CN107646689A - The method that a kind of tissue cultures of hymsleya amabilis and later stage breed - Google Patents
The method that a kind of tissue cultures of hymsleya amabilis and later stage breed Download PDFInfo
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- CN107646689A CN107646689A CN201711084109.8A CN201711084109A CN107646689A CN 107646689 A CN107646689 A CN 107646689A CN 201711084109 A CN201711084109 A CN 201711084109A CN 107646689 A CN107646689 A CN 107646689A
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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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- 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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Abstract
The invention belongs to agricultural high-tech field, and in particular to the method that a kind of tissue cultures of hymsleya amabilis and later stage breed, comprise the following steps：Initial culture base is prepared, the sterilization and processing, inoculation and culture, the preparation of subculture medium and inoculation, culture of rootage, hardening, transplanting of explant.The method of the present invention overcomes the loss for the active ingredient cultivated under the specific environment of prior art laboratory, realize the artificial culture of wild varieties, after testing, dihydrocucurbitacin F, cucurbitacin, hemsloside have detection in the hymsleya amabilis for breeding to obtain with the method for the present invention, and its content is suitable with the content of wild hymsleya amabilis.The method of the present invention improves the planting percent of hymsleya amabilis seedling, the effectiveness for shortening cultivation period, keeping wild hymsleya amabilis, has application prospect in the preservation of hymsleya amabilis wild resource, protection, utilization etc..
Description
Technical field
The present invention relates to a kind of method for tissue culture, and in particular to the side that a kind of tissue cultures of hymsleya amabilis and later stage breed
Method, belong to agricultural high-tech field.
Background technology
Hymsleya amabilis is Curcurbitaceae hymsleya amabilis platymiscium, and the platymiscium mainly originates in Southwestern China according to it is currently understood that there are about kind more than 20
Area.Its is bitter in taste, cold in nature, and traditional medicine usage amount is less, and wild resource is compared with horn of plenty.But with the development of medical science, research
It was found that hymsleya amabilis has the effect of clearing heat and detoxicating, anti-inflammatory analgetic.It is usually used in having a stomachache, canker, the infection of the upper respiratory tract, bronchitis,
Pneumonia, bacillary dysentery, enteritis, urinary infection, septicemia and other multi-infections.Hemsleyadine tablet, hymsleya amabilis stomach and intestine in recent years
The hymsleya amabilis crude extracts such as ball, hymsleya amabilis cellulose capsule, hymsleya amabilis detoxifying pills or compound preparation are widely used in clinic, and hymsleya amabilis resource is continuously available out
Hair is utilized, and wild resource is excessively excavated, while because hymsleya amabilis distributed areas are narrow, it is easily stable by interference from human factor, population
Property is low, and reproductive capacity is not high, and subregion hymsleya amabilis resource is endangered or complete extinction.
The method of prior art artificial propagation hymsleya amabilis mainly has tendril cuttage, seeds cultivation, tissue cultures etc..The group of hymsleya amabilis
It is the nutrient solution using human configuration to knit culture, under suitable temperature environment conditions, fast numerous induction is carried out to hymsleya amabilis, can be shortened
Cultivate the time, but prior art easily causes hymsleya amabilis seedling in fast numerous Induction Process and asexual variation occurs, and it is of the prior art
Fast breeding culture medium exist tissue inductivity it is low, induction seedling it is of poor quality, more weak seedlings and seedling proliferation efficiency are low, seedling replanting survival rate
Low problem.
The content of the invention
For overcome the deficiencies in the prior art, the invention provides the method that a kind of tissue cultures of hymsleya amabilis and later stage are bred.
Its technical scheme is：
The method that a kind of tissue cultures of hymsleya amabilis and later stage breed, specifically includes following steps：
（1）Initial culture base is prepared：Using minimal medium MS as minimal medium, 6-BA, NAA of various dose are added respectively,
It is designated as respectively：
A：The agar of MS+3% sucrose+7%（Control）
B：Agar+the 0.5mg/L6-BA+0.02mg/LNAA of MS+3% sucrose+7%
C：Agar+the 1.0mg/L6-BA+0.02mg/LNAA of MS+3% sucrose+7%
D：Agar+the 1.5mg/L6-BA+0.02mg/LNAA of MS+3% sucrose+7%
（2）The sterilization and processing of explant：Bud section at the top of hymsleya amabilis is positioned in beaker, adds appropriate liquid detergent, running water undershoot
Wash 1 ~ 2h, taking-up is dried and sterilized, and is put in sterile beaker with 70% ethanol immersion 30s, aseptic water washing 3 times, then with 0.1% mercuric chloride
Liquid soaks 15min, jog, finally with aseptic water washing 3 ~ 4 times, is blotted with aseptic filter paper standby；
（3）Inoculation and culture：Material is saved into cutting according to a bud one, morphology lower end is inoculated in above-mentioned steps（1）Described
In Initial culture base, every kind of 20 bottles of culture medium inoculated, 3 bud sections of each bottle of inoculation, the material being inoculated with is placed in illumination cultivation
Cultivated in case, after being inoculated with 30 days, observe growing state of each material in different culture media, selected in the best formula of growing way
Plant as squamous subculture material；
（4）The preparation and inoculation of subculture medium：The stem section of Initial culture plant is subjected to cutting according to one leaf of section, is inoculated in
Minitype cuttage expands numerous group：
E：Agar+the 0.1mg/LIBA of MS+3% sucrose+7%
F：Agar+the 0.3mg/LIBA of MS+3% sucrose+7%
G：Agar+the 0.6mg/LIBA of MS+3% sucrose+7%
And Multiple Buds expand numerous group：
H：Agar+the 0.5mg/L6-BA of MS+3% sucrose+7%
I：Agar+the 1.0mg/L6-BA of MS+3% sucrose+7%
J：Agar+the 1.5mg/L6-BA of MS+3% sucrose+7%
Condition of culture is：26 DEG C of temperature, daily illumination 16h, illuminance 3000Lux, after being inoculated with 30 days, observe each material and exist
Growing state in different culture media, the plant that selection is expanded in the best formula of numerous rate is as follow-up squamous subculture material, subsequently
Squamous subculture culture medium selects the optimal expansion breeding culture medium that first time squamous subculture filters out, subculture 3 times, and each cycle is 30
My god；
（5）Culture of rootage：Expand it is numerous after, carry out culture of rootage, expanding the material of numerous acquisition by more than is cut to the material with 2-3 section
Material, is inoculated in root media：
K：Agar+the 0.5mg/LNAA of 1/MS+3% sucrose+7%
L：Agar+the 1.0mg/LNAA of 1/2MS+3% sucrose+7%
M：Agar+the 1.5mg/LNAA of 1/2MS+3% sucrose+7%
Condition of culture is：27 DEG C of temperature, daily illumination 16h, illuminance 3000Lux, after being inoculated with 30 days, observe each material and exist
Situation of taking root in different culture media, select material of the plant in the best formula of root growth as follow-up acclimatization and transplantses；
（6）Hardening：Blake bottle is placed in the warm canopy of transplanting one week to adapt to planting environment illumination and temperature, after one week, beaten slightly
Abroach 2 days, to adapt to the gaseous environment of planting environment, after hardening, material be placed in clear water to the culture medium washed away in root system,
Immersion 30min is carried out to root system with 800 times of Bordeaux mixture, rotted with reducing during seedlings root cultivation；
（7）Transplanting：Matrix based on the fertile soil of selection pine forest top layer, add the perlite and 0.5 times of volume of 0.2 times of volume
Vermiculite mix after be used as cultivation matrix, matrix is layed on seedbed, sprinkling 800 times of carbendazim, use scuppit within second day
Ditch is outputed, is transplanted according to distance between rows and hills 15cm*10cm, shade density is gradually reduced after seedling root restoration ecosystem, height of seedling is extremely
It can be transplanted during 20cm to plantation field and carry out cool canopy built cultivation.
Further, the step（3）In condition of culture be：26 DEG C of temperature, daily illumination 16h, illuminance are
2000Lux。
Further, the step（6）Before middle transplanting, the shade rate for transplanting warm canopy is controlled more than 0.6 with sunshade net, temperature
Degree control is at 25-32 DEG C.
Further, the step（7）Ensure ambient humidity more than 60% after middle cultivation.
Beneficial effects of the present invention are：
1st, method of the invention overcomes the loss for the active ingredient cultivated under the specific environment of prior art laboratory, realizes open country
The artificial culture of health product kind, after testing, dihydrocucurbitacin F, cucurbitacin, snow in obtained hymsleya amabilis are bred with the method for the present invention
Courage saponin has detection, and its content is suitable with the content of wild hymsleya amabilis.
2nd, method of the invention improves the planting percent of hymsleya amabilis seedling, the effectiveness for shortening cultivation period, keeping wild hymsleya amabilis,
The preservation of hymsleya amabilis wild resource, protection, utilization etc. have application prospect.
Embodiment
Material：Hymsleya amabilis explant is collected in Lincang Hui Kang Chinese medicine plants base in April, 2016, and material is together put with ice cube
Transport is put, kind is big fruit hymsleya amabilis, and selection top tender stem segmentses are as material.
Embodiment 1
The method that a kind of tissue cultures of hymsleya amabilis and later stage breed, specifically includes following steps：
（1）Initial culture base is prepared
Using minimal medium MS as minimal medium, 6-BA, NAA of various dose are added respectively.It is designated as respectively：
1. the agar of MS+3% sucrose+7%（Control）
2. the mg/L 6-BA+0.02mg/L NAA of+7% agar of MS+3% sucrose+0.5
3. the mg/L 6-BA+0.02mg/L NAA of+7% agar of MS+3% sucrose+1.0
4. the mg/L 6-BA+0.02mg/L NAA of+7% agar of MS+3% sucrose+1.5
（2）The sterilization and processing of explant
Bud section at the top of hymsleya amabilis is positioned in beaker, adds appropriate liquid detergent, 1 h is rinsed under running water, taking-up is dried.Handle well
Material carries out disinfection in superclean bench, puts in sterile beaker and soaks 30 s with 70% ethanol, aseptic water washing 3 times, then with 0.1%
Mercuric chloride liquid soaks 15min, jog, finally with aseptic water washing 3 times, is blotted with aseptic filter paper standby.
（3）Inoculation and culture
Material is saved into cutting according to a bud one, morphology lower end is inoculated in above Initial culture base, every kind of culture medium inoculated
20 bottles, 3 bud sections of each bottle of inoculation, the material being inoculated with is placed in illumination box and cultivated.Condition of culture is：Temperature 26
DEG C, the daily h of illumination 16, illuminance is 2000 Lux.After inoculation 30 days, growth of each material in different culture media is observed
Situation, the plant in the best formula of growing way is selected as squamous subculture material.
（4）The preparation and inoculation of subculture medium
The stem section of Initial culture plant is subjected to cutting according to one leaf of section, minitype cuttage is inoculated in and expands numerous group：
5. agar+0.1mg/L the IBA of MS+3% sucrose+7%
6. agar+0.3mg/L the IBA of MS+3% sucrose+7%
7. agar+the 0.6mg/LIBA of MS+3% sucrose+7%
And Multiple Buds expand numerous group：
8. agar+0.5mg/L the 6-BA of MS+3% sucrose+7%
9. agar+1.0mg/L the 6-BA of MS+3% sucrose+7%
10. agar+1.5mg/L the 6-BA of MS+3% sucrose+7%
Condition of culture is：26 DEG C of temperature, the daily h of illumination 16, illuminance are 3000 Lux.
After inoculation 30 days, growing state of each material in different culture media is observed, the best formula of numerous rate is expanded in selection
In plant filtered out from first time squamous subculture optimal as follow-up squamous subculture material, follow-up squamous subculture culture medium
Expand breeding culture medium, subculture 3 times, each cycle is 30 days.
（5）Culture of rootage
Expand it is numerous after, carry out culture of rootage, expanding the material of numerous acquisition by more than is cut to the material with 2 sections, is inoculated in and takes root
In culture medium：
Agar+0.5mg/L the NAA of 11 1/MS+3% sucrose+7%
Agar+1.0mg/L the NAA of 12 1/2MS+3% sucrose+7%
Agar+1.5mg/L the NAA of 13 1/2MS+3% sucrose+7%
Condition of culture is：27 DEG C of temperature, the daily h of illumination 16, illuminance are 3000 Lux.
After inoculation 30 days, take root situation of each material in different culture media is observed, root growth is best matches somebody with somebody for selection
Material of the plant as follow-up acclimatization and transplantses in side.
（6）Hardening and transplanting
Before transplanting, the shade rate for transplanting warm canopy is controlled more than 0.6 with sunshade net, temperature control is within 25 DEG C.
Hardening：Blake bottle is placed in the warm canopy of transplanting one week to adapt to planting environment illumination and temperature, after one week, beaten slightly
Abroach 2 days, to adapt to the gaseous environment of planting environment.
After hardening, material is placed in clear water to the culture medium washed away in root system, root system carried out with 800 times of Bordeaux mixture
30min is soaked, is rotted with reducing during seedlings root cultivation.
Matrix based on the local pine forest top layer fertile soil in selection Lincang, add the perlite and 0.5 of 0.2 times of volume
The vermiculite of times volume be used as cultivation matrix after mixing, and matrix is layed on seedbed, 800 times of carbendazim of sprinkling, second day
Ditch is outputed with scuppit, according to distance between rows and hills（15cm*10cm）Transplanted, ensure ambient humidity more than 60% after cultivation.Treat seedling
Shade density is gradually reduced after root restoration ecosystem, field carries out cool canopy built cultivation to height of seedling to that can be transplanted during 20cm to planting.
Embodiment 2
The method that a kind of tissue cultures of hymsleya amabilis and later stage breed, specifically includes following steps：
（1）Initial culture base is prepared
Using minimal medium MS as minimal medium, 6-BA, NAA of various dose are added respectively.It is designated as respectively：
1. the agar of MS+3% sucrose+7%（Control）
2. the mg/L 6-BA+0.02mg/L NAA of+7% agar of MS+3% sucrose+0.5
3. the mg/L 6-BA+0.02mg/L NAA of+7% agar of MS+3% sucrose+1.0
4. the mg/L 6-BA+0.02mg/L NAA of+7% agar of MS+3% sucrose+1.5
（2）The sterilization and processing of explant
Bud section at the top of hymsleya amabilis is positioned in beaker, adds appropriate liquid detergent, 2h is rinsed under running water, taking-up is dried.Handle well
Material carries out disinfection in superclean bench, puts in sterile beaker and soaks 30 s with 70% ethanol, aseptic water washing 3 times, then with 0.1%
Mercuric chloride liquid soaks 15min, jog, finally with aseptic water washing 4 times, is blotted with aseptic filter paper standby.
（3）Inoculation and culture
Material is saved into cutting according to a bud one, morphology lower end is inoculated in above Initial culture base, every kind of culture medium inoculated
20 bottles, 3 bud sections of each bottle of inoculation, the material being inoculated with is placed in illumination box and cultivated.Condition of culture is：Temperature 26
DEG C, the daily h of illumination 16, illuminance is 2000 Lux.After inoculation 30 days, growth of each material in different culture media is observed
Situation, the plant in the best formula of growing way is selected as squamous subculture material.
（4）The preparation and inoculation of subculture medium
The stem section of Initial culture plant is subjected to cutting according to one leaf of section, minitype cuttage is inoculated in and expands numerous group：
5. agar+0.1mg/L the IBA of MS+3% sucrose+7%
6. agar+0.3mg/L the IBA of MS+3% sucrose+7%
7. agar+the 0.6mg/LIBA of MS+3% sucrose+7%
And Multiple Buds expand numerous group：
8. agar+0.5mg/L the 6-BA of MS+3% sucrose+7%
9. agar+1.0mg/L the 6-BA of MS+3% sucrose+7%
10. agar+1.5mg/L the 6-BA of MS+3% sucrose+7%
Condition of culture is：26 DEG C of temperature, the daily h of illumination 16, illuminance are 3000 Lux.
After inoculation 30 days, growing state of each material in different culture media is observed, the best formula of numerous rate is expanded in selection
In plant filtered out from first time squamous subculture optimal as follow-up squamous subculture material, follow-up squamous subculture culture medium
Expand breeding culture medium, subculture 3 times, each cycle is 30 days.
（5）Culture of rootage
Expand it is numerous after, carry out culture of rootage, expanding the material of numerous acquisition by more than is cut to the material with 3 sections, is inoculated in and takes root
In culture medium：
Agar+0.5mg/L the NAA of 11 1/MS+3% sucrose+7%
Agar+1.0mg/L the NAA of 12 1/2MS+3% sucrose+7%
Agar+1.5mg/L the NAA of 13 1/2MS+3% sucrose+7%
Condition of culture is：27 DEG C of temperature, the daily h of illumination 16, illuminance are 3000 Lux.
After inoculation 30 days, take root situation of each material in different culture media is observed, root growth is best matches somebody with somebody for selection
Material of the plant as follow-up acclimatization and transplantses in side.
（6）Hardening and transplanting
Before transplanting, the shade rate for transplanting warm canopy is controlled more than 0.6 with sunshade net, temperature control is within 32 DEG C.
Hardening：Blake bottle is placed in the warm canopy of transplanting one week to adapt to planting environment illumination and temperature, after one week, beaten slightly
Abroach 2 days, to adapt to the gaseous environment of planting environment.
After hardening, material is placed in clear water to the culture medium washed away in root system, root system carried out with 800 times of Bordeaux mixture
30min is soaked, is rotted with reducing during seedlings root cultivation.
Matrix based on the local pine forest top layer fertile soil in selection Lincang, add the perlite and 0.5 of 0.2 times of volume
The vermiculite of times volume be used as cultivation matrix after mixing, and matrix is layed on seedbed, 800 times of carbendazim of sprinkling, second day
Ditch is outputed with scuppit, according to distance between rows and hills（15cm*10cm）Transplanted, ensure ambient humidity more than 60% after cultivation.Treat seedling
Shade density is gradually reduced after root restoration ecosystem, field carries out cool canopy built cultivation to height of seedling to that can be transplanted during 20cm to planting.
Claims (4)
1. the method that a kind of tissue cultures of hymsleya amabilis and later stage breed, it is characterised in that specifically include following steps：
（1）Initial culture base is prepared：Using minimal medium MS as minimal medium, 6-BA, NAA of various dose are added respectively,
It is designated as respectively：
A：The agar of MS+3% sucrose+7%
B：The mg/L 6-BA+0.02mg/L NAA of+7% agar of MS+3% sucrose+0.5
C：The mg/L 6-BA+0.02mg/L NAA of+7% agar of MS+3% sucrose+1.0
D：The mg/L 6-BA+0.02mg/L NAA of+7% agar of MS+3% sucrose+1.5
（2）The sterilization and processing of explant：Bud section at the top of hymsleya amabilis is positioned in beaker, adds appropriate liquid detergent, running water undershoot
Wash 1 ~ 2 h, taking-up is dried and sterilized, and is put in sterile beaker with 70% ethanol, 30 s of immersion, aseptic water washing 3 times, then with 0.1% liter
Mercury solution soaks 15min, jog, finally with aseptic water washing 3 ~ 4 times, is blotted with aseptic filter paper standby；
（3）Inoculation and culture：Material is saved into cutting according to a bud one, morphology lower end is inoculated in above-mentioned steps（1）Described
In Initial culture base, every kind of 20 bottles of culture medium inoculated, 3 bud sections of each bottle of inoculation, the material being inoculated with is placed in illumination cultivation
Cultivated in case, after being inoculated with 30 days, observe growing state of each material in different culture media, selected in the best formula of growing way
Plant as squamous subculture material；
（4）The preparation and inoculation of subculture medium：The stem section of Initial culture plant is subjected to cutting according to one leaf of section, is inoculated in
Minitype cuttage expands numerous group：
E：Agar+0.1mg/L the IBA of MS+3% sucrose+7%
F：Agar+0.3mg/L the IBA of MS+3% sucrose+7%
G：Agar+the 0.6mg/LIBA of MS+3% sucrose+7%
And Multiple Buds expand numerous group：
H：Agar+0.5mg/L the 6-BA of MS+3% sucrose+7%
I：Agar+1.0mg/L the 6-BA of MS+3% sucrose+7%
J：Agar+1.5mg/L the 6-BA of MS+3% sucrose+7%
Condition of culture is：26 DEG C of temperature, the daily h of illumination 16, illuminance are 3000 Lux, after being inoculated with 30 days, observe each material
Growing state in different culture media, the plant that selection is expanded in the best formula of numerous rate as follow-up squamous subculture material, after
Continuous squamous subculture culture medium selects the optimal expansion breeding culture medium that first time squamous subculture filters out, subculture 3 times, and each cycle is 30
My god；
（5）Culture of rootage：Expand it is numerous after, carry out culture of rootage, expanding the material of numerous acquisition by more than is cut to the material with 2-3 section
Material, is inoculated in root media：
K：Agar+0.5mg/L the NAA of 1/MS+3% sucrose+7%
L：Agar+1.0mg/L the NAA of 1/2MS+3% sucrose+7%
M：Agar+1.5mg/L the NAA of 1/2MS+3% sucrose+7%
Condition of culture is：27 DEG C of temperature, the daily h of illumination 16, illuminance are 3000 Lux, after being inoculated with 30 days, observe each material
Situation of taking root in different culture media, select material of the plant in the best formula of root growth as follow-up acclimatization and transplantses
Material；
（6）Hardening：Blake bottle is placed in the warm canopy of transplanting one week to adapt to planting environment illumination and temperature, after one week, beaten slightly
Abroach 2 days, to adapt to the gaseous environment of planting environment, after hardening, material be placed in clear water to the culture medium washed away in root system,
Immersion 30min is carried out to root system with 800 times of Bordeaux mixture, rotted with reducing during seedlings root cultivation；
（7）Transplanting：Matrix based on the fertile soil of selection pine forest top layer, add the perlite and 0.5 times of volume of 0.2 times of volume
Vermiculite mix after be used as cultivation matrix, matrix is layed on seedbed, sprinkling 800 times of carbendazim, use scuppit within second day
Ditch is outputed, is transplanted according to distance between rows and hills 15cm*10cm, shade density is gradually reduced after seedling root restoration ecosystem, height of seedling is extremely
It can be transplanted during 20cm to plantation field and carry out cool canopy built cultivation.
2. the method that a kind of tissue cultures of hymsleya amabilis according to claim 1 and later stage breed, it is characterised in that the step
Suddenly（3）In condition of culture be：26 DEG C of temperature, the daily h of illumination 16, illuminance are 2000 Lux.
3. the method that a kind of tissue cultures of hymsleya amabilis according to claim 1 and later stage breed, it is characterised in that the step
Suddenly（6）Before middle transplanting, the shade rate for transplanting warm canopy is controlled more than 0.6 with sunshade net, temperature control is at 25-32 DEG C.
4. the method that a kind of tissue cultures of hymsleya amabilis according to claim 1 and later stage breed, it is characterised in that the step
Suddenly（7）Ensure ambient humidity more than 60% after middle cultivation.
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| CN108739123A (en) * | 2018-06-29 | 2018-11-06 | 黔南州司奇中草药种植有限公司 | A kind of implantation methods of hymsleya amabilis |
| CN109287481A (en) * | 2018-10-29 | 2019-02-01 | 云南善源生物科技发展有限公司 | A kind of tissue culture medium of hymsleya amabilis and its mating system |
| CN109315289A (en) * | 2018-10-09 | 2019-02-12 | 云南善源生物科技发展有限公司 | Hymsleya amabilis Novel sterilizing mode |
| CN111411099A (en) * | 2020-05-14 | 2020-07-14 | 云南农业大学 | Hemsleya amabilis acetyl transferase, coding gene thereof and application of hemsleya amabilis acetyl transferase in preparation of cucurbitacin |
| CN111480576A (en) * | 2020-05-20 | 2020-08-04 | 西南林业大学 | Method for inducing hemsleya amabilis polyploid plant by using adventitious buds and application of method |
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2017
- 2017-11-07 CN CN201711084109.8A patent/CN107646689A/en active Pending
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| 王蒙蒙等: "浙江雪胆快速繁殖技术的探讨", 《浙江农业科学》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108739123A (en) * | 2018-06-29 | 2018-11-06 | 黔南州司奇中草药种植有限公司 | A kind of implantation methods of hymsleya amabilis |
| CN109315289A (en) * | 2018-10-09 | 2019-02-12 | 云南善源生物科技发展有限公司 | Hymsleya amabilis Novel sterilizing mode |
| CN109287481A (en) * | 2018-10-29 | 2019-02-01 | 云南善源生物科技发展有限公司 | A kind of tissue culture medium of hymsleya amabilis and its mating system |
| CN111411099A (en) * | 2020-05-14 | 2020-07-14 | 云南农业大学 | Hemsleya amabilis acetyl transferase, coding gene thereof and application of hemsleya amabilis acetyl transferase in preparation of cucurbitacin |
| CN111411099B (en) * | 2020-05-14 | 2022-09-27 | 云南农业大学 | Hemsleya amabilis acetyl transferase, coding gene thereof and application of hemsleya amabilis acetyl transferase in preparation of cucurbitacin |
| CN111480576A (en) * | 2020-05-20 | 2020-08-04 | 西南林业大学 | Method for inducing hemsleya amabilis polyploid plant by using adventitious buds and application of method |
| CN111480576B (en) * | 2020-05-20 | 2021-07-30 | 西南林业大学 | Method for inducing hemsleya amabilis polyploid plant by using adventitious buds and application of method |
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