CN106258979A - A kind of hybridization Paulownia Seedling cultural method - Google Patents
A kind of hybridization Paulownia Seedling cultural method Download PDFInfo
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- CN106258979A CN106258979A CN201610769778.8A CN201610769778A CN106258979A CN 106258979 A CN106258979 A CN 106258979A CN 201610769778 A CN201610769778 A CN 201610769778A CN 106258979 A CN106258979 A CN 106258979A
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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, step includes that aseptic bud is cultivated: be outer implant to hybridize Paulownia tender stem or tender shoots, after sterilized, sterilizing, cleaning, it is seeded in 1/2MS or MS and adds to cultivate in the culture medium of appropriate plant growth regulator and obtain aseptic bud in 12～18 days；Adventitious shoots culture: the aseptic bud of clip, is inoculated into add with MS and cultivates on the adventitious shoots culture base of appropriate plant growth regulator preparation；Strong sprout, root culture: Multiple Buds is inoculated into 1/2MS and/or MS and adds sub-bottle cultivation in the strong sprout of appropriate plant growth regulator, root media after growing to 1 2cm, above-mentioned steps cultivation temperature 15～30 DEG C, light intensity 2000～3000LUX, light application time 6～12 hour/day；Acclimatization and transplants；Transplantation of Regenerated Plantlets of taking root is in garden mould+peat (2:1) substrate, and shade net shelters from heat or light, humidity 60 70%, within 15～30 days, goes out garden.The present invention is greatly improved the hybridization Paulownia Seedling speed of growth and planting benefit, low cost, pollutes little, and the four seasons can produce, and is suitable for extensive nursery.
Description
Technical field
The present invention relates to a kind of cultural method hybridizing Paulownia Seedling.
Background technology
Paulownia Scrophulariaceae Paulownia (Paulownia) plant, distributed more widely in China, for main broad-leaved fast-growing tree
Kind.Paulownia growth is very rapid, becomes a useful person early, among the people have " 1 year bar, 2 years as umbrella, 5 years can saw plate " say.Hybridization
Paulownia " 9501 " is one of Paulownia research institute of the State Administration of Forestry 15 key research project achievements, is the advantage in Paulownia
Kind, it is the famous fast-growing high quality timber seeds of China, strong adaptability, growth is fast, become a useful person early, in the case of condition is suitable, 1 year
Age of tree tree growth up to 7-8 rice, more than footpath, ground 15-20 centimetre, 3-4 can take turns and cut down or cut down together, and when within 5 years, felling, individual plant may be up to
18 meters, the breast timber diameter of a cross-section of a tree trunk 1.3 meters above the ground reaches more than 40 centimeters, volume of timber 0.5-0.6 cubic meter, and the grain of wood is logical straight, and decorative pattern is attractive in appearance, 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 such as plywood, jigsaw, integrated timber excellent
Good timber matter characteristic；Paulownia material insulation excellent insulating property, resonance performance is good, and radiation damping is high, and internal friction is little, is excellent
Stringed musical instrument material, can be equally celebrated for their achievements with famous fiddle butt fish scale PiceameyeriRehd. Et Wils.；The wood fiber content of Paulownia material is up to more than 50%, is plane
The good raw material materials such as card, ecological board, papermaking；Leaf is again good feedstuff etc..Therefore, Paulownia is always broad masses and is liked
Like with one of material and the chief species improving environment, supply falls short of demand for its product, and market prospect is wide, and economic worth is high, paulownia
For native country seeds, it is eco-friendly seeds.Therefore, the industrialization growing and cultivation of hybridization Paulownia becomes of concern and asks
Topic.
At present, the growing and cultivation of hybridization Paulownia mainly uses traditional seedling nursery method to carry out nursery technique or point root moves
Planting, the nursery production cycle is long, and floor space is big, it is difficult to realize the most quickly heavy industrialization growing and cultivation.
Summary of the invention
The technical problem to be solved in the present invention is, it is provided that a kind of hybridization Paulownia Seedling cultural method, overcomes prior art difficult
To realize the defect of the most quickly heavy industrialization growing and cultivation.The present invention solves the technical side that its technical problem is used
Case is:
There is provided one hybridization Paulownia Seedling cultural method, it is characterised in that comprise the steps:
S1, aseptic bud are cultivated:
It is outer implant with hybridization Paulownia tender stem or tender shoots, first through alcohol disinfecting, uses sterile water wash 1 time afterwards, then use
HgCl2After solution carries out sterilizing, through sterile water wash 3-5 time, then it is connected in aseptic bud culture medium and carries out cultivating 12～18 days,
Cultivation temperature is 15～30 DEG C, intensity of illumination 2000～3000LUX, light application time 6～12 hour/day, it is thus achieved that aseptic bud；
Or the hybridization Paulownia seedling just growing the long tender shoots of 1-2.5mm is put in biochemical cultivation case, 38～40 DEG C of conditions
Lower cultivation 6～after 8 days, clip hybridization Paulownia tender shoots is through alcohol disinfecting and HgCl2After solution sterilization, after aseptic washing 3-5 time,
The bud point cutting 1.8-2.2mm length accesses cultivation in newly configured good culture medium, after tender shoots growth 2.5mm length, then cuts 1.8-
The bud point of 2.2mm length accesses in culture medium and cultivates, and more than repeatedly for three times or three times, obtains the aseptic bud of detoxification；
Described medium component includes plant growth regulator, carbon source, 1/2MS and/or MS；
S2. adventitious shoots culture:
The aseptic bud that clip grows, is inoculated on adventitious shoots culture base 15～20 days/generation of cultivation, can subculture 4-5 continuously
Secondary, Shoot propagation coefficient is 10～14.8 times/month, it is thus achieved that substantial amounts of Multiple Buds；Described medium component and the culture medium of step S1
Composition is equal to；
The cultivation temperature of step S2 is 15～30 DEG C, intensity of illumination 2000～3000LUX, light application time 6～12 hours/
My god；
S3, strong sprout, root culture:
During after Multiple Buds grows to 1-2cm length, point Seedling accesses newly configured good culture medium, after cultivating 15～20 days, it is thus achieved that strong
Strong Seedling of taking root；Described medium component is equal to medium component in step S1；
The cultivation temperature of step S3 is 15～30 DEG C, intensity of illumination 2000～3000LUX, light application time 6～12 hours/
My god；
S4, acclimatization and transplants；
Transplanting Paulownia strong sprout step S3 obtained to land for growing field crops seedling exercising, the substrate that described seedling exercising uses is garden mould and peat mixes
Conjunction forms, and shelters from heat or light seedling exercising substrate shade net.
Preferably, the alcohol by volume mark used in described step S1 is 70%～80%, the HgCl used2Solution matter
Amount mark is 0.05%～0.15%.
Preferably, plant growth regulator in the culture medium in described step S1-S3, carbon source, 1/2MS and/or MS
Compound method is: with 1/2MS and/or MS as minimal medium, adds appropriate plant growth regulator and carbon source is formulated；Should
The pH of aseptic 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
Become described 6-BA concentration after culture medium be 0.1mg/L-2.0mg/L, NAA concentration be 0.1mg/L-1.0mg/L, IBA concentration is
0.1mg/L-1.0mg/L。
Preferably, the culture medium in described step S1, S2, S3 also need to add coagulator, agar powder or card can be added
Drawing glue, if selecting agar powder, the addition of agar powder is 5-6g/L；If selection carrageenan, the addition of carrageenan is 8-15g/
L。
Preferably, the optimal condition of culture of culture medium in described step S1, S2, S3 be pH be 6.8, this PH is the suitableeest profit
In Multiple Buds and the acid-base value of root culture, cultivation temperature is 25 ± 2 DEG C, and intensity of illumination is 2800～3000LUX, light application time
10～12 hours/day, this culturing room's temperature and intensity of illumination and time chien shih tissue cultured seedling tiller growth coefficient best, leaf blade size
Moderate, leaf color is dark green, and stem is the most sturdy, can shorten again incubation time 1～2 days.
Preferably, in described step S2, the optimum altitude of the aseptic bud that clip grows is 1-2cm, and such length is beneficial to seedlings picking
Operation, the most beneficially successive transfer culture, too short then Seedling is the least, and culture medium nutrition also has residue, and the nutrition in long culture medium has consumed only,
It is unfavorable for that the growth of Regenerated plant, the speed of growth of the adventitious shoots culture of described step S2 are 15～20 days/generation, cultivates subculture continuously
4-5 time, the Shoot propagation coefficient of described Multiple Buds is 10～14.8 times/month.
Preferably, described carbon source is sucrose or glucose, when using sucrose the culture medium sucrose concentration that is configured to be 18～
30g/L, the culture medium concentration of glucose being configured to when using glucose is 25～40g/L.
Preferably, in the seedling exercising substrate of described step S4, garden mould and peat quality proportioning are (1.5～2.5): 1, seedling exercising district
Humidity is 60-70%.
Implementing the present invention and hybridize Paulownia Seedling cultural method, compared with the prior art, it provides the benefit that:
1, the method applied in the present invention makes Paulownia cultivate the speed of growth and planting benefit is greatly improved；The side of the present invention
One monthly average growth coefficient of method hybridization Paulownia bud reaches 10～14.8, and existing hybridization Paulownia stooling method for transplanting one
Strain at most can only grow up to a young plant；
2, in the method that the present invention uses, aseptic bud is after detoxification treatment, and the speed of growth and planting benefit significantly improve；
The method of the present invention, after detoxification treatment one month, plant grows thickly growth, high uniformity, mean blade width 1.4cm, the number of blade 4～6
Sheet, after transplanting, growth is very fast；Non-detoxification treatment stooling propagation method majority individual plant grows, and height size is uneven, average leaf
Wide 0.4～1.0cm, the number of blade 1～3, grow slower after transplanting；
3, the cost of material that used of method that the present invention uses is low, do not produce in cultivating process environmentally hazardous because of
Element, all uses environmental protection raw material so that environmental pollution is little；
4, the method technique that the present invention uses is relatively simple, the most all can produce, and is suitable for extensive nursery and produces.
Detailed description of the invention
Below in conjunction with embodiment, the invention will be further described.
Embodiment one
One of the present invention hybridization Paulownia Seedling cultural method comprises the steps:
S1, carrying out aseptic bud cultivation: be outer implant to hybridize Paulownia tender stem or tender shoots, first volume ratio is the wine of 70-80%
Essence sterilization, rear by sterile water wash 1 time, then to use mass fraction be 0.05-0.15%HgCl2Solution carry out sterilizing after, warp
Sterile water wash 3-5 time, is then connected in aseptic bud culture medium and cultivates, and obtains aseptic bud after cultivating 16～18 days.
Cultivated days depends mainly on the speed of growth of hybridization Paulownia tender shoots, and general aseptic bud grows to 1 cm and is
Preferably.Certainly, aseptic bud is longer than 1 centimetre (such as growing to 1.5 centimetres) or is 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 compound method is: with 1/2MS and/or MS
For minimal medium, add appropriate plant growth regulator and carbon source is formulated, the pH6.8 of described aseptic bud culture medium.
Described plant growth regulator can use and include but not limited to that 6-BA, NAA or IBA use 6-BA and NAA simultaneously,
IBA.Being configured to the addition of 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA is (0.1mg/L-
1.0mg/L)。
Carbon source can use and include but not limited to sucrose, glucose.When using sucrose, after being configured to culture medium, concentration is
18-30g/L；When using glucose, after being configured to culture medium, concentration is 25-40g/L.
Culture medium coagulator to be added, can add agar powder, carrageenan etc., if selecting agar powder, the addition of agar powder
For 5-6g/L；If selection carrageenan, the addition of carrageenan is 8-15g/L.
The aseptic bud culture medium of hybridization Paulownia can be used by the formula after being configured to culture medium and include but not limited to:
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 described culture medium, must also add carbon source, when carbon source uses sucrose, after being configured to culture medium, addition is 30g/L,
When carbon source uses glucose, after being configured to culture medium, addition is 40g/L.
Note:
MS culture medium: be Murashige and Skoog in 1962 be tobacco cell Training Design, is characterized in inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, and its nitrate content is high, and the quantity of its nutrient and ratio are closed
Suitable, nutrition and the physiological need of plant cell can be met, thus the scope of application is relatively wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named by their name.
1/2MS culture medium: be the culture medium that halves of the amount of the calcium salt in MS culture medium and a great number of elements, be also conventional training
Support base.
Plant growth regulator 6-BA:6-benayl aminopurine, is a kind of basic element of cell division, English general entitled 6-
Benzylaminopurine, molecular formula is C12H11N5。
Plant growth regulator NAA:1-Naphthaleneacetic acid, naphthalene acetic acid, is called for short NAA, is a kind of plant
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula is C12H13NO2, it is a kind of auximone.
The cultivation temperature of step S1 is 15～30 DEG C, intensity of illumination 2000～3000LUX, light application time 6～12 hours/
My god；
S2, adventitious shoots culture:
The aseptic bud that clip grows, is inoculated on adventitious shoots culture base 15～20 days/generation of cultivation, can subculture 4-5 continuously
Secondary, Shoot propagation coefficient is 10～14.8 times/month, it is thus achieved that substantial amounts of Multiple Buds；Described medium component and the culture medium of step S1
Composition is equal to；
The cultivation temperature of step S2 is 15～30 DEG C, intensity of illumination 2000～3000LUX, light application time 6～12 hours/
My god；
S3, strong sprout, root culture:
During after Multiple Buds grows to 1-2cm length, point Seedling accesses newly configured good culture medium, after cultivating 15～20 days, it is thus achieved that strong
Strong Seedling of taking root；Described medium component is equal to medium component in step S1；
The cultivation temperature of step S3 for for 15～30 DEG C, intensity of illumination 2000～3000LUX, light application time 6～12 hours/
My god；
S4, carry out acclimatization and transplants:
Being transplanted by the Paulownia Seedling obtained of step S3, the condition of culture of step S4 is: seedling exercising substrate garden mould+peat matter
Amount proportioning (1.5-2.5): 1, shade net shelters from heat or light, and humidity is 60-70%.
Embodiment two
One of the present invention hybridization Paulownia Seedling cultural method comprises the steps:
S1, carrying out aseptic bud cultivation: be outer implant to hybridize Paulownia tender stem or tender shoots, first volume ratio is the ethanol leaching of 75%
Bubble sterilization, rear by sterile water wash 1 time, then to use mass fraction be 0.15%HgCl2Solution carry out sterilizing after, through sterilized water
Clean 3-5 time, be then connected in aseptic bud culture medium and cultivate, after cultivating 16～18 days, obtain aseptic bud.
Cultivated days depends mainly on the speed of growth of hybridization Paulownia tender shoots, and general aseptic bud grows to 1 cm and is
Preferably.Certainly, aseptic bud is longer than 1 centimetre (such as growing to 1.5 centimetres) or is 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 compound method is: with 1/2MS and/or MS
For minimal medium, add appropriate plant growth regulator and carbon source is formulated, the pH6.8 of described aseptic bud culture medium.
Described plant growth regulator can use and include but not limited to that 6-BA, NAA or IBA use 6-BA and NAA simultaneously,
IBA.Being configured to the addition of 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA is (0.1mg/L-
1.0mg/L)。
Carbon source can use and include but not limited to sucrose, glucose.When using sucrose, after being configured to culture medium, concentration is
30g/L；When using glucose, after being configured to culture medium, concentration is 40g/L.
Culture medium coagulator to be added, can add agar powder, carrageenan etc., if selecting agar powder, the addition of agar powder
For 6g/L；If selection carrageenan, the addition of carrageenan is 15g/L.
The aseptic bud culture medium of hybridization Paulownia can be used by the formula after being configured to culture medium and include but not limited to:
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 described culture medium, must also add carbon source, when carbon source uses sucrose, after being configured to culture medium, addition is 30g/L,
When carbon source uses glucose, after being configured to culture medium, addition is 40g/L.
Note:
MS culture medium: be Murashige and Skoog in 1962 be tobacco cell Training Design, is characterized in inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, and its nitrate content is high, and the quantity of its nutrient and ratio are closed
Suitable, nutrition and the physiological need of plant cell can be met, thus the scope of application is relatively wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named by their name.
1/2MS culture medium: be the culture medium that halves of the amount of the calcium salt in MS culture medium and a great number of elements, be also conventional training
Support base.
Plant growth regulator 6-BA:6-benayl aminopurine, is a kind of basic element of cell division, English general entitled 6-
Benzylaminopurine, molecular formula is C12H11N5。
Plant growth regulator NAA:1-Naphthaleneacetic acid, naphthalene acetic acid, is called for short NAA, is a kind of plant
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula is C12H13NO2, it is a kind of auximone.
The cultivation temperature of step S1 is 25～30 DEG C, intensity of illumination 2500～3000LUX, light application time 6～9 hour/day；
S2, adventitious shoots culture:
The aseptic bud that clip grows, is inoculated on adventitious shoots culture base 15～20 days/generation of cultivation, can subculture 4-5 continuously
Secondary, Shoot propagation coefficient is 10～14.8 times/month, it is thus achieved that substantial amounts of Multiple Buds；Described medium component and the culture medium of step S1
Composition is equal to；
The cultivation temperature of step S2 is 25～30 DEG C, intensity of illumination 2500～3000LUX, light application time 6～9 hour/day；
S3, strong sprout, root culture:
During after Multiple Buds grows to 1-2cm length, point Seedling accesses newly configured good culture medium, after cultivating 15～20 days, it is thus achieved that strong
Strong Seedling of taking root；Described medium component is equal to medium component in step S1；
The cultivation temperature of step S3 is 25～30 DEG C, intensity of illumination 2500～3000LUX, light application time 6～9 hour/day；
S4, carry out acclimatization and transplants:
Being transplanted by the Paulownia Seedling obtained of step S3, the condition of culture of step S4 is: seedling exercising substrate garden mould+peat matter
Amount proportioning 2.5:1, shade net shelters from heat or light, and humidity is 62-65%.
Embodiment three
One of the present invention hybridization Paulownia Seedling cultural method comprises the steps:
S1, carry out aseptic bud cultivation:
The hybridization Paulownia just growing tender shoots is put in biochemical cultivation case, after cultivating 6～8 days under the conditions of 38～40 DEG C,
Clip hybridization Paulownia tender shoots is through volume ratio 70-80% alcohol-pickled 1min sterilization and the HgCl of mass ratio 0.05-0.15%2Solution
After sterilizing, after aseptic washing 3-5 time, the bud point cutting 1.8mm length accesses cultivation in culture medium, after tender shoots growth, then cuts
The bud point of 1.8mm length accesses newly configured good culture medium culturing, more than the most repeatedly for three times or three times, obtains the aseptic of detoxification
Bud；Because can kill certain virus through high-temperature cultivation, the conducting tissue of the bud point growing point of 2mm length is not also grown completely, no
Being beneficial to virus conduction, obtain the aseptic bud of preferable detoxification, such detoxification seedling advantage in planting process is that growth is preferable
Comparatively fast.
Culture medium includes plant growth regulator, carbon source, 1/2MS and/or MS, and compound method is: with 1/2MS and/or MS
For minimal medium, add appropriate plant growth regulator and carbon source is formulated, the pH6.5 of described culture medium.
Described plant growth regulator can use and include but not limited to that 6-BA, NAA or IBA use 6-BA and NAA simultaneously,
IBA.Being configured to the addition of 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA is (0.1mg/L-
1.0mg/L)。
Carbon source can use and include but not limited to sucrose, glucose.When using sucrose, after being configured to culture medium, concentration is
18g/L；When using glucose, after being configured to culture medium, concentration is 25g/L.
Culture medium coagulator to be added, can add agar powder, carrageenan etc., if selecting agar powder, the addition of agar powder
For 5g/L；If selection carrageenan, the addition of carrageenan is 8g/L.
The aseptic bud culture medium of hybridization Paulownia can be used by the formula after being configured to culture medium and include but not limited to:
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 described culture medium, must also add carbon source, when carbon source uses sucrose, after being configured to culture medium, addition is 18g/L,
When carbon source uses glucose, after being configured to culture medium, addition is 25g/L.
Note:
MS culture medium: be Murashige and Skoog in 1962 be tobacco cell Training Design, is characterized in inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, and its nitrate content is high, and the quantity of its nutrient and ratio are closed
Suitable, nutrition and the physiological need of plant cell can be met, thus the scope of application is relatively wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named by their name.
1/2MS culture medium: be the culture medium that halves of the amount of the calcium salt in MS culture medium and a great number of elements, be also conventional training
Support base.
Plant growth regulator 6-BA:6-benayl aminopurine, is a kind of basic element of cell division, English general entitled 6-
Benzylaminopurine, molecular formula is C12H11N5。
Plant growth regulator NAA:1-Naphthaleneacetic acid, naphthalene acetic acid, is called for short NAA, is a kind of plant
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula is C12H13NO2, it is a kind of auximone.
The cultivation temperature of step S1 is 15～20 DEG C, intensity of illumination 2000～2500LUX, light application time 10～12 hours/
My god；
S2, adventitious shoots culture:
The aseptic bud that clip grows, is inoculated on adventitious shoots culture base 15～20 days/generation of cultivation, can subculture 4-5 continuously
Secondary, Shoot propagation coefficient is 10～14.8 times/month, it is thus achieved that substantial amounts of Multiple Buds；Described medium component and the culture medium of step S1
Composition is equal to；
The cultivation temperature of step S2 is 15～20 DEG C, intensity of illumination 2000～2500LUX, light application time 10～12 hours/
My god；
S3, strong sprout, root culture:
During after Multiple Buds grows to 1-2cm length, point Seedling accesses newly configured good culture medium, after cultivating 15～20 days, it is thus achieved that strong
Strong Seedling of taking root；Described medium component is equal to medium component in step S1；
The cultivation temperature of step S3 is 15～20 DEG C, intensity of illumination 2000～2500LUX, light application time 10～12 hours/
My god；
S4, carry out acclimatization and transplants:
Being transplanted by the Paulownia Seedling obtained of step S3, the condition of culture of step S4 is: seedling exercising substrate garden mould+peat
Quality proportioning 1.5:1, shade net shelters from heat or light, and humidity is 60-65%.
Embodiment four
One of the present invention hybridization Paulownia Seedling cultural method comprises the steps:
S1, carry out aseptic bud cultivation: be outer implant to hybridize Paulownia tender stem or tender shoots, first volume ratio be 80% ethanol disappear
Poison, rear by sterile water wash 1 time, then to use mass fraction be 0.05%HgCl2Solution carry out sterilizing after, through sterile water wash
3-5 time, then it is connected in aseptic bud culture medium and cultivates, after cultivating 14～15 days, obtain aseptic bud.
Cultivated days depends mainly on the speed of growth of hybridization Paulownia tender shoots, and general aseptic bud grows to 1 cm and is
Preferably.Certainly, aseptic bud is longer than 1 centimetre (such as growing to 1.5 centimetres) or is 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 compound method is: with 1/2MS and/or MS
For minimal medium, add appropriate plant growth regulator and carbon source is formulated, the pH7.2 of described aseptic bud culture medium.
Described plant growth regulator can use and include but not limited to that 6-BA, NAA or IBA use 6-BA and NAA simultaneously,
IBA.Being configured to the addition of 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA is (0.1mg/L-
1.0mg/L)。
Carbon source can use and include but not limited to sucrose, glucose.When using sucrose, after being configured to culture medium, concentration is
20g/L；When using glucose, after being configured to culture medium, concentration is 30g/L.
Culture medium coagulator to be added, can add agar powder, carrageenan etc., if selecting agar powder, the addition of agar powder
For 6g/L；If selection carrageenan, the addition of carrageenan is 10g/L.
The aseptic bud culture medium of hybridization Paulownia can be used by the formula after being configured to culture medium and include but not limited to:
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 described culture medium, must also add carbon source, when carbon source uses sucrose, after being configured to culture medium, addition is 20g/L,
When carbon source uses glucose, after being configured to culture medium, addition is 30g/L.
Note:
MS culture medium: be Murashige and Skoog in 1962 be tobacco cell Training Design, is characterized in inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, and its nitrate content is high, and the quantity of its nutrient and ratio are closed
Suitable, nutrition and the physiological need of plant cell can be met, thus the scope of application is relatively wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named by their name.
1/2MS culture medium: be the culture medium that halves of the amount of the calcium salt in MS culture medium and a great number of elements, be also conventional training
Support base.
Plant growth regulator 6-BA:6-benayl aminopurine, is a kind of basic element of cell division, English general entitled 6-
Benzylaminopurine, molecular formula is C12H11N5。
Plant growth regulator NAA:1-Naphthaleneacetic acid, naphthalene acetic acid, is called for short NAA, is a kind of plant
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula is C12H13NO2, it is a kind of auximone；
The cultivation temperature of step S1 is 25～30 DEG C, intensity of illumination 2500～3000LUX, light application time 10～12 hours/
My god；
S2, adventitious shoots culture:
The aseptic bud that clip grows, is inoculated on adventitious shoots culture base 15～20 days/generation of cultivation, can subculture 4-5 continuously
Secondary, Shoot propagation coefficient is 10～14.8 times/month, it is thus achieved that substantial amounts of Multiple Buds；Described medium component and the culture medium of step S1
Composition is equal to；
The cultivation temperature of step S2 is 25～30 DEG C, intensity of illumination 2500～3000LUX, light application time 10～12 hours/
My god；
S3, strong sprout, root culture:
During after Multiple Buds grows to 1-2cm length, point Seedling accesses newly configured good culture medium, after cultivating 15～20 days, it is thus achieved that strong
Strong Seedling of taking root；Described medium component is equal to medium component in step S1；
The cultivation temperature of step S3 is 25～30 DEG C, intensity of illumination 2500～3000LUX, light application time 10～12 hours/
My god；
S4, carry out acclimatization and transplants:
The condition of culture of step S4 is: seedling exercising substrate garden mould+peat quality proportioning 2:1, and shade net shelters from heat or light, and humidity is
67-69%.
Embodiment five
One of the present invention hybridization Paulownia Seedling cultural method comprises the steps:
S1, carry out aseptic bud cultivation: be outer implant to hybridize Paulownia tender stem or tender shoots, first volume ratio be 70% ethanol disappear
Poison, rear by sterile water wash 1 time, then to use mass fraction be 0.15%HgCl2Solution carry out sterilizing after, through sterile water wash
3-5 time, then it is connected in aseptic bud culture medium and cultivates, after cultivating 16～18 days, obtain aseptic bud.
Cultivated days depends mainly on the speed of growth of hybridization Paulownia tender shoots, and general aseptic bud grows to 1 cm and is
Preferably.Certainly, aseptic bud is longer than 1 centimetre (such as growing to 1.5 centimetres) or is 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 compound method is: with 1/2MS and/or MS
For minimal medium, add appropriate plant growth regulator and carbon source is formulated, the pH7.0 of described aseptic bud culture medium.
Described plant growth regulator can use and include but not limited to that 6-BA, NAA or IBA use 6-BA and NAA simultaneously,
IBA.Being configured to the addition of 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA is (0.1mg/L-
1.0mg/L)。
Carbon source can use and include but not limited to sucrose, glucose.When using sucrose, after being configured to culture medium, concentration is
25g/L；When using glucose, after being configured to culture medium, concentration is 35g/L.
Culture medium coagulator to be added, can add agar powder, carrageenan etc., if selecting agar powder, the addition of agar powder
For 5.5g/L；If selection carrageenan, the addition of carrageenan is 13g/L.
The aseptic bud culture medium of hybridization Paulownia can be used by the formula after being configured to culture medium and include but not limited to:
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 medium: be Murashige and Skoog in 1962 be tobacco cell Training Design, is characterized in inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, and its nitrate content is high, and the quantity of its nutrient and ratio are closed
Suitable, nutrition and the physiological need of plant cell can be met, thus the scope of application is relatively wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named by their name.
1/2MS culture medium: be the culture medium that halves of the amount of the calcium salt in MS culture medium and a great number of elements, be also conventional training
Support base.
Plant growth regulator 6-BA:6-benayl aminopurine, is a kind of basic element of cell division, English general entitled 6-
Benzylaminopurine, molecular formula is C12H11N5。
Plant growth regulator NAA:1-Naphthaleneacetic acid, naphthalene acetic acid, is called for short NAA, is a kind of plant
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula is C12H13NO2, it is a kind of auximone；
The cultivation temperature of step S1 is 23～27 DEG C, intensity of illumination 2800～3000LUX, light application time 10～12 hours/
My god；
S2, adventitious shoots culture:
The aseptic bud that clip grows, is inoculated on adventitious shoots culture base 15～20 days/generation of cultivation, can subculture 4-5 continuously
Secondary, Shoot propagation coefficient is 10～14.8 times/month, it is thus achieved that substantial amounts of Multiple Buds；Described medium component and the culture medium of step S1
Composition is equal to；
The cultivation temperature of step S2 is 23～27 DEG C, intensity of illumination 2800～3000LUX, light application time 10～12 hours/
My god；
S3, strong sprout, root culture:
During after Multiple Buds grows to 1-2cm length, point Seedling accesses newly configured good culture medium, after cultivating 15～20 days, it is thus achieved that strong
Strong Seedling of taking root；Described medium component is equal to medium component in step S1；
The cultivation temperature of step S3 is 23～27 DEG C, intensity of illumination 2800～3000LUX, light application time 10～12 hours/
My god；
S4, carry out acclimatization and transplants:
The condition of culture of step S4 is: seedling exercising substrate garden mould+peat quality proportioning 2.5:1, shade net shelters from heat or light, humidity
For 66-70%.
Embodiment six
One of the present invention hybridization Paulownia Seedling cultural method comprises the steps:
S1, carry out aseptic bud cultivation: be outer implant to hybridize Paulownia tender stem or tender shoots, first volume ratio be 75% ethanol disappear
Poison, rear by sterile water wash 1 time, then to use mass fraction be 0.15%HgCl2Solution carry out sterilizing after, through sterile water wash
3-5 time, then it is connected in aseptic bud culture medium and cultivates, after cultivating 16～18 days, obtain aseptic bud.
Cultivated days depends mainly on the speed of growth of hybridization Paulownia tender shoots, and general aseptic bud grows to 1 cm and is
Preferably.Certainly, aseptic bud is longer than 1 centimetre (such as growing to 1.5 centimetres) or is 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 compound method is: with 1/2MS and/or MS
For minimal medium, add appropriate plant growth regulator and carbon source is formulated, the pH7.2 of described aseptic bud culture medium.
Described plant growth regulator can use and include but not limited to that 6-BA, NAA or IBA use 6-BA and NAA simultaneously,
IBA.Being configured to the addition of 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA is (0.1mg/L-
1.0mg/L)。
Carbon source can use and include but not limited to sucrose, glucose.When using sucrose, after being configured to culture medium, concentration is
30g/L；When using glucose, after being configured to culture medium, concentration is 40g/L.
Culture medium coagulator to be added, can add agar powder, carrageenan etc., if selecting agar powder, the addition of agar powder
For 6g/L；If selection carrageenan, the addition of carrageenan is 15g/L.
The aseptic bud culture medium of hybridization Paulownia can be used by the formula after being configured to culture medium and include but not limited to:
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 described culture medium, must also add carbon source, when carbon source uses sucrose, after being configured to culture medium, addition is 30g/L,
When carbon source uses glucose, after being configured to culture medium, addition is 40g/L.
Note:
MS culture medium: be Murashige and Skoog in 1962 be tobacco cell Training Design, is characterized in inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, and its nitrate content is high, and the quantity of its nutrient and ratio are closed
Suitable, nutrition and the physiological need of plant cell can be met, thus the scope of application is relatively wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named by their name.
1/2MS culture medium: be the culture medium that halves of the amount of the calcium salt in MS culture medium and a great number of elements, be also conventional training
Support base.
Plant growth regulator 6-BA:6-benayl aminopurine, is a kind of basic element of cell division, English general entitled 6-
Benzylaminopurine, molecular formula is C12H11N5。
Plant growth regulator NAA:1-Naphthaleneacetic acid, naphthalene acetic acid, is called for short NAA, is a kind of plant
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula is C12H13NO2, it is a kind of auximone；
The cultivation temperature of step S1 is 25～30 DEG C, intensity of illumination 2500～3000LUX, light application time 10～12 hours/
My god；
S2, adventitious shoots culture:
The aseptic bud that clip grows, is inoculated on adventitious shoots culture base 15～20 days/generation of cultivation, can subculture 4-5 continuously
Secondary, Shoot propagation coefficient is 10～14.8 times/month, it is thus achieved that substantial amounts of Multiple Buds；Described medium component and the culture medium of step S1
Composition is equal to；
The cultivation temperature of step S2 is 25～30 DEG C, intensity of illumination 2500～3000LUX, light application time 10～12 hours/
My god；
S3, strong sprout, root culture:
During after Multiple Buds grows to 1-2cm length, point Seedling accesses newly configured good culture medium, after cultivating 15～20 days, it is thus achieved that strong
Strong Seedling of taking root；Described medium component is equal to medium component in step S1；
The cultivation temperature of step S3 is 25～30 DEG C, intensity of illumination 2500～3000LUX, light application time 10～12 hours/
My god；
S4, carry out acclimatization and transplants:
The condition of culture of step S4 is: seedling exercising substrate garden mould+peat quality proportioning 2.5:1, shade net shelters from heat or light, humidity
For 66-70%.
Embodiment seven
One of the present invention hybridization Paulownia Seedling cultural method comprises the steps:
S1, carry out aseptic bud cultivation: be outer implant to hybridize Paulownia tender stem or tender shoots, first volume ratio be 78% ethanol disappear
Poison, rear by sterile water wash 1 time, then to use mass fraction be 0.13%HgCl2Solution carry out sterilizing after, through sterile water wash
3-5 time, then it is connected in aseptic bud culture medium and cultivates, after cultivating 16～18 days, obtain aseptic bud.
Cultivated days depends mainly on the speed of growth of hybridization Paulownia tender shoots, and general aseptic bud grows to 1 cm and is
Preferably.Certainly, aseptic bud is longer than 1 centimetre (such as growing to 1.5 centimetres) or is 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 compound method is: with 1/2MS and/or MS
For minimal medium, add appropriate plant growth regulator and carbon source is formulated, the pH6.8 of described aseptic bud culture medium.
Described plant growth regulator can use and include but not limited to that 6-BA, NAA or IBA use 6-BA and NAA simultaneously,
IBA.Being configured to the addition of 6-BA after culture medium is (0.1mg/L-2.0mg/L), and the addition of NAA or IBA is (0.1mg/L-
1.0mg/L)。
Carbon source can use and include but not limited to sucrose, glucose.When using sucrose, after being configured to culture medium, concentration is
28g/L；When using glucose, after being configured to culture medium, concentration is 38g/L.
Culture medium coagulator to be added, can add agar powder, carrageenan etc., if selecting agar powder, the addition of agar powder
For 6g/L；If selection carrageenan, the addition of carrageenan is 15g/L.
The aseptic bud culture medium of hybridization Paulownia can be used by the formula after being configured to culture medium and include but not limited to:
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 described culture medium, must also add carbon source, when carbon source uses sucrose, after being configured to culture medium, addition is 28g/L,
When carbon source uses glucose, after being configured to culture medium, addition is 38g/L.
Note:
MS culture medium: be Murashige and Skoog in 1962 be tobacco cell Training Design, is characterized in inorganic
Salt and ion concentration are higher, are more stable ionic equilibrium solution, and its nitrate content is high, and the quantity of its nutrient and ratio are closed
Suitable, nutrition and the physiological need of plant cell can be met, thus the scope of application is relatively wider, most plants tissue-culturing quick-propagation
Use it as the minimal medium of culture medium.Therefore this culture medium is just named by their name.
1/2MS culture medium: be the culture medium that halves of the amount of the calcium salt in MS culture medium and a great number of elements, be also conventional training
Support base.
Plant growth regulator 6-BA:6-benayl aminopurine, is a kind of basic element of cell division, English general entitled 6-
Benzylaminopurine, molecular formula is C12H11N5。
Plant growth regulator NAA:1-Naphthaleneacetic acid, naphthalene acetic acid, is called for short NAA, is a kind of plant
Auxin.
Plant growth regulator indolebutyric acid IBA molecular formula is C12H13NO2, it is a kind of auximone.
S2, adventitious shoots culture:
The aseptic bud that clip grows, is inoculated on adventitious shoots culture base 15～20 days/generation of cultivation, can subculture 4-5 continuously
Secondary, Shoot propagation coefficient is 10～14.8 times/month, it is thus achieved that substantial amounts of Multiple Buds；Described medium component and the culture medium of step S1
Composition is equal to；
The cultivation temperature of step S2 is 28～30 DEG C, intensity of illumination 2800～3000LUX, light application time 10～12 hours/
My god；
S3, strong sprout, root culture:
During after Multiple Buds grows to 1-2cm length, point Seedling accesses newly configured good culture medium, after cultivating 15～20 days, it is thus achieved that strong
Strong Seedling of taking root；Described medium component is equal to medium component in step S1；
The cultivation temperature of step S3 is 28～30 DEG C, intensity of illumination 2800～3000LUX, light application time 10～12 hours/
My god；
S4, carry out acclimatization and transplants:
Being transplanted by the Paulownia Seedling obtained of step S3, the condition of culture of step S4 is: seedling exercising substrate garden mould+peat matter
Amount proportioning 1.8:1, shade net shelters from heat or light, and humidity is 66-70%.
Claims (10)
1. a hybridization Paulownia Seedling cultural method, it is characterised in that comprise the steps:
S1, aseptic bud are cultivated:
It is outer implant with hybridization Paulownia tender stem or tender shoots, first uses alcohol-pickled sterilization, then use sterile water wash 1 time, then use
HgCl2After solution soaking carries out sterilizing, through sterile water wash 3-5 time, then implant during newly configured good culture medium is cultivated
Row is cultivated 12～18 days, and the temperature of cultivation is 15～30 DEG C, intensity of illumination 2000～3000LUX, and light application time 6～12 hours/
My god；
Described medium component includes plant growth regulator, carbon source, 1/2MS and/or MS；
S2, adventitious shoots culture:
The aseptic bud that clip grows, is inoculated in the newly configured good culture medium with step S1 equivalent components cultivation；Described cultivation
Temperature be 15～30 DEG C, intensity of illumination 2000～3000LUX, light application time 6～12 hour/day；
S3, strong sprout, root culture:
After Multiple Buds grows to 1-2cm length, sub-bottle is cultivated, and trains by the newly configured good culture medium with step S1 equivalent components
Support, obtain the healthy seedling that Paulownia takes root；Described strong sprout, the temperature of root culture are 15～30 DEG C, intensity of illumination 2000～
3000LUX, light application time 6～12 hour/day；
S4, acclimatization and transplants:
The healthy seedling that Paulownia step S3 obtained takes root is transplanted to land for growing field crops seedling exercising, and the substrate that described seedling exercising uses is garden mould and mud
Charcoal mixes, and shelters from heat or light seedling exercising substrate shade net.
2. hybridization Paulownia Seedling cultural method as claimed in claim 1, it is characterised in that can also be by just growing 1-
The hybridization Paulownia of the long tender shoots of 2.5mm is put in biochemical cultivation case, after cultivating 6～8 days under the conditions of 38～40 DEG C, and clip 1.5-
1.8mm hybridization Paulownia tender shoots first through alcohol-pickled sterilization, after by sterile water wash 1 time, use HgCl again2After solution soaking sterilizing,
After aseptic washing 3-5 time, the bud point cutting 1.8-2.2mm length accesses cultivation in newly configured good culture medium, and tender shoots grows
After 2.5mm length, then the bud point cutting 1.8-2.2mm length accesses in culture medium and cultivates, and more than repeatedly for three times or three times, obtains detoxification
Aseptic bud, in described culture medium condition of culture be temperature be 15～30 DEG C, intensity of illumination 2000～3000LUX, light application time 6
～12 hours/day.
3. hybridization Paulownia Seedling cultural method as claimed in claim 1 or 2, it is characterised in that the alcohol by volume of described use divides
Number is 70%～80%, the HgCl used2Liquid quality fraction is 0.05%～0.15%.
4. hybridization Paulownia Seedling cultural method as claimed in claim 1, it is characterised in that the culture medium in described step S1-S3
In plant growth regulator, carbon source, the compound method of 1/2MS and/or MS be: with 1/2MS and/or MS as minimal medium,
Add appropriate plant growth regulator and carbon source is formulated；The pH of described culture medium is 6.5～7.2.
5. hybridization Paulownia Seedling cultural method as claimed in claim 1, it is characterised in that the cultivation in described step S1, S2, S3
The plant growth regulator of base is 6-BA and/or NAA and/or IBA, and after being configured to culture medium, described 6-BA concentration is 0.1mg/L-
2.0mg/L, NAA concentration be 0.1mg/L-1.0mg/L, IBA concentration be 0.1mg/L-1.0mg/L.
6. hybridization Paulownia Seedling cultural method as claimed in claim 1, it is characterised in that the cultivation in described step S1, S2, S3
Base also need to add coagulator, agar powder or carrageenan can be added, if select agar powder, the addition of agar powder is 5-6g/
L；If selection carrageenan, the addition of carrageenan is 8-15g/L.
7. hybridization Paulownia Seedling cultural method as claimed in claim 1, it is characterised in that the cultivation 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, light application time 10～12 hour/day.
8. hybridization Paulownia Seedling cultural method as claimed in claim 1, it is characterised in that the nothing that in described step S2, clip grows
Length 1-2cm of bacterium bud, the speed of growth of the adventitious shoots culture of described step S2 is 15～20 days/generation, cultivates subculture 4-continuously
5 times, the Shoot propagation coefficient of described Multiple Buds is 10～14.8 times/month.
9. hybridization Paulownia Seedling cultural method as claimed in claim 1, it is characterised in that described carbon source is sucrose or glucose, makes
The culture medium sucrose concentration being configured to during with sucrose is 18～30g/L, and the culture medium glucose being configured to when using glucose is dense
Degree is 25～40g/L.
10. hybridization Paulownia Seedling cultural method as claimed in claim 1, it is characterised in that garden in the seedling exercising substrate of described step S4
Soil and peat quality proportioning are (1.5～2.5): 1, and seedling exercising district 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 |
| CN111374056A (en) * | 2020-04-17 | 2020-07-07 | 山东省林业科学研究院 | Proliferation medium formula for culturing stout tetraploid paulownia seedlings and application |
| CN111406647A (en) * | 2020-04-17 | 2020-07-14 | 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 |
| 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 |
| CN116135010A (en) * | 2023-04-10 | 2023-05-19 | 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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| Publication number | Priority date | Publication date | Assignee | Title |
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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 |
| CN111374056A (en) * | 2020-04-17 | 2020-07-07 | 山东省林业科学研究院 | Proliferation medium formula for culturing stout tetraploid paulownia seedlings and application |
| CN111406647A (en) * | 2020-04-17 | 2020-07-14 | 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 |
| 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 |
| CN116135010A (en) * | 2023-04-10 | 2023-05-19 | 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 |
| 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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