CN107173236A - A kind of Lilium Germplasm method for tissue culture - Google Patents
A kind of Lilium Germplasm method for tissue culture Download PDFInfo
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- CN107173236A CN107173236A CN201710592608.1A CN201710592608A CN107173236A CN 107173236 A CN107173236 A CN 107173236A CN 201710592608 A CN201710592608 A CN 201710592608A CN 107173236 A CN107173236 A CN 107173236A
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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 flower cultivation technical field there is provided a kind of Lilium Germplasm method for tissue culture, including：（1）Evoked callus；（2）The squamous subculture of callus；（3）The amplification cultivation of callus.It is an object of the invention to provide a kind of Lilium Germplasm method for tissue culture.The medium component adjustment angle that selection and tissue cultures stage of this method from explant are used, is controlled from the source of tissue cultures and process to pollution rate, so that reduce pollution rate, and this method is easily operated, and it is simple and easy to apply.
Description
Technical field
The invention belongs to flower cultivation technical field, in particular it relates to a kind of Lilium Germplasm method for tissue culture.
Background technology
Lily occupies critical role in world ornamental flower produces, with being continuously increased for market demand, people couple
Liliaceous variety and quality requirement also more and more higher, people to lily wild varieties resource, the tissue culture propagation of Lilium Germplasm and
Test tube seedling, which is grown, has done numerous studies, is produced from the traditional lily ball for cultivating into mother bulb by bulbec by scale cuttage
To lily tissue-culturing rapid propagation.Because lily underground bulb quantity is more, it is easy to obtain, materials are not subject to seasonal restrictions and turn into current lily
The topmost explant material of tissue culture, but bulb growth is in underground, carries disease germs many so that and difficulty is increased in tissue culture disinfecting process.
The acquisition of current tissue-cultured derived plant lily using conventional method for tissue culture, still, often goes out in tissue cultures early stage
The pollution again of endogenetic bacteria is there is also during the fungal contamination of existing large area, subculture, the loss of material is caused, simultaneously
Substantial amounts of manpower and materials are wasted, therefore high pollution rate is to perplex the problem of Lilium Germplasm tissue cultures for a long time.At present, to such as
The existing correlative study report of pollution rate during what reduction Lilium Germplasm tissue culture, is on the one hand that each link of tissue culture process is carried out
Strict control, is on the other hand material to be carried out disinfection such as antibiotic, sodium hypochlorite, mercury chloride using surface disinfectant.But
These disinfectants do not reach good pollution prevention effect, or have injury to material and have harm to operating personnel and environment.
Therefore, how to find a kind of efficiently easily operated and small pollution rate tissue culture technique, be during Lilium Germplasm tissue culture urgently
Technical problem to be solved.
The content of the invention
For defect of the prior art, it is an object of the invention to provide a kind of Lilium Germplasm method for tissue culture.The party
The medium component adjustment angle that selection and tissue cultures stage of the method from explant are used, from the source of tissue cultures
Pollution rate is controlled with process, so that reduce pollution rate, and this method is easily operated, it is simple and easy to apply.
A kind of Lilium Germplasm method for tissue culture provided according to the present invention, comprises the following steps：
（1）Evoked callus：Lilium Germplasm capsule is taken, capsule is cut, robust growth, the obvious seed of embryo is chosen, removed
It is seeded in after kind skin is sterilized on callus inducing medium, 2/3rds of inoculation time control seed length is embedded in culture medium
It is interior, after cultivating 20-40 days, a diameter of 0.3-0.5cm faint yellow, granular embryo callus agglomerate is obtained, it is described to be cured
Injured tissue Fiber differentiation is carried out under conditions of complete darkness, temperature is 20-24 DEG C；
The callus inducing medium formula is：1/2MS, 2,4- dichlorphenoxyacetic acids 0.5-1.3mg/L, α-naphthylacetic acid
0.6-1.0mg/L, 6- benzyl aminoadenine 0.1-0.4mg/L, sodium chloride 0.1-0.3mg/L, sterilization Chinese herbal medicine powder 0.1-2mg/
L, glucose 20-26g/L, agar 6-8g/L, pH value is 5.6-5.8；
（2）The squamous subculture of callus：After the completion of induction of callus, progress subculture training on subculture medium is inoculated into
Support, the squamous subculture is carried out under the conditions of complete darkness, 20-25 DEG C of squamous subculture；
The culture medium prescription of the squamous subculture is：1/2MS, 2,4- dichlorphenoxyacetic acids 0.5-1.1mg/L, 2,4-D 0.6-
1.1mg/L, 6- benzyl aminoadenine 0.1-0.3mg/L, sodium chloride 0.1-0.2mg/L, sterilization Chinese herbal medicine powder 0.1-1.5mg/L,
Fructose 20-25g/L, agar 7-9g/L, pH value is 5.6-5.8；
（3）The amplification cultivation of callus：Squamous subculture is after 4 months on subculture medium for callus agglomerate, by callus group
The fritter that agglomerate cuts into 0.2-0.5cm is knitted, is inoculated on amplification culture medium, dim light culture is carried out, dim light culture refers in light
Carried out according under conditions of 100-700lux, 25 DEG C of temperature；
The amplification cultivation based formulas is：1/2MS, 2,4- dichlorphenoxyacetic acids 0.2-1.1mg/L, 2,4-D 0.3-1.2mg/L,
6- benzyl aminoadenines 0.1-0.3mg/L, sodium chloride 0.1-0.3mg/L, sterilization Chinese herbal medicine powder 0.1-1.1mg/L, maltose 18-
27g/L, mashed potatoes 0.2-2g/L, agar 6-10g/L, pH value is 5.6-5.8.
Preferably, the specific method of the sterilization is to be placed in the salt solution that concentration is 5-7% after the seed for removing kind of skin is cleaned
In embathe 10-20min, then use distilled water flushing 0.5-1h, then with 75% alcohol-pickled 0.5-2min.
Preferably, the composition of the Chinese herbal medicine includes honeysuckle, radix scutellariae, wormwood, betel nut, dandelion.
Preferably, the operation of the evoked callus is completed on superclean bench.
Preferably, after the completion of the induction of callus, before being inoculated on subculture medium, in superclean bench
On cut off callus agglomerate toffee part with scalpel.
Preferably, in the Subculture of the callus, it is inoculated with each culture dish equipped with subculture medium
10-20 block callus agglomerates, a diameter of 0.2-0.4cm of callus agglomerate, every 4 weeks squamous subcultures are once.
Preferably, it is described that callus islands are inoculated into before amplification culture medium, first it is inoculated into containing subculture medium
On culture dish, wrapped up with guarantor's tinfoil after sterilized after three layers, carry out the low-temperature treatment of 10-20 days, low-temperature treatment condition is 1-4
℃。
Preferably, the sterilizing is sterilized using ultraviolet irradiation.
Compared with prior art, the present invention has following beneficial effect：
A kind of Lilium Germplasm method for tissue culture that the present invention is provided, this method takes the seed of Lilium Germplasm as Lilium Germplasm group
The explant of culture is knitted, and explant efficiently solves in induction of callus by sterilization and easily fungi, bacterium occurs
The problem of pollution, pollution rate is reduced from source；It is used as bright spot, training of this method in callus inducing medium, squamous subculture
Support base, amplification culture medium in add sterilization Chinese herbal medicine powder composition, sterilization Chinese herbal medicine with honeysuckle, radix scutellariae, wormwood, betel nut,
Dandelion carries out reasonable compatibility, and mentioned component has different degrees of sterilization, resistant effect to plant.Due to being killed in each culture medium
The addition of bacterium Chinese herbal medicine so that lily lily callus is expanded in a safety, with disease-resistant tissue culture environment, life
It is long, improve the anti-infection ability of lily callus.
Embodiment
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention
Rather than limitation the scope of the present invention.
Embodiment 1
A kind of Lilium Germplasm method for tissue culture that the present embodiment is provided, comprises the following steps：
（1）Evoked callus：Lilium Germplasm capsule is taken, capsule is cut, robust growth, the obvious seed of embryo is chosen, removed
It is seeded in after kind skin is sterilized on callus inducing medium, 2/3rds of inoculation time control seed length is embedded in culture medium
It is interior, after cultivating 40 days, a diameter of 0.3cm faint yellow, granular embryo callus agglomerate is obtained, the callus is lured
Leading culture is carried out under conditions of complete darkness, temperature is 24 DEG C, wherein, the specific method of the sterilization is that will remove kind of a skin
Seed clean after be placed in the salt solution that concentration is 5% and embathe 20min, then use distilled water flushing 0.5h, then with the leaching of 75% alcohol
Steep 2min,
The callus inducing medium formula is：1/2MS, 2,4- dichlorphenoxyacetic acids 1.3mg/L, α-naphthylacetic acid 0.6mg/
L, 6- benzyl aminoadenine 0.4mg/L, sodium chloride 0.1mg/L, sterilization Chinese herbal medicine powder 2mg/L, glucose 20g/L, agar 8g/L,
PH value is 5.6；
（2）The squamous subculture of callus：After the completion of induction of callus, progress subculture training on subculture medium is inoculated into
Support, the squamous subculture is carried out under the conditions of complete darkness, 25 DEG C of squamous subculture；
The culture medium prescription of the squamous subculture is：1/2MS, 2,4- dichlorphenoxyacetic acids 0.5mg/L, 2,4-D 1.1mg/L, 6-
Benzyl aminoadenine 0.1mg/L, sodium chloride 0.2mg/L, sterilization Chinese herbal medicine powder 0.1mg/L, fructose 25g/L, agar 7g/L, pH value
For 5.8；
（3）The amplification cultivation of callus：Squamous subculture is after 4 months on subculture medium for callus agglomerate, by callus group
The fritter that agglomerate cuts into 0.5cm is knitted, is inoculated on amplification culture medium, dim light culture is carried out, dim light culture refers in illumination
Carried out under conditions of 100lux, 25 DEG C of temperature,
The amplification cultivation based formulas is：1/2MS, 2,4- dichlorphenoxyacetic acids 1.1mg/L, 2,4-D 0.3mg/L, 6- benzyl amino
Adenine 0.3mg/L, sodium chloride 0.1mg/L, sterilization Chinese herbal medicine powder 1.1mg/L, maltose 18g/L, mashed potatoes 2g/L, agar
6g/L, pH value is 5.8.
Wherein, the composition of the Chinese herbal medicine includes honeysuckle, radix scutellariae, wormwood, betel nut, dandelion.
Wherein, the operation of the evoked callus is completed on superclean bench.
Wherein, after the completion of the induction of callus, before being inoculated on subculture medium, on superclean bench
With scalpel excision callus agglomerate toffee part.
Wherein, in the Subculture of the callus, 20 are inoculated with each culture dish equipped with subculture medium
Block callus agglomerate, a diameter of 0.2cm of callus agglomerate, every 4 weeks squamous subcultures are once.
Wherein, it is described that callus islands are inoculated into before amplification culture medium, first it is inoculated into the training containing subculture medium
Support on ware, wrapped up with guarantor's tinfoil after sterilized after three layers, carry out the low-temperature treatment of 20 days, low-temperature treatment condition is 1 DEG C.It is described
Sterilizing is sterilized using ultraviolet irradiation.
Embodiment 2
A kind of Lilium Germplasm method for tissue culture that the present embodiment is provided, comprises the following steps：
（1）Evoked callus：Lilium Germplasm capsule is taken, capsule is cut, robust growth, the obvious seed of embryo is chosen, removed
It is seeded in after kind skin is sterilized on callus inducing medium, 2/3rds of inoculation time control seed length is embedded in culture medium
It is interior, after cultivating 20 days, a diameter of 0.5cm faint yellow, granular embryo callus agglomerate is obtained, the callus is lured
Leading culture is carried out under conditions of complete darkness, temperature is 20 DEG C, wherein, the specific method of the sterilization is that will remove kind of a skin
Seed clean after be placed in the salt solution that concentration is 7% and embathe 10min, then use distilled water flushing 1h, then alcohol-pickled with 75%
0.5min,
The callus inducing medium formula is：1/2MS, 2,4- dichlorphenoxyacetic acids 0.5mg/L, α-naphthylacetic acid 1.0mg/
L, 6- benzyl aminoadenine 0.1mg/L, sodium chloride 0.3mg/L, sterilization Chinese herbal medicine powder 0.1mg/L, glucose 26g/L, agar 6g/
L, pH value is 5.8；
（2）The squamous subculture of callus：After the completion of induction of callus, progress subculture training on subculture medium is inoculated into
Support, the squamous subculture is carried out under the conditions of complete darkness, 20 DEG C of squamous subculture,
The culture medium prescription of the squamous subculture is：1/2MS, 2,4- dichlorphenoxyacetic acids 1.1mg/L, 2,4-D 0.6mg/L, 6-
Benzyl aminoadenine 0.3mg/L, sodium chloride 0.1mg/L, sterilization Chinese herbal medicine powder 1.5mg/L, fructose 20g/L, agar 9g/L, pH value
For 5.6；
（3）The amplification cultivation of callus：Squamous subculture is after 4 months on subculture medium for callus agglomerate, by callus group
The fritter that agglomerate cuts into 0.2cm is knitted, is inoculated on amplification culture medium, dim light culture is carried out, dim light culture refers in illumination
Carried out under conditions of 700lux, 25 DEG C of temperature；
The amplification cultivation based formulas is：1/2MS, 2,4- dichlorphenoxyacetic acids 0.2mg/L, 2,4-D 1.2mg/L, 6- benzyl amino
Adenine 0.1mg/L, sodium chloride 0.3mg/L, sterilization Chinese herbal medicine powder 0.1mg/L, maltose 27g/L, mashed potatoes 0.2g/L, agar
10g/L, pH value is 5.6.
Wherein, the composition of the Chinese herbal medicine includes honeysuckle, radix scutellariae, wormwood, betel nut, dandelion.
Wherein, the operation of the evoked callus is completed on superclean bench.
Wherein, after the completion of the induction of callus, before being inoculated on subculture medium, on superclean bench
With scalpel excision callus agglomerate toffee part.
Wherein, in the Subculture of the callus, 10 are inoculated with each culture dish equipped with subculture medium
Block callus agglomerate, a diameter of 0.4cm of callus agglomerate, every 4 weeks squamous subcultures are once.
Wherein, it is described that callus islands are inoculated into before amplification culture medium, first it is inoculated into the training containing subculture medium
Support on ware, wrapped up with guarantor's tinfoil after sterilized after three layers, carry out the low-temperature treatment of 10 days, low-temperature treatment condition is 4 DEG C.It is described
Sterilizing is sterilized using ultraviolet irradiation.
Embodiment 3
A kind of Lilium Germplasm method for tissue culture that the present embodiment is provided, comprises the following steps：
（1）Evoked callus：Lilium Germplasm capsule is taken, capsule is cut, robust growth, the obvious seed of embryo is chosen, removed
It is seeded in after kind skin is sterilized on callus inducing medium, 2/3rds of inoculation time control seed length is embedded in culture medium
It is interior, after cultivating 30 days, a diameter of 0.4cm faint yellow, granular embryo callus agglomerate is obtained, the callus is lured
Leading culture is carried out under conditions of complete darkness, temperature is 22 DEG C, wherein, the specific method of the sterilization is that will remove kind of a skin
Seed clean after be placed in the salt solution that concentration is 6% and embathe 15min, then use distilled water flushing 0.8h, then with the leaching of 75% alcohol
Steep 1.0min,
The callus inducing medium formula is：1/2MS, 2,4- dichlorphenoxyacetic acids 0.9mg/L, α-naphthylacetic acid 0.8mg/
L, 6- benzyl aminoadenine 0.3mg/L, sodium chloride 0.2mg/L, sterilization Chinese herbal medicine powder 0.8mg/L, glucose 26g/L, agar 7g/
L, pH value is 5.7；
（2）The squamous subculture of callus：After the completion of induction of callus, progress subculture training on subculture medium is inoculated into
Support, the squamous subculture is carried out under the conditions of complete darkness, 21 DEG C of squamous subculture；
The culture medium prescription of the squamous subculture is：1/2MS, 2,4- dichlorphenoxyacetic acids 0.5-1.1mg/L, 2,4-D 0.9mg/
L, 6- benzyl aminoadenine 0.2mg/L, sodium chloride 0.2mg/L, sterilization Chinese herbal medicine powder 1.1mg/L, fructose 21g/L, agar 8g/L,
PH value is 5.6；
（3）The amplification cultivation of callus：Squamous subculture is after 4 months on subculture medium for callus agglomerate, by callus group
The fritter that agglomerate cuts into 0.4cm is knitted, is inoculated on amplification culture medium, dim light culture is carried out, dim light culture refers in illumination
Carried out under conditions of 600lux, 25 DEG C of temperature,
The amplification cultivation based formulas is：1/2MS, 2,4- dichlorphenoxyacetic acids 1.1mg/L, 2,4-D 1.2mg/L, 6- benzyl amino
Adenine 0.3mg/L, sodium chloride 0.3mg/L, sterilization Chinese herbal medicine powder 1.1mg/L, maltose 27g/L, mashed potatoes 2g/L, agar
7g/L, pH value is 5.8.
Wherein, the composition of the Chinese herbal medicine includes honeysuckle, radix scutellariae, wormwood, betel nut, dandelion.
Wherein, the operation of the evoked callus is completed on superclean bench.
Wherein, after the completion of the induction of callus, before being inoculated on subculture medium, on superclean bench
With scalpel excision callus agglomerate toffee part.
Wherein, in the Subculture of the callus, 10- is inoculated with each culture dish equipped with subculture medium
20 pieces of callus agglomerates, a diameter of 0.3cm of callus agglomerate, every 4 weeks squamous subcultures are once.
Wherein, it is described that callus islands are inoculated into before amplification culture medium, first it is inoculated into the training containing subculture medium
Support on ware, wrapped up with guarantor's tinfoil after sterilized after three layers, carry out the low-temperature treatment of 12 days, low-temperature treatment condition is 2 DEG C.It is described
Sterilizing is sterilized using ultraviolet irradiation.
Experiment test：
Experiment material：Select the Lilium Germplasm capsule of the suitable robust growth of growing way；
Experiment process：Lilium Germplasm capsule is divided into 4 groups of equal decile, wherein, the 1st group, the 2nd group, the 3rd group, using this implementation
Example 1-3 method carries out tissue cultures, and the 4th group carries out tissue cultures using conventional method, counts pollution condition；
As a result show：Wherein, the 1st group, the 2nd group, the 3rd group it is pollution-free, the 4th group of pollution rate is 10%.As can be seen here, it is of the invention
Lilium Germplasm method for tissue culture, reduce pollution rate.
The specific embodiment of the present invention is described above.It is to be appreciated that the invention is not limited in above-mentioned
Particular implementation, those skilled in the art can make various deformations or amendments within the scope of the claims, this not shadow
Ring the substantive content of the present invention.
Claims (8)
1. a kind of Lilium Germplasm method for tissue culture, it is characterised in that：The method for tissue culture comprises the following steps：
（1）Evoked callus：Lilium Germplasm capsule is taken, capsule is cut, robust growth, the obvious seed of embryo is chosen, removed
It is seeded in after kind skin is sterilized on callus inducing medium, 2/3rds of inoculation time control seed length is embedded in culture medium
It is interior, after cultivating 20-40 days, a diameter of 0.3-0.5cm faint yellow, granular embryo callus agglomerate is obtained, it is described to be cured
Injured tissue Fiber differentiation is carried out under conditions of complete darkness, temperature is 20-24 DEG C；
The callus inducing medium formula is：1/2MS, 2,4- dichlorphenoxyacetic acids 0.5-1.3mg/L, α-naphthylacetic acid
0.6-1.0mg/L, 6- benzyl aminoadenine 0.1-0.4mg/L, sodium chloride 0.1-0.3mg/L, sterilization Chinese herbal medicine powder 0.1-2mg/
L, glucose 20-26g/L, agar 6-8g/L, pH value is 5.6-5.8；
（2）The squamous subculture of callus：After the completion of induction of callus, progress subculture training on subculture medium is inoculated into
Support, the squamous subculture is carried out under the conditions of complete darkness, 20-25 DEG C of squamous subculture；
The culture medium prescription of the squamous subculture is：1/2MS, 2,4- dichlorphenoxyacetic acids 0.5-1.1mg/L, 2,4-D 0.6-
1.1mg/L, 6- benzyl aminoadenine 0.1-0.3mg/L, sodium chloride 0.1-0.2mg/L, sterilization Chinese herbal medicine powder 0.1-1.5mg/L,
Fructose 20-25g/L, agar 7-9g/L, pH value is 5.6-5.8；
（3）The amplification cultivation of callus：Squamous subculture is after 4 months on subculture medium for callus agglomerate, by callus group
The fritter that agglomerate cuts into 0.2-0.5cm is knitted, is inoculated on amplification culture medium, dim light culture is carried out, dim light culture refers in light
Carried out according under conditions of 100-700lux, 25 DEG C of temperature；
The amplification cultivation based formulas is：1/2MS, 2,4- dichlorphenoxyacetic acids 0.2-1.1mg/L, 2,4-D 0.3-1.2mg/L,
6- benzyl aminoadenines 0.1-0.3mg/L, sodium chloride 0.1-0.3mg/L, sterilization Chinese herbal medicine powder 0.1-1.1mg/L, maltose 18-
27g/L, mashed potatoes 0.2-2g/L, agar 6-10g/L, pH value is 5.6-5.8.
2. Lilium Germplasm method for tissue culture according to claim 1, it is characterised in that：The specific method of the sterilization is
It is placed in after the seed for removing kind of skin is cleaned in the salt solution that concentration is 5-7% and embathes 10-20min, then uses distilled water flushing 0.5-
1h, then with 75% alcohol-pickled 0.5-2min.
3. Lilium Germplasm method for tissue culture according to claim 1, it is characterised in that：The composition of the Chinese herbal medicine includes
Honeysuckle, radix scutellariae, wormwood, betel nut, dandelion.
4. Lilium Germplasm method for tissue culture according to claim 1, it is characterised in that：The behaviour of the evoked callus
Make to complete on superclean bench.
5. Lilium Germplasm method for tissue culture according to claim 1, it is characterised in that：The induction of callus
After the completion of, before being inoculated on subculture medium, callus agglomerate toffee portion is cut off with scalpel on superclean bench
Point.
6. Lilium Germplasm method for tissue culture according to claim 1, it is characterised in that：The subculture training of the callus
During supporting, be inoculated with 10-20 block callus agglomerates in each culture dish equipped with subculture medium, callus agglomerate it is straight
Footpath is 0.2-0.4cm, and every 4 weeks squamous subcultures are once.
7. Lilium Germplasm method for tissue culture according to claim 1, it is characterised in that：It is described to connect callus islands
Plant to before amplification culture medium, be first inoculated on the culture dish containing subculture medium, three layers are wrapped up with guarantor's tinfoil after sterilized
Afterwards, the low-temperature treatment of progress 10-20 days, low-temperature treatment condition is 1-4 DEG C.
8. Lilium Germplasm method for tissue culture according to claim 7, it is characterised in that：The sterilizing is to use ultraviolet
Irradiation is sterilized.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109463282A (en) * | 2018-12-14 | 2019-03-15 | 河南城建学院 | A kind of Lilium brownii var viridulum clove numerous rooting method fastly |
| CN113179954A (en) * | 2021-06-18 | 2021-07-30 | 广西金秀瑶族自治县亿草丰茂瑶药有限公司 | Method for cultivating tissue culture seedlings by stem segments of artemisia rupestris L |
| CN115125267A (en) * | 2022-06-21 | 2022-09-30 | 沈阳农业大学 | Method for improving oriental lily 'siberia' genetic transformation efficiency |
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| CN113179954B (en) * | 2021-06-18 | 2022-12-30 | 广西金秀瑶族自治县亿草丰茂瑶药有限公司 | Method for cultivating tissue culture seedlings by stem segments of artemisia rupestris L |
| CN115125267A (en) * | 2022-06-21 | 2022-09-30 | 沈阳农业大学 | Method for improving oriental lily 'siberia' genetic transformation efficiency |
| CN115125267B (en) * | 2022-06-21 | 2023-11-21 | 沈阳农业大学 | Method for improving genetic transformation efficiency of oriental lily' Siberia |
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