example, in the step a), the seed is placed in containing NAA (0.4-0.7mg/L), 6-BA
On the 1/2MS solid medium of (0.1-0.4mg/L), sucrose (30g/L) and agar 6.5g/L, in 23-26 DEG C of sterile culture, institute
It states seed and sprouts the acquisition aseptic seedling.
In another preferred example, in the step a), the seed is placed in containing NAA (0.4-0.6mg/L), 6-BA
On the 1/2MS solid medium of (0.1-0.3mg/L), sucrose (30g/L) and agar 6.5g/L, in 23-26 DEG C of sterile culture, institute
It states seed and sprouts the acquisition aseptic seedling.
In another preferred example, in the step a), the seed is placed in containing NAA (0.5-0.7mg/L), 6-BA
On the 1/2MS solid medium of (0.2-0.4mg/L), sucrose (30g/L) and agar 6.5g/L, in 23-26 DEG C of sterile culture, institute
It states seed and sprouts the acquisition aseptic seedling.
In another preferred example, in the step a), Aseptic seedling culture humidity is 30%-50%, intensity of illumination 1200-
2000lx, light application time 8-12h/d.
In another preferred example, in the step a), seed starts to sprout after culture 8-10 days, and seed is sprouted within 25-30 days
Terminate, germination percentage 93-99%.
In another preferred example, in the step b), the culture of rootage refers in the aminoadenine of benzyl containing 0.1-1mg/L
It is cultivated in the MS culture medium of 1-10mg/L gibberellin.
In another preferred example, in the step b), the culture of rootage refers to fast in the amino gland of benzyl containing 0.3-0.8mg/L
It is cultivated in the MS culture medium of purine and 2-7mg/L gibberellin.
In another preferred example, it in the step b), takes root naturally within 28-32 days.
In another preferred example, 23-26 DEG C of rooted seedling cultivation temperature in the step b), humidity 30%-50%, illumination
Intensity 1200-2000lx, light application time 8-12h/d.
In another preferred example, step c) the adenophora tetraphylla plant obtained is implanted into the step of cultivating in soil.
The present invention establishes the method that the in vitro fast-germination of adenophora tetraphylla seed is taken root, and compared with prior art, sprouts the time
Shorten, germination percentage greatly improves, and seedling physically well develops after being transplanted in soil, and seedling is healthy and strong.The technical effect of the invention
It is embodied in:
(1) present invention establishes adenophora tetraphylla seed asepsis sprouting technology and is trained plant.
(2) method of the invention can realize that adenophora tetraphylla seed is largely efficiently sprouted, and generate root system, aseptic seedling obtained
It can not only be used for adenophora tetraphylla seedling, and provide good material to carry out genetic improvement to adenophora tetraphylla；
(3) the method for the invention budding time shortens, and germination percentage greatly improves.
(4) method of the invention is reproducible, can obtain the axenic germination seedling of adenophora tetraphylla, after root induction,
In transplanting to fast-growth after soil, the purpose for obtaining commercialization plant in a short time is realized.
It should be understood that above-mentioned each technical characteristic of the invention and having in below (eg embodiment) within the scope of the present invention
It can be combined with each other between each technical characteristic of body description, to form a new or preferred technical solution.
Specific embodiment
The feature that the features described above or embodiment that the present invention mentions are mentioned can be in any combination.Disclosed in this case specification
All features can be used in combination with any composition form, each feature disclosed in specification, can by it is any provide it is identical,
The alternative characteristics of impartial or similar purpose replace.Therefore except there is special instruction, revealed feature is only impartial or similar spy
The general example of sign.
It is limited below with reference to specific embodiment technical solution of the present invention is further, but claimed
Range is not only limited to made description.
Unless otherwise defined, it anticipates known to all professional and scientific terms as used herein and one skilled in the art
Justice is identical.In addition, any method similar to or equal to what is recorded and material can be applied to the method for the present invention.Wen Zhong
The preferred implement methods and materials are for illustrative purposes only.
Embodiment 1
(1) it the processing of adenophora tetraphylla fruit pod: takes adenophora tetraphylla maturation not crack fruit pod in tap water flushing 30min, uses dish washing liquid
5min is impregnated, is cleaned up, is placed on spare on superclean bench with tap water.
(2) disinfection of adenophora tetraphylla fruit pod: 30S is impregnated with 75% alcohol, then is impregnated 7 minutes with mercuric chloride (0.1%), use is sterile
Water rinses 5 times, blots fruit pod surface moisture with aseptic filter paper, spare.
(3) it the configuration of germination medium: using 1/2/MS culture medium as basic culture medium, is added into the basal medium
Following additive: pH value is adjusted to by NAA (0.7mg/L), 6-BA (0.1mg/L), sucrose (30g/L) and agar 6.5g/L
6.0, it obtains sprouting root media.
(4) it sows: opening fruit with solution planing tool solution plane, gently pick up seed with tweezers, be seeded in aseptic culture medium, be placed in
It is cultivated in 23-26 DEG C of artificial incubator, humidity 30%, intensity of illumination 1200lx, light application time 8h/d；Routine observation tissue culture
Bottle, starts to sprout after cultivating 10d, and seed sprouting in 28-30 days terminates, germination percentage 98%.
(5) it takes root: aseptic seedling obtained by step (4) is transferred to the aminoadenine of benzyl containing 1mg/L and 10mg/L gibberellin
It is cultivated in MS culture medium, wherein 26 DEG C of cultivation temperature, humidity 30%-50%, intensity of illumination 1200lx, light application time
8h/d；Naturally it takes root within 30-32 days.
(6) it transplants: small transplantation of seedlings obtained by step (5) is transplanted to outdoor after it grows up to plantlet into sterile soil again
Soil makes its continued growth under field conditions (factors), and monthly primary with the fertilising of MS minimal medium, fertilizing method is root application, is survived
Rate is up to 92% or more.
Embodiment 2
(1) it the processing of adenophora tetraphylla fruit pod: takes adenophora tetraphylla maturation not crack fruit pod in tap water flushing 30min, uses dish washing liquid
10min is impregnated, is cleaned up, is placed on spare on superclean bench with tap water.
(2) disinfection of adenophora tetraphylla fruit pod: 20S is impregnated with 75% alcohol, then is impregnated 6 minutes with mercuric chloride (0.1%), use is sterile
Water rinses 3 times, blots fruit pod surface moisture with aseptic filter paper, spare.
(3) it the configuration of germination medium: using 1/2/MS culture medium as basic culture medium, is added into the basal medium
Following additive: pH value is adjusted to by NAA (0.4mg/L), 6-BA (0.4mg/L), sucrose (30g/L) and agar 6.5g/L
5.8, it obtains sprouting root media.
(4) it sows: opening fruit with solution planing tool solution plane, gently pick up seed with tweezers, be seeded in aseptic culture medium, be placed in
It is cultivated in 23-26 DEG C of artificial incubator, humidity 50%, intensity of illumination 2000lx, light application time 8h/d；Routine observation tissue culture
Bottle, starts to sprout after cultivating 10d, and seed sprouting in 25-30 days terminates, germination percentage 95%.
(5) it takes root: aseptic seedling obtained by step (4) is transferred to the aminoadenine of benzyl containing 0.8mg/L and 7mg/L gibberellin
It is cultivated in MS culture medium, wherein 23-26 DEG C of cultivation temperature, humidity 50%, intensity of illumination 1200lx, light application time 12h/
d；Naturally it takes root within 28-31 days.
(6) it transplants: small transplantation of seedlings obtained by step (5) is transplanted to outdoor after it grows up to plantlet into sterile soil again
Soil makes its continued growth under field conditions (factors), and monthly primary with 800 times of liquid fertilisings of a great number of elements in MS, fertilizing method is root
It applies, survival rate is up to 96% or more.
Embodiment 3
(1) it the processing of adenophora tetraphylla fruit pod: takes adenophora tetraphylla maturation not crack fruit pod in tap water flushing 30min, uses dish washing liquid
5min is impregnated, is cleaned up, is placed on spare on superclean bench with tap water.
(2) disinfection of adenophora tetraphylla fruit pod: 10S is impregnated with 75% alcohol, then is impregnated 5 minutes with mercuric chloride (0.1%), use is sterile
Water rinses 3 times, blots fruit pod surface moisture with aseptic filter paper, spare.
(3) it the configuration of germination medium: using 1/2/MS culture medium as basic culture medium, is added into the basal medium
Following additive: pH value is adjusted to by NAA (0.5mg/L), 6-BA (0.2mg/L), sucrose (30g/L) and agar 6.5g/L
5.9, it obtains sprouting root media.
(4) it sows: opening fruit with solution planing tool solution plane, gently pick up seed with tweezers, be seeded in aseptic culture medium, be placed in
It is cultivated in 25 DEG C of artificial incubators, humidity 45%, intensity of illumination 1600lx, light application time 8h/d；Routine observation tissue culture bottle, in
Start to sprout after culture 7d, seed sprouting in 25-27 days terminates, germination percentage 99%.
(5) it takes root: aseptic seedling obtained by step (4) is transferred to the aminoadenine of benzyl containing 0.3mg/L and 7mg/L gibberellin
It is cultivated in MS culture medium, wherein 23-26 DEG C of cultivation temperature, humidity 40%, intensity of illumination 1800lx, light application time 10h/
d；Naturally it takes root within 28-30 days.
(6) it transplants: small transplantation of seedlings obtained by step (5) is transplanted to outdoor after it grows up to plantlet into sterile soil again
Soil makes its continued growth under field conditions (factors), and monthly primary with 800 times of liquid fertilisings of a great number of elements in MS, fertilizing method is root
It applies, survival rate is up to 95% or more.
Embodiment cited by the present invention is most highly preferred embodiment of the invention, only does demonstration and uses, other make the present invention
The equivalent forms such as various changes or modification, equally fall within the range that the application is protected.The adenophora tetraphylla tissue-cultured seedling of culture of the present invention
For axenic germination rate up to 95% or more, the survival rate after transplanting uses members a large amount of in MS up to 92% or more during later period transplanting
The transplanting effect of plain 800 times of liquid fertilising, which is better than, is individually applied fertilizer using MS culture medium；The nothing of adenophora tetraphylla seed provided by the invention
Its germination rate of bacterium Germination Technique and transplanting survival rate are above according to document " Liu Caihong, the cultivation technique of adenophora tetraphylla, base's Chinese medicine
Magazine, 2000,73,26 " and document " auspicious friend of dragon etc., the tissue culture and rapid proliferation of adenophora tetraphylla, Plant Physiology Communications,
In April, 2007, volume 43, the 2nd phase " documented by method cultivation or culture adenophora tetraphylla, taken root, transplant after obtain small-sized plant
Strain fast numerous time is short, is conducive to industrialization promotion.
All references mentioned in the present invention is incorporated herein by reference, independent just as each document
It is incorporated as with reference to such.In addition, it should also be understood that, after reading the above teachings of the present invention, those skilled in the art can
To make various changes or modifications to the present invention, such equivalent forms equally fall within model defined by the application the appended claims
It encloses.
Claims (10)
1. a kind of axenic germination method of adenophora tetraphylla seed, which is characterized in that the method includes the steps:
(a) seed is subjected to axenic germination and obtains aseptic seedling；With
(b) aseptic seedling for obtaining the step a) carries out culture of rootage, obtains rooted seedling；
In another preferred example, the method also includes carrying out disinfection to the seed.
2. the method as described in claim 1, which is characterized in that in the step a), the seed is placed in containing NAA (0.4-
0.7mg/L), on the 1/2MS solid medium of 6-BA (0.1-0.4mg/L), sucrose (30g/L) and agar 6.5g/L, in 23-26
DEG C sterile culture, the seed, which is sprouted, obtains the aseptic seedling.
3. the method as described in claim 1, which is characterized in that in the step b), the culture of rootage refers to containing 0.1-
It is cultivated in the MS culture medium of 1mg/L benzyl aminoadenine and 1-10mg/L gibberellin.
4. method a method according to any one of claims 1-3, which is characterized in that the aseptic seedling and rooted seedling culture humidity be
30%-50%, intensity of illumination 1200-2000lx, light application time 8-12h/d.
5. a kind of propagation method of adenophora tetraphylla seed, which is characterized in that the described method includes:
(a) seed is subjected to axenic germination and obtains aseptic seedling；
(b) aseptic seedling for obtaining the step a) carries out culture of rootage, obtains rooted seedling；With
(c) the step b) rooted seedling obtained is cultivated, takes root and obtains plant.
6. method as claimed in claim 5, which is characterized in that being placed in the seed containing NAA (0.4-0.7mg/L), 6-BA
On the 1/2MS solid medium of (0.1-0.4mg/L), sucrose (30g/L) and agar 6.5g/L, in 23-26 DEG C of sterile culture, institute
It states seed and sprouts the acquisition aseptic seedling.
7. method as claimed in claim 5, which is characterized in that in the step b), the culture of rootage refers to containing 0.1-
It is cultivated in the MS culture medium of 1mg/L benzyl aminoadenine and 1-10mg/L gibberellin.
8. method as claimed in claim 5, which is characterized in that the method also includes the plant for obtaining step c) to be implanted into
The step of being cultivated in soil.
9. method as claimed in claim 5, which is characterized in that in the step c), in the 23-26 DEG C of culture rooted seedling.
10. the method as described in claim 5-7 is any, which is characterized in that the aseptic seedling and rooted seedling culture humidity be
30%-50%, intensity of illumination 1200-2000lx, light application time 8-12h/d.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104145651A (en) * | 2014-07-21 | 2014-11-19 | 宁国千方中药发展有限公司 | Planting method of Adenophora triphylla, perennial herb of the family Campanulaceae |
| CN104273027A (en) * | 2013-07-03 | 2015-01-14 | 中国科学院上海生命科学研究院 | Aseptic germination method of Crassulaceae plant seeds |
| CN107646694A (en) * | 2017-11-13 | 2018-02-02 | 陈培党 | A kind of adenophora tetraphylla Regeneration in Vitro tissue cultures technology |
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2018
- 2018-12-25 CN CN201811587817.8A patent/CN109430058A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104273027A (en) * | 2013-07-03 | 2015-01-14 | 中国科学院上海生命科学研究院 | Aseptic germination method of Crassulaceae plant seeds |
| CN104145651A (en) * | 2014-07-21 | 2014-11-19 | 宁国千方中药发展有限公司 | Planting method of Adenophora triphylla, perennial herb of the family Campanulaceae |
| CN107646694A (en) * | 2017-11-13 | 2018-02-02 | 陈培党 | A kind of adenophora tetraphylla Regeneration in Vitro tissue cultures technology |
Non-Patent Citations (1)
| Title |
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| 龙翔友等: "南沙参的组织培养与快速繁殖", 《植物生理学通讯》 * |
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