CN106797879A - One kind simulation tissue culture method and application - Google Patents
One kind simulation tissue culture method and application Download PDFInfo
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- CN106797879A CN106797879A CN201710042150.2A CN201710042150A CN106797879A CN 106797879 A CN106797879 A CN 106797879A CN 201710042150 A CN201710042150 A CN 201710042150A CN 106797879 A CN106797879 A CN 106797879A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
- A01G13/02—Protective coverings for plants; Coverings for the ground; Devices for laying-out or removing coverings
- A01G13/0231—Tunnels, i.e. protective full coverings for rows of plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
- A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
- A01G9/029—Receptacles for seedlings
- A01G9/0299—Handling or transporting of soil blocks or seedlings
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- Life Sciences & Earth Sciences (AREA)
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- General Health & Medical Sciences (AREA)
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Abstract
A kind of simulation tissue culture method and application of the present invention, comprise the following steps：(1) shed is built, the seedbed of nursery is provided in shed, sealed shed and sterilizing is carried out to its inside；(2) gather and pre-processed for simulating the explant of tissue culture, insert on the seedbed, seal shed；(3) to carrying out sterilizing inside the shed and controlling temperature in shed, relative humidity and the intensity of illumination to carry out nursery maintenance；(4) hardening；(5) transplant to field planting；In the above-mentioned methods, ensure that branch nursery in a relatively aseptic environment is in before taking root of cuttage, it is not easy to by living contaminants and corrosion, seedling raising environment is excellent, takes root fast, fast growth, survival rate is high, and low in investment cost, easily, nursery quantity is big for materials, one shed is in year, can be with nursery three batches, breeding efficiency is high, high financial profit.
Description
Technical field
The invention belongs to seedling-raising technique field, and in particular to it is a kind of using simulate tissue culture technology carry out nursery method and its
Using.
Background technology
At present, conventional seedling-raising technique has two methods of cuttage and seedling culture and tissue culture, wherein, cuttage raise seedling technique is because of it
Reproduction speed slow (1 year generation), low reproduction rate, survival rate are poor (typically there was only 30-60%), so can only be to some easy lifes
The plant of root carries out Sterile culture nursery, and quick breeding then can not be in a short time reached for new varieties, rare seeds
The purpose of listing, much can not meet people's needs；And tissue culture technique can make up the defect of cuttage and seedling culture, have
The advantage that breeding is fast, survival rate is high, but requirement due to tissue culture technology to environmental condition is higher, and need technical merit higher
Technician's operation, management, thus production link is complicated, production cost is high, cost of investment is big, being not suitable for large-scale promotion should
With.Recent domestic ecological construction, establishment of economic forest are developed rapidly and its to the wilderness demand of flowers in addition so that one
The need for the cultivation of a little neies variety of plant does not much catch up with market, this just proposes requirement higher to the development of seedling-raising technique.
For example, Chinese patent literature CN101720658A discloses a kind of method of soilless culture of broussonetia papyrifera seedlings, the method bag
Include following step：1) prepare soilless substrate and sterilize：The soilless substrate is made up of turf and perlite；2) alms bowl is filled：Step 1)
Then the soilless substrate of preparation is piled up the nutritive cube of dress matrix in ridge-up bed or seed plate again in nutritive cube；3) in step
2) the carrying out in soilless substrate nutritive cube to broussonetia papyrifera seedlings is cultivated, and is carried out disinfection, and then lid plastic greenhouse carries out maintenance seedling.It is above-mentioned
By improveing matrix in scheme, strengthen the gas permeability of matrix, addition turf improves nutrition so that seedling cultivation and rooting, germination and growth
Speed is fast, shortens growing-seedling period, and survival rate is high, low cost.But also there is following defect in such scheme：Although its including
The step of disinfection twice, is once that only the matrix is carried out disinfection when matrix is prepared, be for another time cuttage fringe bar it
After carry out disinfection, sterilize twice be all in open environment, due to being carried out disinfection in open environment, rather than
Closed environment, even if being carried out disinfection to matrix, the stronger or energetic germ of the drug resistance in open environment is easily mixed into and disappears
Growth and breeding in matrix after poison, the environment disinfected that causes cuttage fringe bar to grow is sterilized not thoroughly, easily by living contaminants, rot
Fringe bar, such as when root base forms the time to be exceeded 10 days, the fringe bar part for inserting matrix will rot, and cause nursery to fail,
So as to will extension rootage duration, cause seedling growth speed to slow down, seedling cycle stretch-out, survival rate reduction；It is again direct due to its
Cultivated during cuttage fringe bar is inserted into matrix, and cuttage fringe bar is pre-processed, caused the technology and use from culture
Cultivate the tissue culture shoot cultivation for obtaining in bottle to compare, survival rate is low, and the cultivating seedlings time is long, and cost of investment is high.
The content of the invention
Therefore, the technical problem to be solved in the present invention is to overcome the survival rate of seedling existing for existing method for culturing seedlings
Low, growing-seedling period is long and high cost defect, so that providing, a kind of survival rate is high, short and low cost the simulation tissue culture of growing-seedling period is educated
Seedling-growing method and application.
Therefore, the invention provides one kind simulation tissue culture method, comprising the following steps：
(1) build shed and the seedbed of nursery is provided in the shed, the shed is sealed, then to the arch
A sterilizing is carried out inside canopy；
(2) explant for nursery is pre-processed, is then inserted on the seedbed, seal the shed and to institute
Stating carries out re-pasteurization sterilization inside shed；
(3) temperature in the shed, relative humidity and intensity of illumination are controlled to carry out nursery maintenance；
(4) hardening；
(5) transplant to field planting.
Described simulation tissue culture method, in the step (3), the temperature is 25-35 DEG C；The relative humidity
It is 80-95%；The intensity of illumination is 3000-4000Lux, daily illumination 10-12h.
Described simulation tissue culture method, in the step (2), the explant is maternal stiff wood branch, to contain
There is the branch of at least two eyes to cut section for a stem section, and ensure the morphology upper end of the stem section under flat mouth, morphology
It is angle to hold, and the morphology lower end of the stem section is dipped in into root-growing agent, and then the morphology lower end is inserted the base in the seedbed
In matter, depth is 2-3cm；Or
The explant is spray, stem-segment with single bud is cut into, by the morphology lower end root-growing agent of the stem-segment with single bud
Immersion, then inserts the morphology lower end in the matrix in the seedbed, and depth is 2-3cm.
Described simulation tissue culture method, the root-growing agent used when the explant is by maternal stiff wood branch for containing
The mixed liquor of IAA and NAA, the concentration of the IAA is 15-25mg/L, and the concentration of the NAA is 25-35mg/L；The explant
The root-growing agent used during by spray is the mixed liquor containing IAA and NAA, and the concentration of the IAA is 1.5-2.5mg/L, described
The concentration of NAA is 2.5-3.5mg/L.
Preferably, the explant is maternal stiff wood branch, and the root-growing agent for using is the mixed liquor containing IAA and NAA, institute
The concentration of IAA is stated for 20mg/L, the concentration of the NAA is 30mg/L；The explant be spray, the root-growing agent for using for containing
The mixed liquor of IAA and NAA, the concentration of the IAA is 2mg/L, and the concentration of the NAA is 3mg/L.
Described simulation tissue culture method, in the step (1), saturating institute is sprayed using the 700-900 times of bactericide of liquid
The matrix in seedbed is stated to realize a sterilizing to the shed；
In the step (2), the 950-1050 times of bactericide of liquid is prepared to carrying out secondary sterilization sterilization in the shed,
The amount of spraying is the 1/2-2/3 of the saturating seedbed mesostroma of spray.
Preferably, in the step (1), the 750-850 times of bactericide of liquid is prepared to once being disappeared in the shed
Poisoning bacterium.
Preferably, in the step (2), 1000 times of bactericide of liquid are prepared and is killed to carrying out secondary sterilization in the shed
Bacterium, at interval of 3 days once, sprays 4-5 times altogether.
The bactericide is selected from the mixture of any one or more in thiophanate methyl, carbendazim and Bravo.
Described simulation tissue culture method, the matrix in the seedbed includes turf：Perlite：Vermiculite, and turf：Pearl
Rock：Vermiculite mass ratio is 2-4:1:1.Preferably, turf：Perlite：The mass ratio of vermiculite is 3:1:1
Described simulation tissue culture method, the matrix in the seedbed is 8-10 according to mass ratio with water:1 is well mixed
And be fitted into hole tray, by hole tray several described, closely discharge constitutes the seedbed.
Described simulation tissue culture method, in the step (4), treats that the stem section in the step (3) is grown to root
Cauline leaf is various and height is after the seedling of 15~25cm, and ventilating opening is set on the shed top, that is, start hardening, with described
The time of hardening increases, and the ventilating opening gradually increases until throwing off the canopy film completely, the hardening terminates.
Described simulation tissue culture method, the arch span of the shed is 2-2.2m, and sagitta is 1.1-1.3m, the shed
Total length be 22-26m.Preferably, the arch span of the shed is 2.1m, and sagitta is 1.2m, and the total length of the shed is
24m。
The ground and/or top that are additionally included in the shed for building are provided for the minitype nozzle sprayed water or spray
The step of.
The purposes of flowers and trees is being cultivated present invention also offers a kind of above-mentioned simulation tissue culture method, preferably
, described purposes is to cultivate the purposes of tomato, pumila, Chinese wax, Korea Chinese littleleaf box, rose and paper mulberry.
Technical solution of the present invention, has the following advantages that：
(1) a kind of simulation tissue culture method of the present invention, comprises the following steps：(1) shed is built and described
The seedbed of nursery is provided in shed, the shed is sealed, then to carrying out sterilizing inside the shed；(2) to
Pre-processed in the explant of nursery, then inserted on the seedbed, sealed the shed and to being carried out inside the shed
Re-pasteurization is sterilized；(3) controlling the temperature in the shed, relative humidity and intensity of illumination carries out nursery maintenance；(4) hardening；
(5) transplant to field planting；In the above-mentioned methods, the shed of sealing is on the one hand just built by starting, then in the arch of sealing
Sterilized twice in canopy, be once to have built shed and shed inside is carried out disinfection to form one relatively after setting seedbed
Aseptic environment, then carries out cuttage on this basis, is carried out disinfection again after cuttage so that the branch of cuttage is before taking root
The nursery in a relatively aseptic environment, germ is few, it is not easy to by living contaminants and corrosion, takes root fast, fast growth,
Survival rate is high, temperature, relative humidity and intensity of illumination during conserving nursery further through control in shed so that branch is in one
Taken root in the environment of individual advantage nursery, promote it to take root, nursery；On the other hand the tissue culture method of simulating is by cuttage and seedling culture
Method and tissue culture method are organically combined, while needing tissue culture room, autoclaving, aseptic in overcoming tissue culture
In the defect and cuttage and seedling culture of the high standard conditions such as water, blake bottle, agar medium and absolutesterility growing environment reproduction speed it is slow,
The defect of low reproduction rate, survival rate difference, obtains low in investment cost, and materials are easy, and survival rate is high, and nursery quantity is big, one
Shed, can be with nursery three batches in year, and breeding efficiency is high, the beneficial effect such as economic benefit is high.
(2) a kind of simulation tissue culture method of the present invention, by the step (3), controlling the temperature
It it is 25-35 DEG C, the relative humidity is 80-95%, the intensity of illumination is 3000-4000Lux, daily illumination 10-12h, is made
Branch is in the environment of advantage nursery of taking root, take root fast, fast growth, growing-seedling period is short.
(3) a kind of simulation tissue culture method of the present invention, by the step (2), the explant is
Maternal stiff wood branch, is stem section with the branch containing at least two eyes, and the stem section morphology upper end is flat mouth, under morphology
It is angle to hold, and the morphology lower end of the stem section is dipped in into root-growing agent, and then the morphology lower end is inserted the base in the seedbed
In matter；Or the explant is spray, selects current-year branch, stem-segment with single bud is cut into, by under the morphology of the stem-segment with single bud
End is soaked with root-growing agent, is then inserted morphology lower end in the matrix in the seedbed, seals the shed；It is above-mentioned by selecting
Branch or spray are used as cuttage fringe bar, it is ensured that the branch or spray are easily taken root after matrix is inserted, and improve rooting rate, are shortened
Rootage duration, and then shorten growing-seedling period, and hereditary dimensionally stable, can be new in ecological seeds, economic forest, flowers, vegetables
It is widely applied in kind nursery.
(4) a kind of simulation tissue culture method of the present invention, the explant is maternal stiff wood branch, the life of use
Root agent is the mixed liquor containing IAA and NAA, and the concentration of the IAA is 15-25mg/L, and the concentration of the NAA is 25-35mg/L；
The explant is spray, and the root-growing agent for using is the mixed liquor containing IAA and NAA, and the concentration of the IAA is 1.5-2.5mg/
The concentration of L, the NAA is 2.5-3.5mg/L；Root-growing agent is used by by branch or spray so that the branch or spray are more
Easily take root, improve rooting rate, shorten rootage duration.
(5) a kind of simulation tissue culture method of the present invention, the matrix in the seedbed includes turf：Perlite：Leech
The mass ratio of stone is (2-4):1:1 so that the branch or spray are easier growth, fast growth.
(6) a kind of simulation tissue culture method of the present invention, by the step (4), treating the step (3)
In stem section be grown to after seedling reaches garden standard and start hardening, on the shed top, ventilating opening is set, gradually reduce canopy
Interior humidity and temperature, the time with the hardening increase, and the ventilating opening gradually increases until throwing off the canopy film, institute completely
Hardening is stated to terminate；By above-mentioned steps hardening so that the seedling for going out garden can gradually adapt to environment, significantly improve its survival rate.
Brief description of the drawings
In order to illustrate more clearly of the specific embodiment of the invention or technical scheme of the prior art, below will be to specific
The accompanying drawing to be used needed for implementation method or description of the prior art is briefly described, it should be apparent that, in describing below
Accompanying drawing is some embodiments of the present invention, for those of ordinary skill in the art, before creative work is not paid
Put, other accompanying drawings can also be obtained according to these accompanying drawings.
Fig. 1 is by the hybridization paper mulberry figure after hardening described in the embodiment of the present invention 1；
Fig. 2 is by the hybridization paper mulberry figure after hardening described in the embodiment of the present invention 1；
Fig. 3 is the hybridization paper mulberry figure during the hardening described in the embodiment of the present invention 2.
Specific embodiment
Bactericide or root-growing agent for being related in following implementations etc. are commercially available prod, and such as IAA is purchased from Guangzhou woods state
Fertile Co., Ltd；NAA is purchased from Guangzhou Lin Guo chemical fertilizer Co., Ltd；Thiophanate methyl is purchased from Longdeng Chemical Co Ltd, Jiangsu；
Carbendazim is purchased from Guangzhou Lin Guo chemical fertilizer Co., Ltd；Bravo is purchased from Limin Chemical Co., Ltd..
Embodiment 1
A kind of simulation tissue culture method is present embodiments provided, in April, by the miscellaneous of Beijing Huairou area Dongguan Technology Park
1200 plants of paper mulberry seedlings (life, height 150cm in 1 year) is handed over, 60cm is done surely, as stock tree nursery, to gather nursery material use,
Beijing Huairou area Dongguan Technology Park carries out the simulation tissue culture of the paper mulberry, and step is as follows：
(1) shed is built, the shed 26 is set up, the arch span of each shed is 2.1m, and sagitta is 1.2m, described
The total length of shed is 24m, and the seedbed of nursery is provided in the shed, and the matrix in the seedbed is according to turf：Perlite：
The mass ratio of vermiculite is 3:1:1 prepares, and the matrix is 8 according to mass ratio with water:1 it is well mixed be fitted into the hole tray in 72 caves,
By hole tray several described, closely discharge constitutes the seedbed, there is also mounted mini sprinkler in the shed, configure small-sized cistern,
The equipment such as water pump, electrical equipment and light compensating lamp；The shed is covered using plastic greenhouse film, to seal the shed；Then 1000 are prepared
The thiophanate methyl solution of times liquid, is sprayed the sterilization that carries out disinfection by the mini sprinkler, and the amount of spraying is by the base in the hole tray
Matter spray is saturating, accomplishes the purpose of thorough sterilization；
(2) shoot is given birth to then from stock tree collection, stem-segment with single bud is cut into, by its Leaf Pruning into a diameter of 5cm
Peach-shaped leaflet, the morphology lower end of the stem-segment with single bud is soaked 10-20 minute with root-growing agent, in the immersion root-growing agent
Length is 2cm, and the root-growing agent is the mixed liquor containing IAA and NAA, and the concentration of the IAA is 2mg/L, and the NAA's is dense
It is 3mg/L to spend, and then raises canopy film, will be soaked in the matrix in one end insertion seedbed of root-growing agent, and the depth of insertion is
2.5cm, after sticking with, covers the plastic greenhouse film immediately, the shed is sealed, with 1000 times of carbendazim of liquid in the shed
Portion carries out sterilizing, and spraying time 1 minute at interval of 3 days once, is sprayed 4-5 times altogether；
(3) nursery maintenance：Temperature, the relative humidity in the shed are controlled while above-mentioned steps carry out disinfection sterilization
Nursery maintenance is carried out with intensity of illumination；In fine day, by the morning 9:Sunshade net was not covered before 30,9:30-16:When 00
Section covers sunshade net, afternoon 16 on canopy:Sunshade net is removed after 00 to keep and regulate and control temperature in the shed canopy at 25 DEG C
Between, no more than 40 DEG C, the intensity of illumination is 3000Lux, daily illumination 12h to highest, using mini sprinkler water spray regulation institute
Humidity in canopy is stated, general water spray 5-6 time, each time is no more than 50 minutes, controls in canopy relative humidity between 95%, one
In the middle of it nursing, 9:30-16:00 period was the emphasis of nursing, had regulated and controled temperature, illumination, the relation of humidity three, using screening
Screened postive adjusts temperature of shed, using humidity in micro- spray water spray regulation canopy；At at the cloudy day, sunshade net is not covered, water spray number of times is reduced
Or do not spray, it is 25 DEG C to regulate and control the temperature of shed when appropriate using light compensating lamp, and the relative humidity is 95%, the illumination
Intensity is 3000Lux, daily illumination 12h.In maintenance nursery 5-12 angel's stem section root of hair periods, temperature, illumination, wet is regulated and controled
Spend the relation of three；Major part stem section is taken root after 12nd day, itself there is absorption moisture, the ability of nutrient, and resistance is obvious
Enhancing；Almost all is taken root within 17th day, is now transformed into seedling by stem section, and newborn cauline leaf starts growth；Enter life after 20th day
Peak long, seedling has grown to 30cm or so within the 30th day, has reached the standard in garden；
(4) hardening：The above-mentioned cultivation paper mulberry seedling of 30 days is started into hardening, the canopy film on the shed top is thrown off to be formed
Ventilating opening, the middle and lower part of the shed is covered by the canopy film, and the canopy film covers the 2/3 of the shed, during ventilation in first day
Between should the sun is open before not going out in the morning or selects overcast and rainy, humidity and temperature in canopy are gradually reduced, with the hardening
Time increases, and the ventilating opening gradually increases until throwing off the canopy film completely, the hardening terminates, and hardening one week, educates altogether
Into broussonetia papyrifera seedlings as shown in Figure 1-2；
(5) transplant to field planting, plant percent reaches 98%.
Embodiment 2
A kind of simulation tissue culture method is present embodiments provided, in April, by the plantation of Jiangsu Province Changzhou Meng He bases
50000 plants of hybridization paper mulberry nursery stock (life, height 160cm in 2 years), 72cm is done surely, as stock tree, to gather nursery material use,
The simulation tissue culture of the paper mulberry is carried out in Jiangsu Province Changzhou Meng He bases, step is as follows：
(1) shed is built, the shed 26 is set up, the arch span of each shed is 2.2m, and sagitta is 1m, the arch
The total length of canopy is 24m, and big arch shed has also been built outside the shed for rain cover, is provided for educating in the shed
The seedbed of seedling, the matrix in the seedbed is according to turf：Perlite：The mass ratio of vermiculite is 4:1:1 is prepared, and the matrix is pressed with water
It is 10 according to mass ratio:1 it is well mixed is fitted into the hole tray in 72 caves, closely discharged by hole tray several described and constitute the seedbed,
Top in the shed is mounted with mini sprinkler, small-sized cistern, water pump, electrical equipment and light compensating lamp etc. is also configured in shed and is set
It is standby；The shed is covered using plastic greenhouse film, to seal the shed, 850 times of carbendazim solutions of liquid is then prepared, by institute
State mini sprinkler sprinkling irrigation to carry out disinfection sterilization, the amount of spraying is that the matrix spray in the hole tray is saturating, reaches the purpose of thorough sterilization；
(2) stiff wood branch is gathered from the stock tree, is stem section with the branch containing at least two eyes, the stem section shape
State upper end is flat mouth, and morphology lower end is angle, and the morphology lower end of the stem section is dipped into root-growing agent, and the root-growing agent is
Mixed liquor containing IAA and NAA, the concentration of the IAA is 15mg/L, and the concentration of the NAA is 35mg/L, then raises canopy
Film, by the matrix in the morphology lower end insertion seedbed, the depth of insertion is 2.5cm, after sticking with, the plastics is covered immediately
Canopy film, seals the shed, to carrying out sterilizing, the amount of spraying inside the shed is spray with 1050 times of thiophanate methyls of liquid
The 1/2 of saturating matrix, every 1 day once, sprays 4-5 times altogether；
(3) nursery maintenance：By controlling temperature in the shed, relative while above-mentioned steps carry out disinfection sterilization
Humidity and intensity of illumination carry out nursery maintenance；In fine day, by the morning 9:Sunshade net was not covered before 30,9:30-16:
00 period covered sunshade net, afternoon 16 on canopy:Sunshade net is removed after 00 and is existed to keep and regulate and control the temperature in the shed canopy
35 DEG C, no more than 40 DEG C, the intensity of illumination is 4000Lux, daily illumination 10h to highest, using mini sprinkler water spray regulation institute
Humidity in canopy is stated, general water spray 5-6 time, each time is no more than 50 minutes, controls in canopy relative humidity between 80%, one
In the middle of it nursing, 9:30-16:00 period was that the emphasis of nursing has regulated and controled temperature, illumination, the relation of humidity three, using screening
Screened postive adjusts temperature of shed, using humidity in micro- spray water spray regulation canopy；At at the cloudy day, sunshade net is not covered, water spray number of times is reduced
Or do not spray, it is 35 DEG C to regulate and control the temperature of shed when appropriate using light compensating lamp, and the relative humidity is 80%, the illumination
Intensity is 4000Lux, daily illumination 10h.In maintenance nursery 5-12 angel's stem section root of hair periods, temperature, illumination, wet is regulated and controled
Spend the relation of three；Major part stem section is taken root after 12nd day, itself there is absorption moisture, the ability of nutrient, and resistance is obvious
Enhancing；Almost all is taken root within 17th day, is now transformed into seedling by stem section, and newborn cauline leaf starts growth；Enter life after 20th day
Peak long, seedling has grown to 20cm or so within the 27th day, has reached the standard in garden；
(4) hardening：The above-mentioned cultivation paper mulberry seedling of 27 days is started into hardening, the canopy film in the middle part of the shed is opened to be formed
As shown in figure 3, the upper and lower part of the shed is covered by the canopy film, the canopy film covers the 3/4 of the shed to ventilating opening,
Humidity and temperature in canopy are gradually reduced, the time with the hardening increases, and the ventilating opening gradually increases until throwing off completely
The canopy film, the hardening terminates, altogether hardening one week；
(5) transplant to field planting, plant percent reaches 99%.
Embodiment 3
A kind of simulation tissue culture method is present embodiments provided, in April, by the plantation of Jiangsu Province Changzhou Meng He bases
Tomato sapling wood 4000 plants of (life in 2 years), as stock tree, to gather nursery material use, in Jiangsu Province Changzhou Meng He bases
The simulation tissue culture of the tomato tree is carried out, step is as follows：
(1) shed is built, the shed 26 is set up, the arch span of each shed is 2.2m, and sagitta is 1m, the arch
The total length of canopy is 24m, and big arch shed has also been built outside the shed for rain cover, is provided for educating in the shed
The seedbed of seedling, the matrix in the seedbed is according to turf：Perlite：The mass ratio of vermiculite is 2:1:1 is prepared, and the matrix is pressed with water
It is 9 according to mass ratio:1 it is well mixed is fitted into the hole tray in 72 caves, closely discharged by hole tray several described and constitute the seedbed, institute
State shed inner top and there is also mounted mini sprinkler, small-sized cistern, water pump, electrical equipment and light compensating lamp etc. are also configured in the shed and is set
It is standby；The shed is covered using plastic greenhouse film, to seal the shed, 750 times of carbendazim solutions of liquid is then prepared, by institute
State mini sprinkler sprinkling irrigation to carry out disinfection sterilization, the amount of spraying is that the matrix spray in the hole tray is saturating, with thoroughly sterilized；
(2) stiff wood branch is gathered from the stock tree, is stem section with the branch containing at least two eyes, the stem section shape
State upper end is flat mouth, and morphology lower end is angle, and the morphology lower end of the stem section dipped in into root-growing agent, the root-growing agent for
Mixed liquor containing IAA and NAA, the concentration of the IAA is 25mg/L, and the concentration of the NAA is 25mg/L, then raises canopy
Film, by the matrix in the morphology lower end insertion seedbed, the depth of insertion is 3cm, after sticking with, the plastic greenhouse is covered immediately
Film, seals the shed, saturating for spray to carrying out sterilizing, the amount of spraying inside the shed with 950 times of thiophanate methyls of liquid
The 1/2 of matrix, every 1 day once, sprays 4-5 times altogether；
(3) nursery maintenance：By controlling temperature in the shed, relative while above-mentioned steps carry out disinfection sterilization
Humidity and intensity of illumination carry out nursery maintenance；In fine day, by the morning 9:Sunshade net was not covered before 30,9:30-16:
00 period covered sunshade net, afternoon 16 on canopy:Sunshade net is removed after 00 and is existed to keep and regulate and control the temperature in the shed canopy
Between 25 DEG C, the intensity of illumination is 4000Lux, daily illumination 12h, using humidity, one in the mini sprinkler water spray regulation canopy
As spray water 5-6 time, each time be no more than 50 minutes, control canopy in relative humidity between 80%, in the middle of the nursing of a day,
9:30-16:00 period was that the emphasis of nursing has regulated and controled temperature, illumination, the relation of humidity three, using temperature in sunshade net regulation canopy
Degree, using humidity in micro- spray water spray regulation canopy；At at the cloudy day, sunshade net is not covered, water spray number of times is reduced or do not sprayed, makes when appropriate
It is 25 DEG C to regulate and control the temperature of shed with light compensating lamp, and the relative humidity is 80%, and the intensity of illumination is 4000Lux, often
Day illumination 12h.In maintenance nursery 5-12 angel's stem section root of hair periods, temperature, illumination, the relation of humidity three are regulated and controled；The
Major part stem section is taken root after 13 days, itself there is absorption moisture, the ability of nutrient, and resistance is remarkably reinforced；15th day almost
All take root, seedling is now transformed into by stem section, newborn cauline leaf starts growth；Enter summit of growth, the 28th day children after 18th day
Seedling has grown to 25cm, has reached the standard in garden；
(4) hardening：The above-mentioned cultivation tomato tree seedling of 28 days is started into hardening, the canopy film on the shed top is thrown off into shape
Into ventilating opening, the middle and lower part of the shed is covered by the canopy film, and the canopy film covers the 1/4 of the shed, gradually reduces canopy
Interior humidity and temperature, the time with the hardening increase, and the ventilating opening gradually increases until throwing off the canopy film, institute completely
Hardening is stated to terminate；
(5) transplant to field planting, plant percent reaches 98%.
Embodiment 4
A kind of simulation tissue culture method is present embodiments provided, in April, by the plantation of Jiangsu Province Changzhou Meng He bases
4000 plants of hybridization pumila nursery stock (life, height 140cm in 2 years), 50cm is done surely, as stock tree, to gather nursery material use,
The simulation tissue culture of the pumila is carried out in Jiangsu Province Changzhou Meng He bases, step is as follows：
(1) shed is built, the shed 26 is set up, the arch span of each shed is 2.2m, and sagitta is 1m, the arch
The total length of canopy is 24m, and big arch shed has also been built outside the shed for rain cover, is provided for educating in the shed
The seedbed of seedling, the matrix in the seedbed is according to turf：Perlite：The mass ratio of vermiculite is 4:1:1 is prepared, and the matrix is pressed with water
It is 9 according to mass ratio:1 it is well mixed is fitted into the hole tray in 72 caves, closely discharged by hole tray several described and constitute the seedbed, institute
State and mini sprinkler is there is also mounted in shed, configure the equipment such as small-sized cistern, water pump, electrical equipment and light compensating lamp；Covered using plastic greenhouse film
The shed is covered, to seal the shed, 750 times of carbendazim solutions of liquid is then prepared, being sprayed by the mini sprinkler is carried out
Disinfection, the amount of spraying is that the matrix spray in the hole tray is saturating, accomplishes the purpose of thorough sterilization；
(2) stiff wood branch is gathered from the stock tree, is stem section with the branch containing at least two eyes, in the stem section
It is flat mouth to hold, and lower end is angle, and the lower end of the stem section is dipped in into root-growing agent, and the root-growing agent is the mixing containing IAA and NAA
Liquid, the concentration of the IAA is 25mg/L, and the concentration of the NAA is 25mg/L, then raises canopy film, will soak the one of root-growing agent
In the matrix in the end insertion seedbed, the depth of insertion is 3cm, after sticking with, the plastic greenhouse film is covered immediately, seals the arch
Canopy, with 950 times of thiophanate methyls of liquid to carrying out sterilizing inside the shed, the amount of spraying is the 1/2 of the saturating matrix of spray, every
1 day once, sprays 4-5 times altogether；
(3) nursery maintenance：By controlling temperature in the shed, relative while above-mentioned steps carry out disinfection sterilization
Humidity and intensity of illumination carry out nursery maintenance；In fine day, by the morning 9:Sunshade net was not covered before 30,9:30-16:
00 period covered sunshade net, afternoon 16 on canopy:Sunshade net is removed after 00 and is existed to keep and regulate and control the temperature in the shed canopy
Between 35 DEG C, the intensity of illumination is 3000Lux, daily illumination 10h, using humidity, one in the mini sprinkler water spray regulation canopy
As spray water 5-6 time, each time be no more than 50 minutes, control canopy in relative humidity between 95%, in the middle of the nursing of a day,
9:30-16:00 period was that the emphasis of nursing has regulated and controled temperature, illumination, the relation of humidity three, using temperature in sunshade net regulation canopy
Degree, using humidity in micro- spray water spray regulation canopy；At at the cloudy day, sunshade net is not covered, water spray number of times is reduced or do not sprayed, makes when appropriate
It is 35 DEG C to regulate and control the temperature of shed with light compensating lamp, and the relative humidity is 95%, and the intensity of illumination is 3000Lux, often
Day illumination 10h.In maintenance nursery 5-12 angel's stem section root of hair periods, temperature, illumination, the relation of humidity three are regulated and controled；The
Major part stem section is taken root after 13 days, itself there is absorption moisture, the ability of nutrient, and resistance is remarkably reinforced；18th day almost
All take root, seedling is now transformed into by stem section, newborn cauline leaf starts growth；Enter summit of growth, the 30th day children after 22nd day
Seedling has grown to 22cm, has reached the standard in garden；
(4) hardening：The above-mentioned cultivation pumila seedling of 30 days is started into hardening, the canopy film on the shed top is thrown off into shape
Into ventilating opening, the middle and lower part of the shed is covered by the canopy film, and the canopy film covers the 2/3 of the shed, gradually reduces canopy
Interior humidity and temperature, the time with the hardening increase, and the ventilating opening gradually increases until throwing off the canopy film, institute completely
State hardening to terminate, the hardening time is one week；
(5) pumila after above-mentioned hardening is transplanted to field planting, plant percent reaches 95%.Embodiment 5
A kind of simulation tissue culture method is present embodiments provided, in April, by the rose of Beijing Huairou area Dongguan Technology Park
100 plants of rare (life in 1 year), as stock tree nursery, to gather nursery material use, the rose is carried out in Beijing Huairou area Dongguan Technology Park
Rare simulation tissue culture, step is as follows：
(1) shed is built, the shed 26 is set up, the arch span of each shed is 2-2.2m, and sagitta is 1.2m, institute
The total length of shed is stated for 24m, area is 1372.8m altogether2, the seedbed of nursery, the seedbed are provided in the shed
Matrix according to turf：Perlite：The mass ratio of vermiculite is 3:1:1 prepares, and the matrix is 9 according to mass ratio with water:1 mixing
Uniformly it is fitted into the hole tray in 72 caves, by hole tray several described, closely discharge constitutes the seedbed, be there is also mounted in the shed
Mini sprinkler, configures the equipment such as small-sized cistern, water pump, electrical equipment and light compensating lamp；The shed is covered using plastic greenhouse film, to seal
The shed, then prepares 800 times of Bravo solution of liquid, is sprayed by the mini sprinkler and carried out disinfection sterilization, and the amount of spraying is
Matrix spray in the hole tray is saturating, reach the purpose of thorough sterilization；
(2) shoot is given birth to then from stock tree collection, stem-segment with single bud is cut into, by its Leaf Pruning into a diameter of 5cm
Peach-shaped leaflet, the morphology lower end of the stem-segment with single bud is soaked 15 minutes with root-growing agent, the length immersed in the root-growing agent
It is 2cm to spend, and the root-growing agent is the mixed liquor containing IAA and NAA, and the concentration of the IAA is 2.5mg/L, the concentration of the NAA
It is 2.5mg/L, then raises canopy film, will soak in the matrix in one end insertion seedbed of root-growing agent, the depth of insertion is
2.5cm, after sticking with, covers the plastic greenhouse film immediately, the shed is sealed, with 800 times of carbendazim of liquid in the shed
Portion carries out sterilizing, and spraying time 1 minute at interval of 2 days once, is sprayed 4-5 times altogether；
(3) nursery maintenance：By controlling temperature in the shed, relative while above-mentioned steps carry out disinfection sterilization
Humidity and intensity of illumination carry out nursery maintenance；In fine day, by the morning 9:Sunshade net was not covered before 30,9:30-16:
00 period covered sunshade net, afternoon 16 on canopy:Sunshade net is removed after 00 and is existed to keep and regulate and control the temperature in the shed canopy
Between 30 DEG C, the intensity of illumination is 3500Lux, daily illumination 11h, using humidity, one in the mini sprinkler water spray regulation canopy
As spray water 5-6 time, each time be no more than 50 minutes, control canopy in relative humidity between 88%, in the middle of the nursing of a day,
9:30-16:00 period was that the emphasis of nursing has regulated and controled temperature, illumination, the relation of humidity three, using temperature in sunshade net regulation canopy
Degree, using humidity in micro- spray water spray regulation canopy；At at the cloudy day, sunshade net is not covered, water spray number of times is reduced or do not sprayed, makes when appropriate
It is 30 DEG C to regulate and control the temperature of shed with light compensating lamp, and the relative humidity is 88%, and the intensity of illumination is 3500Lux, often
Day illumination 11h；In maintenance the 5-12 days stem section root of hair periods of nursery, temperature, illumination, the relation of humidity three are regulated and controled；13rd
Major part stem section is taken root after it, itself there is absorption moisture, the ability of nutrient, and resistance is remarkably reinforced；16th day almost complete
Portion takes root, and is now transformed into seedling by stem section, and newborn cauline leaf starts growth；Enter summit of growth, the 30th day seedling after 20th day
30cm or so is grown to, has reached the standard in garden；
(4) hardening：The above-mentioned cultivation rose seedling of 30 days is started into hardening, the canopy film on the shed top is thrown off to be formed
Ventilating opening, the middle and lower part of the shed is covered by the canopy film, and the canopy film covers the 2/3 of the shed, during ventilation in first day
Between should the sun is open before not going out in the morning or selects overcast and rainy, humidity and temperature in canopy are gradually reduced, with the hardening
Time increases, and the ventilating opening gradually increases until throwing off the canopy film completely, the hardening terminates, the hardening time is one week；
(5) rose after hardening is transplanted to field planting, plant percent reaches 96%.
Comparative example 1
A kind of method for culturing seedlings is present embodiments provided, it is in April, the hybridization paper mulberry of Beijing Huairou area Dongguan Technology Park is big
1200 plants of seedling (life, height 150cm in 1 year), does 60cm surely, as stock tree nursery, to gather nursery material use, in Beijing Huairou
Area Dongguan Technology Park comprises the following steps according to the method nursery of embodiment 1 in Chinese patent literature CN101720658A：
1st, soilless substrate is prepared：
Turf dry it is dry after crushed with pulverizer, and with the 0.8-1.2 centimetres of sieve in aperture；
To be mixed with the ratio of volume ratio 45: 55 by the turf and perlite of above-mentioned treatment, obtain soilless substrate；
2nd, soilless substrate is disinfected：Soilless substrate prepared by step 1 is with 600 times of thiophanate methyl solution disinfection
Reason, treatment to matrix moistening；
3rd, alms bowl is filled：Sterilized soilless substrate mounted in 6-8 centimetres of bore, in 8-10 centimetres high of nutritive cube, dress 8-9 into
It is full, finally the nutritive cube of dress matrix is piled up in ridge-up bed or seed plate；
4th, plant tissue-cultured seedling：From the stock tree collection then give birth to shoot, be cut into stem-segment with single bud, by its Leaf Pruning into
The peach-shaped leaflet of a diameter of 5cm, is then planted in without in native nutritive cube, and then seedling root is poured and once determined root as tight during plant
Water, finally with after 600 times of first support solution disinfections again lid plastic greenhouse conserve take root, the 18th day rooting rate up to 80%, the 55th day seedling
Can hardening, transplant, plant percent reaches 85%.
Comparative example 2
A kind of method for culturing seedlings is present embodiments provided, it is in April, the hybridization paper mulberry of Beijing Huairou area Dongguan Technology Park is big
1200 plants of seedling (life, height 150cm in 1 year), does 60cm surely, as stock tree nursery, to gather nursery material use, in Beijing Huairou
Area Dongguan Technology Park is according to Chinese patent literature
The method nursery of embodiment 1, comprises the following steps in CN101720658A：
1st, soilless substrate is prepared：
Turf dry it is dry after crushed with pulverizer, and with the 0.8-1.2 centimetres of sieve in aperture；
To be mixed with the ratio of volume ratio 50: 50 by the turf and perlite of above-mentioned treatment, obtain soilless substrate；
2nd, soilless substrate is disinfected：Soilless substrate prepared by step 1 is with 600 times of thiophanate methyl solution disinfection
Reason, treatment to matrix moistening；
3rd, alms bowl is filled：Sterilized soilless substrate mounted in 6-8 centimetres of bore, in 8-10 centimetres high of nutritive cube, dress 8-9 into
It is full, finally the nutritive cube of dress matrix is piled up in ridge-up bed or seed plate；
4th, plant tissue-cultured seedling：Shoot is given birth to then from stock tree collection, stem-segment with single bud is cut into, by its Leaf Pruning
Into the peach-shaped leaflet of a diameter of 5cm, the morphology lower end of the stem-segment with single bud is soaked 10-20 minutes with root-growing agent, immerse institute
The length in root-growing agent is stated for 2cm, the root-growing agent is the mixed liquor containing IAA and NAA, and the concentration of the IAA is 2mg/
The concentration of L, the NAA is 3mg/L, will soak one end insertion of root-growing agent without in native nutritive cube, and the depth of insertion is 2.5cm,
A root water is poured in seedling root then by tight during plant, finally foster in small builder's temporary shed with being covered again after 600 times of first support solution disinfections
Shield, up to 80%, hardening, transplanting by the 50th day seedling, plant percent reach 88% to the 20th day rooting rate.
Comparative example 3
A kind of method for culturing seedlings is present embodiments provided, it is in April, the hybridization paper mulberry of Beijing Huairou area Dongguan Technology Park is big
1200 plants of seedling (life, height 150cm in 1 year), does 60cm surely, as stock tree nursery, to gather nursery material use, in Beijing Huairou
Area Dongguan Technology Park according to the method nursery in Chinese patent literature CN101578962A, comprise the following steps：
A, set up greenhouse and seedbed
The greenhouse of simulated tissue culture test tube microenvironment, by bow member 3, the atomizing mini sprinkler 4 of spray cooling, the He of light compensating lamp 5
Constituted with the water pipe 8 of magnetic valve 7 with timer 6.Mocromembrane composition shed entirety is added a cover on bow member, shed is high 70 centimetres.In shed
Plus with full-shape light compensating lamp 5, light saturation point is 40000Lux.Seedbed is by being easy to the special seedling culture hole plate 1 of the forest of draining and perlite
Matrix 2 is constituted.Seedbed is wide 1 meter, long 3 meters, high 0.7 meter.
B, prepare body material and inorganic nutrient solution
From the stock tree collection then give birth to shoot, be cut into stem-segment with single bud, by its Leaf Pruning into a diameter of 5cm peach
Shape leaflet, sterilizing seals its morphology upper end otch with the solid paraffin of fusing, and its morphology inferior end notch is soaked in into concentration
It is 100mg.kg-10.50h in green plant growth regulator solution, it is standby；
Inorganic nutrient solution is to be by by following weight ratio formula：950 parts of calcium nitrate, 80 parts of potassium nitrate, ammonium dihydrogen phosphate 155
Part, 483 parts of magnesium sulfate, 40 parts of chelated iron, 2.13 parts of manganese sulfate, 2.80 parts of boric acid, 0.22 part of zinc sulfate, 0.08 part of copper sulphate,
0.02 part of ammonium molybdate, it is water-soluble to weight ratio be 0.05%-0.1%.
C, seedling culture
Setting hot-house culture condition is：3000~4000Lux of intensity of illumination, periodicity of illumination is that daily 14h illumination and 10h are black
Secretly, by ventilation, ventilation, it is 22~28 DEG C to reduce relative temperature in seedbed, improves stomatal movement function, reduces physiology and form
Abnormal plant, culture matrix humidity is 70%；
Body material is numerous on seedling culture hole plate；
Fast numerous initial stage (i.e. cell activation phase), sprays once for 24 hours；Callus is taken root the phase, is sprayed twice within 24 hours；
The acclimatization and transplantses phase, spray within 24 hours three times；
Adjusting air humidity is 85~95% in the cell activation phase at fast numerous initial stage, callus take root the phase for 80~
90%, the acclimatization and transplantses phase is 75~85%；
Up to 80%, hardening, transplanting by 55 days seedlings, plant percent reach 82% to 20th day rooting rate.
Obviously, above-described embodiment is only intended to clearly illustrate example, and not to the restriction of implementation method.It is right
For those of ordinary skill in the art, can also make on the basis of the above description other multi-forms change or
Change.There is no need and unable to be exhaustive to all of implementation method.And the obvious change thus extended out or
Among changing still in the protection domain of the invention.
Claims (10)
1. it is a kind of to simulate tissue culture method, it is characterised in that to comprise the following steps：
(1) build shed and the seedbed of nursery is provided in the shed, the shed is sealed, then in the shed
Portion carries out a sterilizing；
(2) explant for nursery is pre-processed, is then inserted on the seedbed, seal the shed and to the arch
Re-pasteurization sterilization is carried out inside canopy；
(3) temperature in the shed, relative humidity and intensity of illumination are controlled to carry out nursery maintenance；
(4) hardening；
(5) transplant to field planting.
2. it is according to claim 1 to simulate tissue culture method, it is characterised in that in the step (3), the temperature
It is 25-35 DEG C；The relative humidity is 80-95%；The intensity of illumination is 3000-4000Lux, daily illumination 10-12h.
3. it is according to claim 1 and 2 to simulate tissue culture method, it is characterised in that described in the step (2)
Explant is maternal stiff wood branch, to cut section as a stem section containing at least two eyes, and is ensured on the morphology of the stem section
Hold as flat mouth, morphology lower end are angle, root-growing agent is dipped in into the morphology lower end of the stem section, then by the morphology lower end
In inserting the matrix in the seedbed；Or
The explant is spray, is cut into stem-segment with single bud, and the morphology lower end of the stem-segment with single bud is soaked with root-growing agent,
Then by the matrix in the morphology lower end insertion seedbed.
4. it is according to claim 3 to simulate tissue culture method, it is characterised in that the explant is maternal stiff wood branch
When the root-growing agent that is used be the mixed liquor containing IAA and NAA, the concentration of the IAA is 15-25mg/L, the concentration of the NAA
It is 25-35mg/L；
The root-growing agent used when the explant is by spray is the mixed liquor containing IAA and NAA, and the concentration of the IAA is
The concentration of 1.5-2.5mg/L, the NAA is 2.5-3.5mg/L.
5. the simulation tissue culture method according to claim any one of 1-4, it is characterised in that in the step (1),
The matrix in the 700-900 times of saturating seedbed of bactericide spray of liquid is used to realize a sterilizing to the shed；
In the step (2), using the 950-1050 times of bactericide of liquid to carrying out re-pasteurization sterilization, institute inside the shed
The amount of spraying for stating bactericide is the 1/2-2/3 of the saturating seedbed mesostroma of spray.
6. the simulation tissue culture method according to claim any one of 1-5, it is characterised in that the seedbed mesostroma
Raw material composition includes turf, perlite and vermiculite, and turf：Perlite：The mass ratio of vermiculite is (2-4):1:1.
7. the simulation tissue culture method according to claim any one of 1-6, it is characterised in that press the matrix and water
It is (8-10) according to mass ratio:1 is well mixed and is fitted into hole tray, and by hole tray several described, closely discharge constitutes the seedbed.
8. the simulation tissue culture method according to claim any one of 1-7, it is characterised in that in the step (4),
After the stem section in the step (3) be grown to root, stem and leaf it is various and height in the seedling of 15~25cm after, on the shed top
Ventilating opening is set, that is, starts hardening, the time with the hardening increases, and the ventilating opening gradually increases until throwing off canopy completely
Film is terminating hardening.
9. the simulation tissue culture method according to claim any one of 1-8, it is characterised in that the arch span of the shed is
2-2.2m, sagitta is 1.1-1.3m, and the total length of the shed is 22-26m；
The ground and/or top that are additionally included in the shed for building are provided for the step of the minitype nozzle sprayed water or spray
Suddenly.
10. it is a kind of to simulate purposes of the tissue culture method in flowers and trees are cultivated as described in power requires any one of 1-9.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115462248A (en) * | 2021-06-11 | 2022-12-13 | 上海市崇明区港沿镇农业综合技术推广服务中心(上海市崇明区港沿镇畜牧兽医站) | Method for obtaining boxwood container seedlings through substrate cuttage |
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Application publication date: 20170606 |