CN106258979B - A kind of hybridization paulownia seedling cultural method - Google Patents
A kind of hybridization paulownia seedling cultural method Download PDFInfo
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- CN106258979B CN106258979B CN201610769778.8A CN201610769778A CN106258979B CN 106258979 B CN106258979 B CN 106258979B CN 201610769778 A CN201610769778 A CN 201610769778A CN 106258979 B CN106258979 B CN 106258979B
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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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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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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Abstract
The present invention provides a kind of hybridization paulownia seedling cultural method, and step includes sterile bud culture：To hybridize paulownia tender stem or tender shoots as explant, after sterilized, sterilizing, cleaning, it is seeded in 1/2MS or MS and adds and cultivate 12~18 days acquisition sterile buds on the culture medium of appropriate plant growth regulator；Adventitious shoots culture：Clip sterile bud is inoculated on the adventitious shoots culture base for adding appropriate plant growth regulator to prepare with MS and cultivates；Strong sprout, culture of rootage：Multiple Buds, which are grown to after 1 2cm, to be inoculated into 1/2MS and/or MS and adds sub-bottle culture in the strong sprout of appropriate plant growth regulator, root media, 15~30 DEG C, 2000~3000LUX of luminous intensity of above-mentioned steps cultivation temperature, 6~12 hour/day of light application time；Acclimatization and transplants；Transplantation of Regenerated Plantlets will be taken root to garden mould+peat (2：1) in matrix, shading net shading, humidity 60 70% is out of the garden for 15~30 days.The present invention greatly improves the hybridization paulownia seedling speed of growth and planting benefit, at low cost, and pollution is small, and the four seasons can produce, and is suitble to extensive nursery.
Description
Technical field
The present invention relates to a kind of cultural methods of hybridization paulownia seedling.
Background technology
Paulownia Scrophulariaceae Paulownia (Paulownia) plant, it is distributed more widely in China, it is main broad-leaved fast-growing tree
Kind.Paulownia growth is very rapid, becomes a useful person morning, it is civil have " 1 year bar, 2 years as umbrella, 5 years can saw plate " say.Hybridization
Paulownia " 9501 " are one of 15 key research project achievements of Paulownia research institute of the State Administration of Forestry, are the advantages in Paulownia
Kind, it is the famous fast-growing high quality timber seeds in China, adaptable, growth is fast, becomes a useful person morning, in the case where condition is suitable, 1 year
Up to 7-8 meters of age of tree tree growth, 15-20 centimeters of ground diameter or more, 3-4, which can take turns, to be cut down or cuts down together, and single plant may be up to when felling within 5 years
18 meters, the chest timber diameter of a cross-section of a tree trunk 1.3 meters above the ground is up to 40 centimeters or more, 0.5-0.6 cubic metres of the volume of timber, and the grain of wood is logical straight, and decorative pattern is beautiful, color and luster
Pleasing, material is light and soft, and density is low, dimensionally stable, does not stick up and does not split, have make veneer wood for example glued board, jigsaw, integrated timber it is excellent
Good timber matter characteristic；Paulownia material insulation excellent insulating property, resonance performance is good, and radiation damping is high, and interior friction is small, is excellent
Stringed musical instrument material, can be eponymous with famous fiddle butt fish scale dragon spruce；The wood-fibred content of paulownia material is up to 50% or more, is plane
The good raw materials material such as card, ecological board, papermaking；Leaf is good feed etc. again.Therefore, paulownia are always that broad masses are liked
One of the chief species with material and improvement environment of love, supply falls short of demand for product, and market prospects are wide, and economic value is high, paulownia
It is eco-friendly seeds for native country seeds.Therefore, hybridize the industrialization growing and cultivation of paulownia to ask as of people's attention
Topic.
Currently, the growing and cultivation of hybridization paulownia is mainly to carry out nursery technique using traditional seedling nursery method or root is divided to move
It plants, the nursery production cycle is long, takes up a large area, it is difficult to realize efficiently quickly heavy industrialization growing and cultivation.
Invention content
The technical problem to be solved in the present invention is, provides a kind of hybridization paulownia seedling cultural method, overcomes the prior art difficult
To realize efficiently the quickly defect of heavy industrialization growing and cultivation.The present invention solves technical side used by its technical problem
Case is：
There is provided a kind of hybridization paulownia seedling cultural method, which is characterized in that include the following steps：
S1, sterile bud culture：
To hybridize paulownia tender stem or tender shoots as explant, first through alcohol disinfecting, sterile water wash is used afterwards 1 time, then use
HgCl2After solution is sterilized, through sterile water wash 3-5 times, then it is connected on sterile bud culture medium and carries out culture 12~18 days,
Cultivation temperature is 15~30 DEG C, 2000~3000LUX of intensity of illumination, and 6~12 hour/day of light application time obtains sterile bud；
Or the hybridization paulownia sapling for just growing 1-2.5mm long tender shoots is put into biochemical cultivation case, in 38~40 DEG C of conditions
After lower culture 6~8 days, clip hybridizes paulownia tender shoots through alcohol disinfecting and HgCl2After solution sterilization, after sterile washing 3-5 times,
It cuts in the new culture medium configured of bud point access of 1.8-2.2mm long and cultivates, after tender shoots grows 2.5mm long, then cut 1.8-
Cultivated in the bud point access culture medium of 2.2mm long, repeatedly for three times or three times more than, obtain the sterile bud of detoxification；
The medium component includes plant growth regulator, carbon source, 1/2MS and/or MS；
S2, adventitious shoots culture：
The sterile bud that clip is grown is inoculated into 15~20 days/generation of culture on adventitious shoots culture base, can continuous subculture 4-5
Secondary, bud growth coefficient is 10~14.8 times/month, obtains a large amount of Multiple Buds；The culture medium of the medium component and step S1
Ingredient is equivalent；
The cultivation temperature of step S2 is 15~30 DEG C, 2000~3000LUX of intensity of illumination, light application time 6~12 hours/
It；
S3, strong sprout, culture of rootage：
In the culture medium that seedling separation access has newly configured after Multiple Buds grow to 1-2cm long, after cultivating 15~20 days, it is good for
Strong rooted seedling；The medium component is equal with medium component in step S1；
The cultivation temperature of step S3 is 15~30 DEG C, 2000~3000LUX of intensity of illumination, light application time 6~12 hours/
It；
S4, acclimatization and transplants；
The paulownia strong sprout that step S3 is obtained is transplanted to crop field hardening, the matrix that the hardening uses is mixed for garden mould and peat
It closes, and is shaded with shading net to hardening matrix.
Preferably, the alcohol by volume score used in the step S1 is 70%~80%, used HgCl2Solution matter
It is 0.05%~0.15% to measure score.
Preferably, the plant growth regulator in the culture medium in the step S1-S3, carbon source, 1/2MS and/or MS
Preparation method is：Using 1/2MS and/or MS as minimal medium, appropriate plant growth regulator is added and carbon source is formulated；It should
The pH of sterile bud culture medium is 6.5~7.2.
Preferably, the plant growth regulator in described step S1, S2, S3 is 6-BA and/or NAA and/or IBA, configuration
At a concentration of 0.1mg/L-2.0mg/L of the 6-BA after culture medium, NAA a concentration of 0.1mg/L -1.0mg/L, IBA is a concentration of
0.1mg/L-1.0mg/L。
Preferably, the culture medium in described step S1, S2, S3 also need to be added coagulator, agar powder or card can be added
Glue is drawn, if selecting agar powder, the addition of agar powder is 5-6g/L；If selecting carragheen, the addition of carragheen is 8-15g/
L。
Preferably, the best condition of culture of culture medium in described step S1, S2, S3 is that pH is 6.8, this PH is most suitable profit
In the acid-base value of Multiple Buds and culture of rootage, cultivation temperature is 25 ± 2 DEG C, and intensity of illumination is 2800~3000LUX, light application time
10~12 hours/day, this culture room temperature and intensity of illumination and time keep tissue-cultured seedling tiller growth coefficient best, leaf blade size
Moderate, leaf color is dark green, and stem is relatively sturdy, and can shorten incubation time 1~2 day.
Preferably, the optimum altitude for the sterile bud that clip is grown in the step S2 is 1-2cm, and such length is conducive to seedlings picking
Operation is also conducive to squamous subculture, and too short then seedling is too small, and culture medium nutrition also has residue, the nutrition in long culture medium to consume only,
It is unfavorable for the growth of Regenerated plant, the speed of growth of the adventitious shoots culture of the step S2 is 15~20 days/generation, continuously cultivates subculture
4-5 times, the bud growth coefficient of the Multiple Buds is 10~14.8 times/month.
Preferably, the carbon source be sucrose or glucose, the culture medium sucrose concentration being configured to when using sucrose be 18~
30g/L, the culture medium concentration of glucose being configured to when using glucose are 25~40g/L.
Preferably, garden mould and peat quality proportioning are (1.5~2.5) in the hardening matrix of the step S4：1, hardening area
Humidity is 60-70%.
Implement present invention hybridization paulownia seedling cultural method, compared with the prior art, advantage is：
1, the method applied in the present invention so that paulownia cultivate the speed of growth and planting benefit greatly improves；The side of the present invention
Method hybridizes a monthly average growth coefficient of paulownia bud up to 10~14.8, and existing hybridization paulownia stooling method for transplanting one
Strain can only at most grow up to a young plant；
2, after detoxification treatment, the speed of growth and planting benefit significantly improve sterile bud in the method that the present invention uses；
The method of the present invention, plant grows thickly growth after detoxification treatment one month, high uniformity, mean blade width 1.4cm, the number of blade 4~6
Piece is grown very fast after transplanting；Non- detoxification treatment stooling propagation method majority single plant growth, height size is uneven, average leaf
Wide 0.4~1.0cm, the number of blade 1~3 are grown relatively slow after transplanting；
3, cost of material used by the method that uses of the present invention is low, do not generated in cultivating process it is environmentally hazardous because
Element all uses environmental protection material so that environmental pollution is small；
4, the method and process that the present invention uses is simpler, can produce throughout the year, is suitble to extensive nursery production.
Specific implementation mode
Below in conjunction with embodiment, the invention will be further described.
Embodiment one
A kind of hybridization paulownia seedling cultural method of the present invention includes the following steps：
S1, sterile bud culture is carried out：To hybridize paulownia tender stem or tender shoots as explant, first volume ratio is the wine of 70-80%
Essence disinfection, uses sterile water wash 1 time afterwards, then uses mass fraction for 0.05-0.15%HgCl2Solution sterilized after, warp
Then sterile water wash 3-5 times is connected on sterile bud culture medium and is cultivated, sterile bud is obtained after cultivating 16~18 days.
Cultivated days depend mainly on the speed of growth of hybridization paulownia tender shoots, and general sterile bud grows to 1 centimetre or so and is
Preferably.Certainly, sterile bud is longer than 1 centimetre (such as growing to 1.5 centimetres) or shorter than 1 centimetre (such as growing to 0.5 centimetre), not shadow
Ring the realization of the object of the invention.
Culture medium includes plant growth regulator, carbon source, 1/2MS and/or MS, and preparation method is：With 1/2MS and/or MS
For minimal medium, appropriate plant growth regulator is added and carbon source is formulated, the pH6.8 of the sterile bud culture medium.
The plant growth regulator can be used including but not limited to 6-BA, NAA or IBA while use 6-BA and NAA,
IBA.The addition for being configured to 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA are (0.1mg/L-
1.0mg/L)。
Including but not limited to sucrose, glucose may be used in carbon source.When using sucrose, it is configured to a concentration of after culture medium
18-30g/L；When using glucose, it is configured to a concentration of 25-40g/L after culture medium.
Coagulator will be added in culture medium, and agar powder, carragheen etc. can be added, if selecting agar powder, the addition of agar powder
For 5-6g/L；If selecting carragheen, the addition of carragheen is 8-15g/L.
The sterile bud culture medium of hybridization paulownia can be used including but not limited to by the formula after being configured to culture medium：
MS+6-BA1.0mg/L
MS+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+IBA0.1mg/L
In the culture medium, carbon source must also be added, when carbon source uses sucrose, addition is 30g/L after being configured to culture medium,
When carbon source uses glucose, addition is 40g/L after being configured to culture medium.
Note：
MS culture mediums：Be Murashige and Skoog in 1962 it is tobacco cell Training Design, its main feature is that inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, its nitrate content is high, and the quantity and ratio of nutrient are closed
It is suitable, the nutrition and physiological requirements of plant cell can be met, thus the scope of application is wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named with their name.
1/2MS culture mediums：It is the culture medium that the amount of the calcium salt and a great number of elements in MS culture mediums halves, and common training
Support base.
Plant growth regulator 6-BA：6-benzyl aminopurine is a kind of basic element of cell division, the general entitled 6- of English
Benzylaminopurine, molecular formula C12H11N5。
Plant growth regulator NAA：1-Naphthaleneacetic acid, methyl α-naphthyl acetate, abbreviation NAA are a kind of plants
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula are C12H13NO2, it is a kind of auximone.
The cultivation temperature of step S1 is 15~30 DEG C, 2000~3000LUX of intensity of illumination, light application time 6~12 hours/
It；
S2, adventitious shoots culture：
The sterile bud that clip is grown is inoculated into 15~20 days/generation of culture on adventitious shoots culture base, can continuous subculture 4-5
Secondary, bud growth coefficient is 10~14.8 times/month, obtains a large amount of Multiple Buds；The culture medium of the medium component and step S1
Ingredient is equivalent；
The cultivation temperature of step S2 is 15~30 DEG C, 2000~3000LUX of intensity of illumination, light application time 6~12 hours/
It；
S3, strong sprout, culture of rootage：
In the culture medium that seedling separation access has newly configured after Multiple Buds grow to 1-2cm long, after cultivating 15~20 days, it is good for
Strong rooted seedling；The medium component is equal with medium component in step S1；
The cultivation temperature of step S3 is 15~30 DEG C, 2000~3000LUX of intensity of illumination, light application time 6~12 hours/
It；
S4, acclimatization and transplants are carried out：
The obtained paulownia seedling of step S3 is transplanted, the condition of culture of step S4 is：Hardening matrix garden mould+peat matter
Amount proportioning (1.5-2.5)：1, shading net shades, humidity 60-70%.
Embodiment two
A kind of hybridization paulownia seedling cultural method of the present invention includes the following steps：
S1, sterile bud culture is carried out：To hybridize paulownia tender stem or tender shoots as explant, the alcohol that first volume ratio is 75% soaks
Bubble disinfection, uses sterile water wash 1 time afterwards, then uses mass fraction for 0.15%HgCl2Solution sterilized after, through sterile water
Cleaning 3-5 times, is then connected on sterile bud culture medium and is cultivated, and sterile bud is obtained after cultivating 16~18 days.
Cultivated days depend mainly on the speed of growth of hybridization paulownia tender shoots, and general sterile bud grows to 1 centimetre or so and is
Preferably.Certainly, sterile bud is longer than 1 centimetre (such as growing to 1.5 centimetres) or shorter than 1 centimetre (such as growing to 0.5 centimetre), not shadow
Ring the realization of the object of the invention.
Culture medium includes plant growth regulator, carbon source, 1/2MS and/or MS, and preparation method is：With 1/2MS and/or MS
For minimal medium, appropriate plant growth regulator is added and carbon source is formulated, the pH6.8 of the sterile bud culture medium.
The plant growth regulator can be used including but not limited to 6-BA, NAA or IBA while use 6-BA and NAA,
IBA.The addition for being configured to 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA are (0.1mg/L-
1.0mg/L)。
Including but not limited to sucrose, glucose may be used in carbon source.When using sucrose, it is configured to a concentration of after culture medium
30g/L；When using glucose, it is configured to a concentration of 40g/L after culture medium.
Coagulator will be added in culture medium, and agar powder, carragheen etc. can be added, if selecting agar powder, the addition of agar powder
For 6g/L；If selecting carragheen, the addition of carragheen is 15g/L.
The sterile bud culture medium of hybridization paulownia can be used including but not limited to by the formula after being configured to culture medium：
MS+6-BA1.0mg/L
MS+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+IBA0.1mg/L
In the culture medium, carbon source must also be added, when carbon source uses sucrose, addition is 30g/L after being configured to culture medium,
When carbon source uses glucose, addition is 40g/L after being configured to culture medium.
Note：
MS culture mediums：Be Murashige and Skoog in 1962 it is tobacco cell Training Design, its main feature is that inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, its nitrate content is high, and the quantity and ratio of nutrient are closed
It is suitable, the nutrition and physiological requirements of plant cell can be met, thus the scope of application is wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named with their name.
1/2MS culture mediums：It is the culture medium that the amount of the calcium salt and a great number of elements in MS culture mediums halves, and common training
Support base.
Plant growth regulator 6-BA：6-benzyl aminopurine is a kind of basic element of cell division, the general entitled 6- of English
Benzylaminopurine, molecular formula C12H11N5。
Plant growth regulator NAA：1-Naphthaleneacetic acid, methyl α-naphthyl acetate, abbreviation NAA are a kind of plants
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula are C12H13NO2, it is a kind of auximone.
The cultivation temperature of step S1 is 25~30 DEG C, 2500~3000LUX of intensity of illumination, 6~9 hour/day of light application time；
S2, adventitious shoots culture：
The sterile bud that clip is grown is inoculated into 15~20 days/generation of culture on adventitious shoots culture base, can continuous subculture 4-5
Secondary, bud growth coefficient is 10~14.8 times/month, obtains a large amount of Multiple Buds；The culture medium of the medium component and step S1
Ingredient is equivalent；
The cultivation temperature of step S2 is 25~30 DEG C, 2500~3000LUX of intensity of illumination, 6~9 hour/day of light application time；
S3, strong sprout, culture of rootage：
In the culture medium that seedling separation access has newly configured after Multiple Buds grow to 1-2cm long, after cultivating 15~20 days, it is good for
Strong rooted seedling；The medium component is equal with medium component in step S1；
The cultivation temperature of step S3 is 25~30 DEG C, 2500~3000LUX of intensity of illumination, 6~9 hour/day of light application time；
S4, acclimatization and transplants are carried out：
The obtained paulownia seedling of step S3 is transplanted, the condition of culture of step S4 is：Hardening matrix garden mould+peat matter
Amount proportioning 2.5：1, shading net shades, humidity 62-65%.
Embodiment three
A kind of hybridization paulownia seedling cultural method of the present invention includes the following steps：
S1, sterile bud culture is carried out：
The hybridization paulownia for just growing tender shoots are put into biochemical cultivation case, after being cultivated 6~8 days under the conditions of 38~40 DEG C,
Clip hybridizes the HgCl that paulownia tender shoots impregnates 1min disinfection and mass ratio 0.05-0.15% through volume ratio 70-80% alcohol2Solution
After sterilizing, after sterile washing 3-5 times, cuts in the bud point access culture medium of 1.8mm long and cultivate, after tender shoots growth, then cut
The new medium culture that has configured of the bud point access of 1.8mm long, repeatedly three times or three times more than, obtain the sterile of detoxification
Bud；Because can kill certain virus by high-temperature cultivation, the conducting tissue of the bud point growing point of 2mm long is not developed also completely, no
It is conducted conducive to virus, obtains preferable detoxification sterile bud, advantage of such detoxification seedling in planting process is that growth is preferable
Comparatively fast.
Culture medium includes plant growth regulator, carbon source, 1/2MS and/or MS, and preparation method is：With 1/2MS and/or MS
For minimal medium, appropriate plant growth regulator is added and carbon source is formulated, the pH6.5 of the culture medium.
The plant growth regulator can be used including but not limited to 6-BA, NAA or IBA while use 6-BA and NAA,
IBA.The addition for being configured to 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA are (0.1mg/L-
1.0mg/L)。
Including but not limited to sucrose, glucose may be used in carbon source.When using sucrose, it is configured to a concentration of after culture medium
18g/L；When using glucose, it is configured to a concentration of 25g/L after culture medium.
Coagulator will be added in culture medium, and agar powder, carragheen etc. can be added, if selecting agar powder, the addition of agar powder
For 5g/L；If selecting carragheen, the addition of carragheen is 8g/L.
The sterile bud culture medium of hybridization paulownia can be used including but not limited to by the formula after being configured to culture medium：
MS+6-BA1.0mg/L
MS+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+IBA0.1mg/L
In the culture medium, carbon source must also be added, when carbon source uses sucrose, addition is 18g/L after being configured to culture medium,
When carbon source uses glucose, addition is 25g/L after being configured to culture medium.
Note：
MS culture mediums：Be Murashige and Skoog in 1962 it is tobacco cell Training Design, its main feature is that inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, its nitrate content is high, and the quantity and ratio of nutrient are closed
It is suitable, the nutrition and physiological requirements of plant cell can be met, thus the scope of application is wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named with their name.
1/2MS culture mediums：It is the culture medium that the amount of the calcium salt and a great number of elements in MS culture mediums halves, and common training
Support base.
Plant growth regulator 6-BA：6-benzyl aminopurine is a kind of basic element of cell division, the general entitled 6- of English
Benzylaminopurine, molecular formula C12H11N5。
Plant growth regulator NAA：1-Naphthaleneacetic acid, methyl α-naphthyl acetate, abbreviation NAA are a kind of plants
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula are C12H13NO2, it is a kind of auximone.
The cultivation temperature of step S1 is 15~20 DEG C, 2000~2500LUX of intensity of illumination, light application time 10~12 hours/
It；
S2, adventitious shoots culture：
The sterile bud that clip is grown is inoculated into 15~20 days/generation of culture on adventitious shoots culture base, can continuous subculture 4-5
Secondary, bud growth coefficient is 10~14.8 times/month, obtains a large amount of Multiple Buds；The culture medium of the medium component and step S1
Ingredient is equivalent；
The cultivation temperature of step S2 is 15~20 DEG C, 2000~2500LUX of intensity of illumination, light application time 10~12 hours/
It；
S3, strong sprout, culture of rootage：
In the culture medium that seedling separation access has newly configured after Multiple Buds grow to 1-2cm long, after cultivating 15~20 days, it is good for
Strong rooted seedling；The medium component is equal with medium component in step S1；
The cultivation temperature of step S3 is 15~20 DEG C, 2000~2500LUX of intensity of illumination, light application time 10~12 hours/
It；
S4, acclimatization and transplants are carried out：
The obtained paulownia seedling of step S3 is transplanted, the condition of culture of step S4 is：Hardening matrix garden mould+peat
Quality proportioning 1.5：1, shading net shades, humidity 60-65%.
Example IV
A kind of hybridization paulownia seedling cultural method of the present invention includes the following steps：
S1, sterile bud culture is carried out：To hybridize paulownia tender stem or tender shoots as explant, the alcohol that first volume ratio is 80% disappears
Poison, uses sterile water wash 1 time afterwards, then uses mass fraction for 0.05%HgCl2Solution sterilized after, through sterile water wash
It 3-5 times, is then connected on sterile bud culture medium and is cultivated, sterile bud is obtained after cultivating 14~15 days.
Cultivated days depend mainly on the speed of growth of hybridization paulownia tender shoots, and general sterile bud grows to 1 centimetre or so and is
Preferably.Certainly, sterile bud is longer than 1 centimetre (such as growing to 1.5 centimetres) or shorter than 1 centimetre (such as growing to 0.5 centimetre), not shadow
Ring the realization of the object of the invention.
Culture medium includes plant growth regulator, carbon source, 1/2MS and/or MS, and preparation method is：With 1/2MS and/or MS
For minimal medium, appropriate plant growth regulator is added and carbon source is formulated, the pH7.2 of the sterile bud culture medium.
The plant growth regulator can be used including but not limited to 6-BA, NAA or IBA while use 6-BA and NAA,
IBA.The addition for being configured to 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA are (0.1mg/L-
1.0mg/L)。
Including but not limited to sucrose, glucose may be used in carbon source.When using sucrose, it is configured to a concentration of after culture medium
20g/L；When using glucose, it is configured to a concentration of 30g/L after culture medium.
Coagulator will be added in culture medium, and agar powder, carragheen etc. can be added, if selecting agar powder, the addition of agar powder
For 6g/L；If selecting carragheen, the addition of carragheen is 10g/L.
The sterile bud culture medium of hybridization paulownia can be used including but not limited to by the formula after being configured to culture medium：
MS+6-BA1.0mg/L
MS+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+IBA0.1mg/L
In the culture medium, carbon source must also be added, when carbon source uses sucrose, addition is 20g/L after being configured to culture medium,
When carbon source uses glucose, addition is 30g/L after being configured to culture medium.
Note：
MS culture mediums：Be Murashige and Skoog in 1962 it is tobacco cell Training Design, its main feature is that inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, its nitrate content is high, and the quantity and ratio of nutrient are closed
It is suitable, the nutrition and physiological requirements of plant cell can be met, thus the scope of application is wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named with their name.
1/2MS culture mediums：It is the culture medium that the amount of the calcium salt and a great number of elements in MS culture mediums halves, and common training
Support base.
Plant growth regulator 6-BA：6-benzyl aminopurine is a kind of basic element of cell division, the general entitled 6- of English
Benzylaminopurine, molecular formula C12H11N5。
Plant growth regulator NAA：1-Naphthaleneacetic acid, methyl α-naphthyl acetate, abbreviation NAA are a kind of plants
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula are C12H13NO2, it is a kind of auximone；
The cultivation temperature of step S1 is 25~30 DEG C, 2500~3000LUX of intensity of illumination, light application time 10~12 hours/
It；
S2, adventitious shoots culture：
The sterile bud that clip is grown is inoculated into 15~20 days/generation of culture on adventitious shoots culture base, can continuous subculture 4-5
Secondary, bud growth coefficient is 10~14.8 times/month, obtains a large amount of Multiple Buds；The culture medium of the medium component and step S1
Ingredient is equivalent；
The cultivation temperature of step S2 is 25~30 DEG C, 2500~3000LUX of intensity of illumination, light application time 10~12 hours/
It；
S3, strong sprout, culture of rootage：
In the culture medium that seedling separation access has newly configured after Multiple Buds grow to 1-2cm long, after cultivating 15~20 days, it is good for
Strong rooted seedling；The medium component is equal with medium component in step S1；
The cultivation temperature of step S3 is 25~30 DEG C, 2500~3000LUX of intensity of illumination, light application time 10~12 hours/
It；
S4, acclimatization and transplants are carried out：
The condition of culture of step S4 is：Hardening matrix garden mould+peat quality proportioning 2：1, shading net shades, and humidity is
67-69%.
Embodiment five
A kind of hybridization paulownia seedling cultural method of the present invention includes the following steps：
S1, sterile bud culture is carried out：To hybridize paulownia tender stem or tender shoots as explant, the alcohol that first volume ratio is 70% disappears
Poison, uses sterile water wash 1 time afterwards, then uses mass fraction for 0.15%HgCl2Solution sterilized after, through sterile water wash
It 3-5 times, is then connected on sterile bud culture medium and is cultivated, sterile bud is obtained after cultivating 16~18 days.
Cultivated days depend mainly on the speed of growth of hybridization paulownia tender shoots, and general sterile bud grows to 1 centimetre or so and is
Preferably.Certainly, sterile bud is longer than 1 centimetre (such as growing to 1.5 centimetres) or shorter than 1 centimetre (such as growing to 0.5 centimetre), not shadow
Ring the realization of the object of the invention.
Culture medium includes plant growth regulator, carbon source, 1/2MS and/or MS, and preparation method is：With 1/2MS and/or MS
For minimal medium, appropriate plant growth regulator is added and carbon source is formulated, the pH7.0 of the sterile bud culture medium.
The plant growth regulator can be used including but not limited to 6-BA, NAA or IBA while use 6-BA and NAA,
IBA.The addition for being configured to 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA are (0.1mg/L-
1.0mg/L)。
Including but not limited to sucrose, glucose may be used in carbon source.When using sucrose, it is configured to a concentration of after culture medium
25g/L；When using glucose, it is configured to a concentration of 35g/L after culture medium.
Coagulator will be added in culture medium, and agar powder, carragheen etc. can be added, if selecting agar powder, the addition of agar powder
For 5.5g/L；If selecting carragheen, the addition of carragheen is 13g/L.
The sterile bud culture medium of hybridization paulownia can be used including but not limited to by the formula after being configured to culture medium：
MS+6-BA1.0mg/L
MS+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+IBA0.1mg/L
Note：
MS culture mediums：Be Murashige and Skoog in 1962 it is tobacco cell Training Design, its main feature is that inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, its nitrate content is high, and the quantity and ratio of nutrient are closed
It is suitable, the nutrition and physiological requirements of plant cell can be met, thus the scope of application is wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named with their name.
1/2MS culture mediums：It is the culture medium that the amount of the calcium salt and a great number of elements in MS culture mediums halves, and common training
Support base.
Plant growth regulator 6-BA：6-benzyl aminopurine is a kind of basic element of cell division, the general entitled 6- of English
Benzylaminopurine, molecular formula C12H11N5。
Plant growth regulator NAA：1-Naphthaleneacetic acid, methyl α-naphthyl acetate, abbreviation NAA are a kind of plants
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula are C12H13NO2, it is a kind of auximone；
The cultivation temperature of step S1 is 23~27 DEG C, 2800~3000LUX of intensity of illumination, light application time 10~12 hours/
It；
S2, adventitious shoots culture：
The sterile bud that clip is grown is inoculated into 15~20 days/generation of culture on adventitious shoots culture base, can continuous subculture 4-5
Secondary, bud growth coefficient is 10~14.8 times/month, obtains a large amount of Multiple Buds；The culture medium of the medium component and step S1
Ingredient is equivalent；
The cultivation temperature of step S2 is 23~27 DEG C, 2800~3000LUX of intensity of illumination, light application time 10~12 hours/
It；
S3, strong sprout, culture of rootage：
In the culture medium that seedling separation access has newly configured after Multiple Buds grow to 1-2cm long, after cultivating 15~20 days, it is good for
Strong rooted seedling；The medium component is equal with medium component in step S1；
The cultivation temperature of step S3 is 23~27 DEG C, 2800~3000LUX of intensity of illumination, light application time 10~12 hours/
It；
S4, acclimatization and transplants are carried out：
The condition of culture of step S4 is：Hardening matrix garden mould+peat quality proportioning 2.5：1, shading net shades, humidity
For 66-70%.
Embodiment six
A kind of hybridization paulownia seedling cultural method of the present invention includes the following steps：
S1, sterile bud culture is carried out：To hybridize paulownia tender stem or tender shoots as explant, the alcohol that first volume ratio is 75% disappears
Poison, uses sterile water wash 1 time afterwards, then uses mass fraction for 0.15%HgCl2Solution sterilized after, through sterile water wash
It 3-5 times, is then connected on sterile bud culture medium and is cultivated, sterile bud is obtained after cultivating 16~18 days.
Cultivated days depend mainly on the speed of growth of hybridization paulownia tender shoots, and general sterile bud grows to 1 centimetre or so and is
Preferably.Certainly, sterile bud is longer than 1 centimetre (such as growing to 1.5 centimetres) or shorter than 1 centimetre (such as growing to 0.5 centimetre), not shadow
Ring the realization of the object of the invention.
Culture medium includes plant growth regulator, carbon source, 1/2MS and/or MS, and preparation method is：With 1/2MS and/or MS
For minimal medium, appropriate plant growth regulator is added and carbon source is formulated, the pH7.2 of the sterile bud culture medium.
The plant growth regulator can be used including but not limited to 6-BA, NAA or IBA while use 6-BA and NAA,
IBA.The addition for being configured to 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA are (0.1mg/L-
1.0mg/L)。
Including but not limited to sucrose, glucose may be used in carbon source.When using sucrose, it is configured to a concentration of after culture medium
30g/L；When using glucose, it is configured to a concentration of 40g/L after culture medium.
Coagulator will be added in culture medium, and agar powder, carragheen etc. can be added, if selecting agar powder, the addition of agar powder
For 6g/L；If selecting carragheen, the addition of carragheen is 15g/L.
The sterile bud culture medium of hybridization paulownia can be used including but not limited to by the formula after being configured to culture medium：
MS+6-BA1.0mg/L
MS+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+IBA0.1mg/L
In the culture medium, carbon source must also be added, when carbon source uses sucrose, addition is 30g/L after being configured to culture medium,
When carbon source uses glucose, addition is 40g/L after being configured to culture medium.
Note：
MS culture mediums：Be Murashige and Skoog in 1962 it is tobacco cell Training Design, its main feature is that inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, its nitrate content is high, and the quantity and ratio of nutrient are closed
It is suitable, the nutrition and physiological requirements of plant cell can be met, thus the scope of application is wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named with their name.
1/2MS culture mediums：It is the culture medium that the amount of the calcium salt and a great number of elements in MS culture mediums halves, and common training
Support base.
Plant growth regulator 6-BA：6-benzyl aminopurine is a kind of basic element of cell division, the general entitled 6- of English
Benzylaminopurine, molecular formula C12H11N5。
Plant growth regulator NAA：1-Naphthaleneacetic acid, methyl α-naphthyl acetate, abbreviation NAA are a kind of plants
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula are C12H13NO2, it is a kind of auximone；
The cultivation temperature of step S1 is 25~30 DEG C, 2500~3000LUX of intensity of illumination, light application time 10~12 hours/
It；
S2, adventitious shoots culture：
The sterile bud that clip is grown is inoculated into 15~20 days/generation of culture on adventitious shoots culture base, can continuous subculture 4-5
Secondary, bud growth coefficient is 10~14.8 times/month, obtains a large amount of Multiple Buds；The culture medium of the medium component and step S1
Ingredient is equivalent；
The cultivation temperature of step S2 is 25~30 DEG C, 2500~3000LUX of intensity of illumination, light application time 10~12 hours/
It；
S3, strong sprout, culture of rootage：
In the culture medium that seedling separation access has newly configured after Multiple Buds grow to 1-2cm long, after cultivating 15~20 days, it is good for
Strong rooted seedling；The medium component is equal with medium component in step S1；
The cultivation temperature of step S3 is 25~30 DEG C, 2500~3000LUX of intensity of illumination, light application time 10~12 hours/
It；
S4, acclimatization and transplants are carried out：
The condition of culture of step S4 is：Hardening matrix garden mould+peat quality proportioning 2.5：1, shading net shades, humidity
For 66-70%.
Embodiment seven
A kind of hybridization paulownia seedling cultural method of the present invention includes the following steps：
S1, sterile bud culture is carried out：To hybridize paulownia tender stem or tender shoots as explant, the alcohol that first volume ratio is 78% disappears
Poison, uses sterile water wash 1 time afterwards, then uses mass fraction for 0.13%HgCl2Solution sterilized after, through sterile water wash
It 3-5 times, is then connected on sterile bud culture medium and is cultivated, sterile bud is obtained after cultivating 16~18 days.
Cultivated days depend mainly on the speed of growth of hybridization paulownia tender shoots, and general sterile bud grows to 1 centimetre or so and is
Preferably.Certainly, sterile bud is longer than 1 centimetre (such as growing to 1.5 centimetres) or shorter than 1 centimetre (such as growing to 0.5 centimetre), not shadow
Ring the realization of the object of the invention.
Culture medium includes plant growth regulator, carbon source, 1/2MS and/or MS, and preparation method is：With 1/2MS and/or MS
For minimal medium, appropriate plant growth regulator is added and carbon source is formulated, the pH6.8 of the sterile bud culture medium.
The plant growth regulator can be used including but not limited to 6-BA, NAA or IBA while use 6-BA and NAA,
IBA.The addition for being configured to 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA are (0.1mg/L-
1.0mg/L)。
Including but not limited to sucrose, glucose may be used in carbon source.When using sucrose, it is configured to a concentration of after culture medium
28g/L；When using glucose, it is configured to a concentration of 38g/L after culture medium.
Coagulator will be added in culture medium, and agar powder, carragheen etc. can be added, if selecting agar powder, the addition of agar powder
For 6g/L；If selecting carragheen, the addition of carragheen is 15g/L.
The sterile bud culture medium of hybridization paulownia can be used including but not limited to by the formula after being configured to culture medium：
MS+6-BA1.0mg/L
MS+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+NAA0.1mg/L
1/2MS or MS+6-BA1.0mg/L+IBA0.1mg/L
In the culture medium, carbon source must also be added, when carbon source uses sucrose, addition is 28g/L after being configured to culture medium,
When carbon source uses glucose, addition is 38g/L after being configured to culture medium.
Note：
MS culture mediums：Be Murashige and Skoog in 1962 it is tobacco cell Training Design, its main feature is that inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, its nitrate content is high, and the quantity and ratio of nutrient are closed
It is suitable, the nutrition and physiological requirements of plant cell can be met, thus the scope of application is wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named with their name.
1/2MS culture mediums：It is the culture medium that the amount of the calcium salt and a great number of elements in MS culture mediums halves, and common training
Support base.
Plant growth regulator 6-BA：6-benzyl aminopurine is a kind of basic element of cell division, the general entitled 6- of English
Benzylaminopurine, molecular formula C12H11N5。
Plant growth regulator NAA：1-Naphthaleneacetic acid, methyl α-naphthyl acetate, abbreviation NAA are a kind of plants
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula are C12H13NO2, it is a kind of auximone.
S2, adventitious shoots culture：
The sterile bud that clip is grown is inoculated into 15~20 days/generation of culture on adventitious shoots culture base, can continuous subculture 4-5
Secondary, bud growth coefficient is 10~14.8 times/month, obtains a large amount of Multiple Buds；The culture medium of the medium component and step S1
Ingredient is equivalent；
The cultivation temperature of step S2 is 28~30 DEG C, 2800~3000LUX of intensity of illumination, light application time 10~12 hours/
It；
S3, strong sprout, culture of rootage：
In the culture medium that seedling separation access has newly configured after Multiple Buds grow to 1-2cm long, after cultivating 15~20 days, it is good for
Strong rooted seedling；The medium component is equal with medium component in step S1；
The cultivation temperature of step S3 is 28~30 DEG C, 2800~3000LUX of intensity of illumination, light application time 10~12 hours/
It；
S4, acclimatization and transplants are carried out：
The obtained paulownia seedling of step S3 is transplanted, the condition of culture of step S4 is：Hardening matrix garden mould+peat matter
Amount proportioning 1.8：1, shading net shades, humidity 66-70%.
Claims (5)
1. a kind of hybridization paulownia seedling cultural method, which is characterized in that include the following steps：
S1, sterile bud culture：
By the way that the hybridization paulownia for just growing 1-2.5mm long tender shoots are put into biochemical cultivation case, 6 are cultivated under the conditions of 38~40 DEG C
After~8 days, clip 1.5-1.8mm hybridization paulownia tender shoots first through alcohol soaking disinfection, after with sterile water wash 1 time, use HgCl again2
Solution impregnate sterilizing after, after sterile washing 3-5 time, the bud point for cutting 1.8-2.2mm long accesses in the new culture medium configured
Culture after tender shoots grows 2.5mm long, then cuts in the bud point access culture medium of 1.8-2.2mm long and cultivates, repeatedly for three times or three times
More than, obtaining the sterile bud of detoxification, it is 15~30 DEG C that condition of culture, which is temperature, in the culture medium, intensity of illumination 2000~
3000LUX, 6~12 hour/day of light application time；
The medium component includes plant growth regulator, carbon source, 1/2MS and/or MS；
S2, adventitious shoots culture：
The sterile bud that clip is grown is inoculated into 15~20 days/generation of culture on adventitious shoots culture base, continuous subculture 4-5 times, bud proliferation
Coefficient is 10~14.8 times/month, obtains a large amount of Multiple Buds；The medium component is equal with the medium component of step S1；
The temperature of the culture is 15~30 DEG C, 2000~3000LUX of intensity of illumination, 6~12 hour/day of light application time；
S3, strong sprout, culture of rootage：
The sub-bottle culture after Multiple Buds grow to 1-2cm long is trained with the new culture medium having configured with step S1 equivalent components
It supports, obtains the healthy seedling that paulownia take root；The strong sprout, the temperature of culture of rootage are 15~30 DEG C, intensity of illumination 2000~
3000LUX, 6~12 hour/day of light application time；
S4, acclimatization and transplants：
The healthy and strong transplantation of seedlings that the obtained paulownia of step S3 are taken root is to crop field hardening, and the matrix that the hardening uses is garden mould and mud
Charcoal mixes, and is shaded with shading net to hardening matrix；
The preparation method of the plant growth regulator in culture medium, carbon source, 1/2MS or MS in the step S1-S3 is：With 1/
2MS or MS is minimal medium, and appropriate plant growth regulator is added and carbon source is formulated；The pH of the culture medium is 6.5
~7.2；
The plant growth regulator of culture medium in described step S1, S2, S3 is 6-BA and/or NAA and/or IBA, is configured to train
A concentration of 0.1mg/L-2.0mg/L of 6-BA, a concentration of 0.1mg/ of NAA a concentration of 0.1mg/L-1.0mg/L, IBA after foster base
L-1.0mg/L；
Agar powder or carragheen is added in the also needing to of culture medium in described step S1, S2, S3, if selecting agar powder, agar powder
Addition be 5-6g/L；If selecting carragheen, the addition of carragheen is 8-15g/L；
The carbon source is sucrose or glucose, and the culture medium sucrose concentration being configured to when using sucrose is 18~30g/L, uses Portugal
The culture medium concentration of glucose being configured to when grape sugar is 25~40g/L.
2. hybridization paulownia seedling cultural method as described in claim 1, which is characterized in that the alcohol by volume score used is
70%~80%, used HgCl2Liquid quality fraction is 0.05%~0.15%.
3. hybridization paulownia seedling cultural method as described in claim 1, it is characterised in that the culture in described step S1, S2, S3
Base pH is 6.8, and cultivation temperature is 25 ± 2 DEG C, and intensity of illumination is 2800~3000LUX, 10~12 hour/day of light application time.
4. hybridization paulownia seedling cultural method as described in claim 1, it is characterised in that the nothing that clip is grown in the step S2
The speed of growth of the length 1-2cm of bacterium bud, the adventitious shoots culture of the step S2 are 15~20 days/generation, continuous to cultivate subculture 4-
5 times, the bud growth coefficient of the Multiple Buds is 10~14.8 times/month.
5. hybridization paulownia seedling cultural method as described in claim 1, it is characterised in that garden in the hardening matrix of the step S4
Soil and peat quality proportioning are (1.5~2.5)：1, hardening area humidity is 60-70%.
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| CN107006376A (en) * | 2017-06-20 | 2017-08-04 | 广西大学 | A kind of paulownia method for plant tissue culture |
| CN111406647B (en) * | 2020-04-17 | 2021-05-07 | 山东省林业科学研究院 | Efficient starting culture medium for directly inducing tetraploid paulownia petioles to regenerate adventitious buds and application |
| CN111374056B (en) * | 2020-04-17 | 2021-05-18 | 山东省林业科学研究院 | Proliferation medium formula for culturing stout tetraploid paulownia seedlings and application |
| CN112690211A (en) * | 2021-01-04 | 2021-04-23 | 海南品优种苗科技有限公司 | Method for virus-free tissue culture and domestication cultivation of paulownia saplings |
| CN113349055A (en) * | 2021-07-07 | 2021-09-07 | 广东粤恬生物科技有限公司 | Tissue culture method of paulownia |
| CN116135010B (en) * | 2023-04-10 | 2024-05-28 | 江苏省农业科学院宿迁农科所(宿迁市农业科学研究院) | Tissue culture and rapid propagation method for paulownia virus-free seedlings |
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| CN103704140A (en) * | 2013-12-31 | 2014-04-09 | 湖北大学 | Method for obtaining polyploidy paulownia tomentosa by performing tissue culture propagation by taking young flowers as explants |
| CN105104210A (en) * | 2015-09-30 | 2015-12-02 | 江苏农林职业技术学院 | Method for efficiently and rapidly reproducing paulownia test-tube plantlets in large scale |
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| CN103704140A (en) * | 2013-12-31 | 2014-04-09 | 湖北大学 | Method for obtaining polyploidy paulownia tomentosa by performing tissue culture propagation by taking young flowers as explants |
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